JP6091067B2 - 細胞賦活剤およびその用途 - Google Patents
細胞賦活剤およびその用途 Download PDFInfo
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- JP6091067B2 JP6091067B2 JP2012064301A JP2012064301A JP6091067B2 JP 6091067 B2 JP6091067 B2 JP 6091067B2 JP 2012064301 A JP2012064301 A JP 2012064301A JP 2012064301 A JP2012064301 A JP 2012064301A JP 6091067 B2 JP6091067 B2 JP 6091067B2
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- food
- cell activator
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- 235000019168 vitamin K Nutrition 0.000 description 1
- 239000011712 vitamin K Substances 0.000 description 1
- 150000003721 vitamin K derivatives Chemical class 0.000 description 1
- 229940046010 vitamin k Drugs 0.000 description 1
- 230000010388 wound contraction Effects 0.000 description 1
- 230000037303 wrinkles Effects 0.000 description 1
- 235000013618 yogurt Nutrition 0.000 description 1
- 239000011701 zinc Substances 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
- 239000002076 α-tocopherol Substances 0.000 description 1
- 235000004835 α-tocopherol Nutrition 0.000 description 1
Description
乾燥させたアーティチョークの可食部600 gに酢酸エチル2300 mLを加え、80℃にて2時間抽出した。抽出液をろ過した後、溶媒を減圧除去し、アーティチョークの酢酸エチル抽出物17.5 gを得た。同様の抽出を再度行い、合計32.3 gの抽出物を得た。得られた抽出物32.3 gを酢酸エチル10 mLに溶解後、シルカゲル30 gを加え、エバポレーターで減圧乾燥し、試料層を調製した。シリカゲル700 gをクロマト管に詰め、先に調製した試料層を積層し、乾式カラムクロマトを作製した。ヘキサン:酢酸エチル混合溶媒(混合比率、2:3)で溶出を開始し、1画分当たり1400 mLずつ溶出液を捕集し、4個の画分に分離した。続いて2番目の画分を乾式カラムクロマトにて、ヘキサン:酢酸エチル:酢酸混合溶媒(混合比率、80:20:1)、ヘキサン:アセトン:メタノール(混合比率、40:8:1)で順次分画し、粗分画物300 mgを得た。続いて、得られた粗分画物約300 mgをジクロロメタン2.0 mLに溶解し、次いでアセトニトリル2.0 mLを加えた。溶液を0.45μmメンブランフィルターにてろ過し、以下のHPLCによる分離操作に付した。
カラム:Capcell Pak C18カラム(UG80, 250 × 10 mm i.d., 株式会社資生堂)
溶媒:アセトニトリル−0.1%ギ酸 70:30
流速:4.0 mL/min
温度:40℃
検出波長:254 nm
負荷量:0.4 mg
本条件で得られたクロマトグラムを図1に示す。図1に示すように、粗分画物には主要ピークが3本観察された。以下、これらをそれぞれピーク1(Rt 10.9)、ピーク2(Rt 11.8)およびピーク3(Rt 12.5)と称する。
カラム:Cosmosil cholester(250 × 10 mm i.d., ナカライテスク株式会社)
溶媒:メタノール−アセトニトリル−0.1%ギ酸 65 : 12 : 23
流速:3.5 mL/min
温度:40℃
検出波長:254 nm
負荷量:1.0 mg
本条件で得られたクロマトグラムを図2に示す。図2から明らかなように、ピーク1は混合物であることが判明し、主要ピークが2本観察された。以下、これらをそれぞれピーク1−1(Rt 19.8)およびピーク1−2(Rt 22.1)と称する。
ヒト正常真皮線維芽細胞を1%ウシ胎児血清(FBS)含有DMEMを用いて、1ウェル当たり2×104個となるように96ウェルプレートに播種した。24時間後、試験試料(上記で単離された各ピークの化合物(計4種))を最終濃度が20μMとなるように添加した。また、コントロールとしては溶媒であるDMSOのみを添加した。試料添加から24時間後、Premix WST-1 Cell Proliferation Assay System(タカラバイオ株式会社製)を用いて、細胞増殖活性を評価した。コントロールの細胞賦活活性(細胞増殖促進作用)を100%としたときの結果を下記の表1に示す。
上記で優れた細胞賦活活性(細胞増殖促進作用)を示した各ピークの化合物(計4種)を、以下のとおり機器分析により同定した。
Claims (4)
- 下記の化合物1〜4の少なくとも1種:
- 前記線維芽細胞がヒト真皮線維芽細胞である、請求項1に記載の線維芽細胞の増殖促進剤。
- 請求項1に記載の前記化合物1〜4の少なくとも1種を含有する、線維芽細胞の増殖促進による細胞賦活剤。
- 請求項3に記載の細胞賦活剤と製薬上許容され得る担体とを含有する薬剤組成物からなる、創傷治癒促進剤。
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