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，广泛的生物学见解为启动药物发现和开发计划提供了机会。避孕药必须高效、可逆、口服生物利用度或适合阴道内使用，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、设计和合成核心，DDS)，为这个跨学科的U54中心提供支持，提供发现、设计、合成和开发男性避孕药的专业知识。在与该中心、其药物开发核心和NICHD的PI高度合作的方式下，我们将优化HIT和LEAD化合物的效力、选择性和药学特性，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，我们选择了几个对精子发生、精子受精、运动和获能重要的药物发现靶点。靶点是分子伴侣Hsp90J3、真核延长因子1a(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U54-中心未来的初级研究人员将致力于其他靶蛋白的研究。核心研究的中心假设是，可以在体外和体内发现、优化和研究每个靶点的小分子抑制剂，最终目标是将一种或多种药物引入临床试验。 研究人员已经为开展这一项目做好了充分准备，因为他们在生殖生物学、药物发现、药物开发方面的研究记录良好，并在男性避孕药的发现和开发方面有成功的合作历史。药物发现的研究环境也很好，提供了大学中不常见的独特设施，如高通量筛选实验室(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)和共同良好制造程序(c-GMP)合成设施(UMN)。 目的1：通过高通量筛选和靶向文库筛选，寻找精子特异性靶点的抑制物。我们将与PIS项目合作，开发适合高通量筛查的检测方法。将筛选包含100,000多种化合物的化合物文库和/或更小的靶向文库，以确定目标蛋白的小分子抑制剂。 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。目标蛋白质将与铅化合物共结晶，蛋白质-HIT复合体(ES)的结构将在原子分辨率下确定。这些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，并用于电子筛选。 目标3：优化筛选HITS的效力和选择性。利用基于结构的药物设计和其他合理设计的药物化学原则，小分子将在效力和选择性方面进行优化。这些研究将与PIS项目和药物开发核心密切合作进行。化合物将被制备、测试，然后进一步优化，目标是产生个位数的纳米分子抑制剂，其选择性约为1000倍，在水中的溶解度为&gt；100µg/ml。 目的4：开发用于临床前和临床评价的先导化合物。我们将与药物开发核心和NICHD合作，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靶点，这些靶点以前从未被探索过用于药物发现。我们正在将最先进的药物发现方法应用于这些项目，并期待着与其他U54中心合作，为他们提供学术机构很少能获得的药物发现和开发资源和专业知识。 在五年中心拨款结束时，我们将为每个靶标确定小分子抑制剂，我们将与目标蛋白质共结晶小分子抑制剂，并将蛋白质-抑制剂相互作用的结构信息用于药物设计。筛选结果将在效力、选择性和药学性能方面进行优化，我们预计至少有一种优化的先导化合物将被选择用于临床试验，并最终将用于男性避孕的商业化。

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, DDS) 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 Hsp90J3, eukaryotic elongation factor 1 a (eEF1A), Na,K-ATPase a4, Doublesex and Mab-3 related transcription factor-1 (Dmrt1), 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 ¿mu¿g/ml. 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.