Inhibition of Prenylated Protein Processing

异戊二烯化蛋白质加工的抑制

• 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
• 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

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，经过翻译后修饰，将其靶向 适当的细胞内位置。 Ras 和其他 -CaaX 蛋白经历三个连续反应： 半胱氨酸的异戊二烯化，特别是法尼基转移酶(FTase)的法呢基化；三者的蛋白水解作用 末端氨基酸（-aaX）；和异戊二烯化半胱氨酸的α-羧甲基化。这个过程是 对于膜定位以及关键 Ras 癌蛋白的活性至关重要。 FTase 抑制剂正在 在临床试验中作为癌症化疗药物进行评估。不幸的是，这些化合物 令人惊讶的是，它对许多 Ras 转化的肿瘤影响很小。现在人们对后续的兴趣越来越大 酶促步骤； Ras 转换酶 (Reel) 进行蛋白水解，异戊二烯半胱氨酸进行 a-羧基甲基化 甲基转移酶 (Icmt) 作为抑制 Ras 蛋白作用的替代靶点。羧甲基化是 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 某些易于合成的、基于类异戊二烯的 Icmt 抑制剂将是有价值的先导化合物 用于开发抗Ras癌症化疗药物。最近发表和未发表的 我们实验室的研究表明，对氨基修饰和 异戊二烯半胱氨酸单元可以提供 Icmt 的小分子抑制剂。本次活动的具体目标 合作、跨学科研究提案如下： 目标 1) 新型法尼醇类似物将 通过固相合成方案，精心设计成相应的异戊二烯半胱氨酸衍生物。这些将 然后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 使用高通量体外测定，将异戊二烯半胱氨酸衍生物作为 Icmt 的潜在抑制剂进行测定 系统和针对 Icmt 活性的特征明确且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错位 Ras、干扰 Ras 的能力。 介导的信号传导，阻断胰腺导管癌的锚定非依赖性生长，并阻断肿瘤 体内生长。