JP2013532700A - 癌を治療するための、レチノール、その前駆体または反応性生物および少なくとも1つのカモミール植物からの植物抽出液を含む組成物 - Google Patents
癌を治療するための、レチノール、その前駆体または反応性生物および少なくとも1つのカモミール植物からの植物抽出液を含む組成物 Download PDFInfo
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Abstract
Description
以下の図面は、本発明をさらに説明することを意図している。それらは、本発明の主題をそれらに限定することを意図するものではない。
以下の実施例は、本発明をさらに詳しく説明することを意図するものである。それらは、本発明の主題をそれらに限定することを意図するものではない。
200gのMatricaria recutita L.の花序からの管状花を、薬物/溶媒比1/7.5(w/w)に対応する1500gの無水エタノールとともに、ターボミキサー(3×1分、8000rpm、Eberbach(登録商標))中で抽出した。得られた混合物を40℃にて1時間攪拌した。次いで、深層セルロースフィルター(AF 15 Filtrox(登録商標))で調製物を2回ろ過した。
固体含量1.05%(m/m)の1266gの透明な茶色がかった抽出液を得た。
200gのMatricaria recutita L.の花序からの管状花を、薬物/溶媒比1/7.5(w/w)に対応する1500gの無水エタノールとともに、ターボミキサー(3×1分、8000rpm、Eberbach(登録商標))中で抽出した。得られた混合物を40℃にて1時間攪拌した。次いで、深層セルロースフィルター(AF15 Filtrox(登録商標))で調製物を2回ろ過した。
固体含量0.99%(m/m)の1262gの透明な茶色がかった抽出液を得た。
実施例1と同様にして得られた1266gのエタノール抽出液(13.3gの天然抽出物を含む)に、26.6gのSpan(登録商標)20(Sigma(登録商標))(ソルビタン(モノ)ラウレート)および2.66gのTranscutol(登録商標)HP(Gattefosse(登録商標))(ジエチレングリコールモノエチルエーテル)を、1/2/0.2の比率で加えた。エタノールを減圧下(100〜25mbar)、40℃の浴温で蒸発させた。沈殿を含む45gの茶色がかった抽出液を得た。
水の含有量は、0.64%(w/w)であることがわかった。
CHARE(AMT2003 Cam)をAuron Healthcare GmbH(Lot:86118G001)によって提供された。それは、固体含量11.68mg/mlであった。以下のとおり、抽出液を調製した:300.000kgの精製水に、15.800kgのMatricaria recutita L.の花序からの管状花を緩やかに攪拌しながら加えた。混合物を90〜95℃に30〜35分間加熱した。続いて、混合物を30〜35℃に冷却し、次いで、深層セルロースフィルターで2回ろ過した。ろ液に1% m/mのMygliol 812 Nを加え、緩やかに10分間攪拌した。続いて、混合物のpHを水酸化ナトリウムで6.3〜6.7に調節した。次いで、最初にろ液を0.22μm膜フィルターを通してマイクロろ過し、得られるろ液を0.1μm膜フィルターを通してろ過し、次いで、得られるろ液を1000kDaウルトラフィルターを通してろ過した。得られた透明な黄色がかった抽出液および0.7% m/mのフェノール purissを加え、フェノールが完全に溶解するまで液体を混合した。
AMT(M)は、4つの成分からなる混合物である:
1)AMT2003 Vit(+):1.027mg/mlのデクスパンテノール、6.845mg/mlのニコチンアミド、0.685mg/mlのピリドキシン-HCL、0.937mg/mlのリボフラビンリン酸ナトリウム、1.712mg/mlのチアミン-HCL、41.280mg/mlのアスコルビン酸ナトリウムおよび付加的に1.79mg/mlのマレイン酸クロルフェニラミンからなり、Auron Healthcare GmbHによって提供された(Ch.-B.:84105G005);
