COMPOUNDS TARGETING THE ALPHA-2 GABA-A RECEPTOR SUBTYPE

描述（由申请人提供）：持续需要有效且安全的治疗焦虑和相关障碍的方法。目前的治疗方法，如针对许多γ -氨基丁酸受体（GABA-A受体）亚型的苯二氮卓类药物是有效的，但患者会遭受许多不想要的副作用。由于缺乏有效的先导物发现和优化所需的结构/功能信息，仅针对抗焦虑α -2 GABA-A受体亚型的化合物的鉴定受到阻碍。本项目一期将应用Neurion的NP2技术和基于结构的药物设计平台，发现α -2 GABA-A受体的选择性变构调节剂。第一阶段的具体目标是：1。我们将生成关于苯二氮卓类药物（BZD）变构调节剂位点配体-受体相互作用的初始结构知识。我们将使用非自然氨基酸诱变和电生理分析来评估在α 2和α 3 GABAAR亚型BZD位点的六个“探针位置”上多达50个先前描述的结构不同的化合物的“调节面板”的结合相互作用。调节剂面板由已知的变构调节剂组成，它们与GABAAR的BZD位点结合，并表现出不同程度的功能亚型特异性。我们将构建GABAAR结合位点的计算模型来获取所有这些信息。因此，我们将通过实验验证α 2和α 3 GABAAR亚型药物受体相互作用的计算机模型。2. 我们将使用Specific Aim 1中开发的计算模型来计算筛选大约200万种商业上可用的化合物以及虚拟组合库，并选择购买或合成大约200种有前途的α 2 GABAAR亚型选择性分子。我们将电生理分析我们的200种化合物对α 1和α 2以及α 3和α 5 GABAAR亚型的效力/功效和选择性。3. 我们将从Specific Aim 2中选择的化合物中选择最有希望的大约30个先导化合物，并进行额外的一轮非自然诱变和建模，如Specific Aim 1中那样，以进一步完善我们的模型。4. 改进后的模型将用于设计和合成10种新型化合物。我们将检测新化合物对α、2、3、5种GABAAR亚型的效力/功效和选择性。

DESCRIPTION (provided by applicant): There continues to be a need for effective yet safe treatments for anxiety and related disorders. Current treatments such as the benzodiazepines which target many of the gamma-aminobutyric acid receptor (GABA-A receptor) subtypes are effective, but patients suffer from a number of unwanted side-effects. The identification of compounds targeting only the anxiolytic alpha-2 GABA-A receptor subtype has been hampered by a lack of structure/function information needed for effective lead discovery and optimization. Phase I of this project will apply Neurion's NP2 technology and structure-based drug design platform to discover selective allosteric modulators for the alpha-2 GABA-A receptor. The phase I specific aims are: 1. We will generate initial structural knowledge about ligand-receptor interactions at the benzodiazepine (BZD) allosteric modulator site. We will use unnatural amino acid mutagenesis and electrophysiological assays to evaluate the binding interactions of a "modulator panel" of up to 50 previously described, structurally diverse compounds at six "probe positions" within the BZD site of both the alpha 2 & alpha 3 GABAAR subtypes. The modulator panel consists of known allosteric modulators that bind to the BZD site of the GABAAR and display varying degrees of functional subtype specificity. We will construct computational models of the GABAAR binding site that capture all this information. We will thus have experimentally validated computer models of drug-receptor interactions at both the alpha 2 & alpha 3 GABAAR subtypes. 2. We will use the computational models developed in Specific Aim 1 to computationally screen approximately 2 million commercially available compounds in addition to virtual combinatorial libraries and to select for purchase or to synthesize approximately 200 promising alpha 2 GABAAR subtype-selective molecules. We will electrophysiologically assay our 200 compounds for potency/efficacy and selectivity against alpha 1 and alpha 2 as well as alpha 3 and alpha 5 GABAAR subtypes. 3. We will choose the most promising approximately 30 leads from the compounds selected in Specific Aim 2, and conduct an additional round of unnatural mutagenesis and modeling as in Specific Aim 1 to further refine our models. 4. The refined model will be used to design and synthesize 10 novel compounds. We will assay the new compounds for potency/efficacy and selectivity against alphal, 2, 3, 5 GABAAR subtypes.