JP2014129267A - Dna損傷抑制剤 - Google Patents
Dna損傷抑制剤 Download PDFInfo
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- JP2014129267A JP2014129267A JP2012287063A JP2012287063A JP2014129267A JP 2014129267 A JP2014129267 A JP 2014129267A JP 2012287063 A JP2012287063 A JP 2012287063A JP 2012287063 A JP2012287063 A JP 2012287063A JP 2014129267 A JP2014129267 A JP 2014129267A
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- extract
- hair
- dna damage
- skin
- dna
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Abstract
【解決手段】 ショウブ科ショウブ属ショウブの抽出物を有効成分とするDNA損傷抑制剤を提供する。前記ショウブ科ショウブ属ショウブの抽出物としてショウブの根の抽出物を用いることができる。
【選択図】なし
Description
細胞内に起因するものとして、正常な代謝に伴って副生する活性酸素及び細胞内pH変動が挙げられる。また、外界に由来するものとして、紫外線、波長の短い電磁波(X線・γ線等)、化学物質、特に多環芳香族化合物(1−ニトロピレン等)、アスベスト、DNA架橋剤(ソラーレン等)、食物やタバコの煙、汚染大気中等に含まれる変異原性物質等、癌の化学療法、放射線療法等が挙げられる。
正常な皮膚においては、基底細胞層で分裂した表皮細胞が形態的・機能的に分化、成熟しながら角化を経て角層となり、順次剥がれ落ちていく。この過程は総称して表皮ターンオーバーと呼ばれるが、各段階が厳格に調節されることにより、定常状態として維持されている。したがって、表皮細胞のDNA損傷は、表皮細胞の増殖・分化や接着剥離に変調をきたし、表皮ターンオーバーが乱れ、皮膚バリア機能の低下や肌あれや、過角化の原因となる。
ここで、DNA損傷には、酸化修飾や脱アミノ化、チミジン2量体の形成、一本鎖切断、二重鎖切断等の様々な形態がある。これらのなかでも二重らせん構造を構成するDNA二重鎖が同時に切断されるいわゆる二重鎖切断は、修復するための鋳型を持たないことからより深刻な損傷であるといえる。
このように、DNA損傷抑制の中でもDNA二重鎖切断を防止できる又は修復できる化合物がDNA損傷抑制物質として望ましく、このような化合物が求められている。
すなわち、本発明は、ショウブ科ショウブ属ショウブの抽出物を有効成分とするDNA損傷抑制剤を提供するものである。
〔1〕ショウブ科ショウブ属ショウブ(Acorus calamus)の抽出物を有効成分とする、DNA損傷抑制剤。
〔2〕ショウブ科ショウブ属ショウブ(Acorus calamus)の根の抽出物を有効成分とする、〔1〕のDNA損傷抑制剤。
〔3〕DNA損傷がDNA二重鎖の切断であることを特徴とする〔1〕又は〔2〕に記載のDNA損傷抑制剤。
