JP2019199410A - Composition for proliferation of human hair papilla cells - Google Patents
Composition for proliferation of human hair papilla cells Download PDFInfo
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- JP2019199410A JP2019199410A JP2018093527A JP2018093527A JP2019199410A JP 2019199410 A JP2019199410 A JP 2019199410A JP 2018093527 A JP2018093527 A JP 2018093527A JP 2018093527 A JP2018093527 A JP 2018093527A JP 2019199410 A JP2019199410 A JP 2019199410A
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- 210000004209 hair Anatomy 0.000 title claims abstract description 34
- 239000000203 mixture Substances 0.000 title claims abstract description 22
- 230000035755 proliferation Effects 0.000 title claims abstract description 9
- 150000001875 compounds Chemical class 0.000 claims abstract description 51
- 239000004480 active ingredient Substances 0.000 claims abstract description 17
- 230000004663 cell proliferation Effects 0.000 claims abstract description 10
- 241000238557 Decapoda Species 0.000 claims description 17
- 230000002062 proliferating effect Effects 0.000 claims description 17
- 239000002994 raw material Substances 0.000 claims description 13
- 239000000126 substance Substances 0.000 claims description 10
- 230000003779 hair growth Effects 0.000 claims description 8
- 230000002500 effect on skin Effects 0.000 claims description 6
- 230000017531 blood circulation Effects 0.000 claims description 3
- 210000003780 hair follicle Anatomy 0.000 claims description 3
- 235000016709 nutrition Nutrition 0.000 claims description 3
- 230000035764 nutrition Effects 0.000 claims description 3
- 210000004761 scalp Anatomy 0.000 claims description 3
- 230000000694 effects Effects 0.000 abstract description 12
- 238000000034 method Methods 0.000 abstract description 8
- RPMOMKUYGKLDSQ-UHFFFAOYSA-N calanthoside Natural products OC1C(O)C(O)C(CO)OC1OC(C1=CC=CC=C1N1)=C1SC1C(O)C(O)C(O)C(CO)O1 RPMOMKUYGKLDSQ-UHFFFAOYSA-N 0.000 abstract description 5
- IBGZNOPZUVZSEJ-UHFFFAOYSA-N glucoindican Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC=2C3=CC=CC=C3NC=2)O1 IBGZNOPZUVZSEJ-UHFFFAOYSA-N 0.000 abstract description 5
- 241001517312 Calanthe Species 0.000 abstract 2
- XEKOWRVHYACXOJ-UHFFFAOYSA-N Ethyl acetate Chemical compound CCOC(C)=O XEKOWRVHYACXOJ-UHFFFAOYSA-N 0.000 description 12
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 12
- 239000002904 solvent Substances 0.000 description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 7
- 239000000401 methanolic extract Substances 0.000 description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- DHVJQUFZGZOFFY-UHFFFAOYSA-N (6-amino-2-imino-4-piperidin-1-ylpyrimidin-1-yl) hydrogen sulfate Chemical compound N=C1N(OS(O)(=O)=O)C(N)=CC(N2CCCCC2)=N1 DHVJQUFZGZOFFY-UHFFFAOYSA-N 0.000 description 4
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 4
- 238000004440 column chromatography Methods 0.000 description 4
- 239000003814 drug Substances 0.000 description 4
- 238000004128 high performance liquid chromatography Methods 0.000 description 4
- 229960000841 minoxidil sulfate Drugs 0.000 description 4
- 239000000284 extract Substances 0.000 description 3
- 239000003960 organic solvent Substances 0.000 description 3
- 208000034656 Contusions Diseases 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 241000233855 Orchidaceae Species 0.000 description 2
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 2
- UIIMBOGNXHQVGW-UHFFFAOYSA-M Sodium bicarbonate Chemical compound [Na+].OC([O-])=O UIIMBOGNXHQVGW-UHFFFAOYSA-M 0.000 description 2
- FEWJPZIEWOKRBE-UHFFFAOYSA-N Tartaric acid Natural products [H+].[H+].[O-]C(=O)C(O)C(O)C([O-])=O FEWJPZIEWOKRBE-UHFFFAOYSA-N 0.000 description 2
- 230000010261 cell growth Effects 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- -1 indole glycosides Chemical class 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 238000003808 methanol extraction Methods 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 2
