Inhibition of Prenylated Protein Processing

异戊二烯化蛋白质加工的抑制

基本信息

批准号：
7336786
负责人：
RICHARD A GIBBS
金额：
$ 26.08万
依托单位：
PURDUE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2007
资助国家：
美国
起止时间：
2007-01-01 至 2009-12-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/7336786
关键词：
Agar Amides Amines Amino Acids Anchorage-Independent Growth Animals Antineoplastic Agents Benchmarking Biological Biological Assay Biological Models Biological Process C-terminal Carboxylic Acids Cell model Cells Chemicals Clinical Trials Collaborations Cysteine Development Diffusion Drug Delivery Systems Ductal Carcinoma Cell Endoplasmic Reticulum Enzyme Inhibitor Drugs Enzyme Inhibitors Enzymes Evaluation Exhibits Family Farnesol Generations Goals Grant Growth Human In Vitro Interdisciplinary Study Intervention Jurkat Cells Laboratories Lead Libraries Ligand Binding Localized Location Malignant Neoplasms Mediating Membrane Methods Methylation Methyltransferase Modeling Modification Mus Numbers Oncogene Proteins Pancreas Pancreatic Ductal Carcinoma Pathway interactions Penetration Phase Plant Resins Principal Investigator Procedures Process Proteins Proteolysis Protocols documentation Publishing Reaction Research Proposals Route Scheme Screening procedure Signal Transduction Soft Agar Assay Solid Staging Sulfonamides Synthesis Chemistry System Time Validation Work Yeasts analog base carboxylate chemotherapeutic agent combinatorial cytotoxicity design farnesylation farnesyltranstransferase geranylgeranylation high throughput screening in vitro Assay in vivo inhibitor/antagonist interest isoprenoid isoprenylation isoprenylcysteine carboxylmethyltransferase member metaplastic cell transformation mutant novel pancreatic neoplasm prenyl prenylation programs protein farnesyltransferase ras Proteins rhoB p20 GDI scale up small molecule tool tumor tumor growth vapor

项目摘要

Members of the Ras family, implicated in many human cancers, are modified post-translationally, targeting them to the appropriate intracellular location. Ras and other -CaaX proteins undergo three sequential reactions: isoprenylation of the cysteine, in particular farnesylation by farnesyltransferase (FTase); proteolysis of the three terminal amino acids (-aaX); and a-carboxyl methylation of the isoprenylated cysteine. This process is crucial for membrane localization and thus activity of the key Ras oncoproteins. FTase inhibitors are being evaluated in clinical trials as cancer chemotherapeutic agents. Unfortunately, these compounds have surprisingly little effect on many Ras-transformed tumors. There is now growing interest in the subsequent enzymatic steps; proteolysis by Ras converting enzyme (Reel) and a-carboxyl methylation by isoprenylcysteine methyltransferase (Icmt) as alternative targets for the inhibition of Ras protein action. Carboxyl methylation is critical for the proper localization of Ras proteins in yeast and mouse cells. Given this important finding, we believe that Icmt represents an excellent target for chemotherapeutic intervention. We hypothesize that certain readily synthesized, isoprenoid-based inhibitors of Icmt will be valuable lead compounds for the development of anti-Ras cancer chemotherapeutic agents. Recent published and unpublished studies from our laboratories have demonstrated that modifications to both the amino modification and the isoprenoid unit ofprenylcysteines can afford small-molecule inhibitors of Icmt. The specific aims of this collaborative, interdisciplinary research proposal are as follows: Aim 1) Novel farnesol analogs will be elaborated to the corresponding prenylcysteine derivatives, via a solid-phase synthetic protocol. These will then undergo further chemical modification to provide libraries of prenylcysteine derivatives. Aim 2) These prenylcysteine derivatives will be assayed as potential inhibitors of Icmt, using high-throughput in vitro assay systems and well-characterized and robust single point assay procedures for Icmt activity. Aim 3) The prenylcysteine analogues will be evaluated for their ability to block prenylcysteine methylation in vivo. Potent cell permeable inhibitors will be evaluated for their ability to mislocalize Ras, interfere with Ras- mediated signaling, block anchorage-independent growth of pancreatic ductal carcinoma, and block tumor growth in vivo.

RAS家族的成员与许多人类癌症有关，在翻译后进行了修改，将其针对适当的细胞内位置。 RAS和其他-CAAX蛋白会经历三个顺序反应：半胱氨酸的异源性化，特别是Farnesyltransferase（FTase）的Farnesylation；三个的蛋白水解末端氨基酸（-aax）;以及异丙肾上腺素的半胱氨酸的A-羧基甲基化。这个过程是对于膜定位至关重要，因此是关键Ras癌蛋白的活性。 Ftase抑制剂正在在临床试验中评估为癌症化学治疗剂。不幸的是，这些化合物具有令人惊讶的是，对许多RAS转换肿瘤的影响很小。现在对随后的兴趣越来越多酶促步骤；通过RAS转化酶（Reel）和A-羧基甲基化的蛋白水解，异位甲基半胱氨酸甲基转移酶（ICMT）作为抑制Ras蛋白作用的替代靶标。羧基甲基化是对于在酵母和小鼠细胞中正确定位RAS蛋白的至关重要。考虑到这一重要发现，我们相信ICMT代表了化学治疗干预的绝佳目标。我们假设这一点某些很容易合成的基于异丙的ICMT抑制剂将是有价值的铅化合物为了开发抗RAS癌化学治疗剂。最近出版和未出版我们实验室的研究表明，对氨基修改和丙基半胱氨酸的类异型单位可以负担ICMT的小分子抑制剂。这个特定的目的协作，跨学科的研究建议如下：目标1）新颖的法尼斯类似物将是通过固相合成方案详细阐述了相应的前半胱氨酸衍生物。这些会然后进行进一步的化学修饰，以提供前甲肾上腺素衍生物的文库。目标2）这些使用高通量体外测定 ICMT活动的系统和良好的单点测定程序。目标3）将评估梅半胱氨酸类似物在体内阻断先半胱氨酸甲基化的能力。有效的细胞渗透抑制剂将被评估其错误定位RA的能力，干扰Ras- 介导的信号传导，胰腺导管癌的锚固无关生长和阻断肿瘤体内生长。