本発明に用いるショウブの抽出物は、一般的な方法で調製することができる。例えば、ショウブを、溶媒に浸漬することにより調製することができる。抽出溶媒としては特に限定されないが、例えば、水、低級アルコール類(メタノール、エタノール、1-プロパノール、2-プロパノール、1-ブタノール、2-ブタノール等)、多価アルコール類(グリセリン、プロピレングリコール、1,3-ブチレングリコール等)、ケトン類(アセトン、メチルケトン等)、エーテル類(ジエチルエーテル、テトラヒドロフラン等)、酢酸エチル等のエステル類;ヘキサン等からなる群より選択される一種又は二種以上を用いることができる。前記アルコール類は、1価アルコール類又は2価アルコール類(例えば、1,3−ブタンジオール、1,4−ブタンジオール等のブタンジオール等)が好ましく、アルコール類の炭素数は1〜5程度であるのが好ましい。さらに、前記溶媒として、水、1,3-ブチレングリコール、エタノール、またはこれらの混合溶液がより好ましく、エタノールまたは水−エタノール混合溶液が特に好ましい。水−エタノール混合溶液においては、エタノールが30〜99体積%であることが好ましく、50〜95体積%であるのが特に好ましい。
DNA損傷の原因には、活性酸素、紫外線照射、電磁波照射、化学物質等が考えられ、本発明の剤はいずれの原因によるものに対しても有効でありうるが、特に活性酸素の発生によるもの、又は紫外線照射によるものに対して有効である。
本発明の剤はいずれの形式の損傷に対しても有効でありうるが、特に、DNA二重鎖の切断の抑制に有効である。
このため、生細胞に添加した場合には「活性酸素」やその他の生体成分との反応生成物が、細胞やDNAを損傷するなど予期せぬ事態を引き起こすこともある(例えば、参考文献1:Biochemical
Pharmacology,66,(2003),1769-1778、参考文献2:Food and
Chemical Toxicology,49,(2011),955-962等参照)。参考文献1及び2には、茶に含まれる強力な抗酸化成分であるカテキンやエピガロカテキンガレードが、生体内に存在する銅(II)イオンや鉄(III)イオンと相互作用し、その反応生成物がDNA損傷を引き起こすことが示されている。
さらに、DNA損傷は生細胞内で起きているという観点で考えた場合、被験物質が化学的な抗酸化作用をどの程度示すかの試験が、生細胞内のDNAの損傷抑制を評価する際に有効な手法とは言い難いものがある。
これに対し、抗γH2AX抗体を用いるDNA損傷抑制試験は、生細胞を用いてDNA二重鎖切断が生じた箇所近傍のヒストンH2AXタンパク質のリン酸化を特異的に検出する手法である。DNA損傷の中でも最も深刻な二重鎖切断の抑制効果を観察及び定量的に判別できることから、より深刻な損傷に対して有効なDNA損傷抑制を評価する際に有効な手法と言える。
肌のキメを整える成分、 皮膚をすこやかに保つ成分、肌荒れを防ぐ成分、肌をひきしめる成分、皮膚にうるおいを与える成分、皮膚の水分、油分を補い保つ成分、皮膚の柔軟性を保つ成分、皮膚を保護する成分、皮膚の乾燥を防ぐ成分、肌を柔らげる成分、肌にはりを与える成分、肌にツヤを与える成分、肌を滑らかにする成分、日やけを防ぐ成分、日やけによるシミ、ソバカスを防ぐ成分、乾燥による小ジワを目立たなくする成分、頭皮・毛髪を清浄にする成分、毛髪、頭皮の不快臭を抑える成分、頭皮、毛髪をすこやかに保つ成分、毛髪にはり・こしを与える成分、頭皮・毛髪にうるおいを与える成分、頭皮・毛髪のうるおいを保つ成分、毛髪をしなやかにする成分、クシどおりをよくする成分、毛髪のつやを保つ成分、毛髪につやを与える成分、フケ・カユミがとれる成分、フケ・カユミを抑える成分、毛髪の水分・油分を補い保つ成分、裂毛・切毛・枝毛を防ぐ成分、髪型を整え、保持する成分、毛髪の帯電を防止する成分である。
ショウブ科ショウブ属ショウブ(Acorus calamus)の根茎100gを細切し、これに90体積%エタノール500mLを加えて加熱抽出した後濾過した。残渣を90体積%エタノール500mLで同様に抽出し全抽出液を合わせ、50℃で減圧濃縮し濃縮液約300mLを得た。この液にエタノールを加えてエタノール濃度を50体積%、全量450mLに調製した。この液を8日間冷所に放置して熟成させた後、濾過した。濾液の溶媒を留去して乾固し、固形分であるショウブ抽出物2.0gを得た。
