JP7291333B2 - 育毛剤 - Google Patents
育毛剤 Download PDFInfo
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- JP7291333B2 JP7291333B2 JP2022519600A JP2022519600A JP7291333B2 JP 7291333 B2 JP7291333 B2 JP 7291333B2 JP 2022519600 A JP2022519600 A JP 2022519600A JP 2022519600 A JP2022519600 A JP 2022519600A JP 7291333 B2 JP7291333 B2 JP 7291333B2
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- hair
- present
- growth
- hair restorer
- palmitoyl dipeptide
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Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
- A61K38/05—Dipeptides
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K8/00—Cosmetics or similar toiletry preparations
- A61K8/18—Cosmetics or similar toiletry preparations characterised by the composition
- A61K8/30—Cosmetics or similar toiletry preparations characterised by the composition containing organic compounds
- A61K8/64—Proteins; Peptides; Derivatives or degradation products thereof
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/04—Peptides having up to 20 amino acids in a fully defined sequence; Derivatives thereof
- A61K38/06—Tripeptides
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P17/00—Drugs for dermatological disorders
- A61P17/14—Drugs for dermatological disorders for baldness or alopecia
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61Q—SPECIFIC USE OF COSMETICS OR SIMILAR TOILETRY PREPARATIONS
- A61Q7/00—Preparations for affecting hair growth
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
- C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
- C07K5/06—Dipeptides
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K5/00—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof
- C07K5/04—Peptides containing up to four amino acids in a fully defined sequence; Derivatives thereof containing only normal peptide links
- C07K5/06—Dipeptides
- C07K5/06086—Dipeptides with the first amino acid being basic
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
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- Medicinal Chemistry (AREA)
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Description
(1)実験動物
C57BL/6Nマウス(オス)およびBalb/c nu/nuマウス(メス)を日本エスエルシー株式会社(日本)より購入し、飼育の後に以下の実験に供した。なお、動物の飼育および試験は、関連法規、省令、および指針を遵守し、理化学研究所実験倫理審査会の承認のもとに実施した。
以下の試薬をそれぞれ用意した。
プラセボ:60%エタノール水溶液
実施例1:パルミトイルジペプチド-5ジアミノヒドロキシ酪酸 0.05%溶液
毛周期の1周期目は、60%エタノール水溶液をプラセボとして塗布する。上記の皮膚試験片を移植したBalb/c nu/nuマウスに、マイクロピペットを用いて25μLの60%エタノールを、皮膚試験片の生着させた左右の背部にそれぞれ塗布した。その後、ドライヤーを用いて冷風をあててエタノールを素早く乾燥させた。この作業を、マウス左右背部に各4回ずつ繰り返した。
