JP5220761B2 - キノア穀物抽出物を含む皮膚科学用途のための組成物 - Google Patents
キノア穀物抽出物を含む皮膚科学用途のための組成物 Download PDFInfo
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- JP5220761B2 JP5220761B2 JP2009543481A JP2009543481A JP5220761B2 JP 5220761 B2 JP5220761 B2 JP 5220761B2 JP 2009543481 A JP2009543481 A JP 2009543481A JP 2009543481 A JP2009543481 A JP 2009543481A JP 5220761 B2 JP5220761 B2 JP 5220761B2
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Description
ケチュア語では、キヌア(quinua)、キウナ(kiuna)であり、(南米には、多数の他の名称がある)
スペイン語では、キノア、キヌア(quinua)、アロス・デル・ペルー(arroz del Peru;ペルーのイネ)であり、
フランス語では、キノア、プティ・リ(petit riz)、リ・ズ・ペルー(riz du Perou)であり、
英語では、キノア、キヌア(quinua)である。
ペルーでは、カンコーラ(Kancolla)、チェウェカ(Cheweca)、ウィツラ(Witulla)、タウァコ(Tahuaco)、カマカニ(Camacani)、ヨカレ(Yocare)、ウィラカユニ(Wilacayuni)、ブランカ・デ・フリ(Blanca de Juli)、アマリラ・デ・マランガニ(Amarilla de Marangani)、パクス(Pacus)、ロサーダ(Rosada)、ブランカ・デ・フニン(Blanca de Junin)、ファルファス(Hualhuas)、ファンカヨ(Huancayo)、マンタロ(Mantaro)、ファカリス(Huacariz)、ファカタス(Huacataz)、アコスタンボ(Acostambo)、ブランカ・アヤクチャナ(Blanca Ayacuchana)およびマリノ(Narino);
ボリビアでは、サハマ(Sajama)、リアル・ブランカ(Real Blanca)、チュカパカ(Chucapaca)、カミリ(Kamiri)、ファランガ(Huaranga)、パサンカラ(Pasancalla)、パンデラ(Pandela)、ツピサ(Tupiza)、ファチャプク(Jachapucu)、ウィラ・コイミニ(Wila Coymini)、ケルー(Kellu)、ウチュサヤ(Uthusaya)、チュルピ(Chullpi)、カスラリ(Kaslali)およびチルピ(Chillpi);
エクアドルでは、インバヤ(Inbaya)、チャウチャ(Chaucha)、INIAP−コチャスキー(INIAP−Cochasqui)、タンラファ(Tanlahua)、ピァルタール(Piartal)、ポロトク(Porotoc)、アマルガ・デル・チンボラソ(Amarga del Chimborazo)、アマルガ・デ・インバブラ(Amarga de Imbabura)およびモラダ(Morada);
コロンビアでは、デュルセ・デ・キトパンパ(Dulce de Quitopampa);
アルゼンチンでは、ブランカ・デ・フフイ(Blanca de Jujuy);
チリでは、バェル(Baer)、リト(Lito)、ファト(Faro)およびピッチャマン(Picchaman)。
機械的プレスでのホットプレス、二軸スクリュー押出機でのプレスのような物理抽出、
有機溶媒(脂肪族アルカン、アルコール、塩素化溶媒、フッ素化溶媒)による化学抽出、
二酸化炭素単独および/または補助溶媒を用いる超臨界媒質での抽出
によって抽出することができる。
a)キノア穀物から出発し、生油およびケーキを抽出し、前記ケーキを回収し、
b)前記ケーキを水または水/アルコール混合物で洗浄してタンパク質部分のみを保持し、次いで
c)タンパク質を可溶化し、
d)タンパク質を濃縮した後、前記タンパク質を加水分解してペプチドとし、
e)ペプチド抽出物を精製して回収する
連続的な工程を含んでなる方法によって得られる。
a)キノア穀物から出発し、生油およびケーキを抽出し、前記ケーキを回収し、
b)前記ケーキを水または水/アルコール混合物で洗浄してタンパク質部分のみを保持し、次いで
c)タンパク質を可溶化し、
d)タンパク質を濃縮した後、前記タンパク質を加水分解してペプチドとし、
e)ペプチド抽出物を精製して回収する
連続的な工程を含んでなる、キノアのペプチドおよび糖類抽出物の製造方法でもある。
皮膚軟化薬、加湿活性剤、ケラチン合成の活性剤、角質調節剤、角質溶解剤、皮膚バリヤー再構成薬(皮膚脂質合成活性剤)、PPAR(ペルオキシゾーム増殖剤応答性受容体)作動薬、RXRまたはLXR作動薬、SERM、ビタミンDまたはコルチコイド受容体の作動薬、ケラチノサイト分化活性剤(レチノイド、カルシドン(calcidone)(商標)、カルシウム)、皮脂調節剤(5−aレダクターゼの阻害剤、特にLaboratoires Expanscienceから発売されている5−a活性Avocuta(商標))、防腐剤、刺激防止薬、鎮静剤、日光フィルターおよび遮断薬、酸化防止薬のような皮膚科学で従来から用いられている活性剤、
抗生物質、プレおよびプロ−バイオティクス(pre- and pro-biotics)、抗菌薬、抗真菌性化合物、抗ウイルス薬、免疫調節薬(タクロリムスまたはピメクロリムス)、オキサゾリン、成長因子、治癒薬または富栄養分子、薬剤、抗炎症薬、色素または低色素薬、脂肪分解薬または脂質形成阻害薬、色素または超微細無機または有機日光フィルターおよび遮断薬、通常または機能性食品: 高または低血糖、抗脂肪または抗蜂巣炎栄養素、閉経の二次徴候に影響する抗コレステロール、酸化防止剤、賦活薬、強壮栄養素のような補助治療作用を有する活性成分、
