CORE--Drug, Discovery, Design and Synthesis

3.核心单元B：药物发现、设计与合成（Georg） a.目的： 非激素男性避孕药的发现和开发尚处于起步阶段。然而，在这方面， 关于男性生殖系统的基础生物学知识已经爆炸性地增长， 提供启动药物发现和开发计划的机会。避孕药必须 高效、可逆、口服生物可利用或适于阴道内施用，并且安全。 为了应对这一机遇和挑战，我们组建了一个核心设施（药物发现，设计和 综合核心，DOS），将支持这个跨学科的U 54中心的专业知识，发现，设计， 合成和开发男性避孕药。以一种与这个中心的私家侦探高度合作的方式， 它的药物开发核心，和NICHD，我们将优化命中和铅化合物的效力，选择性， 和药学性质为临床试验做准备。 该中心的长期目标是开发非激素男性避孕药。为 本申请我们选择了几个对精子发生重要的药物发现靶点， 精子形成、运动和获能。靶点是分子伴侣Hsp 90 p，真核细胞延伸 因子1 oc（eEF 1A）、Na，K-ATP酶a4、Doublexex和Mab-3相关转录因子-1（Dmrt 1）和精子 蛋白酪氨酸激酶。此外，预计U 54中心未来的初级研究员将在 其他靶蛋白。核心的中心研究假设是， 的目标可以被发现，优化，并在体外和体内进行研究，最终目标是 一种或多种药物用于临床试验。 调查人员已经做好了充分的准备来承担这个项目，因为他们有研究的跟踪记录 在生殖生物学，药物发现，药物开发，以及在发现方面的成功合作历史 和男性避孕药的开发。药物发现的研究环境也是 优秀，并提供独特的设施，不经常发现在大学，如高通量筛选 实验室（KU），组合化学实验室（UMN），大规模合成实验室（UMN）， 一个治疗药物发现和开发办公室（KU），以及一个通用的药品生产质量管理程序 （c-GMP）合成设施（UMN）。 目的1：通过高通量筛选和筛选来鉴定精子特异性靶点的抑制剂 目标图书馆。与Pis项目合作，我们将开发适用于高浓度的 通量筛选超过100，000种化合物的化合物库和/或更小的靶向库将被 筛选以鉴定靶蛋白的小分子抑制剂。 目的2：通过蛋白质X射线晶体学确定蛋白质-抑制剂相互作用。靶蛋白 将与铅化合物共结晶，并确定蛋白质-命中复合物的结构 在原子分辨率。结构数据将用于基于结构的药物设计以优化命中 化合物的效力和选择性以及计算机筛选。 目标3：优化效价和选择性的筛选命中。小分子将优化效力 和选择性，采用基于结构的药物设计等合理设计药物化学原理。 这些研究将与Pis项目和药物开发项目密切合作进行。 核心化合物将被制备、测试，然后进一步优化，目标是生成个位数的化合物。 纳摩尔抑制剂具有约1000倍的选择性和>100的水溶性 目标4：开发用于临床前和临床评价的先导化合物。联同 药物开发核心和NICHD，我们将优化先导化合物的药物特性， 准备临床试验。我们还将为动物研究提供大量的先导化合物。 我们的方法对非激素男性避孕药的发现和发展是创新的 因为我们已经确定了几个非常有希望的靶点，这些靶点还没有被探索用于药物发现， 以前我们正在为这些项目带来最先进的药物发现方法， 期待与其他U 54中心合作，并为他们提供药物发现和开发资源 和专业知识，这在学术机构中是很少见的。 在为期五年的中心资助结束时，我们将为每个目标确定小分子抑制剂， 我们将有与靶蛋白共结晶的小分子抑制剂， 关于蛋白质-抑制剂相互作用的信息，用于药物设计。筛选结果将被优化， 效力、选择性和药物性质，我们期望至少一种优化的先导化合物 将被选中进行临床试验，最终将用于男性避孕。

3. Core Unit B: Drug Discovery, Design & Synthesis (Georg) a. Objective: The discovery and development of non-hormonal male contraceptive agents is in its infancy. However, knowledge about the basic biology of the male reproductive system has exploded and extensive biological insights provide the opportunity to initiate drug discovery and development programs. Contraceptive drugs must be highly effective, reversible, orally bioavailable or suitable for intra-vaginal application, and safe. In response to this opportunity and challenge we have assembled a core facility (Drug Discovery, Design & Synthesis Core, DOS) that will support this interdisciplinary U54 center with the expertise to discover, design, synthesize, and develop male contraceptive agents. In a highly collaborative manner with the Pis of this center, its drug development core, and the NICHD we will optimize hit and lead compounds for potency, selectivity, and pharmaceutical properties in preparation for clinical trails. The long-term goal of this center is the development of non-hormonal male contraceptive agents. For the current application we have selected several targets for drug discovery that are important for spermatogenesis, spermiation, motility and capacitation. The targets are the molecular chaperone Hsp90p, eukaryotic elongation factor 1oc (eEF1A), Na,K-ATPase a4, Doublesex and Mab-3 related transcription factor-1 (Dmrtl), and sperm protein tyrosine kinases. In addition, it is anticipated that future junior investigators of the U54-center will work on other target proteins. The central research hypothesis for the core is that small molecule inhibitors of each of the targets can be discovered, optimized, and investigated in vitro and in vivo with the ultimate goal to bring one or more agents towards clinical trials. The investigators