Huth Jeffrey R, Song Danying, Mendoza Renaldo R, Black-Schaefer Candice L, Mack Jamey C, Dorwin Sarah A, Ladror Uri S, Severin Jean M, Walter Karl A, Bartley Diane M, Hajduk Philip J
Global Pharmaceutical Research and Development, Abbott Laboratories, Abbott Park, IL 60064, USA.
Chem Res Toxicol. 2007 Dec;20(12):1752-9. doi: 10.1021/tx700319t. Epub 2007 Nov 15.
We have recently reported on the development of a La assay to detect reactive molecules by nuclear magnetic resonance (ALARM NMR) to detect reactive false positive hits from high-throughput screening, in which we observed a surprisingly large number of compounds that can oxidize or form covalent adducts with protein thiols groups. In the vast majority of these cases, the covalent interactions are largely nonspecific (e.g., affect many protein targets) and therefore unsuitable for drug development. However, certain thiol-reactive species do appear to inhibit the target of interest in a specific manner. The question then arises as to the potential toxicology risks of developing a drug that can react with protein thiol groups. Here, we report on the evaluation of a large set of ALARM-reactive and -nonreactive compounds against a panel of additional proteins (aldehyde dehydrogenase, superoxide dismutase, and three cytochrome P450 enzymes). It was observed that ALARM-reactive compounds have significantly increased risks of interacting with one or more of these enzymes in vitro. Thus, ALARM NMR seems to be a sensitive tool to rapidly identify compounds with an enhanced risk of producing side effects in humans, including alcohol intolerance, the formation of reactive oxygen species, and drug-drug interactions. In conjunction with other toxicology assays, ALARM NMR should be a valuable tool for prioritizing compounds for lead optimization and animal testing.
我们最近报道了一种用于通过核磁共振检测反应性分子的检测方法（ALARM NMR），以检测高通量筛选中出现的反应性假阳性结果，在此过程中我们观察到大量能够氧化或与蛋白质硫醇基团形成共价加合物的化合物。在绝大多数此类情况下，共价相互作用在很大程度上是非特异性的（例如，影响许多蛋白质靶点），因此不适用于药物开发。然而，某些硫醇反应性物质似乎确实以特定方式抑制了感兴趣的靶点。那么问题就出现了，即开发一种能够与蛋白质硫醇基团发生反应的药物可能存在的毒理学风险。在此，我们报告了针对一组其他蛋白质（醛脱氢酶、超氧化物歧化酶和三种细胞色素P450酶）对大量ALARM反应性和非反应性化合物的评估。据观察，ALARM反应性化合物在体外与这些酶中的一种或多种相互作用的风险显著增加。因此，ALARM NMR似乎是一种灵敏的工具，可快速识别在人体内产生副作用风险增加的化合物，包括酒精不耐受、活性氧的形成以及药物相互作用。与其他毒理学检测方法相结合，ALARM NMR应该是一种有价值的工具，可用于对化合物进行优先级排序，以进行先导化合物优化和动物试验。
