Ribonucleotide Reductase Antisense Strategy in AML

AML 中的核糖核苷酸还原酶反义策略

• 批准号: 6801147
• 负责人: GUIDO MARCUCCI
• 金额: $ 31.98万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-15 至 2006-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6801147
• 关键词: SDS polyacrylamide gel electrophoresis; acute myelogenous leukemia; apoptosis; autoradiography; clinical research; clinical trial phase I; combination chemotherapy; dosage; drug resistance; enzyme linked immunosorbent assay; flow cytometry; human subject; neoplasm /cancer chemotherapy; neoplasm /cancer pharmacology; neoplasm /cancer relapse /recurrence; neoplastic cell; patient oriented research; pharmacokinetics; ribonucleotide reductase

DESCRIPTION (provided by applicant): Most patients with acute myeloid leukemia (AML) relapse and ultimately die as a consequence of refractory or resistant disease. Salvage chemotherapy at conventional doses may induce a short-term remission, but this approach is generally not curative. For those patients who are not candidates for allogeneic BMT the prognosis remains dismal and new therapeutic strategies targeting specific leukemogenic mechanisms are highly needed to improve the current clinical results. In the past few years, we have successfully applied the antisense strategy to down-regulate expression of distinct target genes that contribute to chemoresistance in myeloid blasts. Here we propose to explore the activity of a novel ribonucleotide reductase (RR) antisense, GTI-2040, in combination with high-dose ARA-C (cytarabine arabinoside) in refractory or relapsed AML. RR is an enzyme operative in the synthetic pathway of deoxyribonucleotides and a potential key factor in induction of chemoresistance to ARA-C, a nucleoside analog incorporated in a variety of AML chemotherapy regimens. We seek to understand the relation between plasma and intracellular concentration of GTI-2040 and how this correlates to RR down-regulation, patient toxicity and disease response. Specific Aim #1 will conduct a phase I study of GTI-2040 in combination with high-dose ARA-C in refractory or relapsed AML. The ultimate goal is to assess the feasibility of this combination and recommend a dose for future phase II studies in this patient population. Specific Aim #2 will evaluate the plasma pharmacokinetics of GTI-2040 using a sensitive ELISA-based assay that we have developed at OSU. This assay will also allow us to measure intracellular concentrations of GTI-2040 in selected leukemia cells from patients enrolled on this protocol. Specific Aim #3 will examine the correlation between plasma and intracellular concentrations of GTI-2040 with toxicity, disease response and biological endpoints such as down-regulation of the antisense target (i.e., the R2 subunit of RR) at the mRNA and protein levels, changes in RR enzymatic activity and levels of apoptosis in selected leukemia cells collected from patients enrolled in this protocol. It is expected that the analysis of these data will allow us to elucidate the intrinsic mechanisms of the antitumor activity of the antisense compounds and plan their incorporation in future therapeutic strategies for AML patients.

描述（由申请人提供）：大多数急性髓样白血病（AML）复发，最终由于难治性或抗性疾病而死亡。常规剂量的抢救化疗可能会引起短期缓解，但这种方法通常无法治愈。对于那些不是同种异体BMT候选者的患者，预后仍然令人沮丧，针对特定白血病机制的新的治疗策略非常需要改善当前的临床结果。在过去的几年中，我们成功地应用了反义策略来下调不同靶基因的表达，这些基因有助于髓样爆炸中的化学耐药性。在这里，我们建议探索新型核糖核苷酸还原酶（RR）反义的活性，GTI-2040，与耐火或复发AML中的高剂量ARA-C（Cytarabine Arabinoside）结合使用。 RR是脱氧核糖核苷酸合成途径中的酶作用，也是诱导ARA-C化学耐药性的潜在关键因素，ARA-C（一种核苷类似物掺入了各种AML化学疗法方案中。我们试图了解GTI-2040的血浆与细胞内浓度之间的关系，以及与RR下调，患者毒性和疾病反应之间的关系。特定的目标＃1将对GTI-2040进行I期研究，并在难治性或复发AML中结合使用高剂量ARA-C。最终目标是评估这种组合的可行性，并建议对该患者人群进行II期研究的剂量。具体目标＃2将使用我们在OSU开发的基于敏感的ELISA测定法评估GTI-2040的血浆药代动力学。该测定法还将使我们能够从入选该方案的患者中测量选定的白血病细胞中GTI-2040的细胞内浓度。具体目的＃3将检查GTI-2040血浆和细胞内浓度之间的相关性与毒性，疾病反应和生物学终点，例如在MRNA和蛋白质水平上对反义靶标的下调（即R2亚基）的下调（即，RR RR的R2亚基），RR酶活性和蛋白质的蛋白质水平的变化。预计对这些数据的分析将使我们能够阐明反义化合物的抗肿瘤活性的内在机制，并计划将其纳入AML患者的未来治疗策略。

项目成果

Enzyme kinetics of GTI-2040, a phosphorothioate oligonucleotide targeting ribonucleotide reductase.

GTI-2040（一种靶向核糖核苷酸还原酶的硫代磷酸寡核苷酸）的酶动力学。

• DOI: 10.1124/dmd.108.021295
• 发表时间: 2008
• 期刊: Drug metabolism and disposition: the biological fate of chemicals
• 作者: Wei,Xiaohui;Dai,Guowei;Liu,Zhongfa;Cheng,Hao;Xie,Zhiliang;Klisovic,Rebecca;Marcucci,Guido;Chan,KennethK
• 通讯作者: Chan,KennethK