JP7176790B1 - 睡眠改善・記憶力向上のための組成物及びその調製方法と応用 - Google Patents
睡眠改善・記憶力向上のための組成物及びその調製方法と応用 Download PDFInfo
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Abstract
Description
本発明の上記の原料組成は、表1の一般的な中成薬成分とは大きくなり、その処方は発明者が東洋医学の「清養」理論に基づいて研究して確定した処方である。
東洋医学の「清」とは、鎮静と睡眠補助により、体内の睡眠を影響する有害物質要素を除去し、梔子を君薬とする。
漢方の「養」の原則は、補中益気により、主に五臓を調理し、本発明において主に鹿皮膠を君薬とする。
また、本発明において、東洋医学の「清養」の理論を原則とすること以外、梔子と鹿皮膠を「君薬」とし、この2つの薬は本発明の核心薬味である。
乾姜と甘草はいずれも健脾和胃、益気和中できる。
典型的な本発明の組成物の各原料の配合比として、鹿皮膠28部、梔子粉15部、乾姜粉9部、甘草9部の配合が挙げられる。
本発明の組み合わせには、さらに他の漢方薬睡眠補助成分、例えば、酸棗仁、蓮子、百合などを添加してもよいが、これらは必須ではない。
γ-アミノ酪酸(GABA)は、哺乳類の中枢神経系における重要な抑制性神経伝達物質であることがよく知られており、約30%の中枢神経の突出部位はGABAを伝達物質として使用する。十分なGABAは、鎮静安神、神経をリラックスさせる効果がある。L-トリプトファンは精神安定剤として、精神のリズムを調節し、睡眠を改善し、この2つのアミノ酸は「甘睡アミノ酸」と呼ばれている。ビタミンB6は、特定な神経媒体の合成に関与し、脳部機能の正常に必須な物質であり、抗ストレスビタミンでもある。テアニンは、脳興奮性の神経伝達物質であるグルタミン酸とその受容体の結合を阻害し、GABAの分泌を促進し、神経を落ち着かせ、不安を和らげ、鎮静安神の役割を果たし、睡眠率を加速し、睡眠の質を高めることができる。N-アセチルノイラミン酸は、脳の発育を促進し、脳内でガングリオシドを仲介する役割を果たし、実験より、それはラットの行動を変化させ、記憶力を向上させることができることを証明し、母乳で育てた乳児と粉ミルクで育てた乳児の間を比較したところ、前頭皮質の灰白質におけるガングリオシドタンパク質とN-アセチルノイラミン酸が結合する濃度はそれぞれ32%と22%であり、N-アセチルノイラミン酸はシナプスの形成を増やし、神経発育を促進することができることを示す。カゼイン酵素分解物は、チロシン、アルギニン、トリプトファンを含み、その中の特殊なalpha-sl-チロシンは、リラックス効果があり、感情を和らげ、睡眠を補助し、不安を解消する効果があると公認されている。ホスファチジルセリンは、細胞膜の活性物質であり、特に脳細胞に存在し、その機能は主に神経細胞の機能の改善、神経パルスの伝導の調節、脳の記憶機能の向上であり、強い親油性があるため、吸収後に速やかに血液脳関門を通って脳に入ることができ、大脳皮質における神経伝達物質であるアセチルコリンの産生を増加させることもでき、アセチルコリンは、思考、推理、集中力に関連しているため、脳機能向上、集中力や記憶力向上の効果がある。
本発明の組成物の原料薬材は、いずれも直接食用可能な食品であり、直接原料を粉末に研磨して食用してもよいが、加工してより食用しやすい抽出物を得ることが好ましい。本発明の組成物の抽出物の調製方法は特に限定されず、公知の方法を使用でき、例えば、上記の重量比の原料を粉砕して直接食用してもよく、水抽出/エタノール抽出して乾燥後にその抽出物を使用してもよい。溶解性のより高い有機溶剤、例えば、トリクロロメタン、ジクロロメタン、酢酸エチルなどで抽出することも可能であるが、有機溶剤残留による食品安全性問題があるため好ましくない。
本発明の組成物の最も好ましい服用形態は、錠剤及びカプセルである。
