Development of a simple screening system for the discovery of a new drug for diabetes

开发用于发现糖尿病新药的简单筛选系统

• 批准号: 08558074
• 负责人: EBINA Yousuke
• 金额: $ 8.83万
• 依托单位: The University of Tokushima
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 财政年份: 1996
• 资助国家: 日本
• 起止时间: 1996 至 1997
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-08558074/
• 关键词: drug for diabetes; glucose transporter; 糖尿病; GLUT4トランスロケーション; 非アイソトープ測定; 三量体Gタンパク

There are more than 5 million patients of diabetes and a new oral drug must be developed. However, there is not a simple and quantitative method to find the oral diabetic drug. Whole animal was used to find the new drug to estimate the decrease of blood glucose level, but this method is not adeguate toscreen a large number of compound. Translocation of the type 4 glucose transporter (GLUT4) to the cell surface from an intracellular pool is the major mechanism of insulin-stimulated glucose uptake in insulin-target cells. We developed a highly sensitive and quantitative method of detect GLUT4 immunologically on the surface of intact cells, using c-myc epitope-tagged GLUT4 (GLUT4myc). We established stable clones to express GLUT4myc in CHO,L6 myoblasts and 3T3L1 adipocytes. We have developed a simple and new method to screen the compounds for triggering the GLUT4myc translocation in 96 wells using luminessence method. This method is useful for the first screening for developing a new oral drug for diabetes.

目前有500多万糖尿病患者，必须开发一种新的口服药物。然而，目前还没有一种简单、定量的方法来寻找口服糖尿病药物。以往的降血糖新药的筛选是采用整体动物法，但这种方法不适合于大量筛选化合物。4型葡萄糖转运蛋白（GLUT 4）从细胞内转运至细胞表面是胰岛素刺激胰岛素靶细胞葡萄糖摄取的主要机制。我们建立了一种高灵敏度和定量检测GLUT 4在完整细胞表面的免疫学方法，使用c-myc表位标记的GLUT 4（GLUT 4 myc）。我们建立了稳定的克隆表达GLUT 4 myc在CHO，L 6成肌细胞和3 T3 L1脂肪细胞。我们发展了一种简单的新方法，用发光素法在96个威尔斯孔中筛选触发GLUT 4 myc易位的化合物。该方法可用于糖尿病口服新药的初步筛选。

项目成果

Kazuhiko Kishi, Yousuke Ebina, et al.: "Gq-coupled receptors transmit the signal for GLUT4 translocation via an insulin-independent pathway" J. Biol. Chem.271. 26561-26568 (1996)

Kazuhiko Kishi、Yousuke Ebina 等人：“Gq 偶联受体通过不依赖于胰岛素的途径传递 GLUT4 易位信号” J. Biol。

Kozlovsky N., Ebina Y.et.al.: "Transcriptional activation of the glut1 gene in response to oxidative stress in L6 myotubes" J.Biol.Chem.272. 33367-33372 (1997)

Kozlovsky N.、Ebina Y.et.al.：“L6 肌管氧化应激反应中 glut1 基因的转录激活”J.Biol.Chem.272。

Todaka,M.,Ebina.Y.et al.: "Roles of insulin,guanosine 5′-〔γ-thio〕triphosphate and phorbol 12-myristate 13-acetate in signaling pathways of GLUT4 translocation" Biochem.J.315. 875-882 (1996)

Todaka, M., Ebina.Y. 等人：“胰岛素、鸟苷 5-[γ-硫代]三磷酸和佛波醇 12-肉豆蔻酸酯 13-乙酸酯在 GLUT4 易位信号通路中的作用”Biochem.J.315。 -882 (1996)

Kazuhiro Kishi, Yousuke Ebina, et al.: "Gq-coupled receptors transmit the signal for GLUT4 translocation via an insulin-independent pathway" J.Biol.Chem.271. 26561-26568 (1996)

Kazuhiro Kishi、Yousuke Ebina 等人：“Gq 偶联受体通过独立于胰岛素的途径传递 GLUT4 易位信号”J.Biol.Chem.271。

Kishi K., Ebina Y.et.al.: "Bradykinin directly triggers GLUT4 translocation via an insulin-independent pathway" Diabetes. (in press).

Kishi K.、Ebina Y.et.al.：“缓激肽通过不依赖于胰岛素的途径直接触发 GLUT4 易位”糖尿病。