天然植物抽出物(水性または油性相で抽出可能な植物部分:ポリフェノール、フラボノイド、他のペプチドおよび糖など)、植物油の不ケン化物を含む化合物、ステロール不ケン化物、またはそれらを含むことがある生成物(植物油の不ケン化物、特に大豆油の不ケン化物、植物性バターの不ケン化物、またはバター様材料およびそれらの混合物、天然ワックスの不ケン化物、油抽出物の不ケン化物、産業上の油性副生成物の不ケン化物、動物性脂肪抽出物の不ケン化物、魚油の不ケン化物、乳脂肪の不ケン化物、単細胞生物から抽出した脂質の不ケン化物、藻類および海洋生物から抽出した脂質の不ケン化物など)、ステロール、スタノール、フィトステロール、フィトスタノール、トコフェロール、ヒマワリ濃縮物、ナタネおよび/またはヤシ油、オリゴ要素、ビタミン、ω3、6または9脂肪酸、低血糖または高血糖性または甘味植物
からなる群より選択される少なくとも1種類の化合物を含んでなる。
皮膚を着色する薬剤:ジヒドロキシアセトン、メラニン、
天然の着色過程を刺激する薬剤:ソラレン(8−メトキシソラレン、5−メトキシソラレン、4,5’,8−トリメチルソラレンまたはPsorelea corylifoliaおよびAmmi majusの植物抽出物)、カロチノイド(リコペン、カンタキサンチン)、サイクリックAMP経路を刺激する薬剤(1.8−ブロモ−cAMPまたはジブチリル−cAMPのようなcAMP類似体、2.ホルスコリン、3.イソブチル−メチル−キサンチンまたはテオフィリン)、キナーゼCタンパク質の活性剤(ジアセチルグリセロール、特にオレイル−アセチル−グリセロール)、脂肪族または環状ジオール(1,2−プロパンジオール、5−ノルボルナン−2,2−ジメタノール、ノルボルナン−2,2−ジメタノール)、モノテルペン二環性ジオール、チロシン誘導体(L−チロシン、L−DOPA)、ジメチルスルホキシド、リソモトローピック薬(lysomotropic agents)、チミジンジヌクレオチド、DNA断片、メラニン細胞刺激ホルモン類似体、3−イソブチル−l−メチルキサンチン、硝酸ドナー(Brown, Journal of Photochemistry and Photobiology B: biology 63 (2001) 148-161)、
植物抽出物、特にメラニン形成促進作用を示す藻類:Laminaria digitata(Thalitan de Codif)
を挙げることができる。
1) 酪農製品: 例えば、チーズ、バター、ミルクおよび他の乳飲料、乳製品を基剤とした混合物およびスプレッド、アイスクリームおよびヨーグルト、
2) 脂肪を基剤とした製品: マーガリン、スプレッド、マヨネーズ、クーリング・ファッツ(cooling fats)、フライ用油、およびフレンチ・ドレッシング、
3) 穀物からなるシリアルを基剤とする製品: 食材を加熱調理、オーブンで加熱調理または変容させたパンおよびパスタ、
4) 菓子: チョコレート、砂糖菓子、チューインガム、デザート、トッピング、シャーベット、糖衣、および他の付け合わせ、
5) アルコールまたは清涼飲料: 例えばソーダ水および他の清涼飲料、フルーツジュース、補助栄養食品、Boost(商品名)およびEnsure(商品名)の名称で販売されている飲料形態の代替食品、および
6) 様々な製品: 例えばスープ、パスタ用の既製スープ、調理食品および同じ種類の他の製品
に含まれる。
ニキビ、アトピー性皮膚炎、脂漏性皮膚炎、赤鼻、乾癬、脈管傷害、シート皮膚炎(seat dermitis)、糜爛、亀裂、刺傷、特に胸部の亀裂、日射病、任意の種類の光線による炎症、刺激またはアレルギー(化学、物理薬剤(引張応力:妊婦)による)、細菌、真菌またはウイルス、寄生生物(蚤、疥癬、白癬、ダニ、皮膚糸状菌)、放射線または放射物(UV、IR)刺激またはアレルギー、または先天性免疫不全(抗微生物ペプチド)または後天性免疫不全(細胞性、体液性サイトカイン)、皮膚萎縮線条のような、皮膚、および/または
粘膜(歯肉炎(タバコへの耽溺に関する衛生上の新生児の感受性歯肉炎))、パロドントパシー(parodontopathy)、外部または内部雄または雌の性器球(genital spheres)の刺激、および/または
未成熟、正常または成熟した珠皮(脱毛、フケ、多毛症、脂漏性皮膚炎)
のアレルギー性、炎症性、刺激性病理または反応、またはバリヤーまたはホメオスタシスの反応または異常を予防および治療を目的とする。
特に繊維芽細胞の数および活性の減少並びに細胞外マトリックスの過剰崩壊を特徴とする皮膚老化(年代順の外因性老化または光老化および更年期老化)、
皮膚萎縮線条、炎症、フィブロネクチン、IおよびIII型のコラーゲンおよびエラスチンをコードする遺伝子の発現の阻害、機械的膨張の影響下での繊維芽細胞の筋繊維芽細胞への形質転換を特徴とする繊維芽細胞の疾患。このコラーゲン組織の変性により、萎縮性の皮膚瘢痕が形成される。主要な誘発因子は、炎症および機械的ストレスおよびホルモン環境(妊娠中)である。皮膚萎縮線条は、若い本質的に女性集団の約50%を冒している。それらは、一般に妊娠中(妊婦の60〜70%)、思春期中(少年の10%に対して少女の25%)、またはある種の(代謝性、内分泌および感染性)疾患に見られる。これらは、皮膚割線の方法に向けられた直線状の若干くぼんだ狭い外傷であり、折り畳まれた表皮で覆われている。それらの色は進展段階によって変化し、最初は赤色または暗紫色であるが、その後第二期には虹色に輝く白っぽい外観を呈する、
深い創傷は真皮に達し、繊維芽細胞の数の減少およびマトリックスの崩壊を伴う皮膚組織変化を引き起こす。治癒機構を設定して、変化した組織を修復し、繊維芽細胞は増殖し、細胞外マトリックスを改造し、様々な成分の合成
