多羟基芳香族化合物对HIV-1整合酶的抑制作用

STUDIES ON THE INHIBITION OF POLYHYDROXYLATED AROMATIC COMPOUNDS AGAINST HIV-1 INTEGRASE

目的 研究HIV 1整合酶抑制剂 ,为艾滋病的治疗提供新作用靶位的抗HIV药物。方法 用HIV 1整合酶ELISA法检测 3种萘醌类化合物 ,10种白藜芦醇及其衍生物和 7种吡喃香豆素类化合物对整合酶的抑制作用。结果 双羟基 1,4 萘醌 (NQ 2 )对HIV 1整合酶有抑制活性 ,IC50 为 78 5 μmol·L- 1 ,发现萘醌类新化合物NQ 3对HIV 1整合酶的抑制作用优于NQ 2 ,IC50 为 3 7 2 μmol·L- 1 。用分步测定法发现NQ 2主要抑制HIV 1整合酶的链转移活性 ,而NQ 3则对装配和链转移都有较强的抑制。结论 萘醌类化合物 (NQ 2 ,3 )对HIV 1整合酶有抑制作用 ,NQ

AIM Three major enzymes of HIV 1, reverse transcriptase (RT), protease (PR), and integrase (IN), are important targets for anti HIV drugs. Nine RT and five PR inhibitors have been effectively used in treatment of AIDS patients. In order to find active integrase inhibitors, twenty polyhydroxylated aromatic compounds were tested. METHODS ELISA method was used to test the integrase activity. The synthesized donor substrate oligonucleotide representing the HIV 1 U5LTR was immobilized onto Covalink polystyrene microtiter plates, and a synthesized biotinlated 20 bp oligonucleotide was used as the target substrate. The products were detected and quantified by a colorimetric avidin linked alkaline phosphatase reporter system. RESULTS Compound NQ 2 was found to inhibit HIV 1 integrase with the IC 50 of 78 5 μmol·L -1 by ELISA method. Its novel analogue NQ 3 was found to be 2 fold more potent on HIV intrgrase than NQ 2, IC 50 was 37 2 μmol·L -1 . The IC 50 s of NQ 2 and NQ 3 to inhibit the 3′ pro+assembly activity of integrase were 96 94 μmol·L -1 and 8 48 μmol·L -1 ; to inhibit assembly activity were 168 and 6 9 μmol·L -1 and to inhibit strand transfer activity were 49 8 and 1 1 μmol·L -1 , respectively. Compound NQ 2 mostly inhibited the strand transfer activity of HIV 1 integrase. Compound NQ 3 inhibited both the assembly and strand transfer with high activities. CONCLUSION Naphthoquinone compound NQ 3 was found to be a novel HIV integrase inhibitor which warrants further study. Uncoupled ELISA HIV integrase assay is shown to be useful to screen HIV 1 integrase inhibitors.