17beta-Hydroxysteroid Dehydrogenase Type 14: Development of potent and selective inhibitors using crystallization-aided optimization and enzyme characterization

17β-羟基类固醇脱氢酶 14 型：利用结晶辅助优化和酶表征开发有效的选择性抑制剂

• 批准号: 233241443
• 负责人: Professor Dr. Gerhard Klebe
• 金额: --
• 依托单位: Institut für Pharmazeutische Chemie
• 依托单位国家: 德国
• 项目类别: Research Grants
• 财政年份: 2013
• 资助国家: 德国
• 起止时间: 2012-12-31 至 2016-12-31
• 项目状态: 已结题
• 来源: https://gepris.dfg.de/gepris/projekt/233241443?language=en
• 关键词: 17beta; Hydroxysteroid; Dehydrogenase; Type; 14

17beta-Hydroxysteroid dehydrogenase type 14 (17beta-HSD14) is a recently characterized enzyme which is located in the brain, liver and placenta. The apoenzyme has been crystallized. It is able to convert both estradiol and dehydroepiandrosterone into estrone and 5-androstene-3beta, 17beta-diol, respectively but its functional role remains unclear. It has been suggested to be implicated in neuromodulation and in the pathogenesis of neurodegeneration. Potent and selective inhibitors are useful tools to study the role of an enzyme in a disease-oriented model but no inhibitor of this enzyme has been reported to date. In this project, we want to develop highly potent and selective inhibitors of 17beta-HSD14, to provide efficient tool compounds to investigate if this enzyme might be a good drug-target in neuronal diseases, to further structurally characterize the enzyme's active site and gain further understanding into the possible functional role of this enzyme. Starting from reasonably active 17beta-HSD14 inhibitors (identified in a preliminary study or by virtual screening), structural optimization will be performed using crystallography, modeling, chemical synthesis and biological evaluation in an iterative process. Analysis of the crystallized inhibitor-enzyme complex will provide information on the interactions achieved by the ligand with the enzyme and will structurally characterize the active site of the protein. In addition the 3D-structure will suggest other interactions between the enzyme and the inhibitor after structural modification of the latter. Modeling will validate the designed structures. After synthesis, 17beta-HSD14 potency will be evaluated in a NAD/NADH fluorescent-assay, which will be established in house and the potent compounds will enter a new cycle of optimization process. Selectivity toward functionally related enzymes like 17beta-HSD1, 2, 4 and 5 will be addressed using a structure-based or a ligand-based approach. Furthermore, screening for 17beta-HSD14 nonsteroidal substrates might provide an insight into a possible functional role of this enzyme.

17β-羟基类固醇脱氢酶 14 型 (17β-HSD14) 是一种最近鉴定的酶，位于大脑、肝脏和胎盘中。脱辅酶已结晶。它能够将雌二醇和脱氢表雄酮分别转化为雌酮和5-雄烯-3β、17β-二醇，但其功能作用仍不清楚。有人认为它与神经调节和神经变性的发病机制有关。有效的选择性抑制剂是研究酶在疾病模型中的作用的有用工具，但迄今为止尚未报道这种酶的抑制剂。在这个项目中，我们希望开发高效、选择性的17β-HSD14抑制剂，为研究这种酶是否可能成为神经元疾病的良好药物靶点提供有效的工具化合物，进一步从结构上表征该酶的活性位点，并进一步了解该酶可能的功能作用。从具有相当活性的 17β-HSD14 抑制剂（在初步研究或通过虚拟筛选中鉴定）开始，将在迭代过程中使用晶体学、建模、化学合成和生物学评估进行结构优化。对结晶抑制剂-酶复合物的分析将提供配体与酶相互作用的信息，并在结构上表征蛋白质的活性位点。此外，3D 结构将表明在后者的结构修饰后酶和抑制剂之间存在其他相互作用。建模将验证设计的结构。合成后，17beta-HSD14 的效力将在 NAD/NADH 荧光测定中进行评估，该测定将在内部建立，有效的化合物将进入新的优化过程循环。将使用基于结构或基于配体的方法来解决对功能相关酶（如 17beta-HSD1、2、4 和 5）的选择性。此外，筛选 17beta-HSD14 非甾体底物可能有助于深入了解该酶的可能功能作用。