DESIGN AND SYNTHESIS OF INHIBITORS FOR GLYCOENZYMES

糖酶抑制剂的设计与合成

基本信息

批准号：
6285064
负责人：
YOSHITAKA ICHIKAWA
金额：
$ 26.4万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
1996
资助国家：
美国
起止时间：
1996-03-01 至 2002-01-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6285064
关键词：
N acetylglucosamine N acetylglucosaminidase N glycosidase X ray crystallography carbohydrate metabolism chemical synthesis enzyme inhibitors enzyme mechanism glycosylation

项目摘要

DESCRIPTION: (Principal Investigator's Abstract) The long-term goal of this research is to design, synthesize and evaluate specific inhibitors of enzymes involved in carbohydrate metabolism, as a means of elucidating the roles played by these enzymes in biologically and clinically significant processes. Data from this laboratory has already demonstrated that l-N-iminosugars are highly potent and specific inhibitors of beta-glycosidases and that its conjugate with a UDP analog is a specific and potent inhibitor of alpha-galactosyltransferase. This competing continuation will focus on two biologically important carbohydrate metabolic processes: Aim 1. To study the biological role of O-GlcNAc. 0-Linked beta-N-acetylglucosamine (0-GlcNAc) is found in many proteins, and its on-and-off glycosylation is thought to regulate the functions of these proteins. Since glycosylation and phosphorylation often (occur at the same sites in a protein, use of inhibitors of the enzyme that regulate the glycosylation offer a viable alternative to site-directed mutagenesis as an approach to analyzing the role of glycosylation. The biological role of 0-GlcNAc will be elucidated by designing and using specific inhibitors to characterize 0-GIcNAc-specific N-acetylglucosaminidase (0-GlcNAc'ase), the enzyme which removes 0-GlcNAc from proteins. The proposed inhibitors include nagastatin analogs and 1-thio-GIcNAc (S-GlcNAc) derivatives and their peptide-conjugates; one potent and specific inhibitor, a S-GlcNAc derivative has already been synthesized. The most promising inhibitors will be used in several biological systems, including cancer cells, in which 0-GlcNAc is thought to play a key role. Aim 2. To study the reaction mechanism of DNA N glycosylases. DNA base-excision enzymes (N-glycosylases) play a central role in maintaining genetic integrity; however, their reaction mechanisms are still undetermined. This laboratory has recently solved an x-ray crystal structure of the complex of 1-azaribose-containing DNA and one such glycosylase, 3-methyladenine DNA glycosylase (AlkA), and has identified a potential role for the conserved Asp residue in its catalytic process. As a continuation of this work, more specific molecular probes will be developed as a means of gaining additional insight into the mechanisms for the base flipping and N-glycosidic bond-cleaving reactions of other members of this class of enzymes. Given the potential specificity and potency of the proposed inhibitors, these reagents are expected to contribute not only to the study of the biological roles of such important carbohydrate-metabolizing enzymes but also to the development of inhibitor-based drugs with a wide range of clinical applications.

描述：（主要研究者的摘要）本研究的长期目标研究是设计、合成和评估特定的酶抑制剂参与碳水化合物代谢，作为阐明其作用的一种手段这些酶在具有生物学和临床意义的过程中发挥作用。数据该实验室已经证明 L-N-亚氨基糖具有高度 β-糖苷酶的有效和特异性抑制剂及其与 UDP 类似物是 α-半乳糖基转移酶的特异性且有效的抑制剂。这个竞争性的延续将重点关注两个具有重要生物学意义的问题碳水化合物代谢过程：目标 1. 研究碳水化合物的生物学作用 O-GlcNAc。 0-连接 β-N-乙酰氨基葡萄糖 (0-GlcNAc) 存在于许多蛋白质及其开关糖基化被认为可以调节功能这些蛋白质。由于糖基化和磷酸化经常（发生在蛋白质中的相同位点，使用调节酶的抑制剂糖基化为定点突变提供了一种可行的替代方法分析糖基化作用的方法。的生物学作用 0-GlcNAc 将通过设计和使用特定抑制剂来阐明表征 0-GlcNAc 特异性 N-乙酰氨基葡萄糖苷酶 (0-GlcNAc'ase)，从蛋白质中去除 0-GlcNAc 的酶。提议的抑制剂包括那加他汀类似物和1-硫代-GlcNAc (S-GlcNAc)衍生物及其肽缀合物；一种有效且特异性的抑制剂，S-GlcNAc 衍生物已经合成了。最有前途的抑制剂将用于多种生物系统，包括癌细胞，其中 0-GlcNAc 是思想发挥着关键作用。目的2.研究DNA N的反应机理糖基化酶。 DNA 碱基切除酶（N-糖基化酶）在保持遗传完整性；然而，它们的反应机制仍然是未确定。该实验室最近解决了 X 射线晶体结构含有1-氮杂核糖的DNA和一种这样的糖基化酶的复合物， 3-甲基腺嘌呤 DNA 糖基化酶 (AlkA)，并已确定其潜在作用其催化过程中保留的天冬氨酸残基。作为本次活动的延续工作中，将开发更具体的分子探针作为获得对碱基翻转和 N-糖苷机制的更多见解此类酶其他成员的键断裂反应。鉴于所提议的抑制剂、这些试剂的潜在特异性和效力预计不仅有助于研究其生物学作用如此重要的碳水化合物代谢酶，而且还促进了抑制剂类药物具有广泛的临床应用前景。