Dolutegravir (GSK1349572) (IC50=2.7 nM), a two-metal-binding HIV integrase inhibitor, exhibits medium activity against raltegravir-resistant signature mutants Y143R, Q148K, N155H, and G140S/Q148H.

HIV integrase:2.7 nM

S/GSK1349572对9种来自未接触过整合酶抑制剂的HIV-2感染患者的临床分离株显示出强大的抑制效果，EC50值介于0.2 nM至1.4 nM之间。[1] 体外实验中，S/GSK1349572对重组HIV-1整合酶催化的链转移反应的抑制作用表现出IC50值为2.7 nM。此外，S/GSK1349572在以自灭性PHIV慢病毒载体感染的细胞中，如外周血单核细胞（PBMCs）、MT-4细胞和CIP4细胞，对HIV复制的抑制作用表现出EC50值分别为0.51 nM、0.71 nM和2.2 nM。[2] 体外实验中，S/GSK1349572对5种不同的非核苷类逆转录酶抑制剂抵抗性或核苷类逆转录酶抑制剂抵抗性病毒展现出强大的活性，EC50值介于1.3 nM至2.1 nM之间。与对野生型病毒的作用类似，S/GSK1349572对两种蛋白酶抑制剂抵抗病毒也显示出等同的活性，EC50值分别为0.36 nM和0.37 nM。[2]

通过单次静脉注射（IV）给药后，Dolutegravir在大鼠（0.23 mL/min/kg）和猴子（2.12 mL/min/kg）体内的血浆清除率低。大鼠和猴子的半衰期相似，约为6小时，而恒定状态的分布体积（VSS）低。经口服给药，Dolutegravir在空腹状态下的雄性大鼠和单只猴子体内迅速吸收，口服生物利用度高（分别为75.6%和87.0%）。Dolutegravir的暴露水平（Cmax和AUC）随着给予未进食大鼠至多250 mg/kg以及未进食猴子至多50 mg/kg的悬浮液剂量的增加而提高，尽管增加幅度不成比例[3]。

In vitro strand transfer assay: The inhibitory potencies of S/GSK1349572 and other INIs are measured in a strand transfer assay using recombinant HIV integrase. A complex of integrase and biotinylated preprocessed donor DNA-streptavidin-coated Acintillation proximity assay (SPA) beads is formed by incubating 2 μM purified recombinant integrase with 0.66 μM biotinylated donor DNA-4 mg/mL streptavidin-coated SPA beads in 25 mM sodium morpholinepropanesulfonic acid (MOPS) (pH 7.2), 23 mM NaCl, and 10 mM MgCl2 for 5 minutes at 37 °C. These beads are spun down and preincubated with diluted INIs for 60 minutes at 37 °C. Then a 3H-labeled target DNA substrate is added to give a final concentration of 7 nM substrate, and the strand transfer reaction mixture is incubated at 37 °C for 25 to 45 minutes, which allows for a linear increase in the strand transfer of donor DNA to radiolabeled target DNA. The signal is read using a Wallac MicroBeta scintillation plate reader.

MT-4 cells growing exponentially at a density of 500000 or 600000 /mL are infected with HIV-1 strain IIIB at a viral multiplicity of infection of 0.001 or a 50% tissue culture infective dose of 4 to 10. The cells are then aliquoted to 96-well plates in the presence of varying concentrations of S/GSK1349572. After incubation for 4 or 5 days, antiviral activity is determined by a cell viability assay that either measured bioluminescence with a CellTiter-Glo luminescent reagent or measured absorbance at 560 and 690 nm using the yellow tetrazolium MTT reagent [3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyltetrazolium bromide]. (Only for Reference)