Design of Enzymes Mediating Anti-Cancer Drug Activation

介导抗癌药物激活的酶的设计

• 批准号: 6776397
• 负责人: ARNON LAVIE
• 金额: $ 27.4万
• 依托单位: UNIVERSITY OF ILLINOIS AT CHICAGO
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-08-22 至 2006-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6776397
• 关键词: X ray crystallography; allosteric site; antineoplastics; clinical research; deoxycytidine; drug delivery systems; drug design /synthesis /production; enzyme activity; enzyme mechanism; enzyme structure; enzyme substrate; enzyme substrate complex; fludarabine; gemcitabine; human tissue; leukemia; monoclonal antibody; mutant; nucleoside analog; phosphorylation; phosphotransferases; prodrugs; protein structure function

DESCRIPTION (provided by applicant): Despite significant progress in the treatment of hematological malignancies, in particular leukemias, a significant majority of patients do not achieve a remission, or relapse after a prior response. As a result intense research is being directed towards examining methods of improving outcome by increasing the efficacy) of the established agents. Nucleoside analogues, including arabinosylcytosine (AraC), fludarabine, cladribine, pentostatin, and more recently gemcitabine and AraG have been a major part of the arsenal for treating hematological malignancies and solid tumors. The cytotoxic activity of AraC as well as that of other nucleoside analogues, is dependent on their intracellular conversion to an active phosphorylated metabolite. This phosphorylation (activation) is catalyzed by intracellular kinases such as deoxycytidine kinase (dCK). This enzyme is the rate-limiting factor for the activation of prodrugs such as AraC, fludarabine, cladribine, and gemcitabine. We intend to investigate the three dimensional structure as well as the catalytic mechanism of dCK. to determine its substrate specificity to define the factors that limit the efficacy of prodrug phosphorylation by the enzyme. and to apply the information to design and test dCK mutants with enhanced enzymatic (phosphorylation) activity towards these compounds. In addition we will examine the allosteric properties of dCK with the aim of developing an allosteric activator of dCK. Selective delivery of a dCK variant with enhanced activity into leukemic blasts will be an integral part of this project. This will ensure the ultimate goal of increasing the efficacy/toxicity ratio of nucleoside analogues. We will examine the feasibility of using a monoclonal antibody-enzyme delivery system to target leukemic cells with our mutant enzymes. Such a system will have the theoretical advantage of delivering the enzyme selectively into the leukemic blasts, increasing the activity of the drug in these cells only. thereby sparing the normal cells. We will use humanized anti-CD33 antibody conjugated with the mutant dCK to increase the efficacy of AraC in the targeted cells. This Selective Enhanced Enzyme Delivery System (SEEDS) could be applicable in treating other malignancies using the appropriate antibodies, mutant enzymes and drugs.

描述（由申请人提供）：尽管取得了重大进展 血液学恶性肿瘤的治疗，特别是白血病 大多数患者未能缓解或先前的复发 回复。结果，强烈的研究是针对检查的 通过提高已建立的功效来改善结果的方法 代理商。核苷类似物，包括阿拉伯糖基胞嘧啶（ARAC），氟达拉滨， 克拉德替替替替替替替比，五角汀以及最近的吉西他滨和阿拉格一直是 武库的主要部分治疗血液恶性肿瘤和固体 肿瘤。 ARAC以及其他核苷的细胞毒性活性 类似物，取决于它们的细胞内转换为活动 磷酸化的代谢产物。这种磷酸化（激活）由 细胞内激酶，例如脱氧胞苷激酶（DCK）。这种酶是 阿拉克，氟达拉滨等前药激活的速率限制因子， 克拉德替宾和吉西他滨。我们打算研究三维 DCK的结构以及催化机制。确定其底物 定义限制前药功效的因素的特异性 酶的磷酸化。并将信息应用于设计和测试 DCK突变体具有增强的酶促（磷酸化）活性 化合物。此外，我们将检查DCK的变构特性 开发DCK的变构激活剂的目的。 选择性递送DCK变体具有增强活性到白血病爆炸的变体 将是该项目不可或缺的一部分。这将确保最终目标 增加核苷类似物的功效/毒性比。我们将检查 使用单克隆抗体 - 酶递送系统的可行性 白血病细胞与我们的突变酶。这样的系统将具有理论 将酶选择性递送到白血病爆炸中的优势， 仅在这些细胞中增加药物的活性。从而保留 正常细胞。我们将使用与人源化抗CD33抗体与 突变DCK提高了ARAC在靶细胞中的疗效。这 选择性增强的酶输送系统（种子）可以适用 使用适当的抗体，突变酶治疗其他恶性肿瘤 和毒品。