JP7116960B2 - ゲノム安定性増強剤 - Google Patents
ゲノム安定性増強剤 Download PDFInfo
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- JP7116960B2 JP7116960B2 JP2019505908A JP2019505908A JP7116960B2 JP 7116960 B2 JP7116960 B2 JP 7116960B2 JP 2019505908 A JP2019505908 A JP 2019505908A JP 2019505908 A JP2019505908 A JP 2019505908A JP 7116960 B2 JP7116960 B2 JP 7116960B2
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- Prior art keywords
- royal jelly
- peptidase
- gnetum
- acid
- h2ax
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Description
項2.化粧品、飲食品、医薬品、又は医薬部外品である、項1に記載のゲノム安定性増強剤。
項3.酵素分解ローヤルゼリー、グネツム又はその抽出物、及びグネチンCからなる群から選択される少なくとも1種の有効量を哺乳動物に投与する工程を含む、ゲノム安定性を増強させる方法。
項4.ゲノム安定性増強剤の製造における、酵素分解ローヤルゼリー、グネツム又はその抽出物、及びグネチンCからなる群から選択される少なくとも1種の使用。
項5.前記ゲノム安定性増強剤が、化粧品、飲食品、医薬品、又は医薬部外品である、項4に記載の使用。
項6.酵素分解ローヤルゼリー、グネツム又はその抽出物、及びグネチンCからなる群から選択される少なくとも1種を有効成分として含有するゲノム安定性増強用組成物。
項7.化粧品、飲食品、医薬品、又は医薬部外品である、項6に記載のゲノム安定性増強用組成物。
ローヤルゼリーの酵素分解物は、日本国特許第3994120号公報の実施例1に従い調製した。酵素分解ローヤルゼリー及びローヤルゼリーの凍結乾燥粉末の終濃度が100μg/mLとなるように10% FBS添加α-MEM培地に溶解し、フィルター滅菌した。
カラム:東ソー製 TSKgel ODS-100V、5μm、4.6×150 mm
移動層:A液:1.0容量%酢酸含有水、B液:1.0容量%酢酸含有メタノール
グラジェント条件:0分→10分:A:B=65:35(v/v)→63:37(v/v)、10分→20分:A:B=63:37(v/v)→56:44(v/v)、20分→40分:A:B=48:52(v/v)
検出波長:320 nm
流速:0.8 mL/min
ゲルの種類:シリカゲル、Daisogel IR-60-40/63-W
カラムサイズ:φ2×7.48 cm
検出波長:320 nm
移動層:A液:メタノール、B液:クロロホルム
グラジェント条件:0分→2分:A:B=11:89(v/v)、2分→8分:A:B=11:89(v/v)→18:82(v/v)、8分→12分:A:B=18:82(v/v)、12分→14分:A:B=21:79(v/v)、14分→20分:A:B=21:79(v/v)→29:71(v/v)、20分→24分:A:B=29:71(v/v)、24分→30分:A:B=29:71(v/v)→36:64(v/v)、30分→34分:36:64(v/v)
流速: 60 ml/min
正常細胞の増殖停止は、H2AX (DNA損傷修復因子群のDNA損傷部位への集積誘導に必須)レベルの低下に伴って誘導されるため、同時にDNA修復能も低下している。ゲノム不安定性は、この背景で増殖刺激に起因して誘導されることが明確になっている。そこで、酵素分解ローヤルゼリー(酵素処理RJ)及びローヤルゼリー(未処理RJ)のH2AX発現への影響を解析した。
試験例1と同様の方法を用いて、メリンジョ及びトランスレスベラトロールのH2AX発現への影響を解析した。なお、今回は0.26, 0.52, 1.0, 2.1w/v%のメリンジョ種子抽出物又はトランスレスベラトロールを培地に添加し、1時間培養した細胞を回収した。
試験例1と同様の方法を用いて、トランスレスベラトロール、メリンジョ及びグネチンCのH2AX発現への影響を解析した。なお、今回は2.5μM (0.52μg/ml)のトランスレスベラトロール、0.52μg/mlのメリンジョ種子抽出物、又は2.5μMのグネチンCを培地に添加し、0, 1.5, 3, 6, 12, 24時間培養した細胞を回収した。
メリンジョによる生体でのゲノム安定性増強効果を検証するため、ミスマッチ修復因子を欠損したMsh2-KOマウス{ゲノム不安定性の一つのタイプであるマイクロサテライト不安定性(MSI)を生じるリンチ症候群モデル}を用いて実験を行った。このマウスでは、10ヶ月齢頃から、おおよそ半年程度の間に死に至ることが知られている。
メリンジョの摂取が予防的な効果を発揮するのか検証した。8-12週齢のMsh2-KOマウスに0.03w/v%のメリンジョ種子抽出物を投与した群(36 mg/kg体重/日)、又は対照群として滅菌水を投与したマウスに、発癌剤である臭素酸カリウム(0.02%)を含む水を飲料水として与えた。24週後に、腸管に形成された腫瘍の数をカウントし、メリンジョ群と対照群を比較した。ミスマッチ修復因子を欠損したMsh2-KOマウスはゲノム不安定が生じやすく、化学発がん物質であるBrKO3により容易に消化管に腫瘍が形成される。
Claims (2)
- 酵素分解ローヤルゼリー、グネツム又はその抽出物、及びグネチンCからなる群から選択される少なくとも1種を有効成分として含有し、前記酵素がエンドペプチダーゼ作用とエキソペプチダーゼ作用を有するペプチダーゼ、又はエンドペプチダーゼ作用を有するペプチダーゼとエキソペプチダーゼ作用を有するペプチダーゼの組合せである、ゲノム安定性増強剤。
- 化粧品、飲食品、医薬品、又は医薬部外品である、請求項1に記載のゲノム安定性増強剤。
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