JPH1192343A - Agent for prolonging hair growth period - Google Patents
Agent for prolonging hair growth periodInfo
- Publication number
- JPH1192343A JPH1192343A JP27526197A JP27526197A JPH1192343A JP H1192343 A JPH1192343 A JP H1192343A JP 27526197 A JP27526197 A JP 27526197A JP 27526197 A JP27526197 A JP 27526197A JP H1192343 A JPH1192343 A JP H1192343A
- Authority
- JP
- Japan
- Prior art keywords
- acid
- hair
- unsaturated fatty
- hair growth
- growth period
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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- Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
- Cosmetics (AREA)
Abstract
Description
【0001】[0001]
【発明の属する技術分野】本発明は、毛髪成長期延長剤
に関する技術分野に属する発明である。より詳細には、
毛髪伸長の促進をすることによって毛周期における成長
期を維持又は延長する毛髪成長期延長剤に関する。TECHNICAL FIELD The present invention belongs to the technical field related to a hair growth period extending agent. More specifically,
The present invention relates to a hair growth period extending agent that maintains or prolongs the anagen phase in the hair cycle by promoting hair elongation.
【0002】[0002]
【従来の技術】高齢化社会、ストレス社会といわれる現
代社会では、頭部毛髪が様々な原因により脱毛の危機に
さらされる機会がますます多くなってきている。これに
対応して、より優れた「育毛料」を提供すべく様々な試
みがなされている。育毛料が毛髪に与える効果として主
なものに、発毛誘導効果(発毛促進効果,成長期誘導
効果),毛髪を太くする効果,毛髪成長期延長効
果,5α−レダクターゼ阻害効果,血行促進効果,
殺菌効果,フケ防止効果,保湿効果,抗酸化効
果等の効果が挙げられる。2. Description of the Related Art In a modern society referred to as an aging society or a stressed society, there are more and more opportunities for head hair to be exposed to the risk of hair loss due to various causes. In response to this, various attempts have been made to provide a more excellent “hair restoration”. The main effects of hair restoration on hair are hair growth inducing effect (hair growth promotion effect, growth period induction effect), hair thickening effect, hair growth period extension effect, 5α-reductase inhibitory effect, and blood circulation promotion effect. ,
The effects include a bactericidal effect, an anti-dandruff effect, a moisturizing effect, and an antioxidant effect.
【0003】しかしながら、前記のように種々の試みが
なされているにもかかわらず、従来の育毛料では、その
脱毛防止,発毛効果等の育毛作用は必ずしも十分なもの
ではなかった。これはおそらく脱毛の原因がさまざまで
あり、また発毛の機構も非常に複雑であるためと考えら
れている。今まで提供されている「育毛料」は、脱毛を
比較的大雑把な概念、言い換えれば漫然と「脱毛」とい
う現象のみを捉えて開発されており、そのメカニズムに
まで突っ込んで着目して開発されたものは決して多くな
い。その大きな理由が、これらのメカニズムに着目した
育毛効果を簡便に検定することが可能な育毛薬剤検定方
法が十分に提供されていなかったという面を否定できな
いことである。特に上記の毛髪成長期延長効果を検定
する育毛薬剤検定方法の確立は難しく、結果としてこれ
まで提供されてきた育毛料は、毛周期の成長期へと毛髪
を誘導して育毛する上記の発毛誘導効果に着目したも
のや,の5α−レダクターゼ阻害効果に着目したもの
が多かった。[0003] However, despite various attempts as described above, conventional hair restorers have not always had sufficient hair restoring effects such as hair loss prevention and hair growth effects. This is probably due to various causes of hair loss and the very complex mechanism of hair growth. The `` hair restoration '' provided so far has been developed by capturing only the relatively rough concept of hair loss, in other words, simply and simply the phenomenon of `` hair loss '', and was developed by focusing on the mechanism Is not much. The major reason is that it cannot be denied that a hair-growth drug assay method capable of simply testing the hair-growth effect focusing on these mechanisms has not been sufficiently provided. In particular, it is difficult to establish a hair-growth drug assay method for assaying the above-described hair growth period extension effect, and as a result, the hair growth materials provided so far induce the hair to the growth period of the hair cycle, and the hair growth described above. Many focused on the induction effect and many focused on the 5α-reductase inhibitory effect.
【0004】[0004]
【発明が解決しようとする課題】そこで、本発明者はin
vitroで行う簡便な上記の毛髪成長期延長効果を検定
する育毛薬剤検定方法を確立し、その育毛薬剤検定方法
を用いて上記の毛髪成長期延長効果を有する成分を有
効成分とする毛髪成長期延長剤を見出すことを目指し
た。Therefore, the present inventor has proposed in
Establish a simple hair growth drug assay method for assaying the above-described hair growth period extending effect carried out in vitro , and use the hair growth agent assay method to extend the hair growth period using a component having the above-described hair growth period extending effect as an active ingredient. The aim was to find an agent.
【0005】本願は、その一連の課題のうち、特に上記
の毛髪成長期延長効果を有する成分を見出し、これを
有効成分とする毛髪成長期延長剤を提供することをその
課題とする。The object of the present invention is to find a component having the above-mentioned effect of extending the hair growth period, and to provide a hair growth period extending agent containing the same as an active ingredient.
【0006】[0006]
【課題を解決するための手段】本発明者は、種々の物質
における上記の毛髪成長期延長効果を有する成分につ
いて、本発明者が見出した育毛薬剤検定方法を用いて鋭
意検討したところ、不飽和脂肪酸及びその誘導体に、所
望する毛髪成長期延長効果が認められることを見出し、
本発明を完成した。Means for Solving the Problems The present inventors have conducted intensive studies on the components having the above-mentioned effect of extending the hair growth period in various substances using the hair growth drug assay method found by the present inventors. It has been found that a desired hair growth period extending effect is recognized in fatty acids and derivatives thereof,
The present invention has been completed.
【0007】すなわち本発明は、不飽和脂肪酸及び/又
はその誘導体を有効成分とする毛髪成長期延長剤を提供
する。この不飽和脂肪酸は、通常、式 Cn Hm O2 〔炭素原子数nは12〜28,水素原子数m=n+1〜
2n−2〕で表される不飽和脂肪酸である。That is, the present invention provides a hair growth period prolonging agent comprising an unsaturated fatty acid and / or a derivative thereof as an active ingredient. The unsaturated fatty acids are generally of the formula C n H m O 2 [number n of carbon atoms 12 to 28, hydrogen atoms m = n +. 1 to
2n-2].
【0008】そして、さらにこれらの不飽和脂肪酸のう
ち、その二重結合部分がcis 体である不飽和脂肪酸を選
択することが好ましい。これらの条件を満たす不飽和脂
肪酸として、例えばドデセン酸,トリデセン酸,ヘプタ
デセン酸,ペトロセリン酸,パルミトレイン酸,オレイ
ン酸,バクセン酸,リノール酸,α−リノレン酸,γ−
リノレン酸,エイコセン酸,エイコサトリエン酸,エル
カ酸(cis-13-ドコセン酸),ドコサジエン酸,ドコサト
リエン酸,ドコサテトラエン酸,ドコサペンタエン酸,
アラキドン酸,ネルボン酸又はドコサヘキサエン酸から
なる群の不飽和脂肪酸から選ばれる1種又は2種以上の
不飽和脂肪酸を挙げることができる。Further, it is preferable to select, from these unsaturated fatty acids, unsaturated fatty acids whose double bond part is a cis form. As unsaturated fatty acids satisfying these conditions, for example, dodecenoic acid, tridecenoic acid, heptadecenoic acid, petroselinic acid, palmitoleic acid, oleic acid, vaccenic acid, linoleic acid, α-linolenic acid, γ-
Linolenic acid, eicosenoic acid, eicosatrienoic acid, erucic acid (cis-13-docosenoic acid), docosadienoic acid, docosatrienoic acid, docosatetraenoic acid, docosapentaenoic acid,
One or more unsaturated fatty acids selected from the group consisting of arachidonic acid, nervonic acid and docosahexaenoic acid can be mentioned.
【0009】なお、これらの不飽和脂肪酸のうち、ペト
ロセリン酸,ドコサヘキサエン酸又はオレイン酸を選択
することが特に好ましい。[0009] Among these unsaturated fatty acids, it is particularly preferable to select petroselinic acid, docosahexaenoic acid or oleic acid.
【0010】前記したように、本発明において「毛髪成
長期延長剤」は、主に後述する育毛薬剤検定方法によっ
て少なくとも毛包上皮系細胞の分裂増殖活性若しくは毛
幹伸長活性を維持又は促進することで毛髪の成長期を延
長する効果を有する成分を有効成分として配合した、特
に上記の毛髪成長期延長効果に着目した毛髪関連薬剤
であり、いわば「個別効能育毛料」としての特徴を有す
る。As described above, in the present invention, the "hair growth period prolonging agent" is used to maintain or promote at least the mitotic proliferation activity or hair shaft elongation activity of hair follicle epithelial cells by the hair growth drug assay method described below. It is a hair-related drug that has been combined with a component having an effect of extending the growth period of hair as an active ingredient, and particularly focuses on the above-described effect of extending the growth period of hair, and has the characteristics of a so-called “individually effective hair restorer”.
【0011】この「毛髪成長期延長剤」は、例えば毛根
近傍における毛包上皮系細胞の増殖が緩徐であること等
により成長期が短くなって、相対的に成長期毛よりも休
止期毛の割合が多くなってしまうことに起因する脱毛症
に特に有効な薬剤である。また、他の個別効能を有する
育毛料と組み合わせて用いることにより、幅広くの脱毛
症においては,総合的かつ相乗的な効果を上げることが
可能である。すなわち、この「毛髪成長期延長剤」は、
前記した〜の効果を包含する概念を有する一般的な
育毛料用途とは一線を画する用途を有するものである。The "hair growth period extending agent" shortens the anagen phase due to, for example, slow growth of hair follicle epithelial cells in the vicinity of the root of the hair, so that the telogen hair is relatively shorter than the anagen hair. It is a particularly effective drug for alopecia caused by an increased ratio. Also, by using in combination with other hair restorers having individual effects, comprehensive and synergistic effects can be achieved in a wide range of alopecia. That is, this "hair growth period extender"
A general purpose hair restoring application having the concept including the above-mentioned effects (1) to (4) is one that has a distinctive purpose.