に重要な結果を生じることがある。
−ケラチノサイトの移動の刺激:
・ラミニン5(a3β3γ2)を構成する3本の鎖をコードする遺伝子の発現に対する作用;
・遺伝子発現の増加によるMMP−9の合成の作用;
・ケラチノサイトの移動に対する作用、
−ケラチノサイトの増殖の刺激:
・細胞増殖に対する直接作用;
・繊維芽細胞の間接作用:更なるKGFの分泌:ケラチノサイトの細胞分裂を活性化する増殖因子。
精製キノア油を、キノア穀物(1000kg、5.5%収率)を溶媒(n−ヘキサン)で抽出することによっておよびキノア生油(55kg)を得ることによって得る。次に、この油を精製して(70%収率)、精製キノア油(38.5kg)を得る。沈殿物を用いて、黄色の不透明な油が得られ、ヘキサンは全く見られない。
脂肪酸組成:C14 0.1%;C16 8.2%;C16’ 0.2%;C18 0.8%,C18’ 30.4%,C18” 47.2%;C18''' 8.3%;C20 0.6%;C20’ 1.7%;C22 0.7%;C22’ 1.6%;C24 0.2%。
a−トコフェロールの相対%:22.3%;β−トコフェロールの相対%:0.0%;γ−トコフェロールの相対%:61.5%;d−トコフェロールの相対%:16.2%
カンペステロールの相対%:1.53%;スチグマステロールの相対%:3.19%;β−シトステロールの相対%:20.00%;d−5−アベナステロールの相対%:1.7%;d−7−スチグマステロールの相対%:46.35%;d−7−アベナステロールの相対%:8.55%。
スクアレン含量:2.5g/100g
精製キノア油(脱臭物、38.5kg)に分子蒸留工程を施し、濃縮キノア油とも呼ばれる不ケン化物画分で濃縮した精製キノア油を得る(3.85kg、10%収率)。
沈殿物を有する黄色不透明な油;ヘキサンは全く見られない。
a−トコフェロールの相対%:72.3%;β−トコフェロールの相対%:0.7%;γ−トコフェロールの相対%:23.1%;d−トコフェロールの相対%:4.0%。
カンペステロールの相対%:5.57%;スチグマステロールの相対%:3.4%;β−シトステロールの相対%:26.84%;d−5−アベナステロールの相対%:2.3%;d−7−スチグマステロールの相対%:39.48%;d−7−アベナステロールの相対%:5.97%
スクアレン含量:16.8g/100g
キノアのペプチドおよび糖類抽出物を、下記の手順に従って調製した:
出発原料:乾燥材料重量(DM=乾燥材料)に対して10〜12重量%のタンパク質を含むキノアケーキ。このケーキキノアにアルカリ抽出を施す(pHをpH10に調整)。次いで、上清に8kDaのミネラル膜によって限外濾過を施す(タンパク質濃縮)。次いで、保持物に濃縮工程を施した後、酵素加水分解工程を行う。タンパク質の酵素加水分解は、Prolyve 1000を用いて55℃の温度でpH8にて行う。酵素加水分解工程の終了時に、酵素を熱処理によって脱活する。次に、加水分解物に8kDaミネラル膜を用いて限外濾過を行う(濾液のペプチドの回収)。限外濾過物を濃縮した後、場合によっては0.2μm濾過による滅菌および/または凍結乾燥を行う。
防腐剤を含まない橙黄色粉末
粉末の組成(%,w/w)
a−アミノ化窒素(OPA、ロイシン当量):13%±20%
タンパク質(ビューレット、BSA当量):27%±20%
総糖類(アントロン、グルコース当量):24%±20%
実施例2で得た濃縮キノア油の効果を、様々な皮膚マトリックスパラメーターに対して評価した:(a)繊維芽細胞の増殖に対する効果、(b)真皮の細胞外マトリックスに対する効果:コラーゲンI、コラーゲンIIIおよびエラスチンの遺伝子発現、(c)真皮の機械的膨張に対する効果:赤色皮膚萎縮線条に由来する繊維芽細胞によって展開される等尺性力に対する効果。
a.皮膚繊維芽細胞の増殖の検討
MTT[3−(4,5−ジメチルチアゾール−2−イル)−2,5−ジフェニルテトラゾリウムブロミド]試験は、細胞の生育力を測定する比色試験である。MTTは、黄色の水溶性テトラゾリウム塩であり、代謝活性細胞はこれを還元して青色のホルマザン結晶とすることができる。
b.1.同時定量PCRの原理
同時定量PCRまたはQRT−PCR(同時定量ポリメラーゼ連鎖反応)は、増幅により目的の遺伝子発現を特異的かつ定量的に測定できる分子生物学の方法である。定量は、レポーター系としてSYBR Green法、すなわち二本鎖DNA内部に挿入されている蛍光特性を有する分子を用いることによってリアルタイムで遺伝子の増幅を追跡することに基づいている。PCRは、3段階の連続した温度サイクル、すなわち
−変性:2本のDNA鎖の分離、
−ハイブリダイゼーション:特異的プライマーによるターゲット遺伝子に対応するDNA配列の認識、
−伸張:ポリメラーゼの作用による目的配列の伸張
で起こる。
D0に、繊維芽細胞を、10% SVFを添加したRPMI培地で6穴プレートに播種する。D1に、繊維芽細胞を、5ng/mlのTGFβ1または1% SVFを含む0.005%および0.01%DMの濃縮キノア油で1% SVFを含むRPMI培地中で48時間処理した。細胞の処理終了時に、総DNAを抽出した後(抽出キットRNeasyキットMiniKit;Qiagen)、Experionシステム(Experion RNA StdSensキット;Biorad)によってミニチップで定量分析した。次いで、総RNAをcDNA(iScript cDNA Synthesisキット;Biorad)に逆転写する。最後に、目的遺伝子(コラーゲンI、コラーゲンIII、エラスチン)または対照遺伝子(HPRT、GAPDH、YWHAZ、βアクチン=ノーマライザー)に関する新たに合成されたcDNAを、ターゲット配列の特異的プライマーを用いることによってリアルタイムPCR(iQ5,Biorad)で選択的に増幅した。
