Generation of a novel cell-based assay system for screening agents specific to two-pore domain K+ channels

2017 Fiscal Year Final Research Report

• Project/Area Number: 16K15128
• Research Category: Grant-in-Aid for Challenging Exploratory Research
• Allocation Type: Multi-year Fund
• Research Field: Pharmacology in pharmacy
• Research Institution: Nagoya City University
• Principal Investigator: Imaizumi Yuji 名古屋市立大学, 大学院薬学研究科, 教授 (60117794)
• Co-Investigator(Kenkyū-buntansha): 山村 寿男 名古屋市立大学, 大学院薬学研究科, 准教授 (80398362) ; 鈴木 良明 名古屋市立大学, 大学院薬学研究科, 助教 (80707555)
• Project Period (FY): 2016-04-01 – 2018-03-31
• Keywords: イオンチャネル / 薬理学 / ハイスループットスクリーニング / 細胞死 / 創薬 / 薬学 / パッチクランプ / 活動電位

Outline of Final Research Achievements

To develop a new ion channel-targeted drug screening system for high throughput screening (HTS) assay, we have established HEK293-based “test cell” expressing a mutated Na+ channel lacking inactivation and a K+ channel (Kir2.1) that hyperpolarizes the membrane potential (PCT/JP2011/064967). We found that only treatment of the test cells with Ba2+ is enough to induce action potential and cell death. Then two-pore domain potassium (K2P) channels were additionally expressed in the test cells because K2P channels are involved in progression of various disease and thought to be druggable targets. In this system, both activating and inhibitory effects of drugs on K2P channels can be easily and accurately estimated using simple cell death assay. IC50 values of these blockers acquired by both manual and automated process were close to those obtained using patch-clamp recordings. Now drug screenings using compound library are in progress.

Free Research Field

薬理学