Soares Karina M, Blackmon Nicole, Shun Tong Ying, Shinde Sunita N, Takyi Harold K, Wipf Peter, Lazo John S, Johnston Paul A
Pittsburgh Molecular Library Screening Center, Drug Discovery Institute, School of Medicine, University of Pittsburgh, Pittsburgh, Pennsylvania 15260, USA.
Assay Drug Dev Technol. 2010 Apr;8(2):152-74. doi: 10.1089/adt.2009.0247.
We have screened the Library of Pharmacologically Active Compounds (LOPAC) and the National Institutes of Health (NIH) Small Molecule Repository (SMR) libraries in a horseradish peroxidase-phenol red (HRP-PR) H2O2 detection assay to identify redox cycling compounds (RCCs) capable of generating H2O2 in buffers containing dithiothreitol (DTT). Two RCCs were identified in the LOPAC set, the ortho-naphthoquinone beta-lapachone and the para-naphthoquinone NSC 95397. Thirty-seven (0.02%) concentration-dependent RCCs were identified from 195,826 compounds in the NIH SMR library; 3 singleton structures, 9 ortho-quinones, 2 para-quinones, 4 pyrimidotriazinediones, 15 arylsulfonamides, 2 nitrothiophene-2-carboxylates, and 2 tolyl hydrazides. Sixty percent of the ortho-quinones and 80% of the pyrimidotriazinediones in the library were confirmed as RCCs. In contrast, only 3.9% of the para-quinones were confirmed as RCCs. Fifteen of the 251 arylsulfonamides in the library were confirmed as RCCs, and since we screened 17,868 compounds with a sulfonamide functional group we conclude that the redox cycling activity of the arylsulfonamide RCCs is due to peripheral reactive enone, aromatic, or heterocyclic functions. Cross-target queries of the University of Pittsburgh Drug Discovery Institute (UPDDI) and PubChem databases revealed that the RCCs exhibited promiscuous bioactivity profiles and have populated both screening databases with significantly higher numbers of active flags than non-RCCs. RCCs were promiscuously active against protein targets known to be susceptible to oxidation, but were also active in cell growth inhibition assays, and against other targets thought to be insensitive to oxidation. Profiling compound libraries or the hits from screening campaigns in the HRP-PR H(2)O(2) detection assay significantly reduce the timelines and resources required to identify and eliminate promiscuous nuisance RCCs from the candidates for lead optimization.
我们在辣根过氧化物酶-酚红（HRP-PR）H₂O₂检测试验中，对药理活性化合物库（LOPAC）和美国国立卫生研究院（NIH）小分子储存库（SMR）进行了筛选，以鉴定能够在含有二硫苏糖醇（DTT）的缓冲液中产生H₂O₂的氧化还原循环化合物（RCC）。在LOPAC库中鉴定出两种RCC，即邻萘醌β-拉帕醌和对萘醌NSC 95397。从NIH SMR库中的195,826种化合物中鉴定出37种（0.02%）浓度依赖性RCC；3种单结构、9种邻醌、2种对醌、4种嘧啶并三嗪二酮、15种芳基磺酰胺、2种硝基噻吩-2-羧酸盐和2种甲苯酰肼。库中60%的邻醌和80%的嘧啶并三嗪二酮被确认为RCC。相比之下，只有3.9%的对醌被确认为RCC。库中251种芳基磺酰胺中的15种被确认为RCC，并且由于我们筛选了17,868种具有磺酰胺官能团的化合物，我们得出结论，芳基磺酰胺RCC的氧化还原循环活性归因于外围反应性烯酮、芳香族或杂环官能团。匹兹堡大学药物发现研究所（UPDDI）和PubChem数据库的交叉目标查询显示，RCC表现出混杂的生物活性谱，并且与非RCC相比，在两个筛选数据库中具有显著更多的活性标记。RCC对已知易受氧化影响的蛋白质靶标具有混杂活性，但在细胞生长抑制试验中也具有活性，并且对其他被认为对氧化不敏感的靶标也有活性。在HRP-PR H₂O₂检测试验中对化合物库或筛选活动的命中物进行分析，可显著缩短从潜在药物优化候选物中鉴定和消除混杂的有害RCC所需的时间和资源。
