JP7035294B2 - 前立腺肥大症疾患の予防または治療用生薬組成物 - Google Patents
前立腺肥大症疾患の予防または治療用生薬組成物 Download PDFInfo
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- JP7035294B2 JP7035294B2 JP2020531526A JP2020531526A JP7035294B2 JP 7035294 B2 JP7035294 B2 JP 7035294B2 JP 2020531526 A JP2020531526 A JP 2020531526A JP 2020531526 A JP2020531526 A JP 2020531526A JP 7035294 B2 JP7035294 B2 JP 7035294B2
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Description
本特許出願は2017年12月6日付で大韓民国特許庁に提出された大韓民国特許出願第10-2017-0166860号に基づく利益及び優先権を主張する、2018年12月6日付で提出された国際特許出願PCT/KR2018/015446の国内段階移行出願である。前記いずれの特許出願の開示事項も本明細書に参照として挿入される。
単一成分基準重量=単一抽出物製造に用いられた単一成分の重量×(最終複合抽出物製造のために使われた単一抽出物の重量/製造した単一抽出物の重量)。
(a)本発明は、前立腺肥大症疾患の予防または治療用薬剤学的組成物を提供する。
(b)本発明は、前立腺肥大症疾患の改善用食品組成物を提供する。
(c)本発明は、前立腺肥大症疾患の予防または治療方法を提供する。
(d)本発明の薬剤学的組成物を用いる場合、前立腺肥大症を副作用無しで効果的に予防または治療することができる。
韓国の漢方薬流通専門業体である(株)ヒューマンハブから購入した洗浄及び乾燥された兎糸子(Cuscutae Semen)、蓮子肉(Nelumbinis Semen)、及び蒲公英(Taraxaci Herba)を実験に使用した。前記兎糸子、蓮子肉、及び蒲公英の生薬を表1に記載された重量比(w/w)で総240gになるように混合した後、10倍の25%エタノール水溶液を加えて常温で72時間の間よく撹拌んさせて抽出した後、濾過して、これを50-65℃で減圧濃縮した後、凍結乾燥させて総7種類の複合生薬抽出物の粉末を収得しており、これらの歩留まりは表1の通りである。
韓国の漢方薬流通専門業体である(株)ヒューマンハブから購入した洗浄及び乾燥された兎糸子(Cuscutae Semen)、蓮子肉(Nelumbinis Semen)、及び蒲公英(Taraxaci Herba)を実験に使用した。前記兎糸子、蓮子肉、及び蒲公英の生薬を製造例1-6の重量比(w/w)で総240gになるように混合した後、10倍の25%エタノール水溶液を加えて常温で0.5、3、8、24、及び72時間の間よく撹拌させて抽出した後、濾過して、これを50-65℃で減圧濃縮した後、凍結乾燥させて総5種類の複合生薬抽出物の粉末を収得しており、これらの歩留まりは表2の通りである。
韓国の漢方薬流通専門業者である(株)ヒューマンハブから購入した洗浄及び乾燥された兎糸子(Cuscutae Semen)、蓮子肉(Nelumbinis Semen)及び蒲公英(Taraxaci Herba)を実験に使用した。前記兎糸子、蓮子肉、及び蒲公英の生薬を製造例1-6の重量比(w/w)で総240gになるように混合した後、10倍の0、10、25、50、75、及び95%エタノール水溶液を加えて常温で72時間の間よく撹拌させて抽出した後、濾過して、これを50-65℃で減圧濃縮した後、凍結乾燥させて総6種類の複合生薬抽出物粉末を収得しており、これらの歩留まりは表3の通りである。
韓国の漢方薬流通専門業者である(株)ヒューマンハブから購入した洗浄及び乾燥された兎糸子(Cuscutae Semen)、蓮子肉(Nelumbinis Semen)、及び蒲公英(Taraxaci Herba)を実験に使用した。前記兎糸子、蓮子肉、及び蒲公英の生薬を製造例1-6の重量比(w/w)で各30g、30g、及び60gを混合した後、10倍の蒸溜水を加えて95-100℃の温度で3時間の間還流抽出した後、濾過して、これを50-65℃で減圧濃縮した後、凍結乾燥させて複合生薬抽出物の粉末を収得しており、その歩留まりは約11.90%であった。
洗浄及び乾燥された兎糸子(Cuscutae Semen)、蓮子肉(Nelumbinis Semen)、及び蒲公英(Taraxaci Herba)を購入して実験に使用した。各生薬の180gに10倍の25%エタノール水溶液を加えて、常温で72時間の間よく撹拌させて抽出した後、濾過して、これを50-65℃で減圧濃縮した後、凍結乾燥させて総3種類の単一生薬抽出物の粉末を収得しており、これらの歩留まりは表4の通りである。
(実験方法)
