MULTITARGET DRUGS: VERIFICATION-OF-PRINCIPLE STUDIES

多靶点药物：原理验证研究

• 批准号: 6342202
• 负责人: ALEXANDER J VARSHAVSKY
• 金额: $ 15.6万
• 依托单位: CALIFORNIA INSTITUTE OF TECHNOLOGY
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-01-06 至 2001-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6342202
• 关键词: Saccharomyces cerevisiae; chemical binding; chemical structure; chemical structure function; cytotoxicity; drug design /synthesis /production; gene expression; methotrexate; novobiocin; trimetrexate

The insufficient selectivity of drugs is a bane of present-day therapies. This problem is significant for antibacterial drugs, difficult for antivirals, and utterly unsolved for anti-cancer drugs, which remain ineffective against major cancers, and in addition cause severe side effects. T he problem may be solved if therapeutic agents could acquire a multi-target, combinatorial selectivity, killing (or otherwise modifying) a cell if, and only if, it contains a predetermined set of molecular targets and lacks another predetermined set of targets. Recently, a solution to the problem of drug selectivity was proposed that is applicable to small (less than or equal to 1 kD) compounds (Varshavsky, A. PNAS 95, 2094-2099 (1998)). Two ideas, interference/co-dominance (IC) and anti-effectors, should allow a therapeutic regimen to possess combinatorial selectivity, in which the number of positively and negatively sensed macromolecular targets can be two, three, or more. The in vivo effects of such drugs would be analogous to the outcomes of the Boolean operations "and", "or" "not", and combinations thereof. The proposed studies, to be supported by the present R21 application, will verify the principles of multi-target designs using the yeast Saccharomyces cerevisiae as a model organism. Specific Aims 1) The ubiquitin fusion technique will be used to express and analyze, in the yeast S. cerevisiae, a set of multi-moiety linear peptides that act as IC- type compounds in this in vivo setting. 2) Bipartite, IC-type compounds that comprise the anhydrotetracycline (at) moiety and either the methotrexate (mtx) or the trimetrexate (tm) moiety linked to at will be synthesized, and thereafter characterized in vitro (mutual exclusivity of binding to the purified TetR or DHFR) and in vivo (differential toxicity to S. cerevisiae that either lack or express the at- binding E. coli TetR repressor). 3) Bipartite, IC-type compounds containing the novobiocin (nov) moiety and either the methotrexate (mtx) or the trimetrexate (tm) moiety will be synthesized and tested for differential cytotoxicity with S. cerevisiae that either lack or express the nov-binding B-subunit of E. coli DNA gyrates (GyrB). 4) Tripartite, IC-type compounds containing the at moiety, the beta- ecdysone (ec) moiety, and either the methotrexate (mtx) or the trimetrexate (tm) moiety will be synthesized and tested for differential toxicity with S. cerevisiae that lack or express the corresponding macromolecular targets and their pairwise combinations.

药物的选择性不足是当今疗法的祸根。该问题对于抗菌药物很重要，对于抗病毒药而言困难，并且对于抗癌药物完全无法解决，抗癌药仍然对主要癌症无效，此外还会引起严重的副作用。如果治疗剂可以获取多目标，组合选择性，杀死（或其他修改）细胞时，则可以解决问题。最近，提出了一种解决药物选择性问题的解决方案，该问题适用于小（小于或等于1 kD）化合物（Varshavsky，A。PNAS 95，2094-2099（1998））。两种想法，即干涉/共同主持（IC）和抗效应，应允许治疗方案具有组合选择性，其中积极和负面感知的大分子靶标可以是两个，三个或更多。这种药物的体内作用将类似于布尔操作的结果“和”或“不是”，及其组合。拟议的研究将得到当前R21应用的支持，将使用酿酒酵母作为模型有机体来验证多目标设计的原理。具体目的1）泛素融合技术将用于在酵母菌S. cerevisiae中表达和分析，这是一组多动感线性线性肽，在这种体内设置中充当IC类化合物。 2）构成甲基氢环环素（AT）的双分型化合物，以及甲氨蝶呤（MTX）或链接到AT的三甲酸（TM）部分的甲氨蝶呤（MTX）或链接的甲基甲基元素化合物（MTX）或链接的甲氨蝶呤（MTX）或甲基甲基环霉素（MTX）或甲基甲基环霉素（MTX）或甲基苯二酚（MTX）或甲基甲基环霉素（MTX）或链接的甲氨蝶呤（MTX），将在与纤维化（与DHFRIFIED）中的相互排斥性（与DHFRIFIED S.相互的排除性）合成（TM）部分。缺乏或表达结合大肠杆菌Tetr抑制剂的酿酒酵母。 3）含有novobiocin（Nov）部分的IC型化合物以及甲氨蝶呤（MTX）或三甲酸（TM）部分，将合成并用缺乏或表达Nov-Bindbind b-Subunit of E. coli dna gyr的cerevisiae的差异细胞毒性进行测试并测试。 4）三方，包含AT部分的IC型化合物，Beta- ecdysone（EC）部分以及甲氨蝶呤（MTX）或三甲甲酸（TM）部分将被合成并测试与缺乏或表达对应的分子质量的cerevisiae的差异毒性，以使其对相应的corevisiae进行差异毒性，以使其配对组合的组合和他们的配对组合。