【0012】[0012]
【発明の実施の形態】以下、本発明の実施の形態につい
て説明する。本発明毛髪成長期延長剤は、上述したよう
に不飽和脂肪酸及び/又はその誘導体をその有効成分と
する毛髪関連薬剤である。Embodiments of the present invention will be described below. The hair growth period extending agent of the present invention is a hair-related agent containing an unsaturated fatty acid and / or a derivative thereof as an active ingredient as described above.
【0013】本発明毛髪成長期延長剤の有効成分となり
得る不飽和脂肪酸は、人体に対する安全性が担保されて
いる限りにおいて特に限定されるものではないが、炭素
数が12以上28以下で,かつ二重結合たcis 体をとる
不飽和脂肪酸が好ましい。具体的には例えばドデセン
酸,トリデセン酸,ヘプタデセン酸,ペトロセリン酸
(cis-6-オクタデセン酸),パルミトレイン酸(cis-9-
ヘキサデセン酸),オレイン酸,バクセン酸,リノール
酸,α−リノレン酸,γ−リノレン酸,エイコセン酸,
エイコサトリエン酸,エルカ酸(cis-13-ドコセン酸),
ドコサジエン酸,ドコサトリエン酸,ドコサテトラエン
酸,ドコサペンタエン酸,アラキドン酸,ネルボン酸
(cis-15- テトラコセン酸)又はドコサヘキサエン酸等
を挙げることができる。The unsaturated fatty acid which can be used as an active ingredient of the hair growth period prolonging agent of the present invention is not particularly limited as long as its safety against the human body is ensured. Unsaturated fatty acids having a double bond in the cis form are preferred. Specifically, for example, dodecenoic acid, tridecenoic acid, heptadecenoic acid, petroselinic acid (cis-6-octadecenoic acid), palmitoleic acid (cis-9-
Hexadecenoic acid), oleic acid, vaccenic acid, linoleic acid, α-linolenic acid, γ-linolenic acid, eicosenoic acid,
Eicosatrienoic acid, erucic acid (cis-13-docosenoic acid),
Docosadienoic acid, docosatrienoic acid, docosatetraenoic acid, docosapentaenoic acid, arachidonic acid, nervonic acid (cis-15-tetracosenoic acid) or docosahexaenoic acid.
【0014】これらの不飽和脂肪酸のうち、ペトロセリ
ン酸,ドコサヘキサエン酸及び/又はオレイン酸を選択
することが特に好ましい。[0014] Of these unsaturated fatty acids, it is particularly preferred to select petroselinic acid, docosahexaenoic acid and / or oleic acid.
【0015】なお、これらの不飽和脂肪酸の原料が、植
物、例えばコリアンダー,コンフリー等である不飽和脂
肪酸を本発明毛髪成長期延長剤の有効成分として選択す
ることもまた好ましい。[0015] It is also preferable that the unsaturated fatty acid used as a raw material of the plant is, for example, an unsaturated fatty acid such as coriander or comfrey, as an active ingredient of the present invention.
【0016】また本発明毛髪成長期延長剤の有効成分と
して、これらの不飽和脂肪酸の誘導体を選択することも
可能である。It is also possible to select derivatives of these unsaturated fatty acids as active ingredients of the hair growth period extender of the present invention.
【0017】具体的にこの誘導体の態様としては、例え
ば不飽和脂肪酸モノグリセライド、不飽和脂肪酸ジグリ
セライド、不飽和脂肪酸トリグリセライド、製薬学上許
容される不飽和脂肪酸塩,不飽和脂肪酸エステル,不飽
和脂肪酸アミド、不飽和脂肪酸二塩基酸若しくはその製
薬学上許容される塩等が挙げられる。これらの不飽和脂
肪酸又はその誘導体は、それぞれ通常公知の方法で製造
することができる。また、市販品を用いることもできる
のは勿論である。Specific examples of the derivative include unsaturated fatty acid monoglyceride, unsaturated fatty acid diglyceride, unsaturated fatty acid triglyceride, pharmaceutically acceptable unsaturated fatty acid salt, unsaturated fatty acid ester, unsaturated fatty acid amide, Examples include unsaturated fatty acid dibasic acids or pharmaceutically acceptable salts thereof. Each of these unsaturated fatty acids or derivatives thereof can be produced by a generally known method. Of course, a commercially available product can also be used.
【0018】これらの不飽和脂肪酸又はその誘導体は、
強い毛包上皮系細胞の増殖作用及び毛幹の伸長作用を有
しており、これらを単独で又は2種以上を組み合わせ
て、本発明毛髪成長期延長剤の有効成分とすることがで
きる。These unsaturated fatty acids or derivatives thereof are
It has strong hair follicle epithelial cell proliferation activity and hair shaft elongation activity, and these can be used alone or in combination of two or more as active ingredients of the hair growth period extending agent of the present invention.
【0019】本発明毛髪成長期延長剤における上記不飽
和脂肪酸及び/又はその誘導体の配合量は、本発明毛髪
成長期延長剤の具体的形態等に応じて適宜選択し得るも
のであり、特に限定されるべきものではないが、概ね本
剤全体に対して1.0×10-9重量%以上,20.0重
量%以下、好ましくは同0.01重量%以上,10.0
重量%以下となるように配合される。The amount of the unsaturated fatty acid and / or its derivative in the hair growth period extender of the present invention can be appropriately selected according to the specific form of the hair growth period extender of the present invention, and is particularly limited. Although not to be done, it is generally not less than 1.0 × 10 −9 wt% and not more than 20.0 wt%, preferably not less than 0.01 wt% and not more than 10.0 wt% based on the whole agent.
% By weight or less.
【0020】本剤全体に対して1.0×10-9重量%未
満の配合量では、本発明の所期の効果である毛包上皮系
細胞増殖作用に基づく毛髪成長期延長効果が十分に発揮
されず好ましくなく、同20.0重量%を超えて配合し
ても、配合量の増加に見合った効果の増大を見込めない
ばかりではなく、製剤上支障をきたす傾向が顕著となり
好ましくない。When the amount is less than 1.0 × 10 -9 % by weight based on the whole preparation, the effect of extending the hair growth period based on the hair follicle epithelial cell proliferation effect, which is the desired effect of the present invention, is sufficiently achieved. It is not preferred because it is not exhibited, and if it is added in excess of 20.0% by weight, not only is it not possible to expect an increase in the effect corresponding to the increase in the amount added, but also there is a marked tendency to cause problems in the formulation, which is not preferable.
【0021】このようにして、上記不飽和脂肪酸又はそ
の誘導体を有効成分として配合した本発明毛髪成長期延
長剤は、優れた毛包上皮系細胞増殖作用に基づく毛髪成
長期延長効果を有し、前記したように、例えば毛根近傍
における毛包上皮系細胞の増殖が緩徐であること等によ
り成長期が短くなって,相対的に成長期毛よりも休止期
毛の割合が多くなってしまうことに起因する脱毛症に特
に有効な薬剤である。また、他の個別効能を有する育毛
料と組み合わせて用いることにより、特定の脱毛症にお
いては相乗的な効果を上げることもまた可能である。Thus, the hair growth-prolonging agent of the present invention containing the unsaturated fatty acid or its derivative as an active ingredient has a hair growth period-prolonging effect based on an excellent hair follicle epithelial cell proliferation action. As described above, for example, the growth of hair follicle epithelial cells in the vicinity of the hair root is slow, so that the anagen is shortened and the ratio of telogen hair is relatively larger than that of anagen hair. It is a particularly effective drug for alopecia caused by it. It is also possible to increase the synergistic effect in specific alopecia by using it in combination with other hair restorers having individual effects.
【0022】本発明毛髪成長期延長剤の所期の効果であ
る毛髪成長期延長効果における本質的作用である、毛周
期における成長期の維持又は延長作用を特定する手段
は、その特定法自体がその作用を特定するために妥当な
ものである限り特に限定されず、例えばin vitro にお
ける特定法も,in vivo における特定法も用いることが
できるが、その簡便性と有効性を考慮すると、in vitr
o における特定法を用いることが好ましい。The means for specifying the maintenance or prolongation of the growth period in the hair cycle, which is the essential effect of the desired effect of the hair growth period prolonging agent of the present invention, is the method itself. is not particularly limited as far as it is reasonable to identify its action, also specific method in example in vitro, can be used, but a specific method in the in vivo, considering its simplicity and effectiveness, in Vitr
It is preferable to use the identification method in o .
【0023】以下、このin vitro における特定法の一
つである、毛包上皮系培養細胞の増殖効果を検討するこ
とを特徴とする特定法について簡単に説明する(より具
体的には、実施例において記載する。)。The following is a brief description of one of the in vitro identification methods, which is characterized by examining the effect of proliferating cultured follicular epithelial cells (more specifically, in Example 1). Described in.).
【0024】すなわちこの特定法は、「毛包上皮系培養
細胞に無血清培地中で対象物質を接触させて、その細胞
の増殖活性の有無及び/又は強弱を特定することによ
り、その対象物質の毛周期における成長期を延長する効
果を検定する一次スクリーニングを行い、この一次スク
リーニングにより効果の認められた対象物質について、
この対象物質を添加した無血清培地中でヒト毛包を培養
し、毛幹の伸長が認められた対象物質をさらに選択する
二次スクリーニングを行うことを特徴とする育毛薬剤検
定方法」、すなわち毛髪の伸長に直接的に関係する毛包
上皮系細胞に着目し、この培養細胞を用いることによっ
て、所望する毛周期における成長期を延長する効果を特
定する、in vitroの育毛薬剤検定方法である。[0024] That is, this identification method is as follows: "A culture of hair follicle epithelial cells is brought into contact with a target substance in a serum-free medium, and the presence or absence and / or strength of the proliferation activity of the cell is specified. Perform a primary screening to test the effect of prolonging the anagen phase in the hair cycle, and for the target substance that has been confirmed to be effective by this primary screening,
A hair-growth drug assay method comprising culturing a human hair follicle in a serum-free medium to which the target substance has been added, and performing a secondary screening to further select a target substance in which hair shaft elongation has been observed. This is an in vitro hair growth drug assay method that focuses on hair follicle epithelial cells that are directly involved in hair growth and specifies the effect of extending the anagen phase in a desired hair cycle by using the cultured cells.
