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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。
OBJECTIVE: To study the interaction between nucleoside reverse transcriptase inhibitors and calf thymus DNA (ct-DNA) using hollow fiber ultrafiltration. Methods: The hollow fiber was cut into small pieces of about 15 cm, placed in distilled water ultrasonic cleaning 15 s, dry, spare. Take a mixed solution of adefovir (PMEA), lamivudine (LMVD) and emtricitabine (FTC) and DNA molar ratio of 1: 1 0.6 ml in one end closed fine glass tube, glass tube Then, the hollow fiber bent into a U-shape was put into the centrifuge and centrifuged (4 000 r · min -1) for 60 min. After that, the hollow fiber was taken out and about 50 μl of the hollow fiber liquid was extracted by a microinjector. To 3.0 ml were measured after the maximum UV absorption. Results: The average binding rates of PMEA, LMVD, FTC and DNA were 40.7%, 41.0% and 38.6% respectively. Conclusion: The method for determining the binding rate of nucleoside reverse transcriptase inhibitors to DNA has been established, which can characterize the binding strength between drugs and DNA.