Biomolecular Measurement Division, National Institute of Standards and Technology, Gaithersburg, Maryland 20899, United States.
Department of Chemistry, ChEM-H Institute and Stanford Cancer Institute, Stanford University, Stanford, California 94305, United States.
ACS Chem Biol. 2021 Jan 15;16(1):45-51. doi: 10.1021/acschembio.0c00877. Epub 2020 Dec 17.
DNA glycosylases involved in the first step of the DNA base excision repair pathway are promising targets in cancer therapy. There is evidence that reduction of their activities may enhance cell killing in malignant tumors. Recently, two tetrahydroquinoline compounds named SU0268 and SU0383 were reported to inhibit OGG1 for the excision of 8-hydroxyguanine. This DNA repair protein is one of the major cellular enzymes responsible for excision of a number of oxidatively induced lesions from DNA. In this work, we used gas chromatography-tandem mass spectrometry with isotope-dilution to measure the excision of not only 8-hydroxyguanine but also that of the other major substrate of OGG1, i.e., 2,6-diamino-4-hydroxy-5-formamidopyrimidine, using genomic DNA with multiple purine- and pyrimidine-derived lesions. The excision of a minor substrate 4,6-diamino-5-formamidopyrimidine was also measured. Both SU0268 and SU0383 efficiently inhibited OGG1 activity for these three lesions, with the former being more potent than the latter. Dependence of inhibition on concentrations of SU0268 and SU0383 from 0.05 μmol/L to 10 μmol/L was also demonstrated. The approach used in this work may be applied to the investigation of OGG1 inhibition by SU0268 and SU0383 and other small molecule inhibitors in further studies including cellular and animal models of disease.
参与 DNA 碱基切除修复途径第一步的 DNA 糖苷酶是癌症治疗中很有前途的靶点。有证据表明,降低它们的活性可能会增强恶性肿瘤中的细胞杀伤。最近,两种四氢喹啉化合物,分别称为 SU0268 和 SU0383,被报道可以抑制 OGG1 切除 8-羟基鸟嘌呤。这种 DNA 修复蛋白是负责从 DNA 中切除许多氧化诱导损伤的主要细胞酶之一。在这项工作中,我们使用气相色谱-串联质谱法(GC-MS/MS),使用同位素稀释法,不仅测量了 8-羟基鸟嘌呤的切除,还测量了 OGG1 的另一种主要底物,即 2,6-二氨基-4-羟基-5-甲酰胺嘧啶的切除,使用含有多种嘌呤和嘧啶衍生损伤的基因组 DNA。还测量了一个次要底物 4,6-二氨基-5-甲酰胺嘧啶的切除。SU0268 和 SU0383 都能有效地抑制这三种损伤的 OGG1 活性,前者比后者更有效。还证明了抑制作用对 SU0268 和 SU0383 浓度从 0.05 μmol/L 到 10 μmol/L 的依赖性。这项工作中使用的方法可应用于 SU0268 和 SU0383 以及其他小分子抑制剂对 OGG1 抑制的进一步研究,包括疾病的细胞和动物模型。