【0025】この育毛薬剤検定方法の一次スクリーニン
グにおいては、においては、動物(ヒトを含む,以下同
様である)の毛包上皮系細胞を単離して得た培養細胞で
ある「毛包上皮系培養細胞」に対象物質を接触させて、
その増殖の有無及び/又は強弱を特定する。In the primary screening of this method for assaying a hair-growth agent, in the first step, "hair follicle epithelial culture," which is a cultured cell obtained by isolating hair follicle epithelial cells of animals (including humans, the same applies hereinafter) Contact the target substance with the cells,
The presence or absence and / or strength of the proliferation is specified.
【0026】毛包上皮系細胞は、特に毛根近傍の外毛根
鞘細胞とマトリクス細胞等の毛包由来の上皮系細胞のこ
とを指し、内側の毛乳頭細胞は除外される。Hair follicle epithelial cells particularly refer to epidermal cells derived from hair follicles such as outer root sheath cells and matrix cells near the hair root, and the inner dermal papilla cells are excluded.
【0027】毛周期における成長期は、まさにこの毛幹
の伸長,すなわち毛包上皮系細胞が分裂して増殖してい
る時期であり、同退行期及び休止期はこれが鈍化して休
止する時期である。The anagen in the hair cycle is exactly the elongation of the hair shaft, that is, the time when the hair follicle epithelial cells are dividing and proliferating, and the catagen and the telogen are the periods when they slow down and pause. is there.
【0028】つまり、毛周期における成長期を延長又は
維持する物質は、その投与により毛包上皮系細胞の分裂
及び増殖活性を維持することによって、毛髪が毛周期に
おける退行期及び休止期への移行を防ぐ物質、すなわち
毛包上皮系細胞の増殖を促進又は維持し続ける物質であ
ることが結論付けられる。In other words, a substance that prolongs or maintains the anagen phase in the hair cycle maintains the division and proliferative activity of the hair follicle epithelial cells by its administration, whereby the hair shifts to the catagen and telogen phases in the hair cycle. It is concluded that this is a substance that prevents the growth of hair follicle epithelial cells, ie, a substance that continues to promote or maintain the proliferation of hair follicle epithelial cells.
【0029】また、二次スクリーニングにおいては、ヒ
ト毛包を培養し、毛幹の伸長が認められた対象物質をさ
らに選択する。この二次スクリーニングに際しては、ま
ず毛包断片を調製することが必要である。通常は、ヒト
の健毛部の頭髪の中から成長期の頭髪を選択して、この
毛髪の毛球部から約2mm上部を切断して毛包断片を調製
することができる。In the secondary screening, human hair follicles are cultured, and a target substance in which hair shaft elongation is observed is further selected. In this secondary screening, it is necessary to prepare a hair follicle fragment first. Normally, hair in the anagen phase is selected from the hair of human healthy hair, and the upper part of the hair bulb of this hair is cut about 2 mm to prepare a hair follicle fragment.
【0030】この毛包断片は、インシュリンを含む基礎
培地で培養すれば成長期の状態が比較的長期に維持され
るが、インシュリンを含有しない基礎培地で培養すると
1週間程度の短期間のうちに退行期の毛包に類似する状
態に変化する。This hair follicle fragment can maintain its growth phase for a relatively long time when cultured in a basal medium containing insulin, but can be cultured within a short period of about one week when cultured in a basal medium containing no insulin. It changes to a state resembling a catagen follicle.
【0031】従って、インシュリンを含まない基礎培地
中で培養する場合は、1週間程度のうちに評価すること
が好ましく、インシュリンを含む培地を用いる場合には
2週間程度まで培養期間を延長することが可能になる。
どちらの培地においても、この毛包を対象被験物質の存
在下で培養することにより、毛幹の伸長の程度を観察す
ることができる。Therefore, when culturing in a basal medium not containing insulin, it is preferable to evaluate within about one week, and when using a medium containing insulin, it is preferable to extend the culturing period to about two weeks. Will be possible.
In either medium, the degree of hair shaft elongation can be observed by culturing this hair follicle in the presence of the target test substance.
【0032】なお、この二次スクリーニングにおいて用
いる基礎培地は、一般的に用いられている動物細胞培養
用培地を用いることが可能であり、汚染防止のために抗
生物質や抗菌剤を添加することができる。As the basal medium used in this secondary screening, a commonly used medium for culturing animal cells can be used, and antibiotics or antibacterial agents can be added to prevent contamination. it can.
【0033】毛包の培養条件は、5%CO2 ・37℃等
の動物培養における通常の条件で行うことができる。培
養は通常5〜14日間行われる。ヒト毛包における毛幹
の伸長は、例えばミクロメーターを装着した実体顕微鏡
において目視で確認することができる。The hair follicles can be cultured under normal conditions for animal culture such as 5% CO 2 and 37 ° C. The culture is usually performed for 5 to 14 days. Elongation of the hair shaft in the human hair follicle can be visually confirmed, for example, with a stereomicroscope equipped with a micrometer.
【0034】このように上記一次スクリーニングと二次
スクリーニングとを行うことにより、毛周期における成
長期の維持又は延長作用並びに毛幹の伸長作用を特定す
ることができる。As described above, by performing the primary screening and the secondary screening, it is possible to specify the action of maintaining or prolonging the anagen phase and the action of elongating the hair shaft in the hair cycle.
【0035】なおin vitroの育毛薬剤検定方法として
は、他に対象物質を動物の毛乳頭細胞に作用させて、そ
の増殖促進効果を判定する方法も用いることができる。In addition, as an in vitro method for assaying a hair growth drug, a method in which a target substance is allowed to act on hair papilla cells of an animal to determine its growth promoting effect can be used.
【0036】また、in vivo における特定法としては、
例えば「ヌードマウスに対象物質を投与し,このヌード
マウスの体表の発毛部位の状態を特定して,対象物質の
毛周期における成長期を延長する効果を検定する育毛薬
剤検定方法」、すなわち原則的には無毛であるが、その
体表に経時的にその発毛部位が移動する特徴的な発毛を
するヌードマウスにおける発毛部位の広さと発毛部位の
移動速度を特定することによって、毛周期における成長
期の長さを検定する方法等を挙げることができる。In addition, as a specific method in vivo ,
For example, a "hair growth drug assay method in which a target substance is administered to a nude mouse, the state of the hair growth site on the body surface of the nude mouse is specified, and the effect of the target substance on the growth cycle in the hair cycle is assayed" To specify the width of hair growth sites and the speed of hair growth sites in nude mice that are hairless in principle, but whose hair growth sites move over time on the body surface over time. For example, a method of testing the length of the anagen phase in the hair cycle can be mentioned.
【0037】本発明毛髪成長期延長剤が採り得る剤型
は、外皮に適用可能な剤型であれば特に限定されず、例
えば液状,乳液,軟膏等を選択可能である。また、本発
明毛髪成長期延長剤の形態は任意であり、例えばトニッ
ク,ヘアークリーム,ムース,シャンプー,リンス,ク
リーム,乳液,化粧水,パック等の形態を採ることがで
きる。The dosage form that the hair growth period extender of the present invention can take is not particularly limited as long as it is a dosage form applicable to the outer skin, and for example, liquid, emulsion, ointment and the like can be selected. Further, the form of the hair growth period prolonging agent of the present invention is arbitrary, and for example, can be in the form of tonic, hair cream, mousse, shampoo, rinse, cream, milky lotion, lotion, pack and the like.
【0038】本発明毛髪成長期延長剤前記の必須成分に
加えて必要に応じて、かつ本発明の所期の効果を損なわ
ない限りにおいて、化粧品,医薬部外品,医薬品等にお
いて一般的に用いられる各種油性若しくは水性成分.保
湿剤,増粘剤,防腐剤,酸化防止剤,香料,色剤,各種
薬剤等を配合することができる。Hair growth period extender of the present invention Generally used in cosmetics, quasi-drugs, pharmaceuticals, etc., in addition to the above essential components, if necessary and as long as the desired effects of the present invention are not impaired. Various oily or aqueous components used. A humectant, a thickener, a preservative, an antioxidant, a fragrance, a coloring agent, various chemicals and the like can be added.
【0039】例えば、高級飽和脂肪酸,固形パラフィ
ン,流動パラフィン,シリコーン油,スクワラン,モノ
オレイン酸グリセリル,オリーブ油,高級アルコール等
の油分;グリセリン,ヒアルロン酸,プロピレングリコ
ール,マルチトール,アテロコラーゲン,乳酸ナトリウ
ム等の保湿剤;マルメロ粘質物,カルボキシビニルポリ
マー,キサンタンガム等の増粘剤;ニコチン酸アミド,
ニコチン酸ベンジル,ビタミンEアセテート,センブリ
抽出物,塩化カルプロニウム,アセチルコリン誘導体等
の血管拡張剤;セリン,メチオニン,アルギニン等のア
ミノ酸類;ビタミンB6 ,ビタミンE(若しくはその誘
導体),ビオチン,パントテン酸(若しくはその誘導
体)等のビタミン類;ニコチン酸,ニコチン酸メチル,
ニコチン酸トコフェロール等のニコチン酸エステル類;
セファランチン等の皮膚機能亢進剤;エストラジオール
等の女性ホルモン剤;グリチルレチン酸(若しくはその
誘導体)等の消炎剤;ヒノキチオール,ヘキサクロロフ
ェン,ベンザルコニウムクロリド,ビチオノール等の抗
菌剤;メントール等の清涼剤;サリチル酸,亜鉛(若し
くはその誘導体),乳酸(若しくはそのアルキルエステ
ル)等;クエン酸等の有機酸類等を配合することができ
る。本発明毛髪成長期延長剤の具体的処方は後述する。For example, oils such as higher saturated fatty acids, solid paraffin, liquid paraffin, silicone oil, squalane, glyceryl monooleate, olive oil, higher alcohols; glycerin, hyaluronic acid, propylene glycol, maltitol, atelocollagen, sodium lactate Humectant; thickener such as quince, carboxyvinyl polymer, xanthan gum; nicotinamide;
Vasodilators such as benzyl nicotinate, vitamin E acetate, assembly extract, carpronium chloride, and acetylcholine derivatives; amino acids such as serine, methionine, and arginine; vitamin B 6 , vitamin E (or derivatives thereof), biotin, pantothenic acid ( Or derivatives thereof); nicotinic acid, methyl nicotinate,
Nicotinic acid esters such as tocopherol nicotinate;
Skin function enhancers such as cepharanthin; female hormones such as estradiol; anti-inflammatory agents such as glycyrrhetinic acid (or a derivative thereof); antibacterial agents such as hinokitiol, hexachlorophen, benzalkonium chloride, bithionol; , Zinc (or a derivative thereof), lactic acid (or an alkyl ester thereof), and the like; and organic acids such as citric acid, and the like. The specific formulation of the hair growth period extender of the present invention will be described later.