人の前立腺癌から由来した前立腺上皮細胞株であるLNCaP細胞をRPMI培地(Gibco、USA)を用いてウェル当たり1x105細胞数で24ウェル-プレートに準備し、一日間安定化させた。以後、無血清(serum-free)培地で24時間の間培養した後、培養液除去後、テストステロン(Tes)2μMと製造例1-6の複合生薬抽出物、製造例4で製造した複合生薬抽出物を各50、100、及び200μg/mlの濃度で処理した。72時間後、細胞の増殖変化をMTT検査法により確認した。細胞増殖割合はTes処理群に対比した割合で示した。
熱水複合生薬抽出物(製造例4)と25%エタノール水溶液の複合生薬抽出物(製造例1-6)の効果を比較した結果、2つ抽出物全て濃度依存的に前立腺細胞増殖抑制効果が表れた(図1)。
(実験方法)
人の前立腺癌から由来した前立腺上皮細胞株であるLNCaP細胞をRPMI培地(Gibco、USA)を用いてウェル当たり1x105細胞数で24ウェル-プレートに準備し、一日間安定化させた。以後、無血清(serum-free)培地で24時間の間培養した後、培養液除去後、テストステロン(Tes)2μMと製造例3で製造した6種類の複合生薬抽出物を200μg/mlの濃度で処理した。72時間後、細胞の増殖変化をMTT検査法により確認した。細胞増殖割合はTes処理群に対比した割合で示した。
テストステロンによる前立腺上皮細胞株の増殖に対する多様なエタノール水溶液濃度別複合生薬抽出物が及ぼす影響は下記の表5に示した。
(実験方法)
人の前立腺癌から由来した前立腺上皮細胞株のLNCaP細胞をRPMI培地(Gibco、USA)を用いてウェル当たり1x105細胞数で24ウェル-プレートに準備し、一日間安定化させた。以後、無血清(serum-free)培地で24時間の間培養した後、培養液除去後、テストステロン2μMと製造例1-6の複合生薬抽出物または比較例1で製造した3種類の単一生薬抽出物を200μg/mlの濃度で処理した。72時間後、細胞の増殖変化をMTT検査法により確認した。細胞増殖割合はTes処理群に対比した割合で示した。
テストステロンによる前立腺上皮細胞株の増殖に単一生薬抽出物と複合生薬抽出物が及ぼす影響は下記の表6に示した。
(実験方法)
9週齢(体重350g以下)の雄性SDネズミを一週間以上純化飼育した後、体重を基準に、(1)正常群、(2)前立腺肥大症誘導及び蒸溜水投与群(陰性対照群)、(3)前立腺肥大症誘導及び複合生薬抽出物(製造例1-6)投与群、(4)前立腺肥大症誘導及び兎糸子単一生薬抽出物(比較例1-1)投与群、(5)前立腺肥大症誘導及び蓮子肉単一生薬抽出物(比較例1-2)投与群、(6)前立腺肥大症誘導及び蒲公英単一生薬抽出物(比較例1-3)投与群、(7)前立腺肥大症誘導及びフィナステライド(Finasteride)投与群(陽性対照群)に分離した。前立腺肥大症を誘導するためにテストステロン(Testosterone、綿種オイルに溶かして使用)を3mg/kgの容量で1回/3日、総10回皮下注射(subcutaneous injection)した。正常群の場合、同量の綿種オイルを皮下注射した。製造例1-6の複合生薬抽出物及び比較例1の単一生薬抽出物は蒸溜水に溶かして300mg/kgの容量で1回/日、30日間経口投与した。フィナステライド投与群(シグマ、0.2% tween 80に溶かして使用)は10mg/kgの容量で1回/1日、30日間経口投与した。陰性対照群は蒸溜水のみを経口投与した。実験開始日から一週間間隔で総5回体重を測定し、最後の投薬及び処理が終わった翌日にラットを二酸化炭素ガスで犠牲させた後、前立腺組織を摘出して重さを測定した。この測定値から以下の数学式を用いて前立腺増殖抑制効果(%)を示した。
前立腺増殖抑制効果(%)=100-{(複合生薬抽出物群/単一抽出物または陽性対照群前立腺重さ-正常群前立腺重さ)*100/(陰性対照群前立腺重さ-正常群前立腺重さ)}
テストステロンで前立腺肥大症を誘導した結果、正常群の場合、持続的に体重が増加するに反して、テストステロンを処理した群では体重増加が抑制された。しかしながら、陰性対照群、複合生薬抽出物群(製造例1-6)、単一生薬抽出物群(比較例1-1、比較例1-2、及び比較例1-3)、及び陽性対照群の間には有意味な体重の差は観察できなかった(図4)。テストステロンによる前立腺増殖に及ぼす複合生薬抽出物群(製造例1-6)、単一生薬抽出物群(比較例1-1、比較例1-2、及び比較例1-3)、及び陽性対照群の効果は下記の表7に示した。
Claims (7)
- (a)有効成分としての兎糸子、蓮子肉、及び蒲公英の複合抽出物;及び
(b)薬剤学的に許容される担体を含む前立腺肥大症疾患の予防または治療用薬剤学的組成物。 - 前記兎糸子、蓮子肉、及び蒲公英の配合重量比(w/w)は1~4:1~4:1~4である、請求項1に記載の組成物。
- 前記抽出物は、水またはエタノール抽出物である、請求項1に記載の組成物。
- 前記エタノールは、0%超過50%以下のエタノールである、請求項3に記載の組成物。
- 前記エタノールは、25%エタノールである、請求項3に記載の組成物。
- 兎糸子、蓮子肉、及び蒲公英の複合抽出物を有効成分に含む前立腺肥大症疾患の改善用食品組成物。
- 前記兎糸子、蓮子肉、及び蒲公英の配合重量比(w/w)は1~4:1~4:1~4である、請求項6に記載の組成物。
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