JP2019510829A - 細胞培養培地中に有効成分として、アルブミン、ヒアルロン酸またはコラーゲンを含む化粧品組成物 - Google Patents
細胞培養培地中に有効成分として、アルブミン、ヒアルロン酸またはコラーゲンを含む化粧品組成物 Download PDFInfo
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Abstract
Description
細胞培養培地、および有効成分として、アルブミン、ヒアルロン酸およびコラーゲンのいずれか1つを含む化粧品組成物または医薬組成物は、細胞の回復能力を高め、したがって、外的損傷、皮膚疾患、または皮膚状態からの皮膚創傷を改善、予防または治療するための化粧品組成物または医薬組成物として効果的に使用することができ、新規な化粧品または治療剤として使用することができる。また、それは、現在使用されている皮膚軟膏や機能性化粧品に利用することができ、添加剤として容易に開発されうる。
1つの態様では、本発明は、細胞培養培地;およびアルブミン、ヒアルロン酸、およびコラーゲンからなる群から選択されるいずれか1つを含む組成物を提供する。
a)アミノ酸成分は、グリシン、L-アラニン、L-バリン、L-ロイシン、L-イソロイシン、L-トレオニン、L-セリン、L-システイン、L-メチオニン、L-アスパラギン酸、L-アスパラギン、L-グルタミン酸、L-グルタミン、L-リジン、L-アルギニン、L-ヒスチジン、L-フェニルアラニン、L-チロシン、L-トリプトファン、L-プロリン、β-アラニン、γ-アミノ酪酸、オルニチン、シトルリン、ホモセリン、トリヨードチロシン、チロキシンおよびジオキシフェニルアラニンからなる群から選択される少なくとも1つのアミノ酸、またはそれらの組み合わせであり、好ましくは、グリシン、L-アラニン、L-アルギニン、L-システイン、L-グルタミン、L-ヒスチジン、L-リジン、L-メチオニン、L-プロリン、L-セリン、L-トレオニンおよびL-バリンからなる群から選択される少なくとも1つ、またはそれらの組み合わせである;
b)ビタミン成分は、ビオチン、D-パントテン酸カルシウム、葉酸、ナイアシンアミド、塩酸ピリドキシン、リボフラビン、塩酸チアミン、ビタミンB12、塩化コリン、i-イノシトールおよびアスコルビン酸からなる群から選択される少なくとも1つ、またはそれらの組み合わせのビタミンであり、好ましくは、i-イノシトール、塩酸チアミン、ナイアシンアミドおよび塩酸ピリドキシンからなる群から選択される少なくとも1つ、またはそれらの組み合わせである;
c)無機塩成分は、塩化カルシウム(CaCl2)(無水)、硫酸銅五水和物(CuSO4-5H2O)、硫酸第二鉄七水和物(FeSO4-7H2O)、塩化マグネシウム(無水)、硫酸マグネシウム(MgSO4)(無水)、塩化カリウム(KCl)、塩化ナトリウム(NaCl)、リン酸水素二ナトリウム(Na2HPO4)、リン酸二水素ナトリウム一水和物(NaH2PO4-H2O)、硫酸亜鉛七水和物(ZnSO4-7H2O)、硝酸第2鉄九水和物(Fe(NO3)3・9H2O)、および重炭酸ナトリウム(NaHCO3)からなる群から選択される少なくとも1つの無機塩、またはそれらの組み合わせであり、好ましくは、塩化ナトリウム(NaCl)、炭酸水素ナトリウム(NaHCO3)、塩化カリウム(KCl)、塩化カルシウム(CaCl2)(無水)およびリン酸二水素ナトリウム一水和物(NaH2PO4-H2O)からなる群から選択される少なくとも1つの無機塩またはそれらの組み合わせである;
d)その他の成分は、D-グルコース(デキストロース)、ピルビン酸ナトリウム、ヒポキサンチンNa、チミジン、リノール酸、リポ酸、アデノシン、シチジン、グアノシン、ウリジン、2'-デオキシアデノシン、2'-デオキシシチジンHCl、2'-デオキシグアノシン、またはそれらの組み合わせの少なくとも1つであり、好ましくは、ピルビン酸ナトリウムである;および