【0040】[0040]
【実施例】以下、実施例等により本発明をさらに具体的
に説明するが、この実施例等により本発明の技術的範囲
が限定的に解釈されるべきものではない。なお、以下の
実施例等において「%」と表示され,かつ内容量を示す
ものは、特に断らない限り重量%を意味する。まず、本
実施例等において用いた植物の抽出物の毛髪成長期延長
作用を評価するためのin vitroの細胞増殖試験について
説明する。EXAMPLES Hereinafter, the present invention will be described more specifically with reference to Examples and the like, but the technical scope of the present invention should not be construed as being limited by these Examples and the like. In the following examples and the like, "%" and the content are indicated by weight unless otherwise specified. First, an in vitro cell proliferation test for evaluating the effect of the plant extract used in the Examples and the like on extending the hair growth period will be described.
【0041】〔試験例1〕 毛包上皮系培養細胞を用い
た細胞増殖試験 A.ヒト毛包上皮系細胞 1.ヒト毛包上皮系細胞の採取 外科手術の副産物として得られたヒト男性頭皮から毛周
期における成長期の毛包を実体顕微鏡下で機械的に採取
した。この成長期の毛包を1000 U/ml dispase・0.2 %
コラゲナーゼを含むダルベッコの改変MEM(DME
M)で30分間,37℃で処理し、注射針の先を用いて
dermal sheath やdermal papilla、毛球部上皮組織を除
去して、0.05%トリプシン・0.02%EDTAを含むリン
酸緩衝液〔PBS(−):(−)とはカルシウムイオン
やマグネシウムイオンを含まない意味である〕で5分
間,37℃で処理した。Test Example 1 Cell Proliferation Test Using Hair Follicle Epithelial Cultured Cells Human hair follicle epithelial cells 1. Collection of Human Hair Follicle Epithelial Cells Hair follicles during the anagen phase of the hair cycle were mechanically collected from a human male scalp obtained as a by-product of surgery under a stereoscopic microscope. 1000 U / ml dispase 0.2%
Dulbecco's modified MEM containing collagenase (DME
M) for 30 minutes at 37 ° C., using the tip of a syringe needle.
Phosphate buffer solution containing 0.05% trypsin and 0.02% EDTA after removing dermal sheath, dermal papilla and epithelium of hair bulb [PBS (-): (-) means that it does not contain calcium or magnesium ions Yes] for 5 minutes at 37 ° C.
【0042】次にコラーゲン(Type I)コーティングし
た培養皿に毛包を静置し、外殖片培養を行った。なおこ
の際の培地は、無血清培地〔Keratinocyte Growth Medi
um(KGM)〕を用いた(Keratinocyte Serum Free Me
diumを用いることもできる)。この培養の4〜5日後
に、毛包の培養皿への接着及び細胞の増殖が確認できた
時点で培地を交換し、これ以降2日おきに培地交換を行
った。Next, the hair follicle was allowed to stand on a culture dish coated with collagen (Type I), and explant culture was performed. The medium used in this case was a serum-free medium (Keratinocyte Growth Medication).
um (KGM)] (Keratinocyte Serum Free Me
dium can be used). After 4 to 5 days of this culture, when the attachment of the hair follicle to the culture dish and the proliferation of the cells were confirmed, the medium was replaced, and thereafter the medium was replaced every two days.
【0043】このようにして増殖させた細胞を、0.05wt
%トリプシン-0.02 %含有PBS(−)で37℃で5分
間処理した後、等量の0.1 %トリプシンインヒビターで
反応を停止させ、遠心処理(800×g,5 分間) を施して細
胞を回収した。The cells grown in this manner were added to 0.05 wt.
After treatment with PBS (-) containing 5% trypsin-0.02% at 37 ° C for 5 minutes, the reaction was stopped with an equal amount of 0.1% trypsin inhibitor, and the cells were collected by centrifugation (800 xg, 5 minutes). .
【0044】次に、細胞を上記の無血清培地に浮遊させ
て、5000 cells/cm2の密度でコラーゲンコーティング
(Type I)した培養皿に播種し、細胞がsubconfluentに
なるまで2日おきに培地交換を行い、再び0.05wt%トリ
プシン-0.02 %EDTA含有PBS(−)で37℃で5
分間処理した後、等量の0.1 %トリプシンインヒビター
で反応を停止させ、遠心処理(800×g,5 分間) を施し
て、これにより得られたヒト毛包上皮系細胞に細胞凍結
液(セルバンカー:ダイヤトロン製)を添加し、1.0
×106 cell/ml の濃度に調整して、各凍結チューブに
1.0×106 cellずつ入れ、これを凍結保存した。な
お、これらの細胞数は、血球算定板で算出した。Next, cells were resuspended in serum-free medium described above were seeded in culture dishes collagen-coated (Type I) at a density of 5000 cells / cm 2, culture medium every two days until the cells became subconfluent After the exchange, the cells were re-exchanged with PBS (−) containing 0.05 wt% trypsin-0.02% EDTA at 37 ° C. for 5 hours.
After treatment for 1 min, the reaction was stopped with an equal volume of 0.1% trypsin inhibitor, centrifuged (800 xg, 5 min), and the resulting human hair follicle epithelial cells were subjected to cell freezing (Cell Banker). : Diatron) and add 1.0
The concentration was adjusted to × 10 6 cells / ml, and 1.0 × 10 6 cells were added to each cryotube, and the cells were cryopreserved. In addition, these cell numbers were calculated with a blood cell counting plate.
【0045】2.対象物質のアッセイ 上記工程により得た毛包上皮系細胞の線維芽細胞混入率
(FB混入率)を測定(3000倍,5視野)し、その
結果FB混入率が3%以上のものは、アッセイの対象か
ら除外した。そして、この毛包上皮系細胞を培養フラス
コ中に播種後、これを0.25%トリプシン−0.02
%EDTA含有PBS(−)で処理した後、0.1%ト
リプシンインヒビターで反応を停止後、系を1500rp
m で5分間遠心処理を施し、上清を除去し、残渣にKG
M培地20mlを添加して、細胞懸濁液を調製した。2. Assay of target substance The fibroblast contamination rate (FB contamination rate) of the hair follicle epithelial cells obtained by the above steps was measured (3000 times, 5 visual fields). As a result, if the FB contamination rate was 3% or more, the assay was performed. Was excluded from the target. After inoculating the hair follicle epithelial cells into a culture flask, the cells were cultured at 0.25% trypsin-0.02%.
After treatment with PBS (-) containing 0.1% EDTA, the reaction was stopped with 0.1% trypsin inhibitor, and the system was incubated at 1500 rp.
centrifugation at 5 m for 5 minutes, remove the supernatant, and add KG to the residue.
A cell suspension was prepared by adding 20 ml of M medium.
【0046】0.2ml/well の割合で、96well-plate
(I型コラーゲンコーティングプレート:ファルコン社
製)に播種し(3.0×103cell/well)、細胞がウエ
ルの底に沈むまで約20分間室温下で放置した。その
後、37℃,5%CO2 で1日間培養を行い、所望する
ヒト毛包上皮系培養細胞を得た。A 96-well plate at a rate of 0.2 ml / well
(Type I collagen coated plate: manufactured by Falcon) (3.0 × 10 3 cells / well), and allowed to stand at room temperature for about 20 minutes until the cells sank to the bottom of the well. Thereafter, the cells were cultured for 1 day at 37 ° C. and 5% CO 2 to obtain desired human hair follicle epithelial cultured cells.
【0047】B.ラット毛包上皮系細胞 1.ラット毛包上皮系細胞の採取: (1)毛包の採取 新生児(3〜4日令)のラットを二酸化炭素で屠殺した
後、エタノールで消毒し、これらのラットの背部皮膚を
ハサミで採取した。次いで、この採取した背部皮膚を5
%PSF含有PBS(−)に浸漬後、皮膚脂肪層から下
の皮下脂肪や皮膜等を解剖用ハサミで除去した。 次い
で、再びこの背部皮膚を1%PSF含有PBS(−)に
浸し、さらにこれを0.25%トリプシン−0.02%
EDTA含有PBS(−)中に4℃で一晩浸した。B. Rat hair follicle epithelial cells 1. Collection of rat hair follicle epithelial cells: (1) Collection of hair follicles Newborn (3-4 day old) rats were sacrificed with carbon dioxide, disinfected with ethanol, and the back skin of these rats was collected with scissors. . Then, the collected back skin was
After immersion in PBS (-) containing% PSF, the subcutaneous fat, film, and the like below the skin fat layer were removed with scissors for dissection. Then, the back skin was immersed again in PBS (-) containing 1% PSF, and further immersed in 0.25% trypsin-0.02%.
It was immersed in PBS (-) containing EDTA at 4 ° C overnight.
