DESIGN, SYNTHESIS AND NMR CHARACTERIZATION OF FLUORINATED HIV PROTEASE INHIBITOR

这个项目涉及两个不同的工作：最初，我们与 在Burroughs Wellcome的Koszalka和Krenitzky博士并要求提供 核磁共振专业知识，以协助他们在药理学方面的工作 抗击艾滋病的代理人。这些讨论导致了一项合作努力 针对细菌嘌呤活性部位的表征 核苷磷酸化酶，并在阐明酶的基础 药物杜仲素(7-去氮腺苷)的抑制作用。一系列 研究是利用转移的核Overhauser效应设计的 (“TRNOE”)，以表征酶-抑制剂的相互作用。作为以下内容的一部分 这项研究工作，将TRNOE作为函数进行了广泛的建模 酶复合型和非络合型缓蚀剂的核磁共振参数 结合/解离动力学采用松弛矩阵法进行计算 专门为这个问题开发的程序。1因为它是 确定结果一般取决于配体(抑制物) 汇率，我们评估了现有的方法，并开发了几个 决定汇率的新方法。更多 具体而言，制定了涉及以下内容的方法：1)测量 用Carr-Purcell方法计算配体核的横向弛豫速率 Meiom-Gill(CPMG)实验作为脉搏频率的函数，以及2) 作为交换配体的函数的T_1的测量 自旋锁定场的强度2.与交换有关的信息 结节杀菌素与活性部位的结合，以及与PNP络合的结构 结节杀菌素是从这些研究中衍生出来的。 对于该酶在制备过程中的商业应用具有重要意义 核苷类药物。 最近，针对这一发展展开了第二项研究工作。 临床上有用的艾滋病毒蛋白水解酶抑制剂。HIV蛋白水解酶是一口井 已经确定了化疗药物的靶酶，但到目前为止，还没有 已经开发出了完全成功的蛋白酶抑制剂，但没有一种 已在临床上使用。作为起点，我们将使用环状尿素 结构最初是由林和杜邦-默克的柯克提出的。

This project has involved two separate efforts: Initially, we approached Drs. Koszalka and Krenitzky at Burroughs Wellcome and asked to provide NMR expertise to assist their work on the development of pharmacologic agents to combat AIDS. These discussions led to a collaborative effort aimed at the characterization of the active site of bacterial purine nucleoside phosphorylase, and at elucidating the basis for enzyme inhibition by the drug tubercidin (7-deazaadenosine). A series of studies was designed utilizing the transferred nuclear Overhauser Effect ("TRNOE") to characterize the enzyme-inhibitor interaction. As part of this research effort, extensive modeling of the TRNOE as a function of the NMR parameters of the enzyme-complexed and uncomplexed inhibitor and the binding/dissociation kinetics was performed, using relaxation matrix programs developed specifically for this problem.1 Since it was determined that the results depend in general on the ligand (inhibitor) exchange rate, we evaluated existing methodologies and developed several new approaches for the determination of the exchange rate. More specifically, approaches were developed involving: 1) measurements of the transverse relaxation rate for ligand nuclei using a Carr-Purcell- Meiboom-Gill (CPMG) experiment as a function of the pulse rate, and 2) measurement of T1 for the exchanging ligands as a function of the strength of the spin-lock field.2 Information related to the exchange of tubercidin with the active site, and to the structure of PNP-complexed tubercidin was derived from these studies.3 This information is important for the commercial use of this enzyme in the preparation of nucleoside drugs. A second research effort was recently initiated aimed at the development of clinically useful HIV protease inhibitors. HIV protease is a well established target enzyme for chemotherapeutic agents, but to date, no completely successful protease inhibitors have been developed and none is in clinical use. As a starting point, we will use the cyclic urea structure initially proposed by Lam and cowokers at DuPont-Merck.