Experimental study of searching for new antidepressant drugs by applying high throughput screening method

doi:10.3321/j.issn:1673-8225.2004.33.012

Abstract：BACKGROUND: The incidence of depression is increasing. It is of great significance to use new technologies and new methods to develop antidepressant drugs with high efficacy and low toxicity.OBJECTIVE: To establish research system for finding new antidepressant leading compounds by the combination of high throughput screening method and in vivo experiment.DESIGN: A randomized and controlled experiment.SETTING, MATERIALS and INTERVENTIONS: The experiment was carried out in the Drug Institute of Chinese Academy of Medical Science. Seventy Kunming male mice weighing(20 ±2) g, and 10 male Wistar rats weighing(200 ± 20) g were used. Wister rats were killed with decollation, and the 5-Hydroxythryptamine (5-HT) reuptake in rat brain synaptosomes was measured. The mice were randomly divided into control group, fluoxetine group(2.6 mg/kg), J01113 group(30 mg/kg), J01114 group(30 mg/kg)and J10745 group(30 mg/Kg). Continuous gastric lavage and administration lasted for 5 days, once per day, and mice hanging experiment was performed every other day since the second day. The mice were administrated and compared separately in 6 minutes after self-hanging in the administration and control group. Forced swimming test was made every other day and compared in 4 minutes of rats in the administration and control group.MAIN OUTCOME MEASURE: The 5-HT reuptake inhibition in synaptosomes and the immobility time in depression models.RESULTS: The effects of more than 5 000 compounds on 5-HT reuptake were tested with high throughput screening(HTS) in vitro. Three active compounds, J01113, J01114 and J10745 were selected to test their antidepressant effects in the forced swimming test and the tail suspension test in mice. J01113, J01114 and J10745 strongly inhibited 5-HT reuptake. The IC50 of J01113, J01114 and J10745 was 1. 304 × 10-10, 9. 036 × 10-10, and 1. 447 × 10-7 mol/L respectively. After the J01113 was administrated for 3 and 5 days, the immobility time of the hanging mice[(67.2 ±47.33),(95.2 ± 47.5 ) s] was significantly less than that of the control group [(117.8±33.2), (170.2±40.47) s](t=2.77, P ＜0.05; t=3.8, P＜ 0.01 ). Forced swimming test showed that the immobility time after 3 or 5 days of administration for the mice in J01113 [ (9.8 ± 16.59 ), ( 19.4 ± 24.64) s] was significantly lower than those of the control group [(63.7±35.4), (48.7±33.9)s] (t=4.36, P ＜0.01; t=2.21, P＜ 0.05). There was a decrease tendency in the immobility time of J01114 and J10745 group, which showed no significant difference( P ＞ 0.05).CONCLUSION: Large-scale and high-efficiency antidepressant lead compounds canhe found by HTS. J01113 may be a potential antidepressant drug.