Synthetic Probes of Protein Prenylation

蛋白质异戊二烯化的合成探针

• 批准号: 6434588
• 负责人: H Peter Spielmann
• 金额: $ 26.59万
• 依托单位: UNIVERSITY OF KENTUCKY
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-05-01 至 2006-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6434588
• 关键词: Xenopus oocyte; alkyltransferase; analog; antineoplastics; chemical structure function; combinatorial chemistry; drug design /synthesis /production; enzyme inhibitors; farnesyl compound; geranyl compound; isoprenoid; posttranslational modifications; protein localization; pyrophosphates

DESCRIPTION (provided by applicant): Synthetic Probes of Protein Prenylation: Covalent modification by isoprenoid lipids prenyIation) is a critical post-translational event for many proteins involved in cellular signaling. The discovery that the members of the Ras family of protooncogenes are modified by the farnesyl isoprenoid (farnesylation), and that prenylation is required for the oncogenic forms of these proteins to express their transforming potential, has led to intense investigation of protein-farnesyltransferase (FTase) inhibitors (FTIs) as promising cancer chemotherapeutic agents. However, recent developments have made it clear that the mechanism of FTI action is unexpectedly complex, although it involves inhibition of FTase. In addition, the contribution of isoprenoid lipids to the overall biology of Ras is incompletely understood. Therefore, a key to applying FTase-based pharmacological intervention is a thorough understanding of the in vivo farnesylation pathways. Knowledge of the substrate specificity for FTase, and the cellular function of the prenyl moiety are critical to improving the design of future FTIs. The central hypothesis of this study is that the prenyl group plays an active role in directing both post-translational processing and cellular membrane localization of prenylated proteins. An important corollary to this hypothesis is that modifications to the prenyl structure may lead to significant, biologically relevant effects on the activity of the unnaturally prenylated protein. We have synthesized FPP analogs that are transferred to oncogenic Ras but fail to support transformation. These molecules are leads for a unique class of potential anti-cancer: therapeutics we term RFIs (Ras function inhibitors). The specific aims of this project are: 1) specifically substituted unnatural analogs of farnesyl pyrophosphate and the homologous isoprenoid geranylgeranyl pyrophosphate will be synthesized in a combinatorial scheme; 2) these compounds will be screened as substrates or inhibitors of FTase and the closely related enzyme geranylgeranyltransferase I; 3) building upon promising preliminary results in this area, an in vivo isoprenoid structure-function relationship will be established by replacing the H-Ras farnesyl group with a select subset of the analogs available from the studies described in specific aim 1 and 2 and analyzing their biological functions following microinjection into Xenopus oocytes. The results of these experiments will provide further insight into the mechanisms of Ras processing transformation, a greater understanding of the specific functions of the H-Ras farnesyl group in vivo, and fruitful directions for improvements in FTIs as well as novel transferable analogs which might act as RFIs.

描述（由申请人提供）：蛋白质异戊二烯化的合成探针： 类异戊二烯脂质（异戊二烯化）的共价修饰是一个关键 许多参与细胞信号转导的蛋白质的翻译后事件。这 发现原癌基因 Ras 家族的成员被修饰 法呢基异戊二烯（法呢基化），并且需要异戊二烯化 这些蛋白质的致癌形式表达其转化潜力， 引发了对蛋白质法呢基转移酶（FTase）的深入研究 抑制剂（FTIs）作为有前途的癌症化疗药物。然而，最近 事态发展已经表明，FTI 的作用机制是 尽管它涉及 FTase 的抑制，但出乎意料地复杂。此外， 类异戊二烯脂质对 Ras 整体生物学的贡献是 不完全理解。因此，应用基于 FTase 的关键 药物干预是对体内的透彻了解 法尼基化途径。了解 FTase 底物特异性，以及 异戊二烯部分的细胞功能对于改进设计至关重要 未来的 FTI。本研究的中心假设是异戊二烯基团 在指导翻译后处理和 异戊二烯化蛋白质的细胞膜定位。一个重要的推论 这一假设的前提是对异戊二烯结构的修饰可能会导致 对非自然活动的显着的、生物学相关的影响 异戊二烯化蛋白质。我们合成了 FPP 类似物，并将其转移到 致癌 Ras 但不能支持转化。这些分子是 一类独特的潜在抗癌疗法：我们称之为 RFI（Ras 功能抑制剂）。该项目的具体目标是：1）具体 法尼基焦磷酸的取代非天然类似物及其同源物 类异戊二烯香叶基香叶基焦磷酸将以组合方式合成 方案; 2) 这些化合物将被筛选作为底物或抑制剂 FTase 和密切相关的酶香叶基香叶基转移酶 I； 3）建筑 根据该领域有希望的初步结果，体内类异戊二烯 通过替换H-Ras建立结构-功能关系 法呢基团以及研究中可用的类似物的精选子集 具体目标1和2中描述并分析其生物学功能 显微注射到非洲爪蟾卵母细胞后。这些结果 实验将进一步深入了解 Ras 处理机制 转化，更好地了解 H-Ras 的具体功能 体内法呢基团，以及 FTI 改进的富有成果的方向 以及可能充当 RFI 的新型可转移类似物。