e)精製水は、上記のアミノ酸、ビタミン、無機塩およびその他の成分を溶解するために用いられ、少なくとも単回蒸留によって得られた水であってもよいし、あるいはフィルターを介する精製によって得られた水であってもよい。
細胞培養培地、アルブミンおよびI型コラーゲン、または細胞培養培地、アルブミン、III型コラーゲンおよびヒアルロン酸のすべてを含む組成物が、優れた皮膚改善効果を有することが知られている、細胞培養培地およびEGFを含む組成物と比較して、優れた創傷治癒効果を示すことが見出された(図2および図4参照)。
以下、本発明を実施例によって詳細に説明する。以下の実施例は、本発明の範囲を限定することなく本発明をさらに説明することを意図している。
第1表に記載した各成分を超純水に十分に溶解させ、全成分が溶解したことを確認した後、0.2μmのフィルターで濾過した。第1表に列挙された成分のすべては、Sigma-Aldrich、USAから購入した。以下に列挙された22の成分を含む混合組成物である細胞培養培地を調製した(以下BGと称する)。
アルブミン、コラーゲンまたはヒアルロリン酸を、調製例1で調製したBGと、第2表に示す含有量にしたがって混合して、組成物を調製した。
MEMα (Sigma-Aldrich、USA)を、アルブミン、コラーゲンまたはヒアルロン酸と、第3表に示す含有量にしたがって混合して、組成物を調製した。
ヒト皮膚線維芽細胞10%FBS(ウシ胎仔血清)および1%P/S(ペニシリン-ストレプトマイシン)を補充した細胞培養培地DMEM(1X)を用い、ヒト皮膚線維芽細胞株(Hs27)を37℃の恒温恒湿器で5%CO2を維持して培養した。ヒト皮膚線維芽細胞をインサートウェルあたり5.2×104細胞で培養した6ウェルプレートに、イビディカルチャーインサート(ibidi Culture-insert)を付着させた。上記条件下でヒト皮膚線維芽細胞を24時間培養した後、細胞を無血清DMEM(1X)培養液中で18時間再培養して、本実験まで無血清状態を維持した。次いで、イビディカルチャーインサートを除去した。
調製した細胞培養液を、BG(対照群)、BGおよび100μg/mlのアルブミンの混合組成物(実施例1)、BGおよび10μg/mlのI型コラーゲンの混合組成物(実施例2)、または100μg/mlのアルブミンおよび10μg/mlのI型コラーゲンの混合組成物(実施例3)で処理した。
調製した細胞培養溶液を、BG(対照群)、BGおよび100μg/mlのアルブミンの混合組成物(実施例1)、BGおよび0.1mg/mlのヒアルロン酸の混合組成物(実施例4)、BGおよび20μg/mlのIII型コラーゲンの混合組成物(実施例5)、またはBG、100μg/mlのアルブミン、0.1mg/mlのヒアルロン酸および20μg/mlのIII型コラーゲンの混合組成物(実施例9)で処理した。対照群において、BG、100μg/mlのアルブミン、0.1mg/mlのヒアルロン酸、または20μg/mlのIII型コラーゲンを単独で加えた。
実験例1.2と同様の方法で実験を行い、実施例10〜16における細胞の回復能力を調べた。結果として、成分を混合した後にアルブミン、ヒアルロン酸、およびIII型コラーゲンの組み合わせで細胞培養培地を処理した場合、各成分での単独処理と比較して、細胞の回復能力が優れていることが判明した。さらに、上記の3つの物質のすべてを組み合わせて添加すると、最良の回復能力が観察された。
製造例1で調製した細胞培養培地およびアルブミン500μg/mlを含む組成物が、皮膚疾患の一種であるアトピーに有効であるかどうかを調べた。
製造例1で調製した細胞培養培地(BG)およびアルブミン100μg/mlを含む組成物が、角膜びらんに有効であるかどうかを調べた。
