RATIONAL DRUG DESIGN BASED ON CHEMICAL MECHANISMS

在本申请所请求的支持期间， 我们计划继续努力合成有效的和 基于酶作用机制数据的特定酶抑制剂 从我们的实验室和文献中获得。这个 建议的研究涉及合成有机化合物的组合 化学、机械酶学和生化药理学。 我们研究的总体目标是调查 控制细胞生长的特定生化途径。 更具体地说，在研究的第一部分中提出了 在此，我们将继续使用含氟谷氨酸 用叶酸和抗叶酸评估多聚谷氨酸的作用 哺乳动物细胞中的生物合成和水解。例如，我们 将完成伽马-氟叶酸的合成并使用 明确多聚谷氨酸在亚叶酸钙“拯救”中的作用 大剂量甲氨蝶呤化疗后的细胞。我们会 还利用了我们最近的发现，3，3-二氟谷氨酸 (F2Glu)刺激游离细胞内多聚谷氨酸的形成 系统。随着F2Glu合成路线的改进， 我们将合成一系列含F2Glu的叶酸和 为了研究黄连的生化药理作用。 具有增强的形成多谷氨酸能力的化合物 衍生品。 在拟议研究的第二部分中，我们将 采用我们新的合成方法合成络合物 乙酰核苷。我们将完成我们的合成 建议的特定“多底物加合物抑制剂” 我们有多年经验的甲基酶(儿茶酚 O-甲基转移酶、苯乙醇胺N-甲基转移酶)和 研究它们对分离酶的影响。如果这些提议的新 甲基酶抑制剂和预测的一样特异和有效，我们 将用它们来研究儿茶酚胺的生物合成 神经母细胞瘤细胞。与叶酸工作一样，我们的最终目标是 在这项工作中是开发有效的和特定的每种抑制剂 甲基转移酶，以研究它们在细胞生长中的作用 和/或细胞功能。

During the period of support requested in this application, we plan to continue our efforts on the synthesis of potent and specific enzyme inhibitors based on enzyme mechanism data obtained in our laboratory and from the literature. The proposed studies involve a combination of synthetic organic chemistry, mechanistic enzymology, and biochemical pharmacology. The overall goal of our research is to investigate the role of specific biochemical pathways in the control of cell growth. More specifically, in the first portion of the research proposed herein, we will continue our use of fluoroglutamate-containing folates and antifols to assess the role of polyglutamate biosynthesis and hydrolysis in mammalian cells. For example, we will complete the synthesis of gamma-fluoroleucovorin and use it to define the role of polyglutamates in leucovorin "rescue" of cells following high-dose methotrexate chemotherapy. We will also exploit our recent finding that 3,3-difluoroglutamate (F2Glu) stimulates the formation of polyglutamates in cell-free systems. Following improvement of the synthetic route to F2Glu, we will synthesize a series of F2Glu-containing folates and antifols in order to investigate the biochemical pharmacology of compounds with enhanced ability to form polyglutamate derivatives. In the second portion of the proposed research we will employ our new synthetic methodology for the synthesis of complex acetylenic nucleosides. We will complete the synthesis of our proposed specific "multisubstrate adduct inhibitors" of methylases with which we have many years of experience (catechol O-methyltransferase, phenethanolamine N-methyltransferase) and study their effect on isolated enzymes. If these proposed new methylase inhibitors are as specific and potent as predicted, we will use them to study catecholamine biosynthesis in neuroblastoma cells. As with the folate work, our ultimate goal in this work is to develop potent and specific inhibitors of each methyltransferase in order to study their role in cell growth and/or cell function.