【0048】このトリプシン溶液中における浸漬後、背
部皮膚の真皮層と表皮層をピンセットで剥がした後、真
皮層を0.35%コラゲナーゼ含有Ham's F12培
地〔組成(mg/L):l-Alanin(8.9),l-Arginine(HCl:21
1),l-Asparagine(13.2),l-Asparatic acid(13.3),l-Cys
teine (HCl:31.5),l-Glutamic acid(14.7),l-Glutamin
e(146),Glycine(7.5),l-Histidine (HCl:19),l-Isoleu
cine(3.9),l-leucine(13.1),l-Lysine(HCl:36.5),l-Met
hionine(4.5),l-Phenylalanin(5.0),Proline(34.5),l-S
erine(10.5),l-Threonine(11.9),l-Tryptophane(2.0),l
-Tyrosine(5.4),l-Valine(11.7),Biotine(0.0073),Chol
ine(Cl:14.0),VitaminB12(1.36),葉酸(1.32),Inositol
(18.0),Nicotinamide(0.037),パントテン酸(Ca:0.477),
VitaminB6(HCl:0.062),VitaminB2(0.038),VitaminB1(HC
l:0.337),CaCl2(2H2O:44.0),CuSO4・5H2O(0.0025),FeSO
4・7H2O(0.834),KCl(224.0),MgCl2(6H2O:122),"Proc.N
atl.Acad.Sci.USA,53,288(1965)" 以下同様である〕中
で、ハサミで裁断した。この裁断物を含む培地を37℃
で35分間浸透を行った(60rpm )。浸透後、このコ
ラゲナーゼ反応物中に塊状のものが見えなくなるまでピ
ペッティングを行い、DNase (10000 unit) 含有Ha
m's F12培地を添加し、5分間放置した。After immersion in this trypsin solution, the dermis layer and the epidermis layer of the back skin were peeled off with forceps, and the dermis layer was replaced with a Ham's F12 medium containing 0.35% collagenase [composition (mg / L): l- Alanin (8.9), l-Arginine (HCl: 21
1), l-Asparagine (13.2), l-Asparatic acid (13.3), l-Cys
teine (HCl: 31.5), l-Glutamic acid (14.7), l-Glutamin
e (146), Glycine (7.5), l-Histidine (HCl: 19), l-Isoleu
cine (3.9), l-leucine (13.1), l-Lysine (HCl: 36.5), l-Met
hionine (4.5), l-Phenylalanin (5.0), Proline (34.5), lS
erine (10.5), l-Threonine (11.9), l-Tryptophane (2.0), l
-Tyrosine (5.4), l-Valine (11.7), Biotine (0.0073), Chol
ine (Cl: 14.0), VitaminB12 (1.36), folate (1.32), Inositol
(18.0), Nicotinamide (0.037), Pantothenic acid (Ca: 0.477),
VitaminB6 (HCl: 0.062), VitaminB2 (0.038), VitaminB1 (HC
l: 0.337), CaCl 2 (2H 2 O: 44.0), CuSO 4・ 5H 2 O (0.0025), FeSO
4 · 7H 2 O (0.834) , KCl (224.0), MgCl 2 (6H 2 O: 122), "Proc.N
USA, 53, 288 (1965) ", the same applies hereinafter). The medium containing the cut product was cultured at 37 ° C.
For 35 minutes (60 rpm). After infiltration, pipetting was performed until no clumps were seen in the collagenase reaction product, and the DNase (10000 unit) -containing Ha
m's F12 medium was added and left for 5 minutes.
【0049】放置後、得られた懸濁液をさらにピペッテ
ィングした後、ナイロンメッシュ(Nytex 157 mesh) で
濾過した。懸濁液をPBS(−)で希釈し、次いでこの
希釈した懸濁液に遠心処理を施した(4℃,400rpm
,5分間)。遠心後、上清を除いて脂肪分を系から除
去した。After standing, the obtained suspension was further pipetted and filtered through a nylon mesh (Nytex 157 mesh). The suspension was diluted with PBS (−), and the diluted suspension was centrifuged (4 ° C., 400 rpm).
, 5 minutes). After centrifugation, the fat was removed from the system except the supernatant.
【0050】次いで、残渣にPBS(−)を添加して懸
濁後、これにさらに遠心処理を施した〔(4℃,400
rpm ,5分間)×3回〕。この遠心操作により得られた
残渣が、ラットの背部皮膚における毛包である。Next, PBS (-) was added to the residue to suspend it, which was further centrifuged [(4 ° C., 400
rpm, 5 minutes) x 3 times]. The residue obtained by this centrifugation is a hair follicle in the back skin of the rat.
【0051】(2)毛包上皮系細胞の採取 上記操作により得られた毛包に、0.25%トリプシン
−0.02%EDTA含有PBS(−)を5ml添加し
て、細胞懸濁液を37℃で5分間インキュベートした。(2) Collection of hair follicle epithelial cells To the hair follicle obtained by the above operation, 5 ml of PBS (-) containing 0.25% trypsin-0.02% EDTA was added, and the cell suspension was added. Incubate at 37 ° C for 5 minutes.
【0052】インキュベート終了後、等量の0.1%ト
リプシンインヒビターとHam'sF12培地を添加し
て、細胞懸濁液をセルストレーナー(100 μm Nalgene
社製)で濾過後、この細胞懸濁液に遠心処理を施した
(4℃,1500rpm ,5分間)。この系から上清を除
去して、残渣として所望する毛包上皮系細胞を得た。こ
の毛包上皮系細胞に細胞凍結液(セルバンカー:ダイヤ
トロン製)を添加し、1.5×107 cell/ml の濃度に
調整して、各凍結チューブに1.5×107 cellずつ入
れ、これを凍結保存した。After completion of the incubation, equal amounts of 0.1% trypsin inhibitor and Ham's F12 medium were added, and the cell suspension was washed with a cell strainer (100 μm Nalgene).
The suspension was centrifuged (4 ° C., 1500 rpm, 5 minutes). The supernatant was removed from this system to obtain the desired hair follicle epithelial cells as a residue. The hair follicle epithelial cells in cell freezing solution (Cell Banker: Dia Tron Ltd.) was added, was adjusted to a concentration of 1.5 × 10 7 cell / ml, 1.5 × each 10 7 cell to each cryotubes And stored frozen.
【0053】2.毛包上皮系細胞の前培養:系に混入し
ている線維芽細胞を可能な限り系から除去するために、
上記工程により得られた毛包上皮系細胞の前培養を行っ
た。以下、その手順について説明する。2. Preculture of hair follicle epithelial cells: To remove fibroblasts from the system as much as possible,
Preculture of hair follicle epithelial cells obtained by the above steps was performed. Hereinafter, the procedure will be described.
【0054】37℃の恒温槽で、上記工程により得た凍
結細胞を融解した。次いでFAD培地〔Ham's F1
2培地(後述)とMEN培地を容量比で3対1で混合し
たものに、インシュリン(5.0μg/ml),ハイドロコルチ
ゾン(0.45μg/ml),エピダーマルグロウスファクター
(EGF)(10.0ng/ml),コレラトキシン(10-9M)及びウシ
胎児血清(10 %) を含有させた培地、以下同様である〕
を添加し、細胞溶液を希釈して系に遠心処理を施した
(10℃以下,1500rpm ,5分間)。遠心後、上清
を除去し、系にFAD培地を添加して、細胞塊が認めら
れなくなるまでピペッティングを繰り返した。The frozen cells obtained in the above steps were thawed in a thermostat at 37 ° C. Then, FAD medium [Ham's F1
2 medium (to be described later) and MEN medium at a volume ratio of 3: 1 mixed with insulin (5.0 μg / ml), hydrocortisone (0.45 μg / ml), epidermal growth factor
A medium containing (EGF) (10.0 ng / ml), cholera toxin (10 -9 M) and fetal calf serum (10%), and so on.]
Was added to dilute the cell solution, and the system was centrifuged (10 ° C. or lower, 1500 rpm, 5 minutes). After centrifugation, the supernatant was removed, FAD medium was added to the system, and pipetting was repeated until no cell mass was observed.
【0055】次いで、FAD培地で細胞濃度が2.5×
105 cell/ml になるように調整した。I型コラーゲン
でコーティングした75cm3 の培養フラスコに細胞を播
種して、これを37℃,5%CO2 で一晩培養した。培
養後、系をPBS(−)で洗浄し、0.25%トリプシ
ン−0.02%EDTA含有PBS(−)を添加して、
これを37℃,5%CO2 で4分間インキュベートし
た。次に、系にトリプシン溶液と等量の0.1%トリプ
シンインヒビターを添加して、1回軽くゆすった後で上
清を除去して、系に混入している線維芽細胞を除去し
た。Next, the cell concentration was 2.5 × in the FAD medium.
It was adjusted to be 10 5 cells / ml. The cells were seeded in a 75 cm 3 culture flask coated with type I collagen, and cultured overnight at 37 ° C. and 5% CO 2 . After culturing, the system was washed with PBS (-), and PBS (-) containing 0.25% trypsin-0.02% EDTA was added.
This was incubated at 37 ° C., 5% CO 2 for 4 minutes. Next, 0.1% trypsin inhibitor of the same volume as the trypsin solution was added to the system, and the mixture was slightly shaken once, and then the supernatant was removed to remove fibroblasts contaminating the system.
【0056】さらに、系にKGM培地〔表皮角化細胞基
礎培地(Keratinocyto growth medium):Keratinocyto b
asal medium (KBM培地(改変MCDB153培地
(クローネティックス社製)))に,ウシ脳下垂体エキ
ス(BPE)(0.4vol%),インシュリン(0.5μm/ml),ハイドロ
コルチゾン(0.5μm/ml),h-EGF(0.1 ng/ml)を添加した培
地、以下同様である〕を添加し、37℃,5%CO2 で
3日間培養した。Furthermore, KGM medium [Keratinocyto growth medium: Keratinocytob) was added to the system.
bovine pituitary extract (BPE) (0.4 vol%), insulin (0.5 μm / ml), hydrocortisone (0.5 μm / ml) in an asal medium (KBM medium (modified MCDB153 medium (Clonetics))) , h-EGF (0.1 ng / ml), the same applies hereinafter), and cultured at 37 ° C. and 5% CO 2 for 3 days.
