Department of Pharmaceutical Sciences, School of Pharmacy, South University, 709 Mall Boulevard, Savannah, GA 31406, USA.
Department of Biomedical and Pharmaceutical Sciences, School of Pharmacy, University of Rhode Island, Kingston, RI 02881, USA.
Molecules. 2022 May 23;27(10):3352. doi: 10.3390/molecules27103352.
A number of 5′-O-fatty acyl derivatives of 3′-fluoro-2′,3′-dideoxythymidine (FLT, 1) were synthesized. These conjugates were evaluated for their potential as topical microbicides with anti-HIV activity against cell-free (X4 and R5), cell-associated, and multidrug-resistant viruses. Compared to FLT and 3′-azido-2′,3′-dideoxythymidine (AZT), 5′-O-(12-azidododecanoyl) (5), 5′-O-myristoyl (6), and 5′-O-(12-thioethyldodecanoyl) (8) derivatives of FLT were found to be more active against both cell-free viruses (lymphocytotropic and monocytotropic strains) with EC50 values of 0.4 μM, 1.1 μM, and <0.2 μM, respectively, as well as cell-associated virus with EC50 values of 12.6, 6.4, and 2.3 μM, respectively. Conjugates 5, 6, and 8 exhibited >4 and >30 times better antiviral index than FLT and AZT, respectively. Conjugates 5 and 8 were significantly more potent than FLT against many multidrug-resistant strains. A comparison of the anti-HIV activity with the corresponding non-hydrolyzable ether conjugates suggested that ester hydrolysis to FLT and fatty acids is critical to enable anti-HIV activity. Cellular uptake studies were conducted using fluorescent derivatives of FLT attached with 5(6)-carboxyfluorescein through either β-alanine (23) or 12-aminododecanoic acid (24) spacers. The lipophilic fluorescent analog with a long chain (24) showed more than 12 times higher cellular uptake profile than the fluorescent analog with a short chain (23). These studies further confirmed that the attachment of fatty acids improved the cellular uptake of nucleoside conjugates. In addition, 5, 6, and 8 were the least cytotoxic and did not alter vaginal cell and sperm viability compared to the positive control, a commercial topical spermicide (N-9), which significantly decreased sperm and vaginal cell viability inducing the generation of proinflammatory cytokines.
合成了一系列 5′-O-脂肪酰基-3′-氟-2′,3′-二脱氧胸苷（FLT，1）衍生物。这些缀合物被评估为具有抗 HIV 活性的局部杀微生物剂，对无细胞（X4 和 R5）、细胞相关和多药耐药病毒具有活性。与 FLT 和 3′-叠氮-2′,3′-二脱氧胸苷（AZT）相比，FLT 的 5′-O-（12-叠氮十二烷酰基）（5）、5′-O-肉豆蔻酰基（6）和 5′-O-（12-硫代乙基十二烷酰基）（8）衍生物对无细胞病毒（淋巴细胞性和单核细胞性株）更具活性，EC50 值分别为 0.4 μM、1.1 μM 和<0.2 μM，对细胞相关病毒的 EC50 值分别为 12.6、6.4 和 2.3 μM。缀合物 5、6 和 8 的抗病毒指数分别比 FLT 和 AZT 高 4 倍和 30 倍以上。与 FLT 和 AZT 相比，缀合物 5 和 8 对许多多药耐药株的活性更强。与相应的非水解醚缀合物的抗 HIV 活性比较表明，酯水解为 FLT 和脂肪酸对于发挥抗 HIV 活性至关重要。使用通过β-丙氨酸（23）或 12-氨基十二烷酸（24）间隔臂连接 5（6）-羧基荧光素的 FLT 的荧光衍生物进行了细胞摄取研究。带有长链（24）的亲脂性荧光类似物的细胞摄取谱比带有短链（23）的荧光类似物高 12 倍以上。这些研究进一步证实，脂肪酸的连接提高了核苷缀合物的细胞摄取。此外，与阳性对照（一种商业局部杀精子剂 N-9）相比，5、6 和 8 的细胞毒性最低，且不改变阴道细胞和精子活力，而 N-9 显著降低精子和阴道细胞活力，诱导促炎细胞因子的产生。