2)AMT2003Cal:0.5gのグルコン酸カルシウム一水和物および0.8756gのラクトビオン酸カルシウム二水和物からなり、Auron Healthcare GmbHによって提供された(Ch.-B.:0907000134);
3)AMT2003Ins:100 IE/mlのヒトインスリンを含み、Auron Healthcare GmbHによって提供された(Ch.-B.:0902000117);
4)MR15E14(実施例2参照)。
最終AMT濃度は、MR15E14について1%および21μg/mlであった。
AMT(CHARE)は、4つの成分からなる混合物である:
1)AMT2003 Vit(+):1.027mg/mlのデクスパンテノール、6.845mg/mlのニコチンアミド、0.685mg/mlのピリドキシン-HCL、0.937mg/mlのリボフラビンリン酸ナトリウム、1.712mg/mlのチアミン-HCL、41.280mg/mlのアスコルビン酸ナトリウムおよび付加的に1.79mg/mlのマレイン酸クロルフェニラミンからなり、Auron Healthcare GmbHによって提供された(Ch.-B.:84105G005);
2)AMT2003Cal:0.5gのグルコン酸カルシウム一水和物および0.8756gのラクトビオン酸カルシウム二水和物からなり、Auron Healthcare GmbHによって提供された(Ch.-B.:0907000134);
3)AMT2003Ins:100 IE/mlのヒトインスリンを含み、Auron Healthcare GmbHによって提供された(Ch.-B.:0902000117);
4)CHARE(実施例4参照)。
さらに、100% AMT(CHARE)をUPWで希釈して、10% AMT(CHARE)にした。
AMT(UPW)は、4つの成分からなる混合物である:
1)AMT2003 Vit(+):付加的に1.79mg/mlのマレイン酸クロルフェニラミンを有し、Auron Healthcare GmbHによって提供された(Ch.-B.:84105G005);
2)AMT2003Cal:0.5gのグルコン酸カルシウム一水和物および0.8756gのラクトビオン酸カルシウム二水和物からなり、Auron Healthcare GmbHによって提供された(Ch.-B.:0907000134);
3)AMT2003Ins:100 IE/mlのヒトインスリンを含み、Auron Healthcare GmbHによって提供された(Ch.-B.:0902000117);
4)超純水。
さらに、100% AMT(UPW)を超純水UPWで希釈して、10% AMT(UPW)にした。
酢酸レチニル(R7882、ロット:029K12721)は、Sigma Aldrich(ブーフス、スイス)から購入した。1mg/mlのストック溶液を無水エタノールで調製した。実験に用いた最終濃度は、1%エタノール中1μg/mlであった。
全トランスレチノイン酸(95152、ロット:1392337)は、Sigma Aldrich(ブーフス、スイス)から購入した。1mg/mlのストック溶液を無水エタノールで調製した。実験に用いた最終濃度は、1%エタノール中1μg/mlであった。
ヒト神経膠芽腫細胞株A172およびU87MGを、10% FBSを含む培養培地で培養し、加湿インキュベーター中、37℃および5%CO2で保持した。続く実験において用いた培養物を25回未満継代した。細胞をT25cm2培養フラスコに、100,000細胞を播種した(細胞濃度10,000細胞/ml)。それぞれ実施例1および8にしたがって調製したR15E7および酢酸レチニルを、単独で、あるいは一緒に組み合わせて加えた(A172:図1AおよびU87:図1B)。溶媒コントロールは、2%エタノールであった。
式:細胞カウント%=(rd X /rd 溶媒コントロール)×100
を用いて細胞カウント%に変換した。図1Aおよび図1Bを参照。A172(A)およびU87MG(B)細胞株の両方において、酢酸レチニル(1μg/ml)、MR15E7(30μg/ml)および30μg/mlの抽出液と酢酸レチニル(1μg/ml)の組み合わせの潜在的増殖阻害効果が観察された。溶媒コントロールおよび各ウエルの最終エタノール濃度は、2%であった。
