Activity against human immunodeficiency virus type 1, intracellular metabolism, and effects on human DNA Polymerases of 4′-ethynyl-2-fluoro-2′-deoxyadenosine
Activity against human immunodeficiency virus type 1, intracellular metabolism, and effects on human DNA Polymerases of 4′-ethynyl-2-fluoro-2′-deoxyadenosine
复制标题
DOI：
10.1128/aac.00277-07
复制
发表时间：
2007-08-01
影响因子：
4.9
通讯作者：
Mitsuya, Hiroaki
中科院分区：
文献类型：
--
作者：
Nakata, Hirotorao;Amano, Masayuki;Mitsuya, Hiroaki
我们研究了核苷逆转录酶抑制剂4 '-乙炔基-2-氟-2'-脱氧腺苷（EFdA）对人类免疫缺陷病毒1型（HIV-1）的胞浆内抑制和抗病毒活性动力学，EFdA对野生型和多药耐药HIV-1株具有强效活性。当CEM细胞暴露于0.1 μ M [H-3]EFdA或[H-3] 3 '-叠氮基-2'，3 '-双脱氧胸苷（AZT）6 h时，细胞内EFdA-三磷酸（TP）水平为91.6pmol/10（9）个细胞，而AZT为396.5pmol/10（9）个细胞。当CEM细胞暴露于10 μ M [3] HEFdA时，EFdA-TP的量增加22倍（2，090 pmol/10（9）细胞），而[H-3]AZT-TP的量仅适度增加2.4倍（970 pmol/10（9）细胞）。EFdA-TP和AZT-TP的细胞内半衰期值分别与17和3 h相似。当MT-4细胞用0.01 μ M EFdA培养24小时，彻底洗涤以除去EFdA，在没有EFdA的情况下进一步培养不同的时间段，暴露于HIV-1（NL 4 -3），并再培养5天时，在没有药物孵育的情况下培养24和48小时后，保护值分别为75%和47%，而1 μ M AZT组分别为55%和9.2%。EFdA-TP对人聚合酶α、β和γ的50%抑制浓度值分别为>100 μ M、>100 μ M和10 μ M，而ddA-TP的50%抑制浓度值分别为>100 μ M、0.2 μ M和0.2 μ M。这些数据保证了EFdA作为一种潜在的治疗药物，用于那些携带野生型HIV-1和/或多药耐药变异体的患者的进一步开发。
We examined the intracytoplasmic anabolism and kinetics of antiviral activity against human immunodeficiency virus type 1 (HIV-1) of a nucleoside reverse transcriptase inhibitor, 4'-ethynyl-2-fiuoro-2'-deoxyadenosine (EFdA), which has potent activity against wild-type and multidrug-resistant HIV-1 strains. When CEM cells were exposed to 0.1 mu M [H-3]EFdA or [H-3]3'-azido-2',3'-dideoxythymidine (AZT) for 6 h, the intracellular EFdA-triphosphate (TP) level was 91.6 pmol/10(9) cells, while that of AZT was 396.5 pmol/10(9) cells. When CEM cells were exposed to 10 mu M [(3)HEFdA, the amount of EFdA-TP increased by 22-fold (2,090 pmol/10(9) cells), while the amount of [H-3]AZT-TP increased only moderately by 2.4-fold (970 pmol/10(9) cells). The intracellular half-life values of EFdA-TP and AZT-TP were similar to 17 and similar to 3 h, respectively. When MT-4 cells were cultured with 0.01 mu M EFdA for 24 h, thoroughly washed to remove EFdA, further cultured without EFdA for various periods of time, exposed to HIV-1(NL4-3),and cultured for an additional 5 days, the protection values were 75 and 47%, respectively, after 24 and 48 h with no drug incubation, while those with 1 mu M AZT were 55 and 9.2%, respectively. The 50% inhibitory concentration values of EFdA-TP against human polymerases alpha, beta, and gamma were >100 mu M, >100 mu M, and 10 mu M, respectively, while those of ddA-TP were >100 mu M, 0.2 mu M, and 0.2 mu M, respectively. These data warrant further development of EFdA as a potential therapeutic agent for those patients who harbor wild-type HIV-1 and/or multidrug-resistant variants.