JP7388867B2 - 新規なジンセノサイドを含む組成物 - Google Patents
新規なジンセノサイドを含む組成物 Download PDFInfo
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- JP7388867B2 JP7388867B2 JP2019189417A JP2019189417A JP7388867B2 JP 7388867 B2 JP7388867 B2 JP 7388867B2 JP 2019189417 A JP2019189417 A JP 2019189417A JP 2019189417 A JP2019189417 A JP 2019189417A JP 7388867 B2 JP7388867 B2 JP 7388867B2
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- ginsenoside
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- ginsenosides
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Description
分画
高麗人参の種子(Seeds of Panax ginseng)5.5kgをミキサーで細かく粉砕して粉末状にしてメタノールで抽出後、n-ヘキサン、エチルアセテート、n-ブタノールなどを用いて段階的に分画した。脂質(Lipid)の大部分はn-ヘキサンによって除去され、エチルアセテート分画物に残っている脂質はメタノール:水=1:1(v/v)で懸濁後にフリーザーに一晩保管してから上層液だけを取った後、遠心分離機を利用してもう一回除去した。このように前処理が施されたエチルアセテート分画物2.61gとn-ブタノール分画物114.64gをカラム及びHPCCC(High Performance Counter-Current Chromatography)によって下記のように分画した。
n-ブタノール分画物114.64gに対してMPLCにて分画を分け、このときに用いた溶媒はn-ヘキサン/エチルアセテート=10:1→5:1→1:1CHCl3/MeOH=10:1→5:1(v/v)であり、流速は50mL/minとした。前記条件を用いて合計12個の下位分画に分け、各分画をさらにHPCCC、HPLC(High-performance liquid chromatography)、セファデックス(Sephadex)LH-20カラムなどを用いて各分画に含有されている成分を分離し、NMR(Nuclear magnetic resonance)、UV(Ultraviolet rays)、MS(Mass spectrometry)を用いて構造を同定して16種の化合物を究明した。
前記高麗人参の種子抽出物から分離したジンセノサイドの血糖抑制効能を比べるために、次のように多糖類消化酵素の活性の抑制効果を評価した。
前記高麗人参の種子抽出物から分離したジンセノサイドの血糖抑制効能に係る体内糖代謝に及ぼす影響を調べるために、次のように血液中のブドウ糖の細胞吸収(glucose uptake)促進効果を評価した。
3T1-L1脂肪細胞(ATCC)を購入してダルベッコ改変イーグル培地(Dulbecco's Modified Eagle's Medium、Sigma)に10%子ウシ血清(bovine calf serum、Hyclone)及び1%ペニシリン/ストレプトマイシン(Sigma)を添加して5% CO2培養器で培養した。脂肪前駆細胞を脂肪細胞に分化させるために細胞を培養皿にいっぱい満たして培養してから48時間後に、10%ウシ胎児血清(fetal bovine serum(FBS);Hyclone)、0.5mM 3-イソブチル-1-メチルキサンチン(3-isobutyl-1-methylxanthine、Sigma)、1μM デキサメタゾン(dexamethasone、Sigma)、5μg/ml インスリン(Sigma)、1%ペニシリン/ストレプトマイシンが添加されたDMEM培地に取り替え、48時間をさらに培養した。以降、2日おきに10% FBS及び5μg/mlインスリン、1%ペニシリン/ストレプトマイシンが含まれた培地に取り替え、14日間さらに培養して、完全に分化された脂肪細胞を得た。分化された脂肪細胞を低ブドウ糖(low-glucose)DMEM(Sigma)培地を利用して断食(fasting)を誘導した後、血糖吸収分析キット(glucose uptake assay kit、abcam)でブドウ糖吸収能力を観察した。血液中のブドウ糖吸収測定のための陽性対照群としては、糖尿治療剤として用いられているメトホルミン(Sigma;10μM)を用い、高麗人参の種子抽出物から分離した本発明の一実施例である新規なジンセノサイドGS#10と本発明の比較例であるジンセノサイドGS#01-GS#06をそれぞれ10μMずつ処理した。また、ブドウ糖吸収を促進できるインスリン(10nM、Sigma)を一緒に処理することでインスリン依存的/非依存的ブドウ糖輸送能をいずれも測定した。
本発明の一実施例である新規なジンセノサイドGS#10の糖代謝調節効能を、本発明の比較例として紅参指標成分であるジンセノサイドRg1、Rg3及びRb1(Sigma社から購買)と比べた。このとき、前記本発明の比較例であるジンセノサイドRg3の化学構造は、次のとおりである。
ジンセノサイドが細胞毒性活性を通じて人体に有用な効能に影響を及ぼす可能性を排除するために、CCK(Cell Counting Kit)-8を用いて本発明の実施例である新規なジンセノサイドGS#10の存在時の細胞成長を評価した。実験方法は次のとおりである。
前記高麗人参の種子抽出物から分離したジンセノサイドの脂質代謝抑制効能を比べるために脂質合成関連遺伝子の発現抑制効能の実験を次のように実施した。
