In Vitro and In Vivo Analyses of 14-O-Acylanthracyclines

14-O-酰基蒽环类药物的体外和体内分析

• 批准号: 6879685
• 负责人: LEONARD LOTHSTEIN
• 金额: $ 29.93万
• 依托单位: UNIVERSITY OF TENNESSEE HEALTH SCI CTR
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-01 至 2007-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6879685
• 关键词: SCID mouse; anthracyclines; antineoplastics; apoptosis; calcium flux; cell line; cytochrome c; drug resistance; enzyme activity; membrane permeability; membrane potentials; mitochondria; neoplasm /cancer transplantation; pharmacokinetics; protein kinase C

DESCRIPTION (provided by applicant): The 14-O-acyl anthracyclines, represented by N-benzyladriamycin-14-valerate (AD 198) and N-benzyladriamycin-14-pivalate (AD 445), are functionally distinct from doxorubicin (DOX) and other DNA-binding anthracyclines. AD 198 and AD 445 localize in the cytoplasm, specifically bind to the C1 regulatory domain of protein kinase C (PKC) and activate PKC in a manner that rapidly triggers apoptosis in proliferating cells. As we have shown, AD 198 and AD 445 activate mitochondrial-associated PKC-d holoenzyme to trigger mitochondrial membrane depolarization and cytochrome c release in 32D.3 routine myeloid cells. Unlike non-PKC binding anthracyclines, such as DOX, AD 198 and AD 445 cytotoxicity is unaffected by Bcl-2 or Bcl-XL anti-apoptotic protein expression or p53 protein dysfunction in a variety of cell types. AD 198 and AD 445 also circumvent resistance conferred by multidrug transport proteins and altered topoisomerase II activity, In vivo, AD 198 demonstrates tumoricidal activity greater than DOX, with reduced myelosuppression and no significant organ toxicity, including a lack of cardiotoxicity. This proposal describes the next stages of analysis of AD 198 and AD 445 leading to clinical trials. The overall goals of this study are first, to analyze the mechanism by which PKC activation by AD 198 and AD 445 leads to rapid apoptosis. We will begin with 32D.3 cells to determine how drug-induced PKC-d activation leads to mitochondrial depolarization and cytochrome c release in a manner that is unaffected by Bcl-2 or Bcl-XL status. Second, given the clear advantage of these agents over DOX thus far in vitro and in vivo, we will continue these studies by assessing the in vivo efficacy of AD 198 an d AD 445 against implanted HL60 human leukemia cells expressing a variety of resistance mechanisms (Bcl-21P-gplMRPIp53-1-). Completion of these studies will establish in greater detail a mechanism of AD 198 and AD 445 induction of apoptosis and determine whether these novel agents can circumvent multiple, clinically relevant mechanisms of drug resistance in vivo.

描述(申请人提供)：由N-苄基阿霉素-14-戊酸酯(AD 198)和N-苄基阿霉素-14-戊酸酯(AD 445)代表的14-O-酰基蒽环类药物，在功能上不同于阿霉素(DOX)和其他与DNA结合的蒽环类药物。AD 198和AD 445定位于细胞质，与蛋白激酶C(PKC)的C1调节域特异性结合，以一种快速启动增殖细胞凋亡的方式激活PKC。我们发现，在32D.3常规髓系细胞中，AD198和AD445激活线粒体相关的PKC-d全酶，从而触发线粒体膜去极化和细胞色素c的释放。与DOX、AD 198和AD 445等非PKC结合的蒽环类药物不同，在多种细胞类型中，细胞毒作用不受Bcl2或Bclxl抗凋亡蛋白表达或P53蛋白功能障碍的影响。AD 198和AD 445还可以避开多药转运蛋白和改变的拓扑异构酶II活性产生的耐药性。在体内，AD 198显示出比DOX更强的杀瘤活性，减少了骨髓抑制，并且没有明显的器官毒性，包括缺乏心脏毒性。这份提案描述了导致临床试验的公元198和公元445的下一个分析阶段。本研究的总体目标是首先分析由AD 198和AD 445激活的PKC导致快速细胞凋亡的机制。我们将从32D.3细胞开始，确定药物诱导的PKC-d激活如何导致线粒体去极化和细胞色素c释放，而不受Bcl2或Bclxl状态的影响。其次，鉴于到目前为止，这些药物在体外和体内相对于DOX的明显优势 在体内，我们将通过评估AD 198和AD 445对表达多种耐药机制(Bcl-21p-gplMRPIp53-1-)的移植的HL60人白血病细胞的体内疗效来继续这些研究。这些研究的完成将更详细地建立AD 198和AD 445诱导细胞凋亡的机制，并确定这些新药物是否可以在体内规避多种临床相关的耐药机制。