【0057】3.対象物質のアッセイ 上記工程により得た毛包上皮系細胞を播種した培養フラ
スコの線維芽細胞混入率(FB混入率)を測定(300
0倍,5視野)し、その結果FB混入率が2%以上のも
のは、アッセイの対象から除外した。3. Assay of target substance Measure the fibroblast contamination rate (FB contamination rate) of the culture flask in which the hair follicle epithelial cells obtained by the above steps were seeded (300).
(0 times, 5 fields of view), and as a result, those with a FB contamination rate of 2% or more were excluded from the subject of the assay.
【0058】系をPBS(−)で洗浄し、0.25%ト
リプシン−0.02%EDTA含有PBS(−)を添加
して、これを37℃で3分間インキュベートした。次い
で上皮系細胞と線維芽細胞とのトリプシンに対する反応
性の違いを利用して,系から線維芽細胞を除去するため
に、トリプシンを除去し、再び0.25%トリプシン−
0.02%EDTA含有PBS(−)を添加して、37
℃,20rpm で5分間振盪した。The system was washed with PBS (-), PBS (-) containing 0.25% trypsin-0.02% EDTA was added, and this was incubated at 37 ° C for 3 minutes. Taking advantage of the difference in reactivity of epithelial cells and fibroblasts to trypsin, trypsin was removed and 0.25% trypsin-
PBS (-) containing 0.02% EDTA was added, and 37
The mixture was shaken at 20 rpm for 5 minutes.
【0059】次いで、細胞のはがれを顕微鏡下で確認し
た後、10%FBS含有DMEM培地を添加してピペッ
ティングを行い、系を1500rpm で5分間遠心処理を
施した。上清を除去し、KGM培地を添加して、細胞塊
がなくなるまでピペッティングを行った。Next, after confirming the detachment of the cells under a microscope, 10% FBS-containing DMEM medium was added, pipetting was performed, and the system was centrifuged at 1500 rpm for 5 minutes. The supernatant was removed, KGM medium was added, and pipetting was performed until the cell mass disappeared.
【0060】懸濁液をセルストレーナー(100 μm Nalg
ene 社製)で濾過後、懸濁液中の生細胞数を血球算定板
で算出し、系にKGM培地を添加して、系の中の細胞濃
度が2.0×104cell/mlになるように調整した。次い
で、0.2ml/well の割合で、96well-plate(I型コ
ラーゲンコーティングプレート:ファルコン社製)に播
種し(4.0×103cell/well)、細胞がウエルの底に
沈むまで約20分間室温下で放置した。その後、37
℃,5%CO2 で1日間培養を行い、所望するヒト毛包
上皮系培養細胞を得た。The suspension was added to a cell strainer (100 μm Nalg).
After filtration with a hemocytometer, the KGM medium was added to the system to bring the cell concentration in the system to 2.0 × 10 4 cells / ml. It was adjusted to become. Subsequently, the cells were seeded at a rate of 0.2 ml / well on a 96-well-plate (type I collagen coated plate: manufactured by Falcon) (4.0 × 10 3 cells / well), and about 20 × 10 3 cells / well until the cells sank to the bottom of the well. It was left at room temperature for minutes. Then 37
Culturing was performed at 5 ° C. and 5% CO 2 for 1 day to obtain desired human hair follicle epithelial cells.
【0061】C.試験培地の調製: (1)対象物質添加培地の調製 対象物質を培地全体に対して1.0×10-6mol/l ,溶
媒(DMSO)濃度が0.1%となるように、前記KB
M培地に添加した。C. Preparation of test medium: (1) Preparation of culture medium containing target substance The target substance was adjusted to 1.0 × 10 −6 mol / l based on the whole medium, and the KB was adjusted so that the concentration of the solvent (DMSO) became 0.1%.
M medium.
【0062】(2)コントロール培地の調製 ・ネガティブコントロール:KBM培地に、DMSO
(溶媒)を最終濃度が0.1%となるように添加した培
地を調製した。 ・ポジティブコントロール:ネガティブコントロール培
地に、最終濃度が5μg/mlのインシュリン,0.5μg/
mlのハイドロコーチゾンを添加して調製した。(2) Preparation of control medium • Negative control: DMSO was added to KBM medium.
(Solvent) was added to the medium to prepare a final concentration of 0.1%. -Positive control: In a negative control medium, a final concentration of 5 μg / ml insulin, 0.5 μg / ml
It was prepared by adding ml of hydrocortisone.
【0063】D.対象物質培地交換:上記A,Bにおい
てヒト毛包上皮系培養細胞及びラット毛包上皮系培養細
胞を調製した96well-plate中のKGM培地を、対象物
質添加培地及びコントロール培地(200μl/well)と
交換して、交換後37℃,5%CO2 で2日間培養し
た。D. Object substance culture medium exchange: The KGM medium in the 96-well-plate prepared with human hair follicle epithelial cell culture cells and rat hair follicle epithelial cell culture cells in A and B above was replaced with a target substance-added medium and a control medium (200 μl / well). After the exchange, the cells were cultured at 37 ° C. and 5% CO 2 for 2 days.
【0064】なお、この培地の交換はウエル内のKGM
培地を,底面に付着している細胞を傷つけないように留
意しつつマルチピペットで抜いて、その後速やかに対象
物質添加培地等をウエルの両端から添加することにより
行った。The medium was replaced with the KGM in the well.
The medium was removed with a multipipette while taking care not to damage the cells adhering to the bottom surface, and then the medium containing the target substance was immediately added from both ends of the well.
【0065】E.細胞増殖の測定:アラマーブルー(ala
mar blue:アラマーバイオサイエンス社製) を培地量
(容量)に対して、1/10量を添加して、37℃(5
%CO2 )で6時間インキュベートした。インキュベー
ト後、系の595nm及び570nmでの吸光度をマイクロ
プレートリーダー(Micro plate reader:Bio RAD社製)
を用いて測定し、下記計算式に従って,細胞増殖度を算
出した。E. Cell proliferation measurement: alamar blue (ala
mar blue (manufactured by Alamar Biosciences, Inc.) was added at 1/10 the volume (volume) of the medium, and the mixture was added at 37 ° C (5 ° C).
% CO 2 ) for 6 hours. After the incubation, the absorbance of the system at 595 nm and 570 nm was measured using a microplate reader (Micro RAD).
The cell proliferation was calculated according to the following formula.
【0066】[0066]
【数1】 (Equation 1)
【0067】さらに細胞増殖促進指標を、下記計算式に
従って算出した。Further, the cell growth promotion index was calculated according to the following formula.
【数2】 (Equation 2)
【0068】F.結果:測定した上記各対象物質におけ
る、上記細胞増殖促進指標を下記第1表に示す。F. Results: Table 1 below shows the cell growth promotion index of each of the measured target substances.
【表1】 [Table 1]
【0069】この結果より、上記不飽和脂肪酸に毛包上
皮系培養細胞の増殖活性が確かに認められた。すなわ
ち、上記不飽和脂肪酸には毛髪上皮系細胞の分裂増殖活
性の維持による、毛髪における成長期の維持,延長作用
が認められることが明らかになった。From these results, it was confirmed that the unsaturated fatty acids had a growth activity of hair follicle epithelial cells in culture. In other words, it was found that the unsaturated fatty acids exhibited the effect of maintaining and prolonging the anagen phase in hair by maintaining the mitotic proliferation activity of hair epithelial cells.
【0070】G.毛幹の伸長の検討(二次スクリーニン
グ) さらに、上記不飽和脂肪酸について、毛髪伸長効果を検
討した。ヒト毛包の器官培養 実体顕微鏡下でマイクロせん刃を用いて、ヒトの頭皮か
ら成長期の毛包を単離した。単離した毛包は「Williams
E培地(Gibco)にペニシリン,ストレプトマイシ
ン及びファンギゾン(Fungizone)を加えた培地」(以
下、(−)培地ともいう)で洗浄した後に長さを測定
し、この(−)培地にさらに10ng/ml のヒドロコーチ
ゾン,10μg/mlのインスリン,10ng/ml のナトリウ
ムセレナイト及び10μg/mlのトランスフェリンを添加
した培地(以下、(+)培地ともいう)(24穴のマイ
クロプレート使用:1穴当り1ml)中に沈ませて、5%
CO2下,37℃で一晩培養した(前培養)。この前培
養後に再度培養した毛包の長さを測定して、伸長が0.
25mm以上の毛包を選択して、その伸長の程度が均等に
なるように9本ずつの3群に分けた。毛包における毛幹
の伸長は、上記のマイクロプレートをミクロメーターを
装着した倒立顕微鏡を用いて目視で観察した。G. Examination of hair shaft elongation (secondary screening) Further, the hair elongation effect of the above unsaturated fatty acids was examined. Human Hair Follicle Organ Cultures The growing hair follicles were isolated from human scalp using a micro-blade under a stereomicroscope. The isolated hair follicle is "Williams
After washing with an E medium (Gibco) plus a medium containing penicillin, streptomycin and fungizone (hereinafter also referred to as (-) medium), the length was measured, and 10 ng / ml was further added to the (-) medium. In a medium supplemented with hydrocortisone, 10 μg / ml insulin, 10 ng / ml sodium selenite and 10 μg / ml transferrin (hereinafter also referred to as (+) medium) (using a 24-well microplate: 1 ml per well) Sink 5%
The cells were cultured overnight at 37 ° C. under CO 2 (pre-culture). The length of the hair follicles cultured again after this pre-culture was measured to be 0.
Hair follicles of 25 mm or more were selected and divided into three groups of nine hair follicles so that the degree of elongation was uniform. The elongation of the hair shaft in the hair follicle was visually observed on the microplate using an inverted microscope equipped with a micrometer.