- 235000002906 tartaric acid Nutrition 0.000 description 2
- 239000011975 tartaric acid Substances 0.000 description 2
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 1
- ZFMITUMMTDLWHR-UHFFFAOYSA-N Minoxidil Chemical compound NC1=[N+]([O-])C(N)=CC(N2CCCCC2)=N1 ZFMITUMMTDLWHR-UHFFFAOYSA-N 0.000 description 1
- SIKJAQJRHWYJAI-UHFFFAOYSA-N benzopyrrole Natural products C1=CC=C2NC=CC2=C1 SIKJAQJRHWYJAI-UHFFFAOYSA-N 0.000 description 1
- 229940125904 compound 1 Drugs 0.000 description 1
- 229940126214 compound 3 Drugs 0.000 description 1
- 229940125898 compound 5 Drugs 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 229930182470 glycoside Natural products 0.000 description 1
- 230000035876 healing Effects 0.000 description 1
- 235000013402 health food Nutrition 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- PZOUSPYUWWUPPK-UHFFFAOYSA-N indole Natural products CC1=CC=CC2=C1C=CN2 PZOUSPYUWWUPPK-UHFFFAOYSA-N 0.000 description 1
- RKJUIXBNRJVNHR-UHFFFAOYSA-N indolenine Natural products C1=CC=C2CC=NC2=C1 RKJUIXBNRJVNHR-UHFFFAOYSA-N 0.000 description 1
- 231100000053 low toxicity Toxicity 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 229960003632 minoxidil Drugs 0.000 description 1
- 239000012046 mixed solvent Substances 0.000 description 1
- 210000002741 palatine tonsil Anatomy 0.000 description 1
- 229940056360 penicillin g Drugs 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 229910000030 sodium bicarbonate Inorganic materials 0.000 description 1
- 235000017557 sodium bicarbonate Nutrition 0.000 description 1
- VSIVTUIKYVGDCX-UHFFFAOYSA-M sodium;4-[2-(2-methoxy-4-nitrophenyl)-3-(4-nitrophenyl)tetrazol-2-ium-5-yl]benzene-1,3-disulfonate Chemical compound [Na+].COC1=CC([N+]([O-])=O)=CC=C1[N+]1=NC(C=2C(=CC(=CC=2)S([O-])(=O)=O)S([O-])(=O)=O)=NN1C1=CC=C([N+]([O-])=O)C=C1 VSIVTUIKYVGDCX-UHFFFAOYSA-M 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
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- Cosmetics (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
Description
本発明は，ヒト毛乳頭細胞増殖用組成物等に関する。さらに詳しくいうと本発明は，エビネより抽出された化合物であるglucoindicanやcalanthosideを有効成分とするヒト毛乳頭細胞増殖用組成物等に関する。 The present invention relates to a composition for proliferating human dermal papilla cells and the like. More specifically, the present invention relates to a composition for proliferating human hair papilla cells and the like containing glucoindican and calanthoside, which are compounds extracted from shrimp, as active ingredients.
エビネは，日本，中国，台湾，韓国などの山地の陰地に自生するラン科（Orchidaceae）の多年草である。エビネは，その全草または根茎が九子連環草と称され，扁桃腺，痔癒および打撲傷の治療薬として用いられている。
また，エビネの根茎には，ジフェナントレン化合物やインドール配糖体などの有用な成分が含まれていることが知られている（特許文献１から３）。しかしながら，全ての成分について明らかになっているものではなく，未だ知られていない有用成分を含む可能性が秘められているものである。
Ebine is a perennial orchidaceae (Orchidaceae) that grows naturally in the shade of mountains such as Japan, China, Taiwan, and Korea. Ebine, whose whole plant or rhizome is referred to as “Kyokurenkan”, is used as a treatment for tonsils, healing of bruises and bruises.
In addition, it is known that shrimp rhizome contains useful components such as diphenanthrene compounds and indole glycosides (Patent Documents 1 to 3). However, it has not been clarified about all the components, and there is a possibility that useful components not yet known are included.
上記事情を背景として本発明では，エビネの根茎に含まれる有用成分のさらなる探索，ならびに見い出された有用成分の利用方法の発見を課題とする。 In view of the above circumstances, an object of the present invention is to further search for useful components contained in shrimp rhizomes and to find out how to use the found useful components.
発明者らは，鋭意研究の結果，エビネの根茎に含まれる複数の化合物を明らかにした。加えて，これらの化合物のうち，glucoindicanならびにcalanthosideが，毛乳頭細胞増殖効果を有することを見い出し，発明を完成させたものである。 As a result of intensive studies, the inventors have clarified a plurality of compounds contained in the shrimp rhizome. In addition, among these compounds, glucoindican and calanthoside have been found to have a hair papillary cell proliferation effect, and the present invention has been completed.
