NAD(P)H:Quinone oxidoreductase-1-dependent and -independent cytotoxicity of potent quinone Cdc25 phosphatase inhibitors
NAD(P)H:Quinone oxidoreductase-1-dependent and -independent cytotoxicity of potent quinone Cdc25 phosphatase inhibitors
复制标题
DOI：
10.1124/jpet.103.059477
复制
发表时间：
2004-04-01
影响因子：
3.5
通讯作者：
Pan, SS
中科院分区：
文献类型：
--
作者：
Han, YS;Shen, HM;Pan, SS
Cdc25双特异性磷酸酶协调细胞周期进程和细胞信号传导。因此，Cdc25抑制剂代表了潜在的抗癌药物。我们评估了100万种化合物对人Cdc25磷酸酶的抑制作用，并鉴定出许多有效的选择性抑制剂，它们都含有醌。生物还原酶经常解毒或激活醌。因此，我们评估了NAD(P)H：醌氧化还原酶-1 （NQO1）和富含还原酶的微粒体对三种含醌的Cdc25抑制剂的活性的影响：2-（2-羟乙基磺酰）-3-甲基-1,4-萘醌（Cpd 5，化合物5；NSC 672121）， 2,3-二-（2-羟乙基磺酰）-1,4-萘醌（NSC 95397）和6-氯-7-（2-morpholin-4-乙基乙胺）喹啉-5,8-二酮（NSC 663284）。每个抑制剂都被人NQO1 （K-m为0.3-0.5 muM）还原，但没有被微粒体还原。用6种含有不同NQO1量的癌细胞系对化合物进行评价：HT-29（1056 nmol/mg/min）、HCT116 （660 nmol/mg/min）、HCT116- r30a亚系（28 nmol/mg/min）和HCT-116R30A/NQ5亚系（934 nmol/mg/min）、MDA-MB-231/Q2（空NQO1）和MDA-MB-231/Q6亚系（124 nmol/mg/min），但含有相似量的微粒体细胞色素P450还原酶和细胞色素b(5)还原酶。Cpd - 5对HCT-116或HCT-116R30A/NQ5的抑制作用是HCT-116R30A/NQ5的4- 5倍，与nq1水平成正比。相比之下，在所有测试的细胞系中，无论NQO1水平如何，NSC 95375和NSC 663284的生长抑制和G2/M阻滞效果相似（平均IC50分别为1.3+/-0.3和2.6+/-0.4 muM）。NSC 95375和NSC 663284也引起类似的Cdk1过磷酸化，表明类似的Cdc25抑制作用。然而，在NQO1最小的亚系中，需要较低的cpd5浓度才能产生Cdk1过磷酸化。因此，nq01可能通过将cpd5还原为活性较低的对苯二酚来解毒cpd5，而NSC 95397-和NSC 663284产生的细胞毒性不受nq01的影响。
Cdc25 dual-specificity phosphatases coordinate cell cycle progression and cellular signaling. Consequently, Cdc25 inhibitors represent potential anticancer agents. We evaluated >10,000 compounds for inhibition of human Cdc25 phosphatases and identified many potent and selective inhibitors, which all contained a quinone. Bioreductive enzymes frequently detoxify or activate quinones. Therefore, we evaluated the effect of NAD(P)H: quinone oxidoreductase-1 (NQO1) and reductase-rich microsomes on the activity of three quinone-containing Cdc25 inhibitors: 2-(2-hydroxyethylsulfanyl)-3-methyl-1,4-naphthoquinone (Cpd 5, compound 5; NSC 672121), 2,3-bis-(2-hydroxyethylsulfanyl)-1,4-naphthoquinone (NSC 95397), and 6-chloro-7-(2-morpholin-4-ylethylamino) quinoline-5,8-dione (NSC 663284). Each inhibitor was reduced by human NQO1 (K-m of 0.3-0.5 muM) but none by microsomes. Compounds were evaluated with six cancer cell lines containing different amounts of NQO1: HT-29(1056 nmol/mg/min), HCT116 (660 nmol/mg/min), sublines HCT116-R30A (28 nmol/mg/min) and HCT-116R30A/NQ5 (934 nmol/mg/min), MDA-MB-231/Q2 (null NQO1), and subline MDA-MB-231/Q6 (124 nmol/mg/min) but containing similar amounts of microsomal cytochrome P450 reductase and cytochrome b(5) reductase. Growth inhibition and G2/M arrest by Cpd 5 was proportional to NQO1 levels, requiring 4- to 5-fold more Cpd 5 to inhibit HCT-116 or HCT-116R30A/NQ5 compared with HCT-116R30A. In contrast, in all tested cell lines irrespective of NQO1 level, growth inhibition and G2/M arrest by NSC 95375 and NSC 663284 were similar (average IC50 of 1.3+/-0.3 and 2.6+/-0.4 muM, respectively). NSC 95375 and NSC 663284 also caused similar Cdk1 hyperphosphorylation, indicating similar Cdc25 inhibition. However, lower Cpd 5 concentrations were needed to produce Cdk1 hyperphosphorylation in sublines with minimal NQO1. Thus, NQO1 detoxified Cpd 5, probably by reducing it to a less active hydroquinone, whereas NSC 95397- and NSC 663284-generated cytotoxicity was unaffected by NQO1.