結果を、最も安定な対照遺伝子に対して規格化する(geNormアルゴリズムによる):DCt=目的遺伝子Ct−最も安定な対照遺伝子Ct。
c.1.GlaSbox(商標)の提示
繊維芽細胞が含まれているコラーゲンゲルの機械的阻害は、「収縮力」または「等尺性力」と呼ばれる力の発生によって表される。
6容の培地を3容のラット尾コラーゲンI(2mg/ml)および1容の細胞懸濁液(8x105個の細胞/ml)と混合することによって、格子を作成するための培地を調製する。混合物を、GlaSboxの長方形容器に空ける。37℃で数分以内に、ゲルが形成される。活性成分を含むまたは含まない様々な培地を加える。等尺性力を、48時間測定する。処理の終了時に、コラーゲン格子を脱着し、コラゲナーゼ溶液で消化する。37℃で2時間インキュベーションした後、それぞれのコラーゲン格子内の細胞を計数する。力は、処理48時間後の細胞数として表される。
a.繊維芽細胞の増殖: 0.005および0.01% DMの濃縮キノア油による繊維芽細胞の処理は、用量依存的に増殖を有意に刺激する(48時間の処理後には、処理を行わなかったコントロールに対して、それぞれ、+17および+23%の増加)。
図2に示されるように、0.01%濃縮キノア油を培地に添加すると、培養の1時間30分後には繊維芽細胞または赤色皮膚萎縮線条によって生じる収縮力が有意に減少し、これは36時間までに元に戻る。この検討中に得られた曲線は、3つの別個な期からなっている。
第I期:培養の最初の2時間は、等尺性力が弱いままである。この期は、コラーゲンゲルの重合に相当する。
第II期:培養の最初の6〜8時間は、等尺性力は平均値で擬似直線的に最大まで増加する。この期は、繊維芽細胞が伸張してコラーゲン繊維に接着するのに要する時間に相当する。
第III期:等尺性力は、この培養時間中は保持される。この期は、繊維芽細胞によるコラーゲンマトリックスの再配列およびインテグリンa2β1の発現増加に相当する。
ここに示される検討結果により、皮膚組織の調節に対する濃縮キノア油の役割を示すことができた。実際に、濃縮キノア油は、(a)繊維芽細胞の増殖刺激、(b)繊維芽細胞によるコラーゲンI、コラーゲンIIIおよびエラスチン発現の誘導、(c)赤色皮膚萎縮線条の繊維芽細胞の機械特性の変更、すなわち一過性および長期弛緩効果を引き起こすことが示された。
実施例3で得たキノアペプチドの活性を、皮膚の再上皮化に関与する最初の2つの機構について評価した(0.05% DMおよび0.1% DMの2濃度で評価):
ケラチノサイトの移動:(i)ラミニン5を構成する遺伝子の発現に対する効果の評価、(ii)MMP−9の発現および合成に対する効果の評価、および(iii)ケラチノサイトの移動に対する機能上の効果の評価。
ケラチノサイトの増殖:(i)ケラチノサイトの増殖に対する効果の評価、(ii)繊維芽細胞によるKGFの合成に対する効果の評価。
a.ラミニン5およびMMP−9をコードする遺伝子の発現に対するキノアペプチドの効果のRT−PCRによる検討
−リアルタイム定量PCRの原理:実施例4を参照されたい。
−プロトコル:
0日目に、ケラチノサイトを、KGM−2培地で24穴プレートに播種した。D1に、細胞を5ng/mlのTGFβ1または0.05および0.1% DMのキノアペプチドで48時間処理した。細胞の処理を終了したときに、総RNAを抽出した後(RNeasy MiniKit;Qiagen)、Experionシステム(Experion RNA StdSensキット;Biorad)によってミニチップで定量分析した。次いで、総RNAをcDNA(iScript cDNA Synthesisキット;Biorad)に逆転写する。最後に、目的遺伝子(ラミニン5を構成する3本鎖をコードする遺伝子:a3β3d2およびMMP−9)または対照遺伝子(HPRT、GAPDH、YWHAZ、βアクチン=ノーマライザー)に関する新たに合成されたcDNAを、ターゲット配列の特異的プライマーを用いることによってリアルタイムPCR(iQ5,Biorad)で選択的に増幅した。
プラスチック製培養支持体をコラーゲンIでコーティングし、コントロール支持体をゼラチンでコーティングする(ゼラチン上の移動は、天然コラーゲン上より明らかに少ない)。ケラチノサイトをコーティングした皿に播種し、非接着性細胞を37℃および5% CO2で6時間インキュベーションした後に除去した。培地を、次に0.1%キノアペプチドを含むまたは含まない培地に置き換えた。一晩インキュベーションした後、マイトマイシンCの溶液で2時間インキュベーションすることによって細胞分裂を遮断した。人工的な再生瘢痕が細胞マット上に生じ、洗浄後に、処理を再開した。48時間後に、細胞を固定して、核をHoechst製蛍光染料で標識した。
D0に、ケラチノサイトをKGM2培地に播種する。1日目に、細胞を1μMトランス−レチノイン酸(ATRA)または0.05%および0.1% DMのキノアペプチドで24および48時間処理する。処理の終了時に、細胞生育力をMTT試験によって測定する。すなわち、MTTと3時間接触した後、形成したホルマザン結晶をDMSOによって可溶化し、代謝活性細胞、従って生きている細胞の量に比例する光学濃度を、570nmでブランク(細胞の入っていないウェル)に対して読み取る。
ケラチノサイトを24穴プレートに播種し、37℃、5% CO2で24時間インキュベーションした後、細胞を0.05%および0.1% DMのキノアペプチドで処理した。5ng/mlで試験したTGFβをポジティブコントロールとして用いた。処理の48および72時間後、細胞によって分泌されたMMP−9の量を培養物上清でELISAキット(R&D Systems)によって供給業者の推奨する手順に従って測定した。平行して、ウェル当たりの生きている細胞の量を、比色ニュートラルレッド試験によって測定し、生きている細胞の量に比例する光学濃度ODを570nmで読み取った。