10%FBS(ウシ胎仔血清)および1%P/S(ペニシリン-ストレプトマイシン)を補充した細胞培養培地DMEM(1X)を用い、ヒト皮膚線維芽細胞株(Hs27)を37℃の恒温恒湿器で5%CO2を維持して培養した。ヒト皮膚線維芽細胞をインサートウェルあたり5.2×104細胞で培養した6ウェルプレートに、イビディカルチャーインサート(ibidi Culture-insert)を付着させた。上記条件下でヒト皮膚線維芽細胞を24時間培養した後、細胞を無血清DMEM(1X)培養液中で18時間再培養して、本実験まで無血清状態を維持した。次いで、イビディカルチャーインサートを除去した。
Claims (28)
- 細胞培養培地;および
アルブミン、ヒアルロン酸、およびコラーゲンからなる群から選択されるいずれか1つ;を含む組成物。 - 細胞培養培地が、動物細胞培養培地である、請求項1に記載の組成物。
- 細胞培養培地が、アミノ酸成分、ビタミン成分、無機塩成分およびその他の成分から構成される、請求項1に記載の組成物であって、ここで:
a)アミノ酸成分は、グリシン、L-アルギニン、L-システイン、L-グルタミン、L-ヒスチジン、L-リジン、L-メチオニン、L-プロリン、L-セリン、L-トレオニンおよびL-バリンからなる群から選択される少なくとも1つのアミノ酸、またはそれらの組み合わせであり;
b)ビタミン成分は、i-イノシトール、塩酸チアミン、ナイアシンアミドおよび塩酸ピリドキシンからなる群から選択される少なくとも1つのビタミン、またはそれらの組み合わせであり;
c)無機塩成分は、塩化ナトリウム(NaCl)、炭酸水素ナトリウム(NaHCO3)、塩化カリウム(KCl)、塩化カルシウム(CaCl2)(無水)およびリン酸二水素ナトリウム一水和物(NaH2PO4-H2O)からなる群から選択される少なくとも1つの無機塩、またはそれらの組み合わせであり;および
d)その他の成分は、ピルビン酸ナトリウムである;組成物。 - 細胞培養培地が、グリシン、L-アラニン、L-アルギニン塩酸塩、L-システイン塩酸塩一水和物、L-グルタミン、L-ヒスチジン塩酸塩一水和物、L-リジン塩酸塩、L-メチオニン、L-プロリン、L-セリン、L-トレオニン、L-バリン、i-イノシトール、塩酸チアミン、ナイアシンアミド、塩酸ピリドキシン、塩化ナトリウム(NaCl)、炭酸水素ナトリウム(NaHCO3)、塩化カリウム(KCl)、塩化カルシウム(CaCl2)(無水)、リン酸二水素ナトリウム一水和物(NaH2PO4-H2O)、およびピルビン酸ナトリウムから構成される、請求項1に記載の組成物。
- アミノ酸成分が、1重量部のアミノ酸セリンに基づいて、0.2〜1.2重量部のグリシン、0.08〜0.3重量部のL-アラニン、2〜7重量部のL-アルギニン塩酸塩、0.2〜0.9重量部のL-システイン塩酸塩一水和物、10〜18重量部のL-グルタミン、0.8〜2.0重量部のL-ヒスチジン塩酸塩一水和物、1.0〜5.0重量部のL-リジン塩酸塩、0.2〜1.2重量部のL-メチオニン、0.2〜1.2重量部のL-プロリン、1.0〜5.0重量部のL-トレオニン、および1.0〜5.0重量部のL-バリンを含む、請求項3に記載の組成物。
- ビタミン成分が、1重量部のアミノ酸セリンに基づいて、0.2〜0.9重量部のi-イノシトール、0.04〜0.8重量部の塩酸チアミン、0.04〜0.8重量部のナイアシンアミドおよび0.04〜0.8重量部の塩酸ピリドキシンを含む、請求項3に記載の組成物。
- 無機塩成分が、1重量部のアミノ酸セリンに基づいて、50〜200重量部の塩化ナトリウム(NaCl)、50〜120重量部の炭酸水素ナトリウム(NaHCO3)、2.0〜20重量部の塩化カリウム(KCl)、1.0〜10重量部の塩化カルシウム(CaCl2)(無水)および0.5〜5.0重量部のリン酸二水素ナトリウム一水和物(NaH2PO4-H2O)を含む、請求項3に記載の組成物。
- その他の成分が、1重量部のアミノ酸セリンに基づいて、0.5〜5.0重量部のピルビン酸ナトリウムを含む、請求項3に記載の組成物。
