Department of Pharmacology and Toxicology, Faculty of Pharmacy in Hradec Kralove, Charles University, Akademika Heyrovskeho 1203, 500 05 Hradec Kralove, Czech Republic.
Department of Biochemical Sciences, Faculty of Pharmacy in Hradec Kralove, Charles University, Akademika Heyrovskeho 1203, 500 05 Hradec Kralove, Czech Republic.
Biochem Pharmacol. 2019 May;163:290-298. doi: 10.1016/j.bcp.2019.02.035. Epub 2019 Feb 28.
Daunorubicin (DAUN) has served as an anticancer drug in chemotherapy regimens for decades and is still irreplaceable in treatment of acute leukemias. The therapeutic outcome of DAUN-based therapy is compromised by its cardiotoxicity and emergence of drug resistance. This phenomenon is often caused by pharmacokinetic mechanisms such as efflux of DAUN from cancer cells through ATP-binding cassette (ABC) transporters and its conversion to less cytostatic but more cardiotoxic daunorubicinol (DAUN-OL) by carbonyl reducing enzymes (CREs). Here we aimed to investigate, whether two cyclin-dependent kinase inhibitors, AZD5438 and R547, can interact with these pharmacokinetic mechanisms and reverse DAUN resistance. Using accumulation assays, we revealed AZD5438 as potent inhibitor of ABCC1 showing also weaker inhibitory effect to ABCB1 and ABCG2. Combination index analysis, however, shown that inhibition of ABCC1 does not significantly contribute to synergism between AZD5438 and DAUN in MDCKII-ABCC1 cells, suggesting predominant role of other mechanism. Using pure recombinant enzymes, we found both tested drugs to inhibit CREs with aldo-keto reductase 1C3 (AKR1C3). This interaction was further confirmed in transfected HCT-116 cells. Moreover, these cells were sensitized to DAUN by both compounds as Chou-Talalay combination index analysis showed synergism in AKR1C3 transfected HCT-116, but not in empty vector transfected control cell line. In conclusion, we propose AZD5438 and R547 as modulators of DAUN resistance that can prevent AKR1C3-mediated DAUN biotransformation to DAUN-OL. This interaction could be beneficially exploited to prevent failure of DAUN-based therapy as well as the undesirable cardiotoxic effect of DAUN-OL.
柔红霉素(DAUN)作为一种抗癌药物,已在化疗方案中使用了数十年,在治疗急性白血病方面仍然不可替代。基于 DAUN 的治疗的治疗效果因心脏毒性和耐药性的出现而受到影响。这种现象通常是由药代动力学机制引起的,例如 DAUN 通过 ATP 结合盒(ABC)转运蛋白从癌细胞中流出,以及其通过羰基还原酶(CREs)转化为较少细胞抑制但更具心脏毒性的柔红霉素醇(DAUN-OL)。在这里,我们旨在研究两种细胞周期蛋白依赖性激酶抑制剂,AZD5438 和 R547 是否可以与这些药代动力学机制相互作用并逆转 DAUN 耐药性。使用积累测定法,我们发现 AZD5438 是 ABCC1 的有效抑制剂,对 ABCB1 和 ABCG2 也具有较弱的抑制作用。然而,组合指数分析表明,ABCC1 的抑制对 AZD5438 和 DAUN 在 MDCKII-ABCC1 细胞中的协同作用没有显著贡献,表明其他机制起主要作用。使用纯重组酶,我们发现两种测试药物均抑制醛酮还原酶 1C3(AKR1C3)。在转染的 HCT-116 细胞中进一步证实了这种相互作用。此外,正如 Chou-Talalay 组合指数分析所示,这两种化合物使 AKR1C3 转染的 HCT-116 细胞对 DAUN 敏感,而对空载体转染的对照细胞系则没有协同作用。总之,我们提出 AZD5438 和 R547 作为 DAUN 耐药性的调节剂,可防止 AKR1C3 介导的 DAUN 生物转化为 DAUN-OL。这种相互作用可以有益地利用来防止基于 DAUN 的治疗失败以及 DAUN-OL 的不良心脏毒性作用。