繊維芽細胞を、1% SVFのRPMI中24穴プレートに播種し、37℃、5% CO2で24時間インキュベーションした後、細胞を0.05%および0.1% DMのキノアペプチドで処理した。100ng/mlのIL1a(Sigma)をポジティブコントロールとして用いた。処理の24および48時間後に、繊維芽細胞によって塩析されたKGFの量をELISAキット(R&D Systems)で供給業者の推奨する手順に従って測定した。平行して、ウェル当たりの生きている細胞の量を、比色ニュートラルレッド試験によって測定し、生きている細胞の量に比例する光学濃度ODを570nmで読み取った。平行して、ウェル当たりの生きている細胞の量を、MTT比色試験によって測定し、生きている細胞の量に比例する光学濃度ODを570nmで読み取った。
a.ラミニン5のヘテロ三量体a3β3d2の発現
再上皮化過程の遺伝子制御は、いくつかの転写または増殖因子、特に損傷の直後に放出されるTGF−β1を伴う。再上皮化の生理学的条件下では、TGF−β1がラミニン−5の発現を刺激することが明らかにされた。この増殖因子をポジティブコントロールとして用いて、このタンパク質の遺伝子発現を検討し、生理学的条件にアプローチした。
再上皮化に関与するマトリックスプロテアーゼの中でも、MMP−9は特に重要な役割を果たす。これはTGF−β1および炎症促進性サイトカインによって正の調節を受けるが、移動するケラチノサイトにより創傷部位でも発現する。
キノアペプチドの存在下におけるMMP−9の遺伝子発現のプロフィールを検討した後、遺伝子発現の誘導は、タンパク質の分泌も増加することが明らかにされた。従って、48および72時間の処理後のケラチノサイトによるMMP−9の合成および分泌に対するキノアペプチドの影響を評価した。結果を表12に示す。このように、キノアペプチドは、ケラチノサイトによって分泌されるMMP−9の量を有意に増加させる(増加範囲55%以下)。
ケラチノサイトの移動に対するキノアペプチドの機能上の効果を、細胞増殖をブロックした後の人工瘢痕で評価した。
キノアペプチドがケラチノサイトの増殖過程(=再上皮化機構の第二段階)と関連しているかどうかを立証するために、細胞生育力MTT試験を行い、細胞増殖に対するキノアペプチドの直接作用を評価した。表13に示されるように、トランスレチノイン酸(ATRA)=ポジティブコントロールは+36%の増殖率を誘導する。同様に、0.05および0.1% DMで試験したキノアペプチドはケラチノサイトの増殖を極めて有意に刺激し、+20%のオーダーの増加を示す。
再上皮化中に、ケラチノサイト増殖過程を、ケラチノサイトまたは皮膚の繊維芽細胞によって分泌されることがある多数の因子の影響下で行う:KGF(ケラチノサイト増殖因子)の産生。繊維芽細胞によって過剰発現したKGFに応答して、ケラチノサイトが増加することによって外傷を一層速やかに被覆する。皮膚繊維芽細胞によるこの因子の分泌に対するキノアペプチドの効果を、24および48時間の処理後に評価した。表14では、KGFの合成がIL1aの存在下で増加したことが確かめられる。一方、0.05および0.1% MSで試験したキノアペプチドは繊維芽細胞によるKGFの分泌を極めて有意に刺激する。この刺激は用量依存的であり、処理の24時間後には極めて実質的である(未処理コントロールと比較して5および7倍に増加)。
Claims (12)
- キノア穀物抽出物と、必要ならば適当な賦形剤とを含んでなる、化粧品、皮膚科学または栄養補給組成物であって、
前記キノア穀物抽出物が、酵素加水分解により得られるペプチドおよび糖類抽出物である、組成物。 - ペプチドおよび糖類抽出物が、25〜90重量%のペプチドおよび10〜50重量%の糖からなり、ここで百分率は前記ペプチド抽出物の総重量に対して相対的に表されている、請求項1に記載の組成物。
- ペプチドおよび糖類抽出物が、
a)キノア穀物から出発し、生油およびケーキを抽出し、前記ケーキを回収し、
b)前記ケーキを水または水/アルコール混合物で洗浄してタンパク質部分のみを保持し、次いで
c)タンパク質を可溶化し、
d)タンパク質を濃縮した後、前記タンパク質を酵素加水分解してペプチドとし、
e)ペプチドおよび糖類抽出物を精製して回収する
連続的な工程を含んでなる方法によって得られる、請求項1または2に記載の組成物。 - キノア穀物抽出物と、必要ならば適当な賦形剤とを含んでなる、深い創傷のような皮膚組織に関する疾患の予防および/もしくは治療、治癒の促進、または真皮の皮下萎縮の予防および/もしくは治療のための皮膚科学組成物であって、
前記キノア穀物抽出物が、不ケン化物画分において濃縮された油および不ケン化物からなる群より選択されるキノア穀物の脂質抽出物である、組成物。 - キノアのペプチドおよび糖類抽出物の製造方法であって、
a)キノア穀物から出発し、生油およびケーキを抽出し、前記ケーキを回収し、
b)前記ケーキを水または水/アルコール混合物で洗浄してタンパク質部分のみを保持し、次いで
c)タンパク質を可溶化し、
d)タンパク質を濃縮した後、前記タンパク質を酵素加水分解してペプチドとし、
e)ペプチドおよび糖類抽出物を精製して回収する
連続的な工程を含んでなる、方法。 - タンパク質の濃縮(工程d)前に、繊維を除去する、請求項7に記載の方法。
- 酵素加水分解により得られるキノアペプチドおよび糖類抽出物から選択される、キノア抽出物を含んでなる、皮膚科学組成物。
- アレルギー性、炎症性、刺激性病理もしくは反応、または、皮膚および/もしくは粘膜および/もしくは未成熟、正常もしくは成熟珠皮のバリヤーもしくはホメオスタシスの異常の予防および/または治療を目的とする、請求項9に記載の組成物。