ボタン科ボタン属ボタン(Paeonia suffruticosa Andrews)の根100gを細切し、これに50体積%エタノール150mLを加えて室温で5日間抽出を行った。抽出液を濾過後、溶媒を留去して乾固し、固形分であるボタンピ抽出物1.5gを得た。
ヒトメラノサイト(クラボウ社製)を増殖因子HMGS添加254培地(ライフテクノロジーズ社製)を用いてガラスボトムディッシュに10000個/cm2の濃度で播種した。播種翌日に実施例1のショウブ抽出物又は比較例1のボタンピ抽出物を最終濃度5μg/mL又は200μg/mLになるように添加し、37℃5%CO2存在下で1週間培養した。その後、HANKS液に交換した後、過酸化水素を最終濃度0.05μM、実施例1のショウブ抽出物又は比較例のボタンピ抽出物を最終濃度5μg/mL又は200μg/mLになるように添加し、37℃5%CO2存在下1時間培養してDNA損傷を誘導した。またショウブ抽出物又はボタンピ抽出物を添加せずに同様のDNA損傷誘導を行った細胞をコントロールとした。
DNA障害を誘導した細胞をPBSで洗浄し4%パラホルムアルデヒド溶液で細胞の固定を行った。さらに0.1%Triron-A100含有PBS溶液で15分処理した後、1次抗体として抗γH2AX抗体(Phospho-Histone
H2AX, Cell Signaling社製)を含有するPBS溶液を添加し4℃で12時間反応させた。続いて結合しなかった1次抗体溶液をPBSで洗浄除去し、2次抗体(Alexa Flour 546 Anti-rabbit IgG, Molecular Probes社製)を含有するPBSにて室温で2時間処理した。
PBSで洗浄した後に封入剤(DAPI-Fluoromount-G,
SouthernBiotech社製)で処理し蛍光顕微鏡下で細胞を観察した。染色画像を図1に示す。
ショウブ抽出物5μg/mL添加下で培養した場合にはDNA損傷マーカーであるγH2AX抗体染色量が減少し、ショウブ抽出物200μg/mL添加下ではさらにγH2AX抗体染色量が減少していた。ショウブ抽出物が細胞のDNA損傷を抑制していたことが示されている。一方、ボタンピ抽出物を添加して培養した場合には、γH2AX抗体染色量が増加していた。
コントロールでは約40%の細胞がγH2AX陽性つまりDNA損傷を生じていたが、ショウブ抽出物を5μg/mL添加した細胞ではDNA損傷の発生率は約37%、200μg/mL添加では約29%に抑制されており、その効果はショウブ抽出物の濃度依存的であった。一方、ボタンピ抽出物を添加して培養した場合には、ボタンピ抽出物の濃度依存的にDNA損傷が増加していた。
(製法)
A.下記成分(1)〜(8)を混合溶解する。
B.下記成分(9)〜(12)を混合溶解する。
C.AにBを加え混合し、化粧水を得た。
(1)クエン酸 0.05
(2)クエン酸ナトリウム 0.2
(3)ピロリドンカルボン酸ナトリウム(50%)水溶液 0.5
(4)グリチルリチン酸ジカリウム 0.1
(5)グリセリン 3.0
(6)1,3−ブチレングリコール 8.0
(7)精製水 残量
(8)実施例1のショウブ抽出物 0.0001
(9)エタノール 10.0
(10)香料 適量
(11)防腐剤 適量
(12)モノオレイン酸ポリオキシエチレン(20E.O.)
ソルビタン 0.5
(製法)
A.下記成分(1)〜(13)を加熱溶解し、70℃に保つ。
B.下記成分(14)〜(19)を加熱溶解し、70℃に保つ。
C.AにBを加え乳化し、更に下記成分(20)を加え混合する。
D.Cを冷却し、下記成分(21)を加え混合し、乳液を得た。
(1)ステアリン酸 1.0
(2)セタノール 0.5
(3)親油型モノステアリン酸グリセリン 0.5
(4)流動パラフィン 2.0
(5)スクワラン 3.0
(6)ホホバ油 3.0
(7)パルミチン酸セチル 0.2
(8)パルミチン酸レチノール 0.2
(9)酢酸トコフェロール 0.05
(10)防腐剤 適量
(11)モノステアリン酸ソルビタン 0.3
(12)モノオレイン酸ポリオキシエチレン(20E.O.)
