Rational Design of Novel Estrogen Receptor Antagonists

描述（由申请人提供）：本提案的主要目的是开发和优化新型纯雌激素受体（ER）亚型α拮抗剂。尽管在该领域做出了重大努力，但对开发选择性ER拮抗剂的需求仍未得到满足。我们最初提交的一些研究目标已经完成，并为我们的修改方案奠定了坚实的基础。我们的建议涉及多学科的方法，结合有机合成，分子生物学，X射线晶体学和分子建模的专业知识，合理设计和优化的配体是纯拮抗剂的雌激素受体。 我们的具体目标包括：（1）选择性ER α拮抗剂的合成：将合成新提出的分子用于先导优化、增强效力和在受体水平上结合亲和力的选择性。这样一个全面的合成程序应该显着增加我们对新的四氢异喹啉支架的构效关系的理解;（2）雌激素拮抗剂的X射线晶体学分析：与F。Rastinejad（UVA），结合雌激素受体的最有效配体的共晶体的X射线分析将用于探索配体结合和鉴定纯拮抗剂活性所需的结构特征;（3）计算机研究以增强和指导先导物优化：分子模拟方法将用于开发新的药物设计策略，因为结果将展开并定量构效关系（SAR）;（4）筛选雌激素受体拮抗剂和雌激素受体选择性的配体的体外和全细胞测定：将使用各种测定，例如化学发光、基于荧光的竞争性结合、瞬时转染报告子、酵母双杂交和细胞增殖测定来评估所提出的化合物的激动剂/拮抗剂活性和细胞毒性。 该提议的长期目标是产生和优化作为“纯”α-受体拮抗剂的抗雌激素。

DESCRIPTION (provided by applicant): The main objective of this proposal is to develop and optimize novel and pure estrogen-receptor (ER) subtype alpha antagonists. Despite significant efforts in this area, there is an unmet need for developing selective ER antagonists. Some of our originally submitted research goals have been accomplished and lay a solid foundation for our revised proposal. Our proposal involves a multidisciplinary approach that combines expertise in organic synthesis, molecular biology, X-ray crystallography, and molecular modeling to rationally design and optimize ligands that are pure antagonists of estrogen receptors. Our specific aims include: (1) Synthesis of selective ER alpha antagonists: The new proposed molecules will be synthesized for lead optimization, enhancement of potency and selectivity of binding affinity at the receptor level. Such a comprehensive synthesis program should significantly increase our understanding for structure-activity relationships of the new tetrahydroisoquinoline scaffold; (2) X-ray crystallographic analysis of estrogen antagonists: In collaboration with Dr. F. Rastinejad (UVA), x-ray analyses of co-crystals of most potent ligands bound to the estrogen receptor will be used to explore ligand binding and to identify structural features necessary for pure antagonist activity; (3) Computational studies to enhance and guide lead optimization: Molecular modeling methods will be used to develop new drug design strategies as the results unfold and quantitate structure activity relationships (SAR); (4) In vitro and whole cell assays to screen ligands for estrogen receptor antagonism and estrogen receptor selectivity: Various assays such as a chemiluminescence, fluorescence-based competitive binding, a transient transfection reporter, a yeast two hybrid and a cell proliferation assay will be used to evaluate the agonist/antagonist activity and cytotoxicity of proposed compounds. A long-range goal of this proposal is to generate and optimize antiestrogens that are "pure" alpha-receptor antagonists.