- 皮膚の治癒の促進、ニキビの予防および/もしくは治療、色素沈着疾患の調節、多毛症もしくは脂漏性皮膚炎の予防および/もしくは治療、または皮膚および/もしくは粘膜および/もしくは未成熟、正常、もしくは成熟した珠皮のバリヤーもしくはホメオスタシスの異常の予防および/もしくは治療のための、請求項1〜3のいずれか一項に記載の組成物。
- 表面瘢痕、脆弱唇および口唇炎、皮膚老化、皮膚萎縮線条、刺創後の皮膚、皮膚擦過傷、皮膚斑および/または痂皮、および脆弱かつ感受性皮膚からなる群より選択される病理または病状の予防および/または治療のための、請求項11に記載の組成物。
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FR0656001A FR2910815B1 (fr) | 2006-12-28 | 2006-12-28 | Composition comprenant un extrait de graines de quinoa |
FR0656001 | 2006-12-28 | ||
PCT/EP2007/064623 WO2008080974A1 (fr) | 2006-12-28 | 2007-12-28 | Composition comprenant un extrait de graines de quinoa, utilisation dermatologique |
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US (1) | US9125879B2 (ja) |
EP (1) | EP2124982B1 (ja) |
JP (1) | JP5220761B2 (ja) |
KR (1) | KR20090092841A (ja) |
CN (1) | CN101573129B (ja) |
AU (1) | AU2007341237B2 (ja) |
BR (1) | BRPI0720863A2 (ja) |
CA (1) | CA2673990C (ja) |
ES (1) | ES2393413T3 (ja) |
FR (2) | FR2910815B1 (ja) |
HK (1) | HK1132466A1 (ja) |
MX (1) | MX2009007088A (ja) |
WO (1) | WO2008080974A1 (ja) |
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FR2948566B1 (fr) * | 2009-07-30 | 2012-08-10 | Expanscience Lab | Extrait du fruit de schizandra sphenanthera et compositions cosmetiques, dermatologiques et nutraceutiques le comprenant |
FR2948565B1 (fr) * | 2009-07-30 | 2011-10-28 | Expanscience Lab | Composition cosmetique pour le traitement de l'acne comprenant un extrait petidique de schizandra |
FR2949044B1 (fr) * | 2009-08-12 | 2021-05-07 | Expanscience Lab | Composition comprenant une fraction d'insaponifiable |
FR2953136B1 (fr) | 2009-11-30 | 2012-05-11 | Expanscience Lab | Extrait de graines de vigna unguiculata et compositions cosmetiques, pharmaceutiques, dermatologiques, nutraceutiques ou alimentaires le comprenant |
FR2953135B1 (fr) | 2009-11-30 | 2012-05-11 | Expanscience Lab | Extrait de graines d'acacia macrostachya et compositions cosmetiques, pharmaceutiques, dermatologiques, nutraceutiques ou alimentaires le comprenant |
FR2953722B1 (fr) * | 2009-12-16 | 2012-03-09 | Expanscience Lab | Composition comprenant au moins un sucre en c7 pour le traitement de l'alopecie, pour le traitement cosmetique des phaneres, et pour le soin des cheveux, cils ou ongles |
FR2969495B1 (fr) | 2010-12-22 | 2013-10-11 | Expanscience Lab | Extrait de parties aeriennes de maca riche en polyphenols et composition le comprenant |
FR2969496B1 (fr) | 2010-12-22 | 2013-11-08 | Expanscience Lab | Extrait de pulpe et/ou de peau d'avocat riche en polyphenols et compositions cosmetiques, dermatologiques et nutraceutiques le comprenant |
