Stability-activity relationships of a family of G-tetrad forming oligonucleotides as potent HIV inhibitors - A basis for anti-HIV drug design

Recently, we have demonstrated that T30695, a G-tetrad-forming oligonucleot ide, is a potent inhibitor of human immunodeficiency virus, type I (HIV-1) integrase and the K+ induced loop folding of T30695 plays a key role in the inhibition of HIV-1 integrase (Jing, N,, and Hogan, M, E, (1998) J, Biol, Chem. 273, 34992-34999), Here we have modified T30695 by introducing a hydr ophobic bulky group, propynyl dU, or a positively charged group, 5-amino dU , into the bases of T residues of the loops, and by substitution of the T-G r loops by T-T loops. Physical measurements have demonstrated that the subs titution of propynyl dU or 5-amino dU for T in the T residues of the loops did not alter the structure of T30695, and these derivatives also formed an intramolecular G-quartet structure, which is an essential requirement for anti-HIV activity. Measured IC50 and EC50 values show that these substituti ons did not induce an apparent decrease in the ability to inhibit HIV-1 int egrase activity and in the inhibition of HIV-1 replication in cell culture. However, the substitution of T-T loops for T-G loops induced a substantial decrease in both thermal stability and anti-MN activity. The data analysis of T30695 and the 21 derivatives shows a significant, functional correlati on between thermal stability of the G-tetrad structure and the capacity to inhibit HIV-1 integrase activity and between thermal stability of the G-tet rad structure and the capacity to inhibit HIV-1 replication, as assessed wi th the virus strains HIV-1 RF, IIIB, and MN in cell culture. This relations hip between thermostability and activity provides a basis for improving the efficacy of these compounds to inhibit HIV-1 integrase activity and HIV-1 replication in cell culture.