図1Aおよび図1Bに示すように、1μg/mlの酢酸レチニルは、A172およびU87MG膠芽腫細胞細胞増殖に影響を及ぼさないように見えた。しかしながら、実施例10に記載した実験からのデータは、R15E7が、溶媒コントロール(100%)と比較して、A172の細胞数を30%に減少させることを明らかにした。1μg/mlの酢酸レチニルの添加は、細胞増殖をさらに14%に減少させ、化合物単独と比較して、組み合わせによる明らかな有効性の増強が実証された(図1A)。U87MGにおいても同様の効果が見られる(図1B)が、A172において観察される効果ほど強くはない。
ヒト大細胞肺癌細胞株H460を、アメリカン・タイプカルチャー・コレクションから入手した。10% FBSを含むRPMI-1640培地で細胞を培養し、加湿インキュベーター中、37℃および5%CO2で保持した。続く実験において用いた培養物を25回未満継代した。H460細胞を12ウエルプレートに、14375細胞/ウエル(細胞濃度10,000細胞/ml)で播種し、付着させるために一夜放置した。
式:細胞カウント%=(rd X /rd 溶媒コントロール)×100
を用いて細胞カウント%に変換した。
本発明組成物とWO 03101479 A1の教示を比較するために次の実験を行った:ヒト大細胞肺癌細胞株H460を、アメリカン・タイプカルチャー・コレクションから入手した。10% FBSを含むRPMI-1640培地で細胞を培養し、加湿インキュベーター中、37℃および5%CO2で保持した。続く実験において用いた培養物を25回未満継代した。H460細胞を播種した。細胞を12ウエルプレートに、14375細胞/ウエル(細胞濃度10,000細胞/ml)で播種し、付着させるために一夜放置した。実験の1つのセットにおいて、次いで、図3Aに示すように、細胞を、レチノイン酸(1μg/ml)またはビヒクルコントロール(1% エタノール)と組み合わせて、実施例2にしたがって調製したMR15E14(21μg/ml)、実施例5にしたがって調製した1% AMT(M)、および実施例7にしたがって調製した1% AMT(UPW)とともにインキュベートした。
式:細胞カウント%=(rd X /rd 溶媒コントロール)×100
を用いて細胞カウント%に変換した。
この実験において、必要に応じてブルーカモミールの存在下、レチノイン酸(1μg/ml)またはビヒクルコントロール(1%エタノール)と組み合わせたブルーカモミール油(薬局方による:Matricaria aetheroleum)によるH69細胞の処理を行った。この実験のために、ヒト小肺癌細胞ガン細胞株H69を、まず40μmフィルターで篩いにかけて、塊を除去した後、12ウエルプレートに50,000細胞/ウエル/800μl(細胞濃度63,500細胞/ml)を播種した。同日にサンプルを加え、細胞を合計7日間インキュベートした(サンプル:溶媒、1μg/mlのレチノイン酸、ブルーカモミール油1μg/ml+1% EtOH、ブルーカモミール油1μg/ml+1μg/mlのレチノイン酸、ブルーカモミール油3μg/ml+1% EtOH、ブルーカモミール油3μg/ml+1μg/mlのレチノイン酸、ブルーカモミール油10μg/ml+1% EtOH、ブルーカモミール油10μg/ml+1μg/mlのレチノイン酸)。インキュベーション後、細胞をウエルに再懸濁した後、100μlの懸濁液を96ウエルプレートに移した。複製を作成した。次いで、等しい体積のCyQuant検出試薬を加え、37℃にて1時間インキュベートした。生細胞のみが染色される。480/535nmにて蛍光を測定した。実験の最後に得られた生細胞の数を、細胞カウントのパーセントに変換した。溶媒コントロールは、100%で設定した。次いで、各ウエルの生データ(rd)を、
式:細胞カウント%=(rd X /rd 溶媒コントロール)×100
を用いて細胞カウント%に変換した。結果は、レチノイン酸(1μg/ml)またはビヒクルコントロール(1% Ethanol)と組み合わせたブルーカモミール油によるH69細胞の処理を示す図4に見ることができる。図からわかるように、ブルーカモミール油10μg/ml+1μg/mlのレチノイン酸の組み合わせが、H69細胞において最高の阻害効果を示した。