FR2970971B1 (fr) * | 2011-01-31 | 2014-05-30 | Expanscience Lab | Utilisation d'au moins un co-produit de l'industrie du raffinage des huiles vegetales pour obtenir un insaponifiable total purifie d'huile vegetale. |
FR2975004B1 (fr) | 2011-05-13 | 2013-06-28 | Expanscience Lab | Nouvel actif anti-rougeurs et compositions cosmetiques le comprenant |
CL2011003236A1 (es) | 2011-12-21 | 2012-07-20 | Univ Santiago Chile | Metodo para preparar producto alimenticio en formato gel de almidon de quinoa enriquecido en peptidos y maltodextrinas que comprende: extraccion proteica de la harina de quinoa, centrifugación, hidrolisis del sobrenadante con dos o mas enzimas proteoliticas, hidrolisis del precipitado y harina de quinoa para obtener maltodextrina. |
FR2988296B1 (fr) * | 2012-03-23 | 2017-01-27 | Soc D'exploitation De Produits Pour Les Ind Chimiques Seppic | Nouvelle utilisation d'extraits de quinoa comme agent actif pour prevenir et/ou limiter les effets inesthetiques generes par l'hypoxie des cellules endotheliales du corps humain |
FR2999429B1 (fr) | 2012-12-18 | 2015-01-02 | Expanscience Lab | Extrait de graines de passiflore et compositions cosmetiques, pharmaceutiques, dermatologiques ou nutraceutiques le comprenant |
FR3001889B1 (fr) | 2013-02-11 | 2021-02-12 | Expanscience Lab | Utilisation d'une composition comprenant un perseose d'avocat dans la protection des cellules souches epidermiques . |
JP6597992B2 (ja) * | 2013-08-13 | 2019-10-30 | 株式会社シャローム | ポリフェノール含有培養物の製造方法およびポリフェノール含有培養物 |
FR3011008B1 (fr) | 2013-09-24 | 2017-12-29 | Expanscience Lab | Procedes d'evaluation des effets deleteres des uv sur la peau d'enfant |
FR3010906B1 (fr) | 2013-09-25 | 2016-12-23 | Expanscience Lab | Extrait lipidique de graines de passiflores |
FR3016373B1 (fr) | 2014-01-10 | 2018-01-19 | Laboratoires Expanscience | Modele de peau de mammelon reconstitue |
FR3028764B1 (fr) | 2014-11-26 | 2016-12-09 | Expanscience Lab | Extrait peptidique et osidique de fruit de schizandra et amelioration de la reponse du systeme neurosensoriel cutane |
KR101646559B1 (ko) * | 2015-08-07 | 2016-08-08 | 주식회사 코씨드바이오팜 | 피부 건조 유래 가려움증 개선 및 치료용 조성물 |
CN106282282A (zh) * | 2016-08-10 | 2017-01-04 | 华南农业大学 | 一种同时提取辣木籽中油脂和蛋白和/或糖苷的方法 |
CN107496245B (zh) * | 2017-09-15 | 2020-04-03 | 澳宝化妆品(惠州)有限公司 | 一种抗衰老脂质体组合物及其制备方法和应用 |
CN110241163B (zh) * | 2019-06-25 | 2021-07-27 | 江南大学 | 一种碱提-膜法提取藜麦多肽的方法 |
CN111110596A (zh) * | 2020-02-18 | 2020-05-08 | 广州栋方生物科技股份有限公司 | 一种皮脂膜仿生组合物及其制备方法 |
