Development of Eppin as a Male Contraceptive

描述（由申请人提供）： 这项研究计划的长期目标是将eppin发展为男性避孕的靶点。我们一直在研究人精子表面eppin与精液凝固蛋白的相互作用，特别是关于射精后精子活力的恢复。自从我们在2004年的《科学》杂志上发表了关于用eppin高滴度免疫的雄性猴子的完全和可逆避孕的文章以来，我们一直在研究eppin作为一个可药物化的靶点。本申请提供了开发抑制eppin精液凝固蛋白结合的可药用化合物所需的详细步骤。当eppin精凝素结合被阻断时，精子活力被有效抑制。该项目将与NCCU BRITE中心的药物发现研究计划结合。具体目标#1是通过定向文库筛选（命中生成）鉴定eppin-精液凝固蛋白结合的新型、有效和高度特异性抑制剂。基于我们对化合物A4的初步数据，该目的将检验以下假设：可以鉴定eppin-精液凝固蛋白结合的更有效和特异性的抑制剂化合物。为了确定这一特定目标的抑制剂，将建立3个开发单位来协调BRITE中心的研究流程：（1）检测开发单位，（2）检测实施单位，（3）筛选和HTS单位。具体目标#2是通过药物化学和分子建模（命中-先导优化）对具体目标#1中确定的初始命中进行迭代改进，以提高效价、选择性和细胞渗透性并降低细胞毒性。具体目标#2将设立两个工作单位：（1）化学单位和（2）化学信息学单位。具体目标#3将更好地定义精液凝固蛋白和具体目标#1和#2中发现的化合物在eppin上的靶位点。使用eppin-精液凝固蛋白体外测定，重组eppin蛋白片段和具有突变的关键氨基酸残基的eppin将用于更好地定义精液凝固蛋白（或化合物，例如，A4）eppin上的结合位点。这个特定的目标将测试的假设，确切地定义哪些eppin氨基酸是必不可少的精液凝固蛋白和/或目标化合物的结合将使我们能够细化的化合物的特异性。 公众相关性：该研究项目旨在开发一种新的非甾体男性避孕药，使男性比避孕套或输精管结扎术有更多的选择。广泛提供男性避孕药具将加强全世界的计划生育。

DESCRIPTION (provided by applicant): The long-term goals of this research project are to develop eppin as a male contraceptive target. We have been studying the interaction of eppin with semenogelin on the human sperm surface, particularly with regard to the resumption of sperm motility following ejaculation. Since our publication in Science in 2004 on the complete and reversible contraception of male monkeys immunized to a high titer with eppin, we have been investigating eppin as a drugable target. This application presents the detailed steps necessary for developing a drugable compound that inhibits eppin semenogelin binding. When eppin semenogelin binding is blocked, sperm motility is effectively inhibited. This project will be in conjunction (subcontract) with the drug discovery research program at the BRITE center, NCCU. Specific aim #1 is the identification of novel, potent and highly specific inhibitors of eppin-semenogelin binding through directed library screening (hit generation). Based on our preliminary data on compound A4, this aim will test the hypothesis that more potent and specific inhibitor compounds of eppin-semenogelin binding can be identified. To identify inhibitors in this specific aim, 3 development units will be established to coordinate the research flow at the BRITE Center: (1) Assay Development Unit, (2) Assay Implementation Unit (3) Screening and HTS Unit. Specific aim #2 is the iterative improvement of initial hits identified in Specific Aim #1 with medicinal chemistry and molecular modeling (hit-to-lead optimization) to improve the potency, selectivity and cell permeability and to decrease the cell toxicity. Specific aim #2 will establish 2 working units: (1) The Chemistry Unit and (2) The Cheminformatics Unit. Specific aim #3 will better define the target site on eppin for semenogelin and compounds discovered in specific aims #1 and #2. Using the eppin-semenogelin in vitro assays, recombinant eppin protein fragments and eppin with mutated key amino acid residues will be used to better define the semenogelin (or compound, e.g., A4) binding site on eppin. This specific aim will test the hypothesis that defining exactly which eppin amino acids are essential for semenogelin and/or a target compound binding will enable us to refine the specificity of the compound. PUBLIC RELEVANCE: This research project seeks to develop a new non-steroidal male contraceptive that will allow males greater choices than condoms or vasectomy. Wide spread availability of male contraceptives will enhance family planning throughout the world.