Synthetic Probes of Protein Prenylation

蛋白质异戊二烯化的合成探针

• 批准号: 6889530
• 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 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6889530
• 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）：蛋白质异戊烯化的合成探针： 类异戊二烯脂质的共价修饰（异戊二烯化）是一个关键的 许多参与细胞信号传导的蛋白质的翻译后事件。的 发现Ras家族的原癌基因的成员被修饰， 法呢基类异戊二烯（法呢基化）， 这些蛋白质的致癌形式表达其转化潜力， 引起了对蛋白质-法尼基转移酶（FTase）的深入研究 抑制剂（FTIs）作为有前途的癌症化疗剂。但最近的 发展已经清楚地表明，快速道行动的机制是 这是一个出乎意料的复杂过程，尽管它涉及FTase的抑制。此外，本发明还提供了一种方法， 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 药理学干预是对体内 法尼基化途径。了解FTase的底物特异性，以及 异戊二烯基部分的细胞功能对于改进设计是至关重要的 未来的FTIs这项研究的中心假设是， 在指导翻译后加工和 异戊烯化蛋白的细胞膜定位。一个重要的必然 这一假设的另一个原因是，异戊二烯基结构的修饰可能导致 显著的，生物相关的影响，对活动的非自然 异戊二烯化蛋白质。我们已经合成了FPP类似物， 致癌Ras，但不能支持转化。这些分子是 一类独特的潜在抗癌治疗剂，我们称之为RFIs（Ras 功能抑制剂）。该项目的具体目标是：1）具体 焦磷酸法呢酯的取代的非天然类似物和其同源物 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为底物或抑制剂， FT酶和密切相关的酶香叶基香叶基转移酶I; 3）构建 基于在该领域有希望的初步结果， 通过替换H-Ras，建立结构-功能关系 法尼基基团与从研究中获得的类似物的选择子集 在具体目标1和2中描述并分析它们的生物学功能 显微注射到非洲爪蟾卵母细胞中。的结果予以 实验将提供进一步深入了解Ras加工机制 转化，更好地了解H-Ras的具体功能 法呢基基团在体内，和富有成效的方向，改善FTIs， 以及可能作为RFIs的新型可转移类似物。