齐墩果酸衍生物 3,11-二酮基齐墩果-12-烯-28-酸甲酯靶向与 ABCB1 相关的多药耐药。

Oleanolic acid derivative methyl 3,11-dioxoolean-12-en-28-olate targets multidrug resistance related to ABCB1.

机构信息

Department of Clinical Chemistry and Molecular Diagnostics, Poznan University of Medical Sciences, Przybyszewskiego 49, PL 60-355 Poznań, Poland.

出版信息

Pharmacol Rep. 2011;63(6):1500-17. doi: 10.1016/s1734-1140(11)70714-9.

DOI:10.1016/s1734-1140(11)70714-9

PMID:22358098

Abstract

Multidrug resistance (MDR) in leukemia patients is a great incentive to the development of new drugs. In a search for potential multidrug resistance modulators we tested a group of oleanolic acid (OA) analogues modified at C-3, C-11, C-12 and C-28 using an experimental model consisting of three human acute lymphoblastic leukemia cell lines (CCRF-CEM and the multidrug resistant sublines CCRF-VCR1000 and CCRF-ADR5000). The most effective compound, methyl 3,11-dioxoolean-12-en-28-olate (DIOXOL) was more potent in cell viability inhibition than its precursor - OA, and showed similar or even higher activity in the drug resistant than in the wild-type cells. Resistance factor (RF) values obtained for CCRF-VCR1000 and CCRF-ADR-5000 cells using MTT assay were 0.7 and 0.8 (24 h of treatment) and after 72 h of treatment 0.9 and 1.1, respectively. Moreover, 5 μM DIOXOL significantly reduced the expression of the ABCB1 gene in MDR cells by around 30%, and also decreased the level of P-gp protein. Compared to untreated control cells, DIOXOL treatment resulted in a significant P-gp decrease (30% in CCRF-ADR5000 and 50% in CCRF-VCR1000), that was detected by western blot and confirmed by flow cytometry analysis. Moreover, DIOXOL (at 10 μM) significantly inhibited P-gp transport function by more than twofold comparing to control, untreated cells that was demonstrated using rhodamine 123-based functional test. The compound exhibited synergistic activity with ABCB1 substrate - adriamycin in CCRF-VCR1000 cells, indicating partial but significant MDR reversing ability.

摘要

在白血病患者中，多药耐药（MDR）是开发新药的巨大动力。在寻找潜在的多药耐药调节剂的过程中，我们使用包含三个人类急性淋巴细胞白血病细胞系（CCRF-CEM 以及多药耐药亚系 CCRF-VCR1000 和 CCRF-ADR5000）的实验模型测试了一组 3-位、11-位、12-位和 28-位被修饰的齐墩果酸（OA）类似物。最有效的化合物是 3,11-二氧代齐墩果酸 12-烯-28-酸甲酯（DIOXOL），它在抑制细胞活力方面比其前体 OA 更有效，并且在耐药细胞中的活性与在野生型细胞中的活性相似甚至更高。使用 MTT 测定法获得的 CCRF-VCR1000 和 CCRF-ADR-5000 细胞的耐药因子（RF）值分别为 0.7 和 0.8（24 小时处理），72 小时处理后分别为 0.9 和 1.1。此外，5μMDIOXOL 可使 MDR 细胞中 ABCB1 基因的表达减少约 30%，并降低 P-糖蛋白（P-gp）蛋白的水平。与未处理的对照细胞相比，DIOXOL 处理导致 P-gp 显著减少（在 CCRF-ADR5000 中减少 30%，在 CCRF-VCR1000 中减少 50%），这通过蛋白质印迹和流式细胞术分析得到证实。此外，DIOXOL（在 10μM 时）在使用 rhodamine 123 功能测试时，与对照、未经处理的细胞相比，显著抑制 P-gp 转运功能超过两倍。该化合物在 CCRF-VCR1000 细胞中与 ABCB1 底物阿霉素表现出协同活性，表明其具有部分但显著的逆转多药耐药能力。