Graves Tiffany L, Scott John E
Department of Pharmaceutical Sciences, Biomanufacturing Research Institute and Technology Enterprise, North Carolina Central University, 1801 Fayetteville Street, Durham, NC 27707, USA.
Curr Chem Genomics. 2008 Nov 26;2:51-61. doi: 10.2174/1875397300802010051.
PON1 has been demonstrated to be the serum enzyme responsible for detoxifying organophosphate chemical weapons and plays a protective role against atherosclerosis. In order to identify small molecules that modulate PON1 activity in serum, we developed a high throughput kinetic absorbance assay using mouse serum and the organophosphate paraoxon. The IC(50) value obtained for the known PON1 inhibitor, 2-hydroxyquinoline, matched the value reported for purified PON1. A compound library was screened resulting in no confirmed activators, but 12 confirmed inhibitors. Seven of these hits also inhibited purified human PON1. One compound was only two-fold less potent than 2-hydroxyquinoline in the serum assay, but 10-fold more potent against purified PON1. This compound (IC(50) = 420 nM) may be useful towards a chemical probe for PON1. Therefore, this assay has utility as a high throughput assay for discovery of small molecule modulators of PON1 activity that maintain activity in serum.
已证实对氧磷酶1（PON1）是负责解毒有机磷化学武器的血清酶，并对动脉粥样硬化起保护作用。为了鉴定能调节血清中PON1活性的小分子，我们利用小鼠血清和有机磷酸酯对氧磷开发了一种高通量动力学吸光度测定法。已知的PON1抑制剂2-羟基喹啉的半数抑制浓度（IC50）值与纯化的PON1报道值相符。对一个化合物文库进行了筛选，结果未发现确认的激活剂，但有12种确认的抑制剂。其中7种命中化合物也抑制纯化的人PON1。有一种化合物在血清测定中的效力仅比2-羟基喹啉低两倍，但对纯化的PON1的效力高10倍。这种化合物（IC50 = 420 nM）可能对作为PON1的化学探针有用。因此，该测定法可作为一种高通量测定法，用于发现能在血清中保持活性的PON1活性小分子调节剂。