本発明は，以下の構成からなる。
本発明の第一の構成は，下記化７，化８で示される化合物のいずれか又は複数を有効成分とすることを特徴とするヒト毛乳頭細胞増殖用組成物である。
The present invention has the following configuration.
The first constitution of the present invention is a composition for proliferating human hair papilla cells, characterized in that any one or a plurality of compounds represented by the following chemical formulas 7 and 8 is used as an active ingredient.
本発明の第二の構成は，前記有効成分のいずれか又は複数が，天然原料由来である第一の構成に記載のヒト毛乳頭細胞増殖用組成物である。
本発明の第三の構成は，天然原料が，エビネである第二の構成に記載のヒト毛乳頭細胞増殖用組成物である。
本発明の第四の構成は，育毛，発毛，頭皮血行促進，毛包栄養促進のいずれか又は複数の用途として用いられる第一から第三の構成いずれかに記載のヒト毛乳頭細胞増殖用組成物である。
本発明の第五の構成は，ヒト毛乳頭細胞増殖を特徴とする化７で示される化合物の使用方法である。
本発明の第六の構成は，ヒト毛乳頭細胞増殖を特徴とする化８で示される化合物の使用方法である。
The second composition of the present invention is the composition for proliferating human hair papilla cells according to the first composition, wherein one or more of the active ingredients are derived from natural raw materials.
The third composition of the present invention is the composition for proliferating human dermal papilla cells according to the second composition, wherein the natural raw material is shrimp.
The fourth configuration of the present invention is for human hair papilla cell proliferation according to any one of the first to third configurations used for hair growth, hair growth, scalp blood circulation promotion, hair follicle nutrition promotion or a plurality of uses. It is a composition.
The fifth constitution of the present invention is a method of using the compound represented by Chemical formula 7 characterized by proliferation of human hair papilla cells.
The sixth aspect of the present invention is a method of using the compound represented by Chemical Formula 8 characterized by human hair papilla cell proliferation.
本発明により，エビネの根茎に含まれる有用成分のさらなる探索ならびに見い出された有用成分の利用方法の発見が，発明者らによりなされたものである。
すなわち，エビネの根茎に含まれる有用成分として，複数の化合物が抽出され，これらの化合物のうち，glucoindicanならびにcalanthosideは，ヒト毛乳頭細胞増殖効果を有することが発明者らにより明らかとなった。これより，これらを有効成分としてヒト毛乳頭細胞増殖にかかる組成物の提供が可能となったものである。
According to the present invention, the inventors have further searched for useful components contained in shrimp rhizomes and discovered methods for using the found useful components.
That is, a plurality of compounds were extracted as useful components contained in shrimp rhizomes, and among these compounds, it was revealed by the inventors that glucoindican and calanthoside have a human hair papilla cell proliferation effect. Thus, it has become possible to provide a composition relating to proliferation of human hair papilla cells using these as active ingredients.
本発明のヒト毛乳頭細胞増殖用組成物について説明を行う。 The composition for proliferating human hair papilla cells of the present invention will be described.
本発明のヒト毛乳頭細胞増殖用組成物は，化７，化８で示される化合物（以下，これらをまとめて「有効成分化合物」という）のいずれか又は複数を有効成分とすることを特徴とする。 The composition for proliferating human dermal papilla cells of the present invention is characterized in that any one or more of the compounds represented by Chemical Formulas 7 and 8 (hereinafter collectively referred to as “active component compounds”) are used as the active components. To do.
本発明において有効成分化合物は，それぞれ化７がglucoindican，化８がcalanthosideと称される既知の化合物である。
本発明のヒト毛乳頭細胞増殖用組成物は，これらの有効成分化合物の新規用途として，ヒト毛乳頭細胞増殖作用が発明者により見い出されたことに基づくものである。同時に，ヒト毛乳頭細胞増殖を特徴とする化７ないし化８の使用方法にかかる発明が，発明者により完成されたものである。
In the present invention, the active ingredient compounds are known compounds in which Chemical Formula 7 is called glucoindican and Chemical Formula 8 is called calanthoside.
The composition for proliferating human dermal papilla cells of the present invention is based on the fact that the inventor has found a human dermal papilla cell proliferating action as a novel use of these active ingredient compounds. At the same time, the inventors have completed the invention relating to the method of using chemical formulas 7 to 8 characterized by proliferation of human hair papilla cells.
