HOST GUEST METALLOPROTEIN MIMICS

这项研究项目的最终目标是开发 可以通过抑制锌来治疗的疾病的治疗剂 水解酶，包括类风湿和骨关节炎， 牙周炎、青光眼和癌症。这些代理的设计需要 了解(1)酶的作用机理和(2)能量 酶与抑制物之间的结合作用。催化剂 机理和缓蚀剂结合能是许多不同因素的总和 力量。很难衡量个人类型的贡献。 酶的大小和复杂性决定了其相互作用的可能性。这项建议 重点介绍了两种研究最多的锌水解酶， 碳水解酶和羧基肽酶A被认为是许多其他的 锌酶具有相似的结构和作用机制。 我们选择了具体解决酶激活的方法 与活性中心的锌配位的水。碳素的研究 脱水酶表明疏水相互作用、氢键和 锌离子或水附近存在的羧酸盐会影响酶 活性和抑制物结合，但大小和详细 这种影响的物理基础尚不清楚。这项建议旨在 量化影响并确定所述交互作用的方式和原因 做手术吧。在羧基肽酶A中，总的机制途径仍然是 这是一个有争议的问题，更不用说路径的细节了。的确有 目前还没有与锌形成络合物并具有配体和 与CPA中发现的几何形状非常相似的几何图形。这份提案包含 发展这样一种配体的计划。约束性研究将用于确定 无二次离子时缓蚀剂与锌的配位能 酶活性部位的相互作用。一旦这个目标实现了， 将准备一些配基，量化一些 已知的活动站点辅助交互。拟议的行动机制 羧基肽酶A将在精心设计的模型复合体中进行测试。 用来实现这项研究目标的工具包括分子设计 和模型配位络合物的合成。这些综合体将是 用X射线结晶学、电位法、量热法、 动力学研究，以及其他技术。

The ultimate goal of this research project is the development of therapeutic agents for diseases that can be treated by inhibiting zinc hydrolytic enzymes, including rheumatoid and osteoarthritis, periodontitis, glaucoma, and cancer. The design of these agents requires a knowledge of (1) the mechanism of the enzyme and (2) the energies of binding interactions between the enzyme and an inhibitor. The catalytic mechanisms and inhibitor binding energies are the sum of many varied forces. It is difficult to measure the contribution of an individual type of interaction in an enzyme due to its size and complexity. This proposal focuses on two of the most highly studied zinc hydrolytic enzymes, carbonic anhydrase and carboxypeptidase A. It is believed that many other zinc enzymes are similar in structure and utilize similar mechanisms. We have chosen to address specifically the means by which enzymes activate water that is coordinated to zinc in the active site. Studies of carbonic anhydrase suggest that hydrophobic interactions, hydrogen bonds, and the presence of carboxylates near the zinc ion or the water affect enzyme activity and inhibitor binding, but the magnitude and the detailed physical basis for the effects are unknown. This proposal seeks to quantify the effects and establish how and why the stated interactions operate. In carboxypeptidase A, the overall mechanistic path is still a matter of controversy, to say nothing of the details of the path. There is presently no ligand that forms a complex with zinc and has ligands and geometry which closely resemble those found in CPA. This proposal contains a plan to develop such a ligand. Binding studies will be used to determine the energy of inhibitor coordination to zinc in the absence of secondary interactions in the enzyme active site. Once this goal has been achieved, ligands will be prepared which quantify the contribution of some of the known active site secondary interactions. Proposed mechanisms of action of carboxypeptidase A will be tested in carefully designed model complexes. The tools used to achieve the goals of this study include molecular design and synthesis of model coordination complexes. The complexes will be characterized by x-ray crystallography, potentiometry, calorimetry, kinetic studies, and other techniques.