ソルビタン 0.5
(13)ジブチルヒドロキシトルエン 0.1
(14)トリエタノールアミン 0.5
(15)1,3−ブチレングリコール 15.0
(16)グリセリン 3.0
(17)ポリエチレングリコール6000 0.5
(18)実施例1のショウブ抽出物 0.005
(19)精製水 残量
(20)カルボキシビニルポリマー1%水溶液 8.0
(21)香料 適量
(製法)
A.下記成分(1)〜(14)を加熱溶解し、70℃に保つ。
B.下記成分(15)〜(19)を加熱溶解し、70℃に保つ。
C.AにBを加え乳化し、更に下記成分(20)を加え混合する。
D.Cを冷却し、下記成分(21)、(22)を加え混合し、クリームを得た。
(1)ステアリン酸 2.5
(2)セタノール 2.5
(3)親油型モノステアリン酸グリセリン 2.0
(4)ワセリン 2.0
(5)ジペンタエリトリット脂肪酸エステル 2.0
(6)ミリスチン酸イソトリデシル 5.0
(7)流動パラフィン 8.0
(8)スクワラン 5.0
(9)ミツロウ 1.0
(10)パルミチン酸セチル 2.0
(11)セスキオレイン酸ソルビタン 0.5
(12)モノオレイン酸ポリオキシエチレン(20E.O.)
ソルビタン 1.5
(13)コエンザイムQ10 0.1
(14)防腐剤 適量
(15)トリエタノールアミン 1.2
(16)1,3−ブチレングリコール 8.0
(17)グリセリン 2.0
(18)ポリエチレングリコール20000 0.5
(19)精製水 残量
(20)カルボキシビニルポリマー1%水溶液 10.0
(21)実施例1のショウブ抽出物 0.05
(22)香料 適量
(製法)
A.下記成分(1)〜(8)を70℃で加熱混合した。
B.下記成分(9)〜(12)及び(14)〜(15)を50℃で加温混合した。
C.AにBを加えて乳化し、冷却後(13)を添加して油中水型日焼け止めクリームを得た。
(1)ポリオキシアルキレン変性オルガノポリシロキサン(注1) 2.0
(2)パルミチン酸オクチル 15.0
(3)デカメチルシクロペンタシロキサン 20.0
(4)トリベヘン酸グリセリル 1.0
(5)微粒子酸化亜鉛 12.0
(6)微粒子酸化チタン 3.0
(7)パラメトキシケイ皮酸2−エチルヘキシル(注2) 7.0
(8)4−tertブチル−4’−メトキシジベンゾイルメタン
(注3) 1.0
(9)ジプロピレングリコール 5.0
(10)エタノール 5.0
(11)ポリエチレン末 3.0
(12)防腐剤 適量
(13)香料 適量
(14)実施例1のショウブ抽出物 0.0005
(15)精製水 残量
(注1)KF−6017(信越化学工業社製)
(注2)ユビナールMC80(BASF社製)
(注3)PARSOL 1789(L.C.UNITED社製)
(製法)
A.下記成分(1)〜(5)及び(15)を70℃で加熱混合し、室温まで冷却する。
B.Aに下記成分(6)〜(14)を添加混合してパック化粧料を得た。
(1)ポリビニルアルコール 15.0
(2)グリセリン 10.0
(3)ポリオキシエチレン(10)メチルグルコール 3.0
(4)トリオクタン酸グリセリル 5.0
(5)ポリオキシエチレンアルキルエーテルリン酸ナトリウム 1.0
(6)エタノール 20.0
(7)カオリン 2.0
(8)酸化チタン 2.0
(9)グリチルリチン酸ジカリウム 0.1
(10)乳酸(50%水溶液) 0.5
(11)乳酸ナトリウム(50%水溶液) 0.5
(12)防腐剤 適量
(13)香料 適量
(14)実施例1のショウブ抽出物 0.001
(15)精製水 残量
