Potent Anti-R5 Human Immunodeficiency Virus Type 1 Effects of a CCR5 Antagonist, AK602/ONO4128/GW873140, in a Novel Human Peripheral Blood Mononuclear Cell Nonobese Diabetic-SCID, Interleukin-2 Receptor γ-Chain-Knocked-Out AIDS Mouse Model

Abstract

We established human peripheral blood mononuclear cell (PBMC)-transplanted R5 human immunodeficiency virus type 1 isolate JR-FL (HIV-1 JR-FL )-infected, nonobese diabetic-SCID, interleukin 2 receptor␥-chain-knocked-out (NOG) mice, in which massive and systemic HIV-1 infection occurred. The susceptibility of the implanted PBMC to the infectivity and cytopathic effect of R5 HIV-1 appeared to stem from hyperactivation of the PBMC, which rapidly proliferated and expressed high levels of CCR5. When a novel spirodiketopiperazine-containing CCR5 inhibitor, AK602/ONO4128/GW873140 (molecular weight, 614), was administered to the NOG mice 1 day after R5 HIV-1 inoculation, the replication and cytopathic effects of R5 HIV-1 were significantly suppressed. In saline-treated mice (n ‫؍‬ 7), the mean human CD4؉ /CD8 ؉ cell ratio was 0.1 on day 16 after inoculation, while levels in mice (n ‫؍‬ 8) administered AK602 had a mean value of 0.92, comparable to levels in uninfected mice (n ‫؍‬ 7). The mean number of HIV-RNA copies in plasma in saline-treated mice were ϳ10 6 /ml on day 16, while levels in AK602-treated mice were 1.27 ؋ 10 3 /ml (P ‫؍‬ 0.001). AK602 also significantly suppressed the number of proviral DNA copies and serum p24 levels (P ‫؍‬ 0.001). These data suggest that the present NOG mouse system should serve as a small-animal AIDS model and warrant that AK602 be further developed as a potential therapeutic for HIV-1 infection.

Activity against Human Immunodeficiency Virus Type 1, Intracellular Metabolism, and Effects on Human DNA Polymerases of 4′-Ethynyl-2-Fluoro-2′-Deoxyadenosine

“…Briefly, PHA-PBMC (10 6 cells/ml) were exposed to 50 50% tissue culture infectious doses of HIV-1 Ba-L or each primary HIV-1 isolate and cultured in the presence or absence of various concentrations of drugs in 10-fold serial dilutions in 96-well microculture plates (10 5 cells/well). On day 7 of culture, the supernatant was harvested, and the amount of p24 Gag protein was determined by using a fully automated chemiluminescent enzyme immunoassay system (Lumipulse F; Fujirebio, Inc., Tokyo, Japan) (20). The drug concentrations that suppressed the production of p24 Gag protein by 50% (50% effective concentrations [EC 50 s]) were determined by comparison with the level of p24 production in drug-free control cell cultures.…”

Potent Activity of a Nucleoside Reverse Transcriptase Inhibitor, 4′-Ethynyl-2-Fluoro-2′-Deoxyadenosine, against Human Immunodeficiency Virus Type 1 Infection in a Model Using Human Peripheral Blood Mononuclear Cell-Transplanted NOD/SCID Janus Kinase 3 Knockout Mice

“…Pharmacokinetic analysis of EFdA in BALB/c mice. Pharmacokinetic analysis of EFdA in BALB/c mice was performed as previously described (22). In brief, plasma samples were collected periodically for 4 h following a single EFdA administration at a dose of 20 mg/kg of body weight dissolved in 250 l phosphate-buffered saline (PBS).…”

Induction of Robust Cellular and Humoral Virus-Specific Adaptive Immune Responses in Human Immunodeficiency Virus-Infected Humanized BLT Mice

“…A more successful strategy to humanize mice has been to engraft human immune cells and/or tissues into immunodeficient severe combined immunodeficiency (SCID) or nonobese diabetic (NOD)/SCID mice that are unable to reject xenogeneic grafts (39,42,57). Early versions of humanized mice supported productive HIV infec-tion and allowed investigators to begin to address important questions in HIV biology in vivo (23,40,(43)(44)(45). More recently, human cord blood or fetal liver CD34 ϩ cells have been used to reconstitute Rag2 Ϫ/Ϫ interleukin-2 receptor ␥ chaindeficient (␥ c Ϫ/Ϫ ) and NOD/SCID/␥ c Ϫ/Ϫ mice, resulting in higher levels of sustained human immune cell engraftment (27,29,61).…”