本発明で開発された漢方薬組成物は、薬食同源の理論に基づく、効果が確実な、安全性が納得できる睡眠改善の組成物であり、本発明の漢方薬組成物は科学的な動物表現型を採用し、メカニズム探究及び臨床服用結果より確実な効果を検証しており、その効果はベンゾジアゼピン西洋薬の薬効より弱いが、信頼できる安全性を考慮すると、特に不眠症の長期服用、治療又は改善の需要を満たすのに適している。本発明の組成物は、原料の理論基礎がより充実であり、現在市販されている一般的な中枢神経抑制類薬物に比べて、副作用が少なく、副反応がない。伝統的な漢方薬処方に比べて、即効性があり、安全性が高い。東洋医学の「清養」理論に基づいているため、本発明は特に仕事ストレスによる不眠症に適用できる。基礎原料はすべて薬食同源の食品であるため、適用範囲が広く、使用上の禁忌が少ない。
梔子167g(ロット番号:2012021-W、産地:福建、供給元:安徽省亳州市中西薬株式会社)、乾姜100g(ロット番号:1910008-W、産地:山東、供給元:安徽省亳州市中西薬株式会社)、甘草100g(ロット番号:2012011-W、産地:内モンゴル、供給元:安徽潤邦中薬飲片株式会社)を適当に粉砕し、6倍量の水を加えて100℃水温で1時間抽出し、ろ過して抽出液を得、上記の抽出を3回繰り返し、抽出液を合わせ、ろ過、濃縮して流エキスを得、得られた流エキスを熱いうちに鹿皮膠310gと均一に混合し、70℃のオーブンで2日間乾燥させ、漢方薬組成物粉末N405gを得た。
2.1 実験動物
野生型AB系統ゼブラフィッシュ(杭州環特生物提供)、自然交配繁殖の方法で行い、計180匹、各実験組30匹、年齢は受精後5日目(5dpf)であった。ゼブラフィッシュはいずれも28℃の魚養殖用水(水質:逆浸透水1Lあたりに200mgインスタント海塩を加え、導電率450~550μS/cm、pH6.5~8.5、硬度50~100 mg/L CaCO3)で飼育され、実験動物使用許可証番号はSYXK(浙)2012-0171である。飼育管理は国際AAALAC認証の要求に従う。
分散分析及びDunnett’s T-テストにより統計学分析を行い、p<0.05は有意差ありを示し、代表的な実験プロットを提供する。
毒理学及び最大検出濃度(MTC)の測定
実験終了時に、漢方薬組成物粉末N の125~2000μg/mL濃度組のいずれも顕著な異常は認められなかった。よって、漢方薬組成物粉末Nの不眠症ゼブラフィッシュに対するMTCは2000μg/mLであると確定した。詳細は表1に示されている。
漢方薬組成物粉末Nは、500、1000、2000μg/mLの濃度組の条件で、PTZに誘発されたゼブラフィッシュ不眠症における覚醒活動量、覚醒総時間のいずれに対しても明らかに改善効果を示し、詳細は図1~4に示されている。
具体的な結果の統計図は図1~4に参照し、モデル対照組に比べて、*はp<0.05、**はp<0.01、***はp<0.001を示している。
3.1 実験動物
野生型AB系統ゼブラフィッシュ(杭州環特生物提供)、自然交配繁殖の方法で行い、計360匹、各実験組は30匹、年齢は5dpfであった。
5dpf野生型AB系統ゼブラフィッシュ360匹を6ウェルプレートにランダムに選び、各ウェル30匹処理し、漢方薬組成物粉末Nを500、1000μg/mL濃度で水溶液で与え、陽性対照組のオキサゼパムの濃度は29μg/mLであり、同時に正常対照組及びモデル対照組を設け、各ウェルの容量は3mLであった。28℃のインキュベーターで24hインキュベートした後、正常対照組以外の他の実験組は691μg/mL PTZでゼブラフィッシュを誘導し、不眠症モデルを確立し、3つの並行実験を行った。実験終点になった後、RNA快速抽出キットを用いて各実験組のゼブラフィッシュの全RNAを抽出し、紫外-可視光分光光度計で総RNA濃度と純度を測定した。ゼブラフィッシュサンプルの全RNAを2μg取り、cDNA第一鎖合成キットの説明に従って操作し、20μLのcDNAを合成して-20℃で保存し、q-PCRによりβ-actin、gabra1、mtnr1aa遺伝子のRNA相対発現量を検測した。β-actinを遺伝子発現の内部参照として、gabra1とmtnr1aa遺伝子のRNA相対発現量を計算した。
