A HTS assay for voltage-gated ion channel targets

描述(由申请人提供)：膜分析公司(MAI)致力于开发和商业化电压门控离子通道靶标的高通量筛选(HTS)分析。这种分析被称为离子通道分析(ICA)。ICA使用电场来刺激电压门控离子通道。细胞的反应是使用荧光技术进行光学检测的。ICA是一种增强型96孔微孔板。在96个井的底面上都有微电极。生长在井底的细胞与微电极电接触。当在两个电极之间施加电压时，细胞膜上的局部电势会改变，从而触发电压门控离子通道的响应。然后可以测量药物阻断、增加或降低这种反应的作用。 电压门控离子通道在人体心脏跳动、肌肉收缩和荷尔蒙释放等生物过程中起着关键作用。因此，这些渠道也是重要的药物靶点。不存在快速测试药物对电压门控离子通道的作用的有效方式。传统的膜片钳速度太慢，甚至连目前处于测试阶段的新型自动化膜片钳产品也没有所需的吞吐量。结果是错过了好的药物。此外，药物对心脏中一种特殊的电压门控离子通道HERG通道的未知副作用已导致5种药物从美国市场下架。因此，即使药物的目标分子不是电压门控离子通道电势，新药仍然必须针对电压门控离子通道HERG通道进行测试，以揭示副作用。 MAI建议为电压门控离子通道靶标开发一种急需的HTS分析方法。该检测方法易于使用，并与大多数大型制药公司目前使用的设备兼容。一期资金将用于制造100个96孔板器件。首批100个设备的制造将验证拟议制造工艺的技术可行性，并为正在进行的生物测试和化验开发提供试板。还将进行电气测试。

DESCRIPTION (provided by applicant): Membrane Assays, Inc. (MAI) seeks to develop and commercialize a high throughput screening (HTS) assay for voltage-gated ion channel targets. The assay is called the Ion Channel Assay (ICA). The ICA uses an electric field to stimulate voltage-gated ion channels. The cellular response is detected optically using fluorescence technology. The ICA is an enhanced 96-well microwell plate. On the bottom surface of each of the 96 wells are microelectrodes. Cells grown on the bottom of the well make electrical contact with the microelectrodes. When a voltage is applied between the electrodes, the local electrical potential across the cellular membrane is altered triggering a response by the voltage-gated ion channels. The action of a pharmaceutical to block, increase or decrease this response can then be measured. Voltage-gated ion channels play a critical role in such biological processes as the beating of the human heart, the contraction of muscles, and the release of hormones. Therefore, these channels are also important drug targets. No effective manner of rapidly testing the action of a pharmaceutical against a voltage-gated ion channel exists. Traditional patch clamping is too slow and even the new automated patch clamp products, currently in beta testing, do not have the throughput required. The result is missed good drugs. In addition, unknown side-effects of pharmaceuticals on a particular voltage-gated ion channel in the heart, the hERG channel, have led to the removal of 5 drugs from U.S. market. Thus, even if the target molecular of a pharmaceutical is not a voltage-gated ion channel potential new drugs must still be tested against a voltage-gated ion channel, the hERG channel, to uncover side-effects. MAI proposes to develop a critically needed HTS assay for voltage-gated ion channel targets. The assay is easy to use and compatible with equipment currently in place at most large pharmaceutical companies. Phase I funds will be used to manufacture 100, 96-well plate devices. The fabrication of these first 100 devices will verify the technical feasibility of the proposed manufacturing process and provide plates for ongoing biological testing and assay development. Electrical testing will also be performed.