【0071】対象物質の評価 対象物質である上記不飽和脂肪酸を最終濃度が1.0×
10-5mol/l (DMSO0.1%)となるように、
(−)培地に対象物質のDMSO溶液を添加して調製し
た。また、同時に(−)培地に有機溶媒(DMSO)を
最終濃度が0.1%になるように添加した培地(コント
ロール培地)を調製した。The above unsaturated fatty acid, which is a target substance for evaluation , is added at a final concentration of 1.0 ×
10 -5 mol / l (DMSO 0.1%)
(-) The medium was prepared by adding a DMSO solution of the target substance to the medium. At the same time, a medium (control medium) was prepared by adding an organic solvent (DMSO) to the (-) medium to a final concentration of 0.1%.
【0072】前述した3つの毛包群の培地を、上記の培
地にそれぞれ交換し、さらに5日間5%CO2 下,37
℃で培養した。培養開始5日目のそれぞれの毛包の長さ
を測定し、コントロール群と対象物質添加群との毛包に
おける毛幹の伸長を比較した。その結果を第2表に表
す。The medium of the three hair follicle groups described above was replaced with the above medium, respectively, and further cultured for 5 days under 5% CO 2 under 37%.
Incubated at ℃. The length of each hair follicle on the 5th day from the start of culture was measured, and the elongation of the hair follicles in the hair follicles of the control group and the control substance-added group was compared. The results are shown in Table 2.
【0073】[0073]
【表2】 [Table 2]
【0074】この結果より、不飽和脂肪酸においては毛
包上皮系細胞の増殖を促進するのみならず、毛包におけ
る毛幹の伸長を促進し、毛包器官として調和のとれた毛
髪成長期延長効果が認められることが明らかになった。
以下、本発明毛髪成長期延長剤の処方を実施例として示
し、さらにこれらの育毛効果の検討を行った。From these results, it is found that unsaturated fatty acids not only promote the growth of hair follicle epithelial cells, but also promote the growth of hair shafts in hair follicles, and have a prolonged hair growth period as a hair follicle organ. Was found to be recognized.
Hereinafter, the formulations of the hair growth period prolonging agent of the present invention are shown as examples, and the hair growth effects thereof were further studied.
【0075】〔実施例1〕 液状毛髪成長期延長剤の調
製 ペトロセリン酸0.1%及びオレイン酸0.1%を、7
0%エタノール90%、ドデシルベンゼンスルホン酸
0.49%、硬化ヒマシ油エチレンオキシド(40モ
ル)付加物0.5%及びイオン交換水(残余)と攪拌混
合して溶解させた。さらにイオン交換水(10%)を添
加混合して、液状の毛髪成長期延長剤を得た。この液状
の毛髪成長期延長剤の処方において、ペトロセリン酸と
オレイン酸とを除去して調製した液状の剤を対照として
調製した(比較例1)。[Example 1] Preparation of liquid hair growth period prolonging agent Petroseric acid 0.1% and oleic acid 0.1%
The mixture was dissolved by stirring and mixing with 0% ethanol 90%, dodecylbenzenesulfonic acid 0.49%, hydrogenated castor oil ethylene oxide (40 mol) adduct 0.5%, and ion-exchanged water (residual). Further, ion-exchanged water (10%) was added and mixed to obtain a liquid hair growth period extender. In the formulation of the liquid hair growth period prolonging agent, a liquid agent prepared by removing petroseric acid and oleic acid was prepared as a control (Comparative Example 1).
【0076】〔実施例2〕 乳液状毛髪成長期延長剤の
調製 以下の処方により、乳液状の毛髪成長期延長剤を調製し
た。 配合成分 配合量(重量%) (A相) ペトロセリン酸 1.0 ポリオキシエチレン(60モル)付加硬化ヒマシ油 2.0 グリセリン 10.0 ジプロピレングリコール 10.0 1,3−ブチレングリコール 5.0 ポリエチレングリコール1500 5.0 (B相) セチルイソオクタネート 10.0 スクワラン 5.0 ワセリン 2.0 プロピルパラベン 2.0 (C相) カルボキシビニルポリマー1%水溶液 30.0 ヘキサメタリン酸ソーダ 0.03 イオン交換水 8.35 (D相) イオン交換水 4.5 (E相) KOH 0.12 イオン交換水 5.0Example 2 Preparation of Emulsion Hair Growth Prolonging Agent An emulsion liquid hair growth prolonging agent was prepared according to the following formulation. Ingredients Ingredients Amount (% by weight) (A phase) Petroseric acid 1.0 Polyoxyethylene (60 mol) addition-hardened castor oil 2.0 Glycerin 10.0 Dipropylene glycol 10.0 1,3-butylene glycol 5.0 Polyethylene glycol 1500 5.0 (B phase) Cetyl isooctanoate 10.0 Squalane 5.0 Vaseline 2.0 Propyl paraben 2.0 (C phase) Carboxyvinyl polymer 1% aqueous solution 30.0 Sodium hexametaphosphate 0.03 ion Exchanged water 8.35 (D phase) Ion exchanged water 4.5 (E phase) KOH 0.12 Ion exchanged water 5.0
【0077】<製造法>A相、B相をそれぞれ60℃で
加熱溶解し、混合してホモミキサー処理しゲルを作り、
これにD相を徐々に添加しホモミキサーで分散した。次
にこれに溶解したC相を加え、最後に溶解したE相を添
加しホモミキサーで乳化してO/W乳液型の毛髪成長期
延長剤を得た。<Production Method> A phase and a B phase are each heated and dissolved at 60 ° C., mixed, and homogenized to form a gel.
Phase D was gradually added to this and dispersed with a homomixer. Next, the dissolved C phase was added thereto, and finally, the dissolved E phase was added and emulsified with a homomixer to obtain an O / W emulsion type hair growth period extender.
【0078】〔実施例3〕 クリーム状毛髪成長期延長
剤の調製 以下の処方により、クリーム状の毛髪成長期延長剤を調
製した。 配合成分 配合量(重量%) (A相) 流動パラフィン 5.0 セトステアリルアルコール 5.5 グリセリルモノステアレート 3.0 EO(20モル)−2−オクチルドデシルエーテル 8.0 プロピルパラベン 0.3 香料 0.1 (B相) ペトロセリン酸 5.0 グリセリン 8.0 ジプロピレングリコール 20.0 ポリエチレングリコール4000 5.0 ドデシル硫酸ナトリウム 0.1 ヘキサメタリン酸ソーダ 0.005 イオン交換水 39.995Example 3 Preparation of Creamy Hair Growth Prolonging Agent A creamy hair growth extending agent was prepared according to the following formulation. Ingredients Ingredients Amount (% by weight) (A phase) Liquid paraffin 5.0 cetostearyl alcohol 5.5 glyceryl monostearate 3.0 EO (20 mol) -2-octyldodecyl ether 8.0 propylparaben 0.3 Fragrance 0.1 (B phase) Petroselinic acid 5.0 Glycerin 8.0 Dipropylene glycol 20.0 Polyethylene glycol 4000 5.0 Sodium dodecyl sulfate 0.1 Sodium hexametaphosphate 0.005 Ion-exchanged water 39.995
【0079】<製造法>A相、B相をそれぞれ加熱溶解
し混合し、ホモミキサーで乳化してクリーム状の毛髪成
長期延長剤を得た。<Production Method> The phases A and B were each dissolved by heating, mixed, and emulsified by a homomixer to obtain a creamy hair growth period extender.
【0080】〔試験例2〕 本発明毛髪成長期延長剤の
育毛作用の検討 本発明毛髪成長期延長剤の脱毛防止、発毛効果等の育毛
作用を調べるために、以下の方法でヒトに対してトリコ
グラム試験を実施した。被験試料及び対照試料は、実施
例1〜3の本発明毛髪成長期延長剤、比較例1の剤及び
70%エタノールである。[Test Example 2] Examination of hair growth effect of hair growth period extender of the present invention To perform a trigram test. The test sample and the control sample are the hair growth period extending agent of the present invention of Examples 1 to 3, the agent of Comparative Example 1, and 70% ethanol.
【0081】試験方法 上記試料の使用前と使用後の抜去毛髪の毛根を顕微鏡下
で観察し、毛根の形態から,成長の止まった毛の毛根で
ある「休止期毛根」数(脱毛を訴える人は正常な人より
もこの休止期毛根の割合が多いことが認められている)
を計数し、その割合の増減によってこれらの試料の育毛
作用を比較した。 Test Method The hair roots of the extracted hair before and after use of the above sample were observed under a microscope, and the number of “resting hair roots”, which are the hair roots of hair that had stopped growing, was determined based on the shape of the hair root (for those who report hair loss). Has been found to have a higher proportion of this telogen root than a normal person.)
Was counted, and the hair growth effect of these samples was compared by increasing or decreasing the ratio.
【0082】すなわち、被験試料及び対照試料をそれぞ
れ男性被験者10名の頭皮に1日2回,1回2mlずつ6
カ月間連続して塗布し、塗布直前及び6カ月間塗布終了
直後に被験者1名につき100本ずつ毛髪を抜去し、そ
れぞれの毛根を顕微鏡下で観察した。また、上記試料に
おける育毛効果が有効か無効かに関する実使用テストを
行った。これらの試験の結果を、下記第3表に示す。That is, each of the test sample and the control sample was applied to the scalp of 10 male subjects twice a day, 2 ml at a time, for 6 times.
The hair was applied continuously for months, and immediately after the application and immediately after the completion of the application for six months, 100 hairs were removed from each subject and each hair root was observed under a microscope. In addition, a practical use test was performed to determine whether the hair growth effect of the sample was effective or ineffective. The results of these tests are shown in Table 3 below.
【0083】[0083]
【表3】 [Table 3]
【0084】この第3表の結果から、不飽和脂肪酸であ
るペトロセリン酸を有効成分として配合した本発明毛髪
成長期延長剤には、このペトロセリン酸の毛髪成長期延
長効果に基づく育毛効果が認められた。また、上記した
ペトロセリン酸以外の不飽和脂肪酸等においても、ペト
ロセリン酸と同様に毛髪成長期延長効果が認められたこ
とから、これらの植物抽出物等を有効成分として配合し
た本発明毛髪成長期延長剤においても、上記の実施例と
同様に育毛効果が認められることは明らかである。From the results shown in Table 3, the hair growth-prolonging agent of the present invention containing petrothenic acid, which is an unsaturated fatty acid, as an active ingredient has a hair-growth effect based on the hair growth-prolonging effect of petroselinic acid. Was. In addition, the unsaturated fatty acids other than petroselinic acid described above also exhibited a hair growth period extending effect similar to that of petroselinic acid. It is clear that the hair growth effect can be observed in the same manner as in the above Examples.