分散分析及びDunnett’s T-テストにより統計学分析を行い、p<0.05は有意差ありを示す。
遺伝子相対発現量の公式計算によれば、モデル対照組のgabra1相対発現量は1.00であり、正常対照組(1.21)に比べてp<0.01であり、モデル確立の成功を示す。陽性対照組のオキサゼパムの29μg/mL gabra1相対発現量は1.23であり、モデル対照組(1.00)に比べてp<0.01であり、オキサゼパムがgabra1遺伝子の発現を顕著に増加できることを示す。漢方薬組成物粉末Nが500と1000μg/mL濃度であるとき、gabra1相対発現量はそれぞれ1.25、1.14であり、モデル対照組(1.00)に比べて、p<0.01 & p<0.05である。漢方薬組成物粉末Nはgabra1遺伝子の発現を顕著に増加できることを示す。
4.1 被験者と診断基準
中国精神疾病分類方案および診断基準第二版(CCMD-2-R)の不眠症の診断基準に従って、睡眠障碍を有する20~45歳の被験者60人を選び、前記被験者の症状は以下の基準を満たす。(1)身体疾患又は精神障害に継発する症状のいずれでもない。(2)睡眠障害が最も重要な症状であり、他の症状はすべて不眠症に継発し、これらの症状は、夢が多い、早朝覚醒、目覚めやすい、寝つきが悪い、睡眠が浅い、目が覚めると寝付けない、目が覚めても気分が悪い、倦怠感又は日中の眠気を含む。(3)上記の睡眠障害の症状は週に少なくとも3回発生し、30日以上続く。(4)不眠症により気分のイライラ又は憂鬱が著しく、精神活動の効率が低下し、日常生活と仕事に支障をきたす。
被験者をランダムに3組に分け、各組に被験者20人とした。ビタミンCを空白とし、メラトニンを陽性対象として比較を行い、それぞれ就寝前の30分間~1時間に服用し、空白組は1錠/回、対照組は1錠/回(3mg/粒)、実験組は漢方薬組成物粉末N 3g/回服用した。就寝前の眠りの改善状況、深い睡眠の長さ、睡眠の長さ、及び目覚め後の精神状態、体力状況の評価結果を記録することで統計を行った。
眠りの改善状況は、通常と比べて、顕著に改善した場合、わずかに改善した場合、改善しなかった場合に分けられる。睡眠状況は連続睡眠時間を評価レベルとし、目覚めた後の精神状態はエネルギーが満ちているかどうかを評価レベルとし、体力状況は気力が欠如しているかどうかを評価レベルとし、それぞれ優、中、悪の3つのレベルを設け、いずれも優のレベルに達する場合、睡眠質を一級と判定し、2つが優のレベルに達する場合、二級と判定し、1つ又はそれ以下が優のレベルに達する場合、三級と判定する。結果は表3にまとめた。
実施例1で得られた漢方薬組成物粉末Nは、それぞれヒトの推奨量の5倍、10倍、20倍で動物実験用量を設定し、即ち、空白対照組を83mg/kg、167mg/kg、333mg/kgとし、連続的にサンプルを14日間強制経口投与した後に水迷路実験を行い、空白組に同体積の生理食塩水を投与した。
実験結果は以下の通りである。
本明細書で引用されたすべての出版物及び特許文献は、各出版物又は特許がそれぞれ明確に本願に参照により引用されるように、参照により本願に引用される。本願に開示された実質と範囲を逸脱しない場合、本願に開示された各実施形態に対して様々な変更および均等物の置き換えを行うことができる。文中に特に説明しない限り、本開示の実施形態のすべての特徴、ステップ又は実施形態は、他の任意の特徴、ステップ又は実施形態と組み合わせて使用することができる。
Claims (10)
- 梔子粉14~18部、乾姜粉7~11部、甘草7~11部を取り、粉砕し、重量で5~9倍量の水を加え、80~100℃の水温で0.5~3時間抽出し、ろ過して抽出液を得、上記の抽出を2~4回繰り返し、抽出液を合わせ、ろ過、濃縮して流エキスを得、得られた流エキスを熱いうちに鹿皮膠20~30部と均一に混合し、乾燥させ、抽出物を得るステップを含むことを特徴とする睡眠改善・記憶力向上のための組成物の調製方法。