- 細胞培養培地が、無血清培地である、請求項1に記載の組成物。
- 細胞培養培地が、DMEM/F-12(ダルベッコ改変イーグル培地/栄養素混合物 F-12)、DMEM、MEMα(最小必須培地α)、BME(イーグル基礎培地)、GMEM (グラスゴー最小必須培地)、RPMI-1640培地、およびそれらの組み合わせからなる群から選択される、請求項2に記載の組成物。
- アルブミンが、ヒト血清アルブミンまたはウシ血清アルブミンである、請求項1に記載の組成物。
- アルブミンが、10〜1000μg/mlの濃度で含まれる、請求項1に記載の組成物。
- ヒアルロン酸が、0.01〜1mg/mlの濃度で含まれる、請求項1に記載の組成物。
- コラーゲンが、I型コラーゲンまたはIII型コラーゲンである、請求項1に記載の組成物。
- I型コラーゲンが、1〜100μg/mlの濃度で含まれる、請求項14に記載の組成物。
- III型コラーゲンが、2〜200μg/mlの濃度で含まれる、請求項14に記載の組成物。
- 有効成分として、請求項1に記載の組成物を含む、皮膚状態を改善するための化粧品組成物。
- 皮膚細胞の機能の低下または喪失から皮膚を保護するか、または皮膚状態を改善する、請求項17に記載の組成物であって、皮膚状態の改善が、しわの発生の阻害、皮膚の老化の阻害、皮膚の弾力性の改善、皮膚再生、損傷または創傷治癒、角膜再生、皮膚刺激緩和、およびそれらの組み合わせからなる群から選択される、組成物。
- 有効成分として、請求項1に記載の組成物を含む、皮膚疾患を予防または治療するための医薬組成物。
- 皮膚疾患が、アトピー性皮膚炎、アレルギー、皮膚湿疹、にきび、乾癬、掻痒、およびそれらの組み合わせからなる群から選択される、請求項19に記載の組成物。
- 皮膚状態を改善するための化粧品組成物の製造における、請求項17に記載の化粧品組成物の使用。
- 請求項17に記載の化粧品組成物を用いる、皮膚状態を改善するための方法。
- 該化粧品組成物を、それを必要とする対象の皮膚に塗布することを含む、請求項22に記載の方法。
- 対象が、哺乳動物である、請求項23に記載の方法。
- 請求項19に記載の医薬組成物を用いる、皮膚状態を予防または治療するための方法。
- 該医薬組成物を、それを必要とする対象の皮膚に塗布することを含む、請求項25に記載の方法。
- 対象が、哺乳動物である、請求項26に記載の方法。
- 皮膚疾患を予防または治療するための医薬品の製造における、請求項19に記載の医薬組成物の使用。
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KR102553943B1 (ko) | 2023-07-12 |
EP3437628A4 (en) | 2019-11-27 |
US11071706B2 (en) | 2021-07-27 |
JP6934682B2 (ja) | 2021-09-15 |
CN115350115B (zh) | 2024-03-22 |
CN109195579B (zh) | 2022-09-16 |
KR20220030226A (ko) | 2022-03-10 |
KR102368465B1 (ko) | 2022-03-02 |
EP3437628A1 (en) | 2019-02-06 |
US20200297613A1 (en) | 2020-09-24 |
CN115350115A (zh) | 2022-11-18 |
CN109195579A (zh) | 2019-01-11 |
WO2017171497A1 (ko) | 2017-10-05 |
US20210353519A1 (en) | 2021-11-18 |
KR20170113460A (ko) | 2017-10-12 |
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