食べ物から摂取した脂肪は、腸で膵臓リパーゼ(pancreatic lipase)によって分解され吸収される。このため、血中脂質濃度を減少させるためには、肝での脂質代謝だけではなく小腸での脂肪分解もまた抑制される必要があり、高麗人参の種子由来のジンセノサイドが脂肪の消化にも影響を及ぼすかを調べるために、次のような実験を実施した。
本発明の一実施例である新規なジンセノサイドGS#10(高麗人参の種子抽出物から分離)の脂質代謝抑制効能を、本発明の比較例として紅参の指標成分であるジンセノサイド3種(Rg1、Rg3、Rb1;Sigma)と比べるために、前記試験例5及び6と同様な方法にて実験を行った。
前記高麗人参の種子抽出物から分離したジンセノサイドのコレステロール合成関連遺伝子の活性調節効能を比較するために、次のような実験を実施した。
本発明の一実施例である新規なジンセノサイドGS#10のコレステロール合成関連遺伝子の活性調節効能を、本発明の比較例として紅参の指標成分であるジンセノサイド3種(Rg1、Rg3、Rb1;Sigma)と比べるために、前記試験例8と同様な方法にて実験を行った。このとき、HepG2細胞に紅参の指標成分(Ginsenoside Rg1、Rg3、Rb1;Sigma)及び新規なジンセノサイドGS#10を各1、10μMの濃度で24時間処理した。
前記高麗人参の種子抽出物から分離したジンセノサイドの抗肥満効能を比べるために、脂肪細胞分化抑制効能実験を次のように実施した。
前記試験例10と同様な方法にて脂肪細胞分化抑制効能を比べており、本発明の一実施例である新規なジンセノサイドGS#10(高麗人参の種子抽出物から分離)を、本発明の比較例として紅参の指標成分であるジンセノサイド3種(Rg1、Rg3、Rb1;Sigma)と比べた。このとき、前記各ジンセノサイドの濃度は1、10μMであった。
血管内皮細胞で生成される酸化窒素(nitric oxide;NO)は血管を拡張して血流量を増加させる役割をするので、血管内皮細胞での酸化窒素生成量を増加させることができたら血流量の増加を通じて血行も改善することができることを意味する。
血管内皮細胞の生存率を増加させることで血管の老化が防止又は遅延できたら、血管の機能を維持し且つ血管の弾力低下も防止又は改善できるので、血行を改善させることができる。
本発明の一実施例である新規なジンセノサイドGS#10の血流量増加及び血管老化防止効能を、本発明の比較例として紅参指標成分であるジンセノサイドRg1、Rg3及びRb1(Sigmaから購買)と比べた。
下記の表に記載された組成にて通常の方法に従い柔軟化粧水を製造した。
下記の表に記載された組成にて通常の方法に従い栄養化粧水を製造した。
下記の表に記載された組成にて通常の方法に従いマッサージクリームを製造した。
ジンセノサイドPG-RT8 100mg、ラクトース400mg、とうもろこし澱粉400mg及びステアリン酸マグネシウム2mgを混合した後、通常の錠剤の製造方法に従い打錠して錠剤を製造した。
ジンセノサイドPG-RT8 100mg、ラクトース400mg、とうもろこし澱粉400mg及びステアリン酸マグネシウム2mgを混合した後、通常のカプセル剤の製造方法に従いゼラチンカプセルに充填してカプセル剤を製造した。
ジンセノサイドPG-RT8 50mg、無水結晶ブドウ糖250mg及び澱粉550mgを混合し、流動層造粒機を用いて顆粒に成形した後、分包に充填した。
ジンセノサイドPG-RT8 50mg、ブドウ糖10g、クエン酸0.6g、及び液状オリゴ糖25gを混合した後、精製水300mlを加えて、各瓶に200mlずつ充填する。瓶に充填した後、130℃で4~5秒間殺菌してドリンク剤を製造した。
ジンセノサイドPG-RT8 50mg、とうもろこしシロップ(corn syrup)1.8g、脱脂牛乳0.5g、大豆レシチン0.5g、バター0.6g、植物性硬化油0.4g、砂糖1.4g、マーガリン0.58g、及び食塩20mgを混合し、キャラメル成形した。
下記の表に記載された組成にて通常の方法に従い注射剤を製造した。
Claims (14)
- (20S,24R)-6-O-β-D-グルコピラノシル(1→2)-β-D-グルコピラノシド-ダンマル-3-オン-20,24-エポキシ-6a,12b,25-トリオール((20S,24R)-6-O-β-D-glucopyranosyl(1→2)-β-D-glucopyranoside-dammar-3-one-20,24-epoxy-6a,12b,25-triol)、その薬学的に許容可能な塩、その水和物又はその溶媒和物を有効成分として含む、代謝調節及び血行改善の一つ以上の用途の組成物であって、
前記代謝調節は、血糖調節、脂質代謝調節又は抑制、コレステロール調節、抗肥満の一つ以上を含み、
前記組成物は、食品組成物又は薬学組成物である、
組成物。 - 前記有効成分は、高麗人参の種子から抽出したものである、請求項1又は2に記載の組成物。
- 血糖降下用である、請求項1~3のいずれか一項に記載の組成物。
- 血液中の糖分解抑制用である、請求項1~4のいずれか一項に記載の組成物。
- 血液中のブドウ糖の細胞吸収促進用である、請求項1~5のいずれか一項に記載の組成物。
- 糖尿又は糖尿合併症の予防又は治療用である、請求項1~6のいずれか一項に記載の組成物。
- 高血圧、糖尿病、高脂血症、心筋こうそく、狭心症、高中性脂肪血症、動脈硬化症、及び高コレステロール血症からなる群より選ばれた一つ以上の予防又は改善用である、請求項1~7のいずれか一項に記載の組成物。
- 脂肪細胞分化抑制用である、請求項1~8のいずれか一項に記載の組成物。
- 体内脂質合成又は蓄積抑制用である、請求項1~9のいずれか一項に記載の組成物。
- 血流量増加用である、請求項1~10のいずれか一項に記載の組成物。
- 皮膚血色改善用である、請求項1~11のいずれか一項に記載の組成物。
- 前記有効成分を組成物の総質量に対して0.0001~99.9質量%の範囲で含む、請求項1~12のいずれか一項に記載の組成物。
- 前記組成物の投与量は、0.05mg/kg/日~10g/kg/日の範囲である、請求項1~13のいずれか一項に記載の組成物。
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