- 梔子粉14~18部、乾姜粉7~11部、甘草7~11部を取り、粉砕し、それぞれ重量で5~9倍量の水を加え、80~100℃の水温で0.5~3時間抽出し、ろ過して抽出液を得、上記の抽出を2~4回繰り返し、抽出液を合わせ、ろ過、濃縮して流エキスを得、得られた梔子粉の流エキス、乾姜粉の流エキス、甘草の流エキスを真空乾燥又は噴霧乾燥させ、得られた梔子粉の流エキス乾燥粉末、乾姜粉の流エキス乾燥粉末及び甘草の流エキス乾燥粉末を、鹿皮膠20~30部と混合し、粉砕し、篩にかけて、均一に混合し、抽出物を得るステップを含むことを特徴とする睡眠改善・記憶力向上のための組成物の調製方法。
- 梔子粉14~16部、乾姜粉8~10部、甘草8~10部を取り、粉砕し、重量で5~9倍量の水を加え、80~100℃の水温で0.5~3時間抽出し、ろ過して抽出液を得、上記の抽出を2~4回繰り返し、抽出液を合わせ、ろ過、濃縮して流エキスを得、得られた流エキスを熱いうちに鹿皮膠26~30部と均一に混合し、乾燥させ、抽出物を得るステップを含むことを特徴とする睡眠改善・記憶力向上のための組成物の調製方法。
- 梔子粉14~16部、乾姜粉8~10部、甘草8~10部を取り、粉砕し、それぞれ重量で5~9倍量の水を加え、80~100℃の水温で0.5~3時間抽出し、ろ過して抽出液を得、上記の抽出を2~4回繰り返し、抽出液を合わせ、ろ過、濃縮して流エキスを得、得られた梔子粉の流エキス、乾姜粉の流エキス、甘草の流エキスを真空乾燥又は噴霧乾燥させ、得られた梔子粉の流エキス乾燥粉末、乾姜粉の流エキス乾燥粉末及び甘草の流エキス乾燥粉末を、鹿皮膠26~30部と混合し、粉砕し、篩にかけて、均一に混合し、抽出物を得るステップを含むことを特徴とする睡眠改善・記憶力向上のための組成物の調製方法。
- 梔子粉15部、乾姜粉9部、甘草9部を取り、粉砕し、重量で5~9倍量の水を加え、80~100℃の水温で0.5~3時間抽出し、ろ過して抽出液を得、上記の抽出を2~4回繰り返し、抽出液を合わせ、ろ過、濃縮して流エキスを得、得られた流エキスを熱いうちに鹿皮膠28部と均一に混合し、乾燥させ、抽出物を得るステップを含むことを特徴とする睡眠改善・記憶力向上のための組成物の調製方法。
- 梔子粉15部、乾姜粉9部、甘草9部を取り、粉砕し、それぞれ重量で5~9倍量の水を加え、80~100℃の水温で0.5~3時間抽出し、ろ過して抽出液を得、上記の抽出を2~4回繰り返し、抽出液を合わせ、ろ過、濃縮して流エキスを得、得られた梔子粉の流エキス、乾姜粉の流エキス、甘草の流エキスを真空乾燥又は噴霧乾燥させ、得られた梔子粉の流エキス乾燥粉末、乾姜粉の流エキス乾燥粉末及び甘草の流エキス乾燥粉末を、鹿皮膠28部と混合し、粉砕し、篩にかけて、均一に混合し、抽出物を得るステップを含むことを特徴とする睡眠改善・記憶力向上のための組成物の調製方法。
- 請求項1~6のいずれか一項に記載の睡眠改善・記憶力向上のための組成物の調製方法に、さらに、酸棗仁、蓮子、百合の少なくとも一種の漢方薬成分を加えることを特徴とする睡眠改善・記憶力向上のための組成物の調製方法。
- 請求項1~6のいずれか一項に記載の睡眠改善・記憶力向上のための組成物の調製方法に、さらに、テアニン、カゼイン酵素分解物、N-アセチルノイラミン酸、ビタミンB6、及びホスファチジルセリンから選択される成分の少なくとも一種を加えることを特徴とする睡眠改善・記憶力向上のための組成物の調製方法。
- 請求項1~6のいずれか一項に記載の睡眠改善・記憶力向上のための組成物の調製方法に、さらに、薬学的に許容できる補助材料を加えて睡眠改善・記憶力向上のための薬物製剤を調製する方法。
- 請求項1~6のいずれか一項に記載の睡眠改善・記憶力向上のための組成物の調製方法に、さらに、食用可能な補助材料を加えて機能性食品を製造する方法。
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