薬理学的実施例10〜12に示されるように、実施例1〜4にしたがって調製されたカモミール抽出液は、様々な起源の癌細胞増殖を阻害するのに効果的である。実施例10〜12にさらに示されるように、レチノイン酸または酢酸レチニルと合せたカモミール抽出液と腫瘍細胞のインキュベーションは、この抗腫瘍活性を目覚しく増強し、少なくとも腫瘍細胞死をもたらした。実施例10〜12では、ビタミンA誘導体単独で投与される場合に、緩やかで比較的弱い抗増殖効果しかもたらさないが(該効果は、主にビタミンの分化作用を介して媒介されるように見える)、カモミール抽出液とレチノールまたは誘導体の組み合わせが、事実、相乗的に作用しており、抗腫瘍活性の著しい増加を導くと言うことができる。さらに、AMTに見出される成分などのさらなる成分を含むビタミンA/カモミールペアの組み合わせは、むしろ逆効果であり、不利であると推定することができる。
実施例1と同様にして得られた350ga エタノール抽出液に、108.26gのルトロールL44(BASF、Art. No.50143751、バッチNo.WPYF580B)を1/2.2の比率で加えた。エタノールを減圧(100から25mbar)下および浴温40℃にて蒸発させて、茶色がかった抽出液を得た。水の含有量は、0.4%(w/w)であるのがわかった。このように、本願の実施例3で用いたSpan 20の代わりに、本実施例においてはルトロールL44を用いる。
本研究の目的は、同系同所PAN02腫瘍マウスモデルにおけるカモミール抽出液(MR31E25C1)およびレチノイン酸(RetAcid)の経口併用の有効性についての情報を得ることであった。
グループ 1:非接種、非処理コントロールグループ。
グループ 2:接種、非処理コントロールグループ。
グループ 3:接種、ビヒクルSpan(登録商標)20;1日3回。
グループ 4:接種、MR31E25C1;484.5mg/kg、1日3回。
グループ 5:接種、レチノイン酸;25mg/kg 1日2回 最初の2日間;10mg/kg 1日1回 処理機関の残り。
グループ 6:接種、レチノイン酸;25mg/kg 1日2回 最初の2日間;10mg/kg 1日1回 処理機関の残り;およびMR31E25C1;484.5mg/kg、1日3回。
Claims (13)
- 癌または腫瘍疾患の予防、治療またはにおける使用のための、(i)ビタミンA、その反応生成物、代謝物または前駆体;および
(ii)カモミールの植物抽出液またはその有効成分;
を含む組成物。 - 成分(i)および(ii)が、有効成分のみを表す請求項1に記載の組成物。
- 該ビタミンAまたは誘導体が、ビタミンA、レチノイン酸、全トランスレチノイン酸、13-cis-レチノイン酸、レチニルパルミテートなどのビタミンAのエステル、レチノール、レチナール、酢酸レチニル、アリトレチノイン、ビタミンAの前駆体として、β-カロテン、α-カロテン、γ-カロテンまたはβ-クリプトキサンチン、およびビタミンAの代謝産物として、レチノイン酸から選ばれる請求項1または2に記載の組成物。
- 植物抽出液が、好ましくはカモミールの揮発油、好ましくはMatricaria recutitaである請求項1〜3のいずれかに記載の組成物。
- 植物抽出液が、Matricaria recutita L.の管状花、好ましくはMatricaria recutita L.の花序からの管状花から製造されるカモミール抽出液である請求項1〜4のいずれかに記載の組成物。
- 組成物が、約50mg〜約1500mg、好ましくは約50mg〜約1000mgのMatricaria recutita L.の管状花からのカモミール抽出液、および/または典型的に約3mg〜約90mg、好ましくは約3mg〜約90mgのビタミンAの反応生成物、代謝物または前駆体のいずれかを含む請求項1〜5のいずれかに記載の組成物。
- 組成物または組成物の製造前の植物抽出液が、クロスポビドン(架橋ポビドン)と硫酸ナトリウムに接触させることよって精製されている請求項1〜6のいずれかに記載の組成物。
- 癌疾患が、