本発明に用いられる有効成分化合物は，そのままの形で有効成分として用いることができるが，本発明の趣旨に鑑み，生体内における代謝等を受けて化合物の構造を変化させ有効成分化合物として作用するなどの，いわゆるDDS化された有効成分化合物を含むものとして用いることもできる。 The active ingredient compound used in the present invention can be used as an active ingredient as it is, but in view of the gist of the present invention, it undergoes in vivo metabolism and the like to change the structure of the compound and act as an active ingredient compound. It can also be used as a compound containing an active ingredient compound converted to DDS.
本発明において有効成分化合物は，化学的に合成したものを用いてもよいし，天然原料由来のものを用いてもよい。天然原料の場合は，エビネを原料として用いることができる。 In the present invention, a chemically synthesized compound may be used as the active ingredient compound, or a compound derived from a natural raw material may be used. In the case of a natural raw material, shrimp can be used as a raw material.
エビネを原料として用いる場合，エビネの全草，好ましくは地下部（根茎）を用いればよく，これらをそのまま用いてもよいし，乾燥させた後，原料として用いてもよい。このエビネ原料について，溶媒を用いて抽出を行う。用いる溶媒としては，エビネ原料からの有効成分化合物の抽出が可能な限り特に限定する必要はなく，水ないし種々の有機溶媒を用いることができる。
溶媒として，水ないしメタノールやエタノールなどの水溶性アルコール，又はこれらの混合溶媒を用いることが好ましい。これにより，安価かつ効率的に，有効成分の抽出を行うことが可能となり，エビネ溶媒抽出物の製造コストを減少させる効果を有する。
有機溶媒として，最も好ましくは，エタノールを用いることができる。エタノールは，人体への毒性が低い有機溶媒であるため，エビネ溶媒抽出物や，これを原料に製造される医薬の安全性を向上させる効果を有する。
When using shrimp as a raw material, the whole plant of shrimp, preferably the underground part (rhizome) may be used, and these may be used as they are or may be used as a raw material after drying. The shrimp raw material is extracted using a solvent. The solvent to be used is not particularly limited as long as the active ingredient compound can be extracted from the raw material of shrimp, and water or various organic solvents can be used.
As the solvent, it is preferable to use water or a water-soluble alcohol such as methanol or ethanol, or a mixed solvent thereof. This makes it possible to extract the active ingredient at low cost and efficiently, and has the effect of reducing the manufacturing cost of the prawn solvent extract.
Most preferably, ethanol can be used as the organic solvent. Since ethanol is an organic solvent having low toxicity to the human body, it has the effect of improving the safety of the ebine solvent extract and pharmaceuticals produced from it.
本発明において有効成分化合物を含む組成物とすることにより，ヒト毛乳頭細胞増殖効果を発揮するものである。また，ヒト毛乳頭細胞増殖を通じて，育毛，発毛，頭皮血行促進，毛包栄養促進などの効果が期待できるものである。
本発明において組成物とは，本発明の趣旨に基づきヒト毛乳頭細胞増殖効果の発揮が期待される，医薬品，医薬部外品，サプリメント，健康食品など様々な形態の製品を指すものとして定義される。
In the present invention, a composition containing an active ingredient compound exhibits a human hair papilla cell proliferation effect. In addition, the effects of hair growth, hair growth, scalp blood circulation promotion, and hair follicle nutrition promotion can be expected through the proliferation of human hair papilla cells.
In the present invention, the composition is defined as referring to various forms of products such as pharmaceuticals, quasi-drugs, supplements, and health foods that are expected to exert human hair papilla cell proliferation effects based on the spirit of the present invention. The
以下，本発明について，実施例を挙げて詳述するが，本発明を実施例に基づき限定的に解釈すべきでないことはいうまでもない。 EXAMPLES Hereinafter, although an Example is given and explained in full detail about this invention, it cannot be overemphasized that this invention should not be interpreted limitedly based on an Example.
＜＜実験例１，溶媒抽出物の作製＞＞
１．図１，ないし図２の手順に順じて，化合物の探索・抽出を行った。
２．まず，宮崎産エビネの地下部について，メタノールによる抽出を３回繰り返し行い，メタノール抽出液を得た。得られたメタノール抽出液を減圧下溶媒留去してメタノール抽出エキスを得た。
<< Experimental Example 1, Preparation of Solvent Extract >>
1. The compounds were searched and extracted in accordance with the procedures shown in FIGS.
2. First, extraction with methanol was repeated three times for the underground part of Miyazaki Ebine, and a methanol extract was obtained. The obtained methanol extract was evaporated under reduced pressure to obtain a methanol extract.
