D'Cruz O J, Venkatachalam T K, Zhu Z, Shih M J, Uckun F M
Drug Discovery Program, Wayne Hughes Institute, St. Paul, Minnesota 55113, USA.
Biol Reprod. 1998 Sep;59(3):503-15. doi: 10.1095/biolreprod59.3.503.
Detergent-based vaginal microbicides, in addition to their high contraceptive failure rates, cause mucosal erosion and local inflammation that might increase the risk of heterosexual human immunodeficiency virus (HIV) transmission. In a systematic effort to identify a microbicide contraceptive potentially capable of preventing the sexual transmission of HIV as well as providing fertility control, a series of novel aryl phosphate derivatives of 5-bromo-6-methoxy-3'-azido-3'-deoxythymidine (AZT; zidovudine) were synthesized and examined for dual anti-HIV and sperm-immobilizing activity (SIA). Whereas AZT displayed potent anti-HIV activity (IC50 = 0.006 microM) but lacked SIA (EC50 > 300 microM), two 5-bromo-6-methoxy-aryl phosphate derivatives of AZT, compounds WHI-05 and WHI-07, exhibited potent anti-HIV activity as well as SIA. The IC50 (HIV) and EC50 (SIA) values for WHI-07 were 439-fold and 13.5-fold lower, respectively, than those for the detergent-based virucidal spermicide, nonoxynol-9 (N-9). Sperm motion kinematics using computer-assisted sperm motion analysis combined with confocal laser scanning microscopy, high-resolution low-voltage scanning, and transmission electron microscopy demonstrated that both WHI-05 and WHI-07 cause a complete and irreversible loss of sperm motility in a concentration- and time-dependent fashion without concomitantly affecting the sperm acrosomal membrane integrity. In experiments designed to assess the fertilizing capacity of treated sperm, preincubation of sperm with either compound resulted in a concentration-dependent loss of the ability to adhere to and penetrate zona-free hamster eggs as well as inhibition of binding to human zona. WHI-07 applied intravaginally prior to artificial insemination of epididymal sperm drastically reduced fertility in hormonally primed CD-1 mice. Unlike the intravaginal application of N-9, repetitive intravaginal application of WHI-07 did not damage the vaginal epithelium or cause local inflammation. Structure-function relationship analyses showed that the addition of bromo-methoxy functional groups to AZT was essential for, and the aryl phosphate derivatization contributory to, the SIA of both compounds. Compounds WHI-05 and WHI-07 may be useful as dual-function vaginal contraceptives for women who are at high risk for acquiring HIV/acquired immunodeficiency virus syndrome by heterosexual vaginal transmission.
基于洗涤剂的阴道杀微生物剂，除了避孕失败率高之外，还会导致粘膜糜烂和局部炎症，这可能会增加异性间人类免疫缺陷病毒（HIV）传播的风险。为了系统地寻找一种有可能预防HIV性传播并提供生育控制的杀微生物剂避孕药，合成了一系列5-溴-6-甲氧基-3'-叠氮基-3'-脱氧胸苷（AZT；齐多夫定）的新型芳基磷酸酯衍生物，并检测其双重抗HIV和精子固定活性（SIA）。虽然AZT显示出强大的抗HIV活性（IC50 = 0.006 microM）但缺乏SIA（EC50 > 300 microM），但AZT的两种5-溴-6-甲氧基芳基磷酸酯衍生物，即化合物WHI-05和WHI-07，表现出强大的抗HIV活性以及SIA。WHI-07的IC50（HIV）和EC50（SIA）值分别比基于洗涤剂的杀病毒杀精剂壬苯醇醚-9（N-9）低439倍和13.5倍。使用计算机辅助精子运动分析结合共聚焦激光扫描显微镜、高分辨率低电压扫描和透射电子显微镜进行的精子运动运动学研究表明，WHI-05和WHI-07均以浓度和时间依赖性方式导致精子运动完全且不可逆地丧失，同时不影响精子顶体膜的完整性。在旨在评估处理后精子受精能力的实验中，精子与任何一种化合物预孵育都会导致其粘附并穿透无透明带仓鼠卵的能力以及与人透明带结合的能力呈浓度依赖性丧失。在附睾精子人工授精前经阴道施用WHI-07可显著降低激素预处理的CD-1小鼠的生育力。与经阴道施用N-9不同，重复经阴道施用WHI-07不会损害阴道上皮或引起局部炎症。结构-功能关系分析表明，向AZT添加溴甲氧基官能团对于两种化合物的SIA至关重要，而芳基磷酸酯衍生化对其有促进作用。对于通过异性阴道传播感染HIV/获得性免疫缺陷综合征风险较高的女性，化合物WHI-05和WHI-07可能可用作双功能阴道避孕药。
