D'Cruz O J, Zhu Z, Yiv S H, Chen C L, Waurzyniak B, Uckun F M
Department of Reproductive Biology, Hughes Institute, St. Paul, Minnesota 55113, USA.
Contraception. 1999 May;59(5):319-31. doi: 10.1016/s0010-7824(99)00041-4.
Heterosexual transmission of HIV to women is the fastest-growing mode of transmission. In a systematic effort to develop a microbicide capable of preventing HIV transmission as well as providing fertility control, novel phenyl phosphate derivatives of 3'-azido-3'-deoxythymidine (zidovudine, ZDV) have been identified that exhibit potent anti-HIV and spermicidal activities. This study reports the synthesis, characterization, and preclinical formulation of compound WHI-05, 5-bromo-6-methoxy-5,6-dihydro-3'-azidothymidine-5'-(p-methoxyphenyl) methoxyalaninyl phosphate. The anti-HIV activities of WHI-05 and ZDV were compared by measuring p24 antigen production and reverse transcriptase activity as markers of viral replication using human peripheral blood mononuclear cells (PBMC) infected with both ZDV-sensitive and ZDV-resistant strains of HIV. The sperm immobilizing activity (SIA) of WHI-05 was compared with that of ZDV and nonoxynol-9 (N-9) by computer-assisted sperm analysis (CASA). The effect of WHI-05 on sperm membrane integrity was examined by high resolution, low voltage scanning electron microscopy (HR-LVSEM). The in vitro cytotoxicity profile of WHI-05 versus N-9 were compared using normal human vaginal, ectocervical, and endocervical epithelial cells. The in vivo vaginal tolerance, absorption, and toxicity of a 2% WHI-05 gel-microemulsion was tested in the rabbit. Whereas ZDV displayed potent anti-HIV activity but lacked SIA, WHI-05 elicited both potent anti-HIV activity and SIA. WHI-05 inhibited the replication of ZDV-sensitive as well as ZDV-resistant strains of HIV in PBMC. CASA combined with HR-LVSEM demonstrated that WHI-05-induced SIA was not associated with membrane damage. Unlike, N-9, the spermicidal activity of WHI-05 was not associated with cytotoxicity to reproductive tract epithelial cells. Repetitive intravaginal application of a 2% WHI-05 gel-microemulsion did not damage the vaginal epithelium or cause local inflammation in the rabbit model. As a potent anti-HIV agent that has spermicidal activity and is devoid of mucosal toxicity, WHI-05 shows a unique clinical potential to become the active ingredient for a vaginal contraceptive for women who are at high risk for acquiring HIV by heterosexual vaginal transmission.
艾滋病毒向女性的异性传播是增长最快的传播方式。为了系统地研发一种既能预防艾滋病毒传播又能控制生育的杀微生物剂，已鉴定出新型的3'-叠氮基-3'-脱氧胸苷（齐多夫定，ZDV）苯基磷酸酯衍生物，它们具有强大的抗艾滋病毒和杀精活性。本研究报告了化合物WHI-05（5-溴-6-甲氧基-5,6-二氢-3'-叠氮胸苷-5'-(对甲氧基苯基)甲氧基丙氨酰磷酸酯）的合成、表征及临床前制剂研究。通过测量p24抗原产生量和逆转录酶活性作为病毒复制的标志物，使用感染了对ZDV敏感和耐药的艾滋病毒株的人外周血单核细胞（PBMC），比较了WHI-05和ZDV的抗艾滋病毒活性。通过计算机辅助精子分析（CASA）比较了WHI-05与ZDV及壬苯醇醚-9（N-9）的精子固定活性（SIA）。通过高分辨率、低电压扫描电子显微镜（HR-LVSEM）检查了WHI-05对精子膜完整性的影响。使用正常人阴道、宫颈外口和宫颈内膜上皮细胞比较了WHI-05与N-9的体外细胞毒性特征。在兔体内测试了2% WHI-05凝胶微乳剂的阴道耐受性、吸收情况及毒性。ZDV显示出强大的抗艾滋病毒活性，但缺乏SIA，而WHI-05既具有强大的抗艾滋病毒活性又具有SIA。WHI-05抑制了PBMC中对ZDV敏感及耐药的艾滋病毒株的复制。CASA与HR-LVSEM相结合表明，WHI-05诱导的SIA与膜损伤无关。与N-9不同，WHI-05的杀精活性与对生殖道上皮细胞的细胞毒性无关。在兔模型中，重复阴道内应用2% WHI-05凝胶微乳剂不会损伤阴道上皮或引起局部炎症。作为一种具有杀精活性且无黏膜毒性的强效抗艾滋病毒药物，WHI-05显示出独特的临床潜力，有望成为通过异性阴道传播感染艾滋病毒风险较高的女性阴道避孕药的活性成分。