３．図１に順じて，下記一連の操作を行った。
(1) メタノール抽出エキスについて，酢酸エチル／3% 酒石酸溶液にて分液を行った。
(2) 酢酸エチル層について，溶媒を留去し，さらに順相および逆相カラムクロマトグラフィー，次いでHPLCにて精製を行い，化合物１，化合物４を得た。
(3) 3% 酒石酸溶液層に炭酸水素ナトリウムを添加してpH9.0に調整後，クロロホルムにより，分液操作を行った。得られたクロロホルム層について，溶媒を留去し，さらに順相および逆相カラムクロマトグラフィー，次いでHPLCにて精製を行い，化合物２，３を得た。
3. The following series of operations were performed in accordance with FIG.
(1) The methanol extract was separated with an ethyl acetate / 3% tartaric acid solution.
(2) About the ethyl acetate layer, the solvent was distilled off, and further purified by normal phase and reverse phase column chromatography and then by HPLC to obtain Compound 1 and Compound 4.
(3) Sodium bicarbonate was added to the 3% tartaric acid solution layer to adjust the pH to 9.0, followed by liquid separation with chloroform. About the obtained chloroform layer, the solvent was distilled off, and further purified by normal phase and reverse phase column chromatography and then by HPLC to obtain compounds 2 and 3.
４．また，図２に準じて，下記一連の操作を行った。
(1) 実験例１に記載の方法と同様の操作により，メタノール抽出エキスを得た。さらに，得られたメタノール抽出エキスについて，酢酸エチル／水にて分液を行った。
(2) 酢酸エチル層について，溶媒溶媒を留去し，さらに順相および逆相カラムクロマトグラフィー，次いでHPLCにて精製を行い，化合物５（不図示）を得た。
(3) 水層について，Diaion HP-20 カラムに通した後，水，メタノールの順で溶出を行い，メタノール溶出液を濃縮後，さらに順相および逆相カラムクロマトグラフィー，次いでHPLCにて精製を行い，化合物６，７，８を得た。
4). In addition, the following series of operations were performed according to FIG.
(1) A methanol extract was obtained in the same manner as in the method described in Experimental Example 1. Furthermore, the obtained methanol extract was subjected to liquid separation with ethyl acetate / water.
(2) For the ethyl acetate layer, the solvent was distilled off, and further purified by normal phase and reverse phase column chromatography, followed by HPLC to obtain Compound 5 (not shown).
(3) After passing through the Diaion HP-20 column, the water layer is eluted in the order of water and methanol. After concentrating the methanol eluate, it is further purified by normal phase and reverse phase column chromatography followed by HPLC. This gave compounds 6, 7, and 8.
＜＜実験例２．ヒト毛乳頭細胞増殖促進試験＞＞
実験例１により見いだされた複数の化合物について，ヒト毛乳頭細胞にどのような影響を及ぼすかを調べることを目的に実験を行った。
<< Experimental Example 2. Human hair papilla cell growth promotion test >>
An experiment was conducted for the purpose of investigating the influence of a plurality of compounds found in Experimental Example 1 on human hair papilla cells.
＜実験方法＞
１．ペニシリンGおよびストレプトマイシンを含むDMEM培地により細胞の培養を行った。ヒト毛乳頭細胞（HFDPC）は，96穴マイクロプレートを用い，1.0×104cells/100μL/wellにて，37℃，5%CO2下，24時間培養を行った。
２．さらに，各化合物を含む培養液を，100μL/well加え，37℃，5%CO2下，4日間，培養を行った。なお，医薬品として，育毛，発毛等を目的に使用されているミノキシジル硫酸塩を陽性対象として同様の操作を行った。
３．培養後，各wellにWST-8溶液（株式会社同仁化学研究所）を加え，37℃，5%CO2下，培養を行い，細胞増殖の指標としてO.D.値（450-655nm）の測定を行った。
<Experiment method>
1. Cells were cultured in DMEM medium containing penicillin G and streptomycin. Human hair papilla cells (HFDPC) were cultured for 24 hours at 37 ° C. and 5% CO 2 at 1.0 × 10 4 cells / 100 μL / well using a 96-well microplate.
