SUPPRESSION OF HUMAN-IMMUNODEFICIENCY-VIRUS TYPE-1 ACTIVITY IN-VITRO BY OLIGONUCLEOTIDES WHICH FORM INTRAMOLECULAR TETRADS

An oligonucleotide (I100-15) composed of only deoxyguanosine and thymidine was able to inhibit human immunodeficiency virus type-1 (HIV-1) in culture assay systems. I100-15 did not block virus entry into cells but did reduce viral-specific transcripts. As assessed by NMR and polