Wang Peiyuan, Rachakonda Suguna, Zennou Veronique, Keilman Meg, Niu Congrong, Bao Donghui, Ross Bruce S, Furman Phillip A, Otto Michael J, Sofia Michael J
Pharmasset, Inc., Princeton, NJ, USA.
Antivir Chem Chemother. 2012 May 14;22(5):217-38. doi: 10.3851/IMP2079.
Nucleoside reverse transcriptase inhibitors (NRTIs) are an effective class of agents that has played a vital role in the treatment of HIV infections. (-)-β-D-(2R,4R)-dioxolane-thymine (DOT) is a thymidine analogue that is active against wild-type and NRTI-resistant HIV-1 mutants. It has been shown that the anti-HIV activity of DOT is limited due to poor monophosphorylation.
To further enhance the anti-HIV activity of DOT, an extensive structure-activity relationship analysis of phosphoramidate prodrugs of DOT monophosphate was undertaken. These prodrugs were evaluated for anti-HIV activity using Hela CD4 β-gal reporter cells (P4-CCR5 luc cells).
Among the synthesized prodrugs, the 4-bromophenyl benzyloxy l-alanyl phosphate derivative of DOT was the most potent, with a 50% effective concentration of 0.089 μM corresponding to a 75-fold increase in activity relative to the parent nucleoside DOT with no increased cytotoxicity. The metabolic stability of a selected number of potent DOT phosphoramidates was also evaluated in simulated gastric fluid, simulated intestinal fluid, human plasma and liver S9 fractions.
A series of new phosphoramidate prodrugs of DOT were prepared and evaluated as inhibitors of HIV replication in vitro. Metabolic stability studies indicated that these DOT phosphoramidate derivatives have the potential to show acceptable stability in the gastrointestinal tract, but they metabolize rapidly in the liver.
核苷类逆转录酶抑制剂（NRTIs）是一类有效的药物，在治疗HIV感染中发挥了至关重要的作用。（-）-β-D-（2R，4R）-二氧戊环胸腺嘧啶（DOT）是一种胸腺嘧啶类似物，对野生型和耐NRTI的HIV-1突变体具有活性。已表明，由于单磷酸化效果不佳，DOT的抗HIV活性受到限制。
为进一步增强DOT的抗HIV活性，对DOT单磷酸的氨基磷酸酯前药进行了广泛的构效关系分析。使用Hela CD4β-半乳糖苷酶报告细胞（P4-CCR5 luc细胞）评估这些前药的抗HIV活性。
在合成的前药中，DOT的4-溴苯基苄氧基-L-丙氨酰磷酸衍生物活性最强，50%有效浓度为0.089μM，相对于母体核苷DOT，活性增加了75倍，且细胞毒性未增加。还在模拟胃液、模拟肠液、人血浆和肝脏S9组分中评估了选定的几种强效DOT氨基磷酸酯的代谢稳定性。
制备了一系列新的DOT氨基磷酸酯前药，并评估了它们作为HIV复制体外抑制剂的活性。代谢稳定性研究表明，这些DOT氨基磷酸酯衍生物在胃肠道中可能具有可接受的稳定性，但在肝脏中代谢迅速。