(製法)
A.下記成分(1)〜(7)を70℃で加熱混合し、この混合物に下記成分(13)〜(18)を加えて混合し70℃に保つ。
B.下記成分(8)〜(12)を70℃で加熱混合する。
C.BにAを加えて乳化し、冷却後、下記成分(19)〜(20)を添加してリキッドファンデーションを得た。
(1)ジペンタエリトリット脂肪酸エステル 2.0
(2)流動パラフィン 5.0
(3)ステアリン酸 2.0
(4)セタノール 1.0
(5)自己乳化型モノステアリン酸グリセリル 1.0
(6)パラメトキシケイ皮酸2−エチルヘキシル 8.0
(7)防腐剤 適量
(8)グリセリン 5.0
(9)トリエタノールアミン 1.0
(10)カルボキシメチルセルロース 0.2
(11)ベントナイト 0.5
(12)精製水 残量
(13)酸化チタン 6.0
(14)微粒子酸化チタン 2.0
(15)微粒子酸化亜鉛 4.0
(16)マイカ 2.0
(17)タルク 4.0
(18)着色顔料 適量
(19)実施例1のショウブ抽出物 0.1
(20)香料 適量
(製法)
A.成分(1)〜(3)を加熱混合し、75℃に保つ。
B.成分(4)〜(9)を混合し、75℃に保つ。
C.AにBを徐々に加え、軟膏剤を得た。
(1)ステアリン酸 18.0
(2)セタノール 4.0
(3)酢酸dl−α―トコフェロール(注4) 0.2
(4)トリエタノールアミン 2.5
(5)グリセリン 5.0
(6)グリチルリチン酸ジカリウム(注5) 0.5
(7)実施例1のショウブ抽出物 1.0
(8)パラオキシ安息香酸メチル 0.1
(9)精製水 残量
(注4)エーザイ社製
(注5)和光純薬工業社製
(製法)
A.成分(1)〜(5)を混合溶解する。
B.成分(6)〜(10)を混合溶解する。
C.AとBを混合して均一にし、養毛剤を得た。
(1)エタノール 50.0
(2)ポリオキシエチレン硬化ヒマシ油(80E.O.) 0.5
(3)メントール 0.01
(4)カンファ 0.005
(5)フェノキシエタノール 0.05
(6)精製水 残量
(7)実施例1のショウブ抽出物 0.01
(8)オタネニンジン抽出物 注6 0.5
(9)パントテニルアルコール 注7 0.1
(10)グリセリン 5.0
注6 一丸ファルコス社製
注7 関東化学社製
(製法)
A.成分(1)〜(11)を常温にて均一混合し、シャンプーを得た。
1.ヤシ油脂肪酸メチルタウリンナトリウム 10.0
2.テトラデセンスルホン酸ナトリウム 5.0
3.ヤシ油脂肪酸アミドプロピルベタイン 5.0
4.ヤシ油脂肪酸時エタノールアミド 4.0
5.塩化ナトリウム 0.5
6.精製水 残量
7.カチオン化セルロース 0.1
8.実施例1のショウブ抽出物 0.05
9.エタノール 1.0
10.メチルパラベン 0.1
11.香料 適量
エアゾール原液処方
(製法)
A:成分1〜15を均一に混合溶解して、エアゾール原液を得た。
B:Aのエアゾール原液と噴射剤(液化石油ガス)の質量比が97:3になるようにエアゾール缶に充填し、泡沫状ヘアトリートメントを得た。
1.塩化ベヘニルトリメチルアンモニウム 0.5
2.アモジメチコン 注8 1.0
3.ジメチルポリシロキサン(10mPa・s) 2.0
4.N−ラウロイル−L−グルタミン酸ジ(フィトステリル
・ベヘニル・2−オクチルドデシル) 注9 0.5
5.1,3−ブチレングリコール 5.0
6.実施例1のショウブ抽出物 0.2
7.精製水 5.0
8.ポリクオタニウム−51 注10 0.5
9.ポリメタクリロイルオキシエチルホスホリルコリン