【0085】[0085]
【発明の効果】本発明により、毛髪伸長の促進をするこ
とによって毛周期における成長期を維持又は延長する毛
髪成長期延長剤が提供される。Industrial Applicability According to the present invention, there is provided a hair growth-prolonging agent that maintains or prolongs the anagen in the hair cycle by promoting hair elongation.
─────────────────────────────────────────────────────
────────────────────────────────────────────────── ───
【手続補正書】[Procedure amendment]
【提出日】平成10年1月30日[Submission date] January 30, 1998
【手続補正1】[Procedure amendment 1]
【補正対象書類名】明細書[Document name to be amended] Statement
【補正対象項目名】0047[Correction target item name] 0047
【補正方法】変更[Correction method] Change
【補正内容】[Correction contents]
【0047】B.ラット毛包上皮系細胞 1.ラット毛包上皮系細胞の採取: (1)毛包の採取 新生児(3〜4日令)のラットの背部皮膚を採取し、こ
の採取した背部皮膚を5%PSF含有PBS(−)に浸
漬後、皮膚脂肪層から下の皮下脂肪や皮膜等を解剖用ハ
サミで除去した。次いで、再びこの背部皮膚を1%PS
F含有PBS(−)に浸し、さらにこれを0.25%ト
リプシン−0.02%EDTA含有PBS(−)中に4
℃で一晩浸した。B. Rat hair follicle epithelial cells 1. Collection of rat hair follicle epithelial cells: (1) Collection of hair follicle The back skin of a newborn (3 to 4 day old) rat was collected, and the collected back skin was immersed in PBS (-) containing 5% PSF. Then, the subcutaneous fat, skin, and the like below the skin fat layer were removed with scissors for dissection. Then, again this back skin was 1% PS
F-containing PBS (-), and further immersed in PBS (-) containing 0.25% trypsin-0.02% EDTA.
Soaked overnight at ° C.
フロントページの続き (72)発明者 鈴木 順 神奈川県横浜市金沢区福浦2−12−1 株 式会社資生堂第2リサーチセンター内 (72)発明者 相馬 勤 神奈川県横浜市金沢区福浦2−12−1 株 式会社資生堂第2リサーチセンター内 (72)発明者 尾郷 正志 神奈川県横浜市金沢区福浦2−12−1 株 式会社資生堂第2リサーチセンター内 (72)発明者 田島 正裕 神奈川県横浜市港北区新羽町1050番地 株 式会社資生堂第1リサーチセンター内Continued on the front page (72) Inventor Jun Suzuki 2-12-1 Fukuura, Kanazawa-ku, Yokohama, Kanagawa Prefecture Inside Shiseido Second Research Center Co., Ltd. (72) Tsutomu Soma 2-12-, Fukuura, Kanazawa-ku, Yokohama, Kanagawa 1 Shiseido 2nd Research Center Co., Ltd. (72) Inventor Masashi Ogo 2-12-1 Fukuura, Kanazawa-ku, Yokohama-shi, Kanagawa Prefecture 2nd Shiseido 2nd Research Center Co., Ltd. (72) Inventor Masahiro Tajima Kohoku, Yokohama-shi, Kanagawa 1050 Nippa-cho, Tokyo-ku Shiseido First Research Center
Claims (5)
成分とする毛髪成長期延長剤。1. A hair growth period extending agent comprising an unsaturated fatty acid and / or a derivative thereof as an active ingredient.
2n−2〕で表される不飽和脂肪酸である、請求項1記
載の毛髪成長期延長剤。Wherein the unsaturated fatty acid has the formula C n H m O 2 [number of carbon atoms and n is 12 to 28, the number of hydrogen atoms m = n +. 1 to
The hair growth prolonging agent according to claim 1, which is an unsaturated fatty acid represented by 2n-2].
求項1又は請求項2記載の毛髪成長期延長剤。3. The hair growth period extending agent according to claim 1, wherein the double bond of the unsaturated fatty acid is a cis form.
酸,ヘプタデセン酸,ペトロセリン酸,パルミトレイン
酸,オレイン酸,バクセン酸,リノール酸,α−リノレ
ン酸,γ−リノレン酸,エイコセン酸,エイコサトリエ
ン酸,エルカ酸(cis-13-ドコセン酸),ドコサジエン
酸,ドコサトリエン酸,ドコサテトラエン酸,ドコサペ
ンタエン酸,アラキドン酸,ネルボン酸及びドコサヘキ
サエン酸からなる群の不飽和脂肪酸から選ばれる1種又
は2種以上の不飽和脂肪酸である、請求項1乃至請求項
3のいずれかの請求項記載の毛髪成長期延長剤。4. The unsaturated fatty acid is dodecenoic acid, tridecenoic acid, heptadecenoic acid, petroselinic acid, palmitoleic acid, oleic acid, vaccenic acid, linoleic acid, α-linolenic acid, γ-linolenic acid, eicosenoic acid, eicosatrienoic acid. One or two unsaturated fatty acids selected from the group consisting of, erucic acid (cis-13-docosenoic acid), docosadienoic acid, docosatrienoic acid, docosatetraenoic acid, docosapentaenoic acid, arachidonic acid, nervonic acid and docosahexaenoic acid The hair growth period prolonging agent according to any one of claims 1 to 3, which is at least one unsaturated fatty acid.
キサエン酸及び/又はオレイン酸である請求項4記載の
毛髪成長期延長剤。5. The hair growth extender according to claim 4, wherein the unsaturated fatty acid is petroselinic acid, docosahexaenoic acid and / or oleic acid.
Priority Applications (6)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP27526197A JPH1192343A (en) | 1997-09-22 | 1997-09-22 | Agent for prolonging hair growth period |
TW087104279A TW592719B (en) | 1997-03-26 | 1998-03-23 | Hair growth phase extender composition containing unsaturated fatty acid and/or its derivatives or an extract of a plant belonging to the Coriandrum |
US09/047,447 US6139852A (en) | 1997-03-26 | 1998-03-25 | Extract composition as hair growth phase extender |
DE69826547T DE69826547T2 (en) | 1997-03-26 | 1998-03-25 | Composition for extending the hair growth phase |
EP98400690A EP0867169B1 (en) | 1997-03-26 | 1998-03-25 | Hair growth phase extender composition |
KR1019980010424A KR100503695B1 (en) | 1997-03-26 | 1998-03-26 | Hair Growth Extension Composition |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP27526197A JPH1192343A (en) | 1997-09-22 | 1997-09-22 | Agent for prolonging hair growth period |
Publications (1)
Publication Number | Publication Date |
---|---|
JPH1192343A true JPH1192343A (en) | 1999-04-06 |
Family
ID=17552959
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP27526197A Withdrawn JPH1192343A (en) | 1997-03-26 | 1997-09-22 | Agent for prolonging hair growth period |
Country Status (1)
Country | Link |
---|---|
JP (1) | JPH1192343A (en) |
Cited By (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2003505491A (en) * | 1999-07-30 | 2003-02-12 | ユニリーバー・ナームローゼ・ベンノートシヤープ | Skin care composition |
WO2005059114A1 (en) * | 2003-12-17 | 2005-06-30 | Idemitsu Kosan Co., Ltd. | Fat composition for promoting animal cell growth, medium for culturing animal cells containing the composition and external skin preparation |
JP2005198648A (en) * | 2003-12-17 | 2005-07-28 | Idemitsu Kosan Co Ltd | Oil and fat composition of animal cell growth promoting agent and animal cell culture medium containing the composition |
JP2015105265A (en) * | 2013-12-02 | 2015-06-08 | 共栄化学工業株式会社 | Composition for hair |
JP2015532920A (en) * | 2012-09-26 | 2015-11-16 | タンジェント リプロファイリング リミテッド | Androgen synthesis modulator |
JP2019085345A (en) * | 2017-11-02 | 2019-06-06 | ロート製薬株式会社 | Hair growth factor production promoter and/or cell adhesion molecule production promoter |
CN115054618A (en) * | 2022-08-08 | 2022-09-16 | 深圳大学 | Application of schizochytrium limacinum oil extract in promoting hair growth or preventing and treating alopecia |
-
1997
- 1997-09-22 JP JP27526197A patent/JPH1192343A/en not_active Withdrawn
Cited By (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP2003505491A (en) * | 1999-07-30 | 2003-02-12 | ユニリーバー・ナームローゼ・ベンノートシヤープ | Skin care composition |
WO2005059114A1 (en) * | 2003-12-17 | 2005-06-30 | Idemitsu Kosan Co., Ltd. | Fat composition for promoting animal cell growth, medium for culturing animal cells containing the composition and external skin preparation |
JP2005198648A (en) * | 2003-12-17 | 2005-07-28 | Idemitsu Kosan Co Ltd | Oil and fat composition of animal cell growth promoting agent and animal cell culture medium containing the composition |
JP2015532920A (en) * | 2012-09-26 | 2015-11-16 | タンジェント リプロファイリング リミテッド | Androgen synthesis modulator |
JP2019052159A (en) * | 2012-09-26 | 2019-04-04 | タンジェント リプロファイリング リミテッド | Modulators of androgen synthesis |
JP2015105265A (en) * | 2013-12-02 | 2015-06-08 | 共栄化学工業株式会社 | Composition for hair |
JP2019085345A (en) * | 2017-11-02 | 2019-06-06 | ロート製薬株式会社 | Hair growth factor production promoter and/or cell adhesion molecule production promoter |
CN115054618A (en) * | 2022-08-08 | 2022-09-16 | 深圳大学 | Application of schizochytrium limacinum oil extract in promoting hair growth or preventing and treating alopecia |
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