are well prepared to undertake this project because they have a track record of research in reproductive biology, drug discovery, drug development, and a successful history of collaborating on the discovery and development of male contraceptive agents. The research environment for drug discovery is also excellent and provides unique facilities that are not often found at universities such as high throughput screening laboratories (KU), a combinatorial chemistry laboratory (UMN), a large-scale synthesis laboratory (UMN), an Office for Therapeutics Discovery and Development (KU), and a common Good Manufacturing Procedures (c-GMP) synthesis facility (UMN). Objective 1 : Identify inhibitors for sperm-specific targets by high throughput screening and by screening of targeted libraries. In collaboration with the project Pis, we will develop assays that are suitable for high throughput screening. A compound library of over 100,000 compounds and/or smaller targeted libraries will be screened to identify small molecule inhibitors of the target proteins. Objective 2: Determine protein-inhibitor interactions by protein X-ray crystallography. The target proteins will be co-crystallized with lead compounds and the structure of the protein-hit complex(es) will be determined at atomic resolution. The structural data will be used for structure-based drug design to optimize hit compounds for potency and selectivity and for in silico screening. Objective 3: Optimize screening hits for potency and selectivity. Small molecules will be optimized for potency and selectivity, using structure- based drug design and other rational design medicinal chemistry principles. These studies will be carried out in close collaboration with the project Pis and the Drug Development Core. Compounds will be prepared, tested and then further optimized with the goal to generate single digit nanomolar inhibitors that posses about 1000-fold selectivity and aqueous solubility of >100 Objective 4: Develop lead compounds for preclinical and clinical evaluation. In collaboration with the Drug Development Core and the NICHD we will optimize the pharmaceutical properties of lead compounds in preparation for clinical trails. We will also provide large-scale amounts of lead compounds for animal studies. Our approach toward the discovery and development of non-hormonal male contraceptive agents is innovative because we have identified several highly promising targets that have not been explored for drug discovery before. We are bringing state-of-the-art drug discovery methods to bear on these projects and are looking forward to working with the other U54 centers and providing them with drug discovery and development resources and expertise that are rarely available at academic institutions. At the end of the five-year center grant we will have identified small molecule inhibitors for each of the targets, we will have co-crystallized small molecule inhibitors with the target proteins and will have used structural information about protein-inhibitor interactions for drug design. The screening hits will have been optimized for potency, selectivity, and pharmaceutical properties and we expect that at least one optimized lead compound will have been selected for clinical trials and will ultimately be commercialized for male contraception.