注11 0.5
10.ポリクオタニウム−65 注12 0.5
11.ポリクオタニウム−64 注13 0.5
12.ポリクオタニウム−61 注14 0.5
13.香料 0.1
14.メチルパラベン 0.3
15.精製水 残量
注8 SM 8904 COSMETIC EMALSION(東レ・ダウコーニング社製)
注9 エルデュウPS−304(味の素社製)
注10 LIPIDURE−PMB(日油社製)
注11 LIPIDURE−HM−600(日油社製)
注12 LIPIDURE−A(日油社製)
注13 LIPIDURE−C(日油社製)
注14 LIPIDURE−S(日油社製)
(製法)
A.成分1〜7を均一に混合し、常法に従って錠剤を得た。
(1)乳糖 24.0
(2)結晶セルロース 20.0
(3)コーンスターチ 15.0
(4)実施例1のショウブ抽出物 0.1
(5)デキストリン 残量
(6)グリセリン脂肪酸エステル 5.0
(7)二酸化ケイ素 1.0
(製法)
A.成分1〜5を均一に混合し、常法に従って清涼飲料を得た。
(1)果糖ブドウ糖液糖 30.0
(2)乳化剤 0.5
(3)実施例1のショウブ抽出物 0.001
(4)香料 適量
(5)精製水 残量
〔1〕ショウブ科ショウブ属ショウブ(Acorus calamus)の抽出物を有効成分とする、DNA損傷抑制剤。
〔2〕ショウブ科ショウブ属ショウブ(Acorus calamus)の根の抽出物を有効成分とする、〔1〕のDNA損傷抑制剤。
〔3〕DNA損傷がDNA二重鎖の切断であることを特徴とする〔1〕又は〔2〕に記載のDNA損傷抑制剤。
Claims (3)
- ショウブ科ショウブ属ショウブ(Acorus calamus)の抽出物を有効成分とする、DNA損傷抑制剤。
- ショウブ科ショウブ属ショウブ(Acorus calamus)の根の抽出物を有効成分とする、請求項1のDNA損傷抑制剤。
- DNA損傷がDNA二重鎖の切断であることを特徴とする請求項1又は2に記載のDNA損傷抑制剤。
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JP2015044788A (ja) * | 2013-07-29 | 2015-03-12 | 株式会社コーセー | ショウブ抽出液を含む表皮幹細胞性維持剤 |
JP2018080127A (ja) * | 2016-11-15 | 2018-05-24 | ポーラ化成工業株式会社 | ケラタン硫酸産生促進剤 |
WO2020203217A1 (ja) * | 2019-03-29 | 2020-10-08 | サントリーホールディングス株式会社 | Ltbp-1発現促進用組成物及びltbp-1発現促進作用を有する物質のスクリーニング方法 |
JP2021008439A (ja) * | 2019-07-02 | 2021-01-28 | 株式会社ミルボン | 多剤式化粧料、及び多剤式化粧料を用いた美容方法 |
JP7352489B2 (ja) | 2020-02-26 | 2023-09-28 | 久光製薬株式会社 | マッサージ用エアゾール製剤 |
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JP2015044788A (ja) * | 2013-07-29 | 2015-03-12 | 株式会社コーセー | ショウブ抽出液を含む表皮幹細胞性維持剤 |
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