Dewald B, Baggiolini M
Biochem Pharmacol. 1987 Aug 1;36(15):2505-10. doi: 10.1016/0006-2952(87)90523-5.
This paper describes a testing model for the detection and evaluation of PAF antagonists, based on the inhibition of PAF-elicited elastase release by human neutrophils. Incubations are performed in microtiter plates in the presence of a specific fluorogenic elastase substrate allowing direct measurement of the exocytosis response by means of a 96-well fluorescence reader. Determinations of the IC50 values for five established PAF antagonists, Ro 19-3704, BN 52021, CV-3988, 48740 RP and kadsurenone, showed that the new model is comparable in sensitivity and discriminative capacity to other in vitro assays. From the effect of antagonists on the PAF concentration-response curve pA2 values could be calculated and information on the type of antagonism obtained. BN 52021 was found to behave as a competitive antagonist while Ro 19-3704 showed a more complex type of inhibition. As a one-plate system, the test is simple to handle and highly reproducible, and appears therefore particularly useful for large drug screening programs.
本文描述了一种基于抑制人中性粒细胞PAF诱导的弹性蛋白酶释放来检测和评估PAF拮抗剂的测试模型。在微量滴定板中进行孵育，同时存在一种特定的荧光弹性蛋白酶底物，可通过96孔荧光读数器直接测量胞吐反应。对五种已确立的PAF拮抗剂Ro 19 - 3704、BN 52021、CV - 3988、48740 RP和海风藤酮的IC50值测定表明，新模型在灵敏度和鉴别能力方面与其他体外试验相当。根据拮抗剂对PAF浓度 - 反应曲线的影响，可以计算出pA2值，并获得有关拮抗类型的信息。发现BN 52021表现为竞争性拮抗剂，而Ro 19 - 3704表现出更复杂的抑制类型。作为一种单板系统，该测试操作简单且具有高度可重复性，因此对于大型药物筛选项目显得特别有用。