WO2022173974A1 (en) * | 2021-02-10 | 2022-08-18 | TRI-K Industries, Inc. | A novel composition and method containing pea and quinoa natural peptides and niacinamide to enhance skin glow, radiance and even skin tone |
CN113462632B (zh) * | 2021-08-13 | 2023-08-29 | 徐州医科大学 | 一种苦瓜外泌体、提取方法及在制备治疗烧烫伤皮肤药物中的应用 |
CN114767595B (zh) * | 2022-05-13 | 2023-02-10 | 山东福瑞达生物股份有限公司 | 一种舒缓修护组合物的制备方法及其应用 |
CN115228133B (zh) * | 2022-07-28 | 2024-04-12 | 黑龙江八一农垦大学 | 一种连续复合酶法提取杂粮中结合态多酚的方法 |
CN116036247B (zh) * | 2023-02-03 | 2024-04-05 | 江中药业股份有限公司 | 用于抑制炎症反应、促进血管生成及伤口愈合的组合物及应用 |
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AU3589899A (en) | 1998-04-17 | 1999-11-08 | Her Majesty The Queen In Right Of Canada As Represented By The Department Of Agriculture And Agri-Food Canada | Process for recovery and purification of saponins and sapogenins from quinoa ((chenopodium quinoa)) |
FR2798591B1 (fr) | 1999-09-22 | 2001-10-26 | Pharmascience Lab | Utilisation d'un produit d'huile vegetale pour augmenter la synthese des lipides cutanes en cosmetique, pharmacie ou dermatologie et en tant qu'additif alimentaire |
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US20090191243A9 (en) | 2000-01-03 | 2009-07-30 | Hill John C | High unsaponifiables and methods of using the same and its derivatives and uses thereof |
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WO2006053415A1 (en) * | 2004-11-18 | 2006-05-26 | Biopharmacopae Design International Inc. | Plant extract having matrix metalloprotease inhibiting activity and dermatological uses thereof |
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FR2869541B1 (fr) | 2004-04-30 | 2007-12-28 | Expanscience Sa Lab | Utilisation d'une composition comprenant du d-mannoheptulose et/ou du perseitol dans le traitement et la prevention des maladies liees a une modification de l'immunite innee |
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AU2007341237A1 (en) | 2008-07-10 |
US20100136144A1 (en) | 2010-06-03 |
CN101573129A (zh) | 2009-11-04 |
CA2673990A1 (fr) | 2008-07-10 |
AU2007341237B2 (en) | 2013-05-09 |
CA2673990C (fr) | 2016-04-12 |
EP2124982B1 (fr) | 2012-08-22 |
FR2910815A1 (fr) | 2008-07-04 |
FR2910815B1 (fr) | 2010-10-29 |
WO2008080974A1 (fr) | 2008-07-10 |
HK1132466A1 (en) | 2010-02-26 |
CN101573129B (zh) | 2012-02-22 |
MX2009007088A (es) | 2009-07-08 |
EP2124982A1 (fr) | 2009-12-02 |
JP2010514740A (ja) | 2010-05-06 |
BRPI0720863A2 (pt) | 2014-03-04 |
ES2393413T3 (es) | 2012-12-21 |
US9125879B2 (en) | 2015-09-08 |
KR20090092841A (ko) | 2009-09-01 |
FR2942960A1 (fr) | 2010-09-17 |
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