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）， 调节多胺稳态并有效抑制肿瘤生长， 临床前模型系统和另一种，多胺抑制剂，CGP-48664， 靶向多胺生物合成酶S-腺苷蛋氨酸 脱羧酶消耗多胺库并抑制体内肿瘤生长。 这两种药物都是在两个独立的药物发现中合理开发的 在实验室的积极参与下。尽管未 顺序相连，本提案的具体目标由 优化这些有前途的新的临床开发的共同目标， 实体瘤治疗。 在这方面， 拟定：目的1，验证以下药物的临床前适应症作用方式 DENSPM和CGP-48664在从接受治疗的患者获得的实体瘤中的作用 与DENSPM或CGP 48664;目的2，检查潜在的监管 参与有效和异质诱导的机制， DENSPM人黑色素瘤细胞系的多胺分解代谢酶 从临床前研究中可以看出， 类似物抗肿瘤活性的决定因素;技术上可行时 目标3：跟踪最近观察到的 多胺稳态的明显紊乱和多药 在人类黑色素瘤细胞系的耐药表型-具体来说，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。方法 用于这些研究的包括人细胞培养技术， 生化酶测定，多胺及其类似物的HPLC检测， 罗丹明保留测定，流式细胞术北方印迹，南方印迹， 核连续试验、mRNA半衰期测定和体外 翻译测定。

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.