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：
7213662
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/7213662
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

DESCRIPTION (provided by applicant): 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 3 sequential reactions: isoprenylation of the cysteine, in particular farnesylation by farnesyltransferase (FTase); proteolysis of the 3 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 of prenylcysteines 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.

描述（由申请人提供）：与许多人类癌症有关的 Ras 家族成员经过翻译后修饰，将其靶向适当的细胞内位置。 Ras 和其他 CaaX 蛋白经历 3 个连续反应：半胱氨酸的异戊二烯化，特别是法尼基转移酶 (FTase) 的法尼基化； 3 个末端氨基酸 (-aaX) 的蛋白水解；和异戊二烯化半胱氨酸的α-羧甲基化。这一过程对于膜定位以及关键 Ras 癌蛋白的活性至关重要。 FTase 抑制剂作为癌症化疗药物正在临床试验中进行评估。不幸的是，这些化合物对许多 Ras 转化的肿瘤几乎没有什么作用。现在人们对随后的酶促步骤越来越感兴趣； Ras 转换酶 (Reel) 的蛋白水解作用和异戊二烯半胱氨酸甲基转移酶 (Icmt) 的 a-羧基甲基化作用作为抑制 Ras 蛋白作用的替代靶标。羧甲基化对于 Ras 蛋白在酵母和小鼠细胞中的正确定位至关重要。鉴于这一重要发现，我们相信 Icmt 代表了化疗干预的绝佳目标。我们假设某些容易合成的、基于类异戊二烯的 Icmt 抑制剂将成为开发抗 Ras 癌症化疗药物的有价值的先导化合物。我们实验室最近发表和未发表的研究表明，对异戊二烯半胱氨酸的氨基修饰和类异戊二烯单元进行修饰可以提供 Icmt 的小分子抑制剂。这项跨学科合作研究计划的具体目标如下：目标 1) 通过固相合成方案，将新型法尼醇类似物精细化为相应的异戊二烯半胱氨酸衍生物。然后这些将进行进一步的化学修饰以提供异戊二烯半胱氨酸衍生物文库。目标 2) 这些异戊二烯半胱氨酸衍生物将作为 Icmt 的潜在抑制剂进行测定，使用高通量体外测定系统和针对 Icmt 活性的良好表征和稳健的单点测定程序。目标 3) 将评估异戊二烯半胱氨酸类似物在体内阻断异戊二烯半胱氨酸甲基化的能力。将评估有效的细胞渗透性抑制剂的 Ras 错误定位、干扰 Ras 介导的信号传导、阻断胰腺导管癌锚定非依赖性生长以及阻断体内肿瘤生长的能力。