CORE--Drug, Discovery, Design and Synthesis

3.核心单位B：药物发现、设计和合成(Georg) A.目标： 非荷尔蒙男性避孕药的发现和发展还处于初级阶段。然而， 关于男性生殖系统基本生物学的知识已经爆炸式增长，并具有广泛的生物学洞察力 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、口服生物利用度或适合阴道内应用，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U54中心，拥有发现、设计、 合成、开发男性避孕药。以与该中心的PIS高度合作的方式， 它的药物开发核心，以及NICHD我们将优化HIT和先导化合物的效力，选择性， 和药物特性，为临床试验做准备。 该中心的长期目标是开发非荷尔蒙男性避孕药。对于 目前的应用我们已经为药物发现选择了几个对精子发生重要的靶点， 受精、运动和获能。靶标是分子伴侣Hsp90p，真核延长 1oC因子(EEF1a)、Na，K-ATPase A4、双性和单抗相关转录因子-1(Dmrt1)和精子 蛋白酪氨酸激酶。此外，预计未来U54中心的初级调查人员将工作 在其他目标蛋白上。对核心的中心研究假设是，每个核心的小分子抑制剂 的目标可以在体外和体内被发现、优化和研究，最终目标是带来 一种或多种药物用于临床试验。 调查人员已经做好了承担这一项目的准备，因为他们有研究的记录 在生殖生物学、药物发现、药物开发以及在这一发现上合作的成功历史 以及男性避孕药的开发。药物发现的研究环境也是 非常好，并提供了大学中不常见的独特设施，如高通量筛查 实验室(KU)、组合化学实验室(UMN)、大型合成实验室(UMN)、 治疗药物发现和开发办公室(KU)和共同产品制造程序 (C-GMP)合成设施(UMN)。 目标1：通过高通量筛选和筛选确定精子特异性靶点的抑制物 目标库。与PIS项目合作，我们将开发适合于HIGH的分析方法 吞吐量筛选。一个包含100,000多个化合物的化合物库和/或更小的目标库将是 筛选鉴定靶蛋白的小分子抑制物。 目的：用蛋白质X-射线结晶学方法确定蛋白质与抑制物的相互作用。靶蛋白 将与铅化合物共结晶，并将确定蛋白质-HIT复合体的结构 在原子分辨率下。结构数据将用于基于结构的药物设计，以优化HIT 化合物的效力和选择性以及用于电子筛选。 目标3：优化筛选HITS的效力和选择性。小分子的效力将得到优化 并有选择性，运用基于结构的药物设计等合理设计药物化学的原则。 这些研究将与PIS项目和药物开发密切合作进行 核心。将准备、测试化合物，然后进一步优化，目标是产生个位数 具有约1000倍选择性和&gt；100水溶解性的纳米分子缓蚀剂 目的4：开发用于临床前和临床评价的先导化合物。与 药物开发核心和NICHD我们将优化先导化合物的药学性能 为临床试验做准备。我们还将为动物研究提供大规模的先导化合物。 我们发现和开发非荷尔蒙男性避孕药的方法是创新的 因为我们已经确定了几个非常有希望的目标，这些目标还没有被探索用于药物发现 在此之前。我们正在将最先进的药物发现方法应用于这些项目，并正在寻找 期待与其他U54中心合作，为它们提供药物发现和开发资源 以及学术机构难得的专业知识。 在五年中心拨款结束时，我们将为每个靶点确定小分子抑制剂， 我们将有与目标蛋白共结晶的小分子抑制剂，并将使用结构 药物设计中蛋白质-抑制物相互作用的信息。点映点击率将经过优化， 效力、选择性和药物性质，我们预计至少有一种优化的先导化合物 将被选中进行临床试验，并最终将被商业化用于男性避孕。

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.