2. Furthermore, the culture solution containing each compound was added at 100 μL / well and cultured at 37 ° C. under 5% CO 2 for 4 days. In addition, the same operation was performed for minoxidil sulfate, which is used for the purpose of hair growth and hair growth as a positive drug.
3. After incubation, add WST-8 solution (Dojindo Laboratories) to each well, incubate at 37 ° C, 5% CO 2 , and measure OD value (450-655nm) as an index of cell proliferation It was.
＜実験結果＞
１．各化合物の検討結果を図３に示す。
(1) メタノール抽出エキスについて，10μg/mLまではほとんど変化は見られなかったが，それ以降濃度に応じて，ヒト毛乳頭細胞の増殖効果が確認された。また，100μg/mLの濃度では，有意な細胞増殖の促進が観察された。
(2) 化合物１ならびに２については，ヒト毛乳頭細胞の増殖効果は認められず，それぞれ高濃度において細胞毒性が認められた。
(3) 化合物３について，100μMにて，増加傾向が確認された。
(4) 化合物６について，検討を行った濃度範囲において，ほとんど変化は見られなかった。
(5) 化合物７について，100μMにて，有意なヒト毛乳頭細胞の増殖効果が確認された。
(6) 化合物８について，一貫して100%を越えており，さらに100μMで，検討を行った化合物の中で最も強く，かつ有意なヒト毛乳頭細胞の増殖効果が確認された。
<Experimental result>
1. The examination result of each compound is shown in FIG.
(1) Methanol extract showed almost no change up to 10 μg / mL, but the growth effect of human hair papilla cells was confirmed depending on the concentration thereafter. In addition, significant enhancement of cell growth was observed at a concentration of 100 μg / mL.
(2) With regard to compounds 1 and 2, no proliferative effect on human hair papilla cells was observed, and cytotoxicity was observed at high concentrations.
(3) For Compound 3, an increasing trend was confirmed at 100 μM.
(4) For compound 6, there was almost no change in the concentration range examined.
(5) For compound 7, a significant proliferation effect of human hair papilla cells was confirmed at 100 μM.
(6) Compound 8 consistently exceeded 100%, and at 100 μM, the strongest and significant proliferative effect of human hair papilla cells was confirmed among the compounds examined.
２．ミノキシジル硫酸塩の結果を図４に示す。ミノキシジル硫酸塩の10μMにおいて，約130%のヒト毛乳頭細胞の有意な増殖効果が確認された。 2. The results for minoxidil sulfate are shown in FIG. A significant growth effect of about 130% of human hair papilla cells was confirmed at 10 μM of minoxidil sulfate.
３．これらの結果から，エビネより抽出された化合物は，ミノキシジル塩酸塩と濃度は異なるものの，化合物７についてはミノキシジル硫酸塩にせまるほどの，化合物８においてはこれを超えるヒト毛乳頭細胞の増殖効果が確認された。
４．よって，化合物７ならびに化合物８が，ヒト毛乳頭細胞増殖をもたらす化合物として有用であることが示された。
3. From these results, though the concentration of the compound extracted from shrimp is different from that of minoxidil hydrochloride, compound 8 is confirmed to have a proliferative effect on human hair papilla cells that exceeds that in compound 8, but in compound 8 It was done.
4). Therefore, it was shown that Compound 7 and Compound 8 are useful as compounds that cause human hair papilla cell proliferation.
Claims (6)
A composition for proliferating human dermal papilla cells, comprising any one or more of the compounds represented by Chemical formulas 7 and 8 as an active ingredient:
The composition for proliferating human hair papilla cells according to claim 1, wherein one or more of the active ingredients are derived from natural raw materials.
The composition for proliferating human hair papilla cells according to claim 2, wherein the natural raw material is shrimp.
The composition for proliferating human hair papilla cells according to any one of claims 1 to 3, which is used for hair growth, hair growth, scalp blood circulation promotion, hair follicle nutrition promotion or a plurality of uses.
Use of a compound represented by Chemical Formula 7 characterized by human hair papilla cell proliferation
Use of a compound represented by Chemical formula 8 characterized by proliferation of human hair papilla cells
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| JPH05294813A (en) * | 1992-02-17 | 1993-11-09 | Manabu Nomura | Hair producing and hair growing agent |
| JPH10229978A (en) * | 1997-02-18 | 1998-09-02 | Shiseido Co Ltd | Method of detecting hair growth |
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