McQuade T J, Tomasselli A G, Liu L, Karacostas V, Moss B, Sawyer T K, Heinrikson R L, Tarpley W G
Infectious Disease Research Unit, Upjohn Company, Kalamazoo, MI 49001.
Science. 1990 Jan 26;247(4941):454-6. doi: 10.1126/science.2405486.
A synthetic peptidemimetic substrate of the human immunodeficiency virus 1 (HIV-1) protease with a nonhydrolyzable pseudodipeptidyl insert at the protease cleavage site was prepared. The peptide U-81749 inhibited recombinant HIV-1 protease in vitro (inhibition constant Ki of 70 nanomolar) and HIV-1 replication in human peripheral blood lymphocytes (inhibitory concentration IC50 of 0.1 to 1 micromolar). Moreover, 10 micromolar concentrations of U-81749 significantly inhibited proteolysis of the HIV-1 gag polyprotein (p55) to the mature viral structural proteins p24 and p17 in cells infected with a recombinant vaccinia virus expressing the HIV-1 gag-pol genes. The HIV-1 like particles released from inhibitor-treated cells contained almost exclusively p55 and other gag precursors, but not p24. Incubation of HIV-like particles recovered from drug-treated cultures in drug-free medium indicated that inhibition of p55 proteolysis was at least partially reversible, suggesting that U-81749 was present within the particles.
制备了一种人免疫缺陷病毒1（HIV-1）蛋白酶的合成拟肽底物，该底物在蛋白酶切割位点处具有不可水解的假二肽插入序列。肽U-81749在体外抑制重组HIV-1蛋白酶（抑制常数Ki为70纳摩尔），并在人外周血淋巴细胞中抑制HIV-1复制（抑制浓度IC50为0.1至1微摩尔）。此外，10微摩尔浓度的U-81749显著抑制感染表达HIV-1 gag-pol基因的重组痘苗病毒的细胞中HIV-1 gag多蛋白（p55）向成熟病毒结构蛋白p24和p17的蛋白水解。从抑制剂处理的细胞中释放的HIV-1样颗粒几乎只含有p55和其他gag前体，而不含有p24。在无药物培养基中培养从药物处理培养物中回收的HIV样颗粒表明，对p55蛋白水解的抑制至少部分是可逆的，这表明U-81749存在于颗粒内。
