Synthetic Probes of Protein Prenylation

蛋白质异戊二烯化的合成探针

• 批准号: 6745114
• 负责人: H Peter Spielmann
• 金额: $ 28.96万
• 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/6745114
• 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.

说明书(申请人提供)：蛋白质预基化的合成探针： 类异戊二烯类脂的共价修饰)是一个关键 参与细胞信号传递的许多蛋白质的翻译后事件。这个 原癌基因RAS家族成员被修饰的发现 法尼基异戊二烯(法尼化)，以及异戊二烯基法呢化是必需的 这些蛋白质的致癌形式来表达它们的转化潜力， 引起了对蛋白质法尼基转移酶(FTase)的深入研究 抑制物(FTI)是一种很有前途的癌症化疗药物。然而，最近 事态发展表明，快速道行动的机制是 出乎意料的复杂，尽管它涉及到FTase的抑制。此外, 类异戊二烯类脂对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 法尼化途径。了解FTase的底物专一性，以及 戊烯基部分的细胞功能是改进设计的关键 未来的FTI。这项研究的中心假设是戊烯基团 在指导翻译后处理和 前烯基化蛋白的细胞膜定位。一个重要的推论 对这一假设的看法是，对戊烯基结构的修饰可能会导致 对非自然生物的活动有重大的、生物相关的影响 戊烯基化的蛋白质。我们已经合成了FPP类似物，这些类似物被转移到 致癌RAS，但不支持转化。这些分子是引线 一类独特的潜在抗癌药物：我们称之为RFI(RAS)的治疗学 功能抑制剂)。本项目的具体目标是：1)具体 金合欢基焦磷酸盐的取代非天然类似物及其同系物 异戊二烯类香叶基香叶基焦磷酸酯的合成 方案；2)这些化合物将被筛选为底物或抑制剂 FTase和密切相关的香叶基香叶基转移酶I；3)构建 在这一领域有希望的初步结果之后，体内的类异戊二烯 将通过取代H-RAS来建立结构-功能关系 具有可从研究中获得的类似物的精选子集的法尼基 在具体目标1和2中描述并分析它们的生物学功能 在非洲爪哇卵母细胞显微注射后。这些研究的结果 实验将进一步深入了解RAS处理的机制 转型，更好地了解H-RAS的具体功能 体内的法尼基，以及FTIs的丰硕方向 以及可作为RFI的新型可转让类似物。