Identifying modulators of hERG channel activity using the PatchXpress® planar patch clamp
Identifying modulators of hERG channel activity using the PatchXpress® planar patch clamp
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DOI：
10.1177/1087057104272295
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发表时间：
2005-03-01
影响因子：
--
通讯作者：
Mathes, C
中科院分区：
文献类型：
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作者：
Dubin, AE;Nasser, N;Mathes, C
作者使用PatchXpress（（R））7000 A系统，使用模拟“金标准”常规全细胞膜片钳的程序来测量hERG通道的化合物活性。使用旨在鉴定单一浓度下的化合物活性或从累积浓度依赖性范例获得化合物效力的方案，在hERG 302-HEK细胞上测试一组70种化合物，包括效力跨越3个数量级的hERG拮抗剂。在暴露于化合物和随后冲洗威尔斯孔以确定阻断的可逆性后，用参比化合物阻断作为质量控制。电参数和电压依赖性是类似的，使用传统的全细胞膜片钳。化合物效力的等级顺序也与通过常规方法测定的等级顺序相当。一个例外是氟桂利嗪，一种特别亲脂的化合物。PatchXpress（R）准确地鉴定了29种中等效力的拮抗剂的活性，这些拮抗剂仅微弱地取代放射性标记的阿司咪唑，并且在结合测定中为假阴性。最后，从一组相对不活跃的化合物中没有观察到假命中。PatchXpress（R）的高质量数据采集应该有助于加速离子通道调节剂的二次筛选和药物发现过程。
The authors used the PatchXpress((R)) 7000A system to measure compound activity at the hERG channel using procedures that mimicked the "gold-standard" conventional whole-cell patch clamp. A set of 70 compounds, including hERG antagonists with potencies spanning 3 orders of magnitude, were tested on hERG302-HEK cells using protocols aimed at either identifying compound activity at a single concentration or obtaining compound potency from a cumulative concentration dependence paradigm. After exposure to compounds and subsequent washout of the wells to determine reversibility of the block, blockade by a reference compound served as a quality control. Electrical parameters and voltage dependence were similar to those obtained using a conventional whole-cell patch clamp. Rank order of compound potency was also comparable to that determined by conventional methods. One exception was flunarizine, a particularly lipophilic compound. The PatchXpress((R)) accurately identified the activity of 29 moderately potent antagonists, which only weakly displace radiolabeled astemizole and are false negatives in the binding assay. Finally, no false hits were observed from a collection of relatively inactive compounds. High-quality data acquisition by PatchXpress((R)) should help accelerate secondary screening for ion channel modulators and the drug discovery process.