FACILITATING THE CLINICAL EVALUATION OF DENSPM

两种多胺拮抗剂计划在RPCI进行临床试验-- 一种是多胺类似物，N1，N11-二乙基去甲精胺(DENSPM)，它 解除多胺动态平衡并有效抑制肿瘤生长 临床前模型系统和另一个，多胺抑制剂，CGP-48664， 其靶标是多胺生物合成酶S-腺苷甲硫氨酸 脱羧酶，耗尽多胺库，抑制体内肿瘤生长。 这两种药物在两个独立的药物发现中得到了合理的开发 在本实验室的积极参与下，开展了多项举措。虽然不是 顺序相连，本提案的具体目标由 优化这些有希望的新技术的临床开发的共同目标 实体肿瘤疗法。在这方面，下列活动包括 建议：目标1，验证临床前指征的作用模式 DNSPM和CgP-48664在治疗后实体瘤中的表达 使用DENSPM或CGP48664；目标2，检查潜在的监管 参与强效和异质诱导的机制 DENSPM人黑色素瘤细胞株的多胺分解酶 来自临床前研究的反应似乎是体内的关键 类似物抗肿瘤活性的决定因素；在技术上可行的情况下 发现；目标3跟进最近观察到的 多胺动态平衡的不同紊乱与多药 人类黑色素瘤细胞系的耐药表型--具体地说，我们将 检查这种明显联系的基础和治疗潜力 多胺拮抗剂在解决多药耐药性方面的作用。方法论 在这些研究中使用的技术包括人类细胞培养技术， 生化酶分析、高效液相色谱检测多胺及其类似物， 罗丹明滞留试验、流式细胞术Northern印迹、Southern印迹、 核连续试验、信使核糖核酸半衰期测定和体外试验 翻译分析。

Two polyamine antagonists are scheduled for clinical trial here at RPCI-- one, a polyamine analog, N1 ,N11-diethylnorspermine (DENSPM), which deregulates polyamine homeostasis and potently inhibits tumor growth in preclinical model systems and the other, a polyamine inhibitor, CGP-48664, which target the polyamine biosynthetic enzyme, S-adenosylmethionine decarboxylase, depletes polyamine pools an inhibits tumor growth in vivo. Both agents have been rationally developed in two separate drug discover initiatives with the active involvement of this laboratory. Although not sequentially linked, the Specific Aims of this proposal are unified by the common goal of optimizing the clinical development of these promising new solid tumor therapies. In this context, the following activities are proposed: Aim 1, to validate the preclinically-indicated mode of action of both DENSPM and CGP-48664 in solid tumors obtained from patients treated with DENSPM or CGP48664; Aim 2, to examine the underlying regulatory mechanisms involved in the potent and heterogeneous induction of a polyamine catabolizing enzyme by DENSPM human melanoma cell lines--a response which appears from preclinical studies to be a critical in vivo determinant of antitumor activity of the analog; when technically possible findings ; Aim 3 to follow up on a recently observed association between distinct disturbances in polyamine homeostasis and the multidrug resistance phenotype in human melanoma cell lines--specifically, we will examine the basis for this apparent linkage and the therapeutic potential of polyamine antagonists in addressing multidrug resistance. Methodologies to be used in these studies include human cell culture techniques, biochemical enzyme assays, HPLC detection of polyamines and their analogs, rhodamine retention assays, flow cytometry Northern blots, Southern blots, nuclear run-on assays, mRNA half-life determinations and in vitro translation assays.