Department of Refractory Viral Infections, National Center for Global Health and Medicine Research Institute, Tokyo, Japan.
Experimental Retrovirology Section, HIV and AIDS Malignancy Branch, National Cancer Institute, National Institutes of Health, Bethesda, Maryland, USA.
Antimicrob Agents Chemother. 2019 May 24;63(6). doi: 10.1128/AAC.02635-18. Print 2019 Jun.
We generated two novel nonpeptidic HIV-1 protease inhibitors (PIs), GRL-001-15 and GRL-003-15, which contain unique crown-like tetrahydropyranofuran (Crn-THF) and P2'-cyclopropyl-aminobenzothiazole (Cp-Abt) moieties as P2 and P2' ligands, respectively. GRL-001-15 and GRL-003-15 have -monofluorophenyl and -monofluorophenyl at the P1 site, respectively, exert highly potent activity against wild-type HIV-1 with 50% effective concentrations (ECs) of 57 and 50 pM, respectively, and have favorable cytotoxicity profiles with 50% cytotoxic concentrations (CCs) of 38 and 11 μM, respectively. The activity of GRL-001-15 against multi-PI-resistant HIV-1 variants was generally greater than that of GRL-003-15. The EC of GRL-001-15 against an HIV-1 variant that was highly resistant to multiple PIs, including darunavir (DRV) (HIV-1), was 0.17 nM, and that of GRL-003-15 was 3.3 nM, while DRV was much less active, with an EC of 216 nM. The emergence of HIV-1 variants resistant to GRL-001-15 and GRL-003-15 was significantly delayed compared to that of variants resistant to selected PIs, including DRV. Structural analyses of wild-type protease (PR) complexed with the novel PIs revealed that GRL-001-15's -fluorine atom forms halogen bond interactions (2.9 and 3.0 Å) with Gly49 and Ile50, respectively, of the protease flap region and with Pro81' (2.7 and 3.2 Å), which is located close to the protease active site, and that two fluorine atoms of GRL-142-13 form multiple halogen bond interactions with Gly49, Ile50, Pro81', Ile82', and Arg8'. In contrast, GRL-003-15 forms halogen bond interactions with Pro81' alone, suggesting that the reduced antiviral activity of GRL-003-15 is due to the loss of the interactions with the flap region.
我们生成了两种新型非肽类 HIV-1 蛋白酶抑制剂（PIs），GRL-001-15 和 GRL-003-15，它们分别含有独特的冠醚四氢呋喃（Crn-THF）和 P2'-环丙基-氨基苯并噻唑（Cp-Abt）部分作为 P2 和 P2'配体。GRL-001-15 和 GRL-003-15 在 P1 位分别含有 -单氟苯基和 -单氟苯基，对野生型 HIV-1 具有高度有效的活性，50%有效浓度（ECs）分别为 57 和 50 pM，细胞毒性谱良好，50%细胞毒性浓度（CCs）分别为 38 和 11 μM。GRL-001-15 对多种 PIs 耐药的 HIV-1 变异体的活性通常大于 GRL-003-15。对包括达芦那韦（DRV）（HIV-1）在内的多种 PIs 耐药的 HIV-1 变异体的 GRL-001-15 的 EC 为 0.17 nM，而 GRL-003-15 的 EC 为 3.3 nM，而 DRV 的活性则低得多，EC 为 216 nM。与对包括 DRV 在内的选定 PIs 耐药的变异体相比，GRL-001-15 和 GRL-003-15 耐药变异体的出现明显延迟。野生型蛋白酶（PR）与新型 PIs 复合物的结构分析表明，GRL-001-15 的 -氟原子分别与蛋白酶瓣区的 Gly49 和 Ile50 以及靠近蛋白酶活性位点的 Pro81'形成卤键相互作用（2.9 和 3.0 Å），而 GRL-142-13 的两个氟原子与 Gly49、Ile50、Pro81'、Ile82'和 Arg8'形成多个卤键相互作用。相比之下，GRL-003-15 仅与 Pro81'形成卤键相互作用，这表明 GRL-003-15 的抗病毒活性降低是由于与瓣区的相互作用丧失所致。
