中空纤维超滤法研究核苷类逆转录酶抑制药与DNA的结合率

目的:采用中空纤维超滤法研究核苷类逆转录酶抑制剂类药物与小牛胸腺DNA（ct-DNA）的相互结合率。方法:将中空纤维切成约15 cm小段,放入重蒸水中超声清洗15 s,晾干、备用。分别取阿德福韦（PMEA）、拉米夫定（LMVD）和恩曲他滨（FTC）与DNA摩尔比为1:1的混合液0.6 ml于一端封闭的细玻璃管中,玻璃管中再放入弯曲成U型的中空纤维,置离心机中离心(4 000 r·min^-1)60 min后,取出中空纤维,用微量进样器抽出中空纤维内液体约50μl,稀释至3.0 ml后分别测其最大紫外吸收。结果:PMEA、LMVD、FTC与DNA的平均结合率分别为40.7%、41.0%、38.6%。结论:确定了核苷类逆转录酶抑制剂类药物与DNA结合率的测定方法,可表征药物与DNA的结合强度。

Study on the Binding Rate of Nucleoside Reverse Transcriptase Inhibitors with DNA by Hollow Fiber Ultrafiltration

Objective ： To study the binding rate of the reverse transcriptase inhibitors with calf thymus DNA （ct-DNA） by hollow fiber ultrafihration. Method ：The hollow fiber was cut into 15 cm each piece, then put into distilled water and treated by ultrasound for 15 s and used after dring. The solution （adefovir/lamivudine/emtricitabine： DNA = 1：1 ） of 0. 6 ml was put into a thin glass tube with one side sealed. The glass tube with hollow fiber with U-shape was centrifuged with 4 000 r·min^ -1 for 60 mins. Then the hollow fiber was taken out and 50 μl of the filtrate was drew with the syringe. The filtrate was diluted to 3.0 ml for UV determination. Result： The average binding rates for adefovir, lamivudine and emtricitabine with DNA were 40. 7% ,41.0% and 38. 6%, respectively. Conclusion： A method has been established for determination of binding rate as well as binding stability between reverse transcriptase inhibitors and DNA.