神経原性、間葉または上皮起源の固形腫瘍とその転移癌、結腸癌、黒色腫、腎癌、リンパ腫、急性骨髄性白血病(AML)、急性リンパ性白血病(ALL)、慢性骨髄性白血病(CML)、慢性リンパ性白血病(CLL)、消化管腫瘍、肺癌、神経膠腫、甲状腺腫瘍、乳癌、前立腺癌、肝癌;パピローマウイルス誘発癌(子宮頸癌など)、腺癌、ヘルペスウイルス誘発腫瘍(バーキットリンパ腫、EBV誘発B細胞リンパ腫など)、B型肝炎誘発腫瘍(肝細胞癌)、HTLV-1およびHTLV-2誘発リンパ腫、聴神経腫/聴神経鞘腫、子宮頸がん、肺癌、咽頭癌、肛門癌、神経膠芽腫、リンパ腫、直腸癌、星状細胞腫、脳腫瘍、胃癌、網膜芽細胞腫、基底細胞腫、脳転移癌、髄芽腫、膣癌、膵臓癌、精巣癌、黒色腫、甲状腺癌、膀胱癌、ホジキン症候群、髄膜腫、シュネーベルガー疾患、気管支癌、下垂体腫瘍、菌状息肉腫、食道癌、乳癌、カルチノイド、神経鞘腫棘細胞癌、バーキットリンパ腫、喉頭癌、腎癌、胸腺腫、コーパス癌、骨癌、骨肉腫、非ホジキンリンパ腫、尿道癌、CUP症候群、頭部/頚部腫瘍、乏突起膠腫、外陰部癌、腸癌、結腸癌、食道癌、いぼ類、小腸の腫瘍、頭蓋咽頭腫、卵巣癌、軟組織腫瘍/肉腫、卵巣癌、肝臓癌、膵臓癌、子宮頸癌、子宮体癌、肝転移癌、陰茎癌、舌癌、胆嚢癌、白血病、形質細胞腫、子宮癌、蓋腫瘍、前立腺癌などの様々なウイルス誘発腫瘍、特に神経原性、間葉または上皮起源の固形腫瘍とその転移癌から選ばれる癌または腫瘍疾患から選ばれる請求項1〜7のいずれかに記載の組成物。 - 請求項1〜8のいずれかに記載の、(i)ビタミンA、その反応生成物、代謝物または前駆体;および
(ii)カモミールの植物抽出液またはその有効成分;ならびに
さらに医薬的に許容しうる担体を含む医薬組成物。 - 医薬組成物が、約50mg〜約1500mg、好ましくは約50mg〜約1000mgのMatricaria recutita L.の管状花からのカモミール抽出液、および/または典型的に約3mg〜約90mg、好ましくは約3mg〜約90mgのビタミンAの反応生成物、代謝物または前駆体のいずれかを含む請求項9に記載の医薬組成物。
- 医薬組成物がさらに、ジメチルスルホキシド(DMSO)および/またはジエチレングリコールモノエチルエーテルなどの浸透促進剤を含む請求項9または10に記載の医薬組成物。
- 医薬組成物が、気道を介して、経口投与で、注射で、非経口注入で、好ましくは経口的、経直腸的、局所的もしくは非経口的に投与される請求項9〜11のいずれかに記載の医薬組成物。
- 医薬組成物が、(従来の)癌療法の前、同時および/または後に投与される請求項9〜12のいずれかに記載の医薬組成物。
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PCT/EP2011/003923 WO2012016706A1 (en) | 2010-08-05 | 2011-08-04 | Composition comprising retinol, a precursor or a reaction product of it and a plant extract from at least one chamomilla plant for the treatment of cancer |
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KR20130135226A (ko) | 2013-12-10 |
US10028989B2 (en) | 2018-07-24 |
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EP2600856A1 (en) | 2013-06-12 |
RU2013109416A (ru) | 2014-09-10 |
US20140037771A1 (en) | 2014-02-06 |
EP2420228A1 (en) | 2012-02-22 |
CN103037855A (zh) | 2013-04-10 |
CA2805075A1 (en) | 2012-02-09 |
US20140234452A1 (en) | 2014-08-21 |
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ZA201300084B (en) | 2013-08-28 |
US20190358281A1 (en) | 2019-11-28 |
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US20180117108A1 (en) | 2018-05-03 |
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