Balb/c nu/nuマウスの皮膚試験片の移植部位から3領域をそれぞれ選択し、それら領域からそれぞれ5本の毛を選択して、発毛の状態を確認し記録した。観察と記録は、目視およびデジタルマイクロスコープ(株式会社キーエンス、日本)により行った。
(1)太毛化の計測方法
上記の試験例1において2周期目が終了した毛幹を用いて、太毛化の計測を行った。太毛化の計測には、採取した毛幹のうち、Zigzag毛の3本を用いた。その中心部分の太くなっている範囲を1辺 100μmの正方形で3か所を選択した。各選択した範囲ごとに異なる5か所をさらに選び、そこのZigzag毛の毛幹径を計測し、太毛化の程度を評価した。
毛幹径を計測した結果を図2および表2に示す。ここで、図2および表2中の**はp<0.01で有意であることを示す。
(1)ヒト毛乳頭細胞及び培地について
ヒト毛乳頭細胞(カタログ番号:CA602t05a、白色人種、29歳男性由来、東洋紡株式会社(日本))を購入し、プロトコールに記載されるようにして細胞を維持・培養して試験評価を行った。
試験用薬剤として、以下の各濃度(終濃度)のパルミトイルジペプチド-5ジアミノヒドロキシ酪酸を含むヒト毛乳頭細胞用培地からなる薬剤溶液を調製し、それを使用した。
実施例1:11.54709375μM
実施例2:23.0941875μM
実施例3:46.188375μM
実施例4:92.37675μM
実施例5:184.7535μM
実施例6:369.507μM
ヒト毛乳頭細胞を1×103個/ウェルとなるように、96ウェルプレートに播種した。CO2インキュベーター(5%CO2、37℃)内で1日間培養した後、ヒト毛乳頭細胞の培地を、上記の各濃度のパルミトイルジペプチド-5ジアミノヒドロキシ酪酸を含むヒト毛乳頭細胞用培地からなる薬剤溶液に置換した。その後、細胞プレートをCO2インキュベーターに戻し、さらに24時間、48時間、及び72時間培養した。培養後、培養の上澄み液を除去し、リン酸緩衝生理食塩水(略称:PBS)で細胞を洗浄した。PBSで洗浄した後、生細胞数測定試薬SF(ナカライテスク株式会社(日本))を10%含む培地を1ウェルあたり100μL添加した。添加後、培養上澄み液の吸光度(測定波長450nm、参照波長620nm)を測定した。この値をもとに、無添加対照群の細胞増殖率を100%とした際の各ウェルの細胞増殖率をそれぞれ算出した。
ヒト毛乳頭細胞にパルミトイルジペプチド-5ジアミノヒドロキシ酪酸を作用させた後の経時的な生細胞率の変化を測定し、その結果を、図3に示した。
(1)ヒト毛乳頭細胞及び培地について
上記試験例3と同様にして、ヒト毛乳頭細胞を維持・培養して試験評価を行った。
試験用薬剤として、以下の各濃度(終濃度)の薬剤溶液を調製し、使用した。
比較例1:アデノシン 100μM
実施例1:パルミトイルジペプチド-5ジアミノヒドロキシ酪酸 1μM
実施例2:パルミトイルジペプチド-5ジアミノヒドロキシ酪酸 10μM
実施例3:パルミトイルジペプチド-5ジアミノヒドロキシ酪酸 300μM
ヒト毛乳頭細胞を6×103個/ウェルとなるように、24ウェルプレートに播種した。CO2インキュベーター(5%CO2、37℃)内で、1日間培養後、各試験用薬剤を含む培地に置換した。その後、細胞プレートをCO2インキュベーターに戻し、さらに24時間培養した。
FGF-7遺伝子発現検出用プライマー
順方向:gagagaaaatccttctgcctgttg(配列番号1)
逆方向:cctggtgcaacttgagcctt(配列番号2)
GAPDH遺伝子発現検出用プライマー
順方向:catccctgcctctactggcgctgcc(配列番号3)
逆方向:ccaggatgcccttgagggggccctc(配列番号4)
各遺伝子の増幅曲線と閾値線との交点より、Ct値(PCRサイクル数)を算出した。目的遺伝子のCt値より内部標準GAPDH遺伝子のCt値で除した値が相対発現量となる。
ヒト毛乳頭細胞にパルミトイルジペプチド-5ジアミノヒドロキシ酪酸を24時間作用させた後のFGF-7遺伝子の発現量の変化を測定し、その結果を図4に示した。ここで、図4中の**はp<0.01で有意であることを示す。
Claims (11)
- パルミトイルジペプチド-5ジアミノヒドロキシ酪酸を含み、パルミトイルジペプチド-5ジアミノブチロイルヒドロキシスレオニンを含まない、外用剤である育毛剤。
- パルミトイルジペプチド-5ジアミノヒドロキシ酪酸の含有量が全体に対して0.001~20重量%である、請求項1に記載の育毛剤。
- パルミトイルジペプチド-5ジアミノヒドロキシ酪酸の含有量が全体に対して0.005~10重量%である、請求項1または請求項2に記載の育毛剤。
- 毛幹成長促進または発毛に用いるための、請求項1~請求項3のいずれか1項に記載の育毛剤。
- 毛幹伸長速度を向上させるために使用する、請求項1~請求項4のいずれか1項に記載の育毛剤。
- 毛幹最大長を向上させるために使用する、請求項1~請求項4のいずれか1項に記載の育毛剤。
- 毛幹径を増大させるために使用する、請求項1~請求項4のいずれか1項に記載の育毛剤。
- 毛数を増加させるために使用する、請求項1~請求項4のいずれか1項に記載の育毛剤。
- 溶液である、請求項1~請求項8のいずれか1項に記載の育毛剤。
- 頭髪、須毛、眉毛および/または睫毛用の、請求項1~請求項9のいずれか1項に記載の育毛剤。
- 請求項1~請求項10のいずれか1項に記載の育毛剤を非ヒト動物に投与することを含む育毛方法。
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