Ace Inhibitors And 5 - Lox Inhibitor In Vitro Screening Model Research

In this study, taking test compounds from the three-dimensional pharmacophore model as a research object, in vitro screening technology was used to find the active compound. Main research contents and the results are as follows:UV was used to establish the screening model in vitro of ACE inhibitors. The IC50 of captopril tested was similar with the values in the literature. The results showed that the model was accurate, reliable, and could be applied to in vitro drug screening.HPLC was used to establish the screening model in vitro of ACE inhibitors. The IC50 of captopril tested was similar with the values in the literature. The results showed that the model was accurate, reliable, and could be applied to in vitro drug screening. The content of enzymolysis product hippuric acid was determined by RP-HPLC method. RP-HPLC was performed on a Brava BDS C18 column (250 mm×4.6 mm,5 um) using a mobile phase consisting of acetonitrile-0.01% formic acid (25:75) at 30℃with the flow rate of 1.0 mL·min-1 and detection wavelength at 228 nm. The calibration curve was linear in the range of 1.12-37.42μg·mL-1 (r=0.9999) for determination of hippuric acid.HPLC was used to establish the screening model in vitro of 5-LOX inhibitors. The IC50 of zileuton tested was similar with the values in the literature. The results showed that the model was accurate, reliable, and could be applied to in vitro drug screening. The content of LTB4 and 5-HETE, the main products of 5-LOX pathway of arachidonic acid metabolism, was determinded by RP-HPLC method. RP-HPLC was performed on a Brava BDS C18 column (250 mm×4.6 mm,5 um) using a mobile phase consisting of 0.02% acetic acid-methanol at 30℃with the flow rate of 0.8 mL·min-1 and detection wavelength at 275 nm and 235 nm. The calibration curve was linear in the range of 2.0-50.0μg·mL-1 (r=0.9999) for determination of LTB4, 5.0-125.0μg·mL-1 (r=1.0000) for determination of 5-HETE. The average recoveries of LTB4 and 5-HETE were 100.0%(RSD=1.1%) and 97.7%(RSD=1.5%), respectively. The intra-day and inter-day RSDs were 0.7%,1.1%and 0.3%,1.3%.The model of rabbit platelet aggregation which can be induced to release ofβ-glucosidase by PAF was established. The IC50 of Ginkgolide B tested was similar as the report values in the literature. The results showed that the model was accurate, reliable, and could be applied to in vitro drug screening.The model of P450 activity was established. The results showed 1.60×10-6-2.00×10-5 mol·L-1 Zileuton solution can induce CYP3A.