AEGiS-11IAC: Development of an anti-HIV oligodeoxynucleotide gp12A: inhibition of multiple HIV strains and in vivo pharmacokinetic properties.

11th International AIDS Conference

Vancouver, British Columbia — July 7-12, 1996

Development of an anti-HIV oligodeoxynucleotide gp12A: inhibition of multiple HIV strains and in vivo pharmacokinetic properties.

Int Conf AIDS 1996 Jul 7-12; 11:30 (abstract no. LB.B.6039)
Anazodo MI, Liu C, Powers C, Duta E, Friesen A, Wainberg M, Wright J; Manitoba Institute of Cell Biology, Winnipeg, MB, Canada. Fax: (204) 787-2190.

OBJECTIVE: Preclinical and clinical development of an antisense oligodeoxynucleotide drug against HIV and its application to the treatment of HIV/AIDS disease.

METHODS: A partially thiolated antisense oligodeoxynucleotide sequence (GP12A) was synthesized that targets a conserved region of the HIV-1 gag sequence. Initial studies to screen for the antisense activity of GP12A was performed during the B4.14 cell line that constitutively expresses the HIV proteins. To determine if GP12A inhibits replication of multiple HIV strains, human peripheral blood mononuclear cells (PBMC) were infected with three established strains of HIV (IIIB, NL43 and JRFL). Viral replication was measured by reverse transcriptase activity in the culture supernatant and p24 production in the cell lysate by immunoblot analysis. Cellular uptake and stability of GP12A was characterized using B4.14 cells, while the pharmacokinetic properties were characterized by administering (i.v., s.c. & i.p.) labeled GP12A into CD-1 mice.

RESULTS: GP12A treatment (1 micromolar) of B4.14 cells or infected cultures inhibited gene expression as well as replication of all the viral strains tested (up to 100%) relative to untreated cultures, and cultures treated with inverse sequence of GP12A as a control. No cytotoxic effects have been observed in cell culture. GP12A has been found to distribute to most tissues including the brain, with relatively high concentrations in the thymus after a single 30 mg/kg body weight injection. Traces of GP12A persisted in blood and plasma for 9 hours and in the tissues for 7 days post injection. Single doses of up to 250 mg/kg body weight has been tolerated in CD-1 mice for 2 months without any death or observable changes.

CONCLUSION: We conclude that GP12A is a potent inhibitor of HIV gene expression and replication of many HIV strains in culture and that partial thiolation of GP12A protected the antisense sequence from nuclease destruction without significant changes in the localization of GP12A to its target. Pharmacokinetic studies showed distribution into tissues without noticeable toxicity. These results suggest that GP12A will be a very useful molecular reagent for analyzing HIV proliferation and has therapeutic potential against HIV/AIDS disease. This work was supported by a research grant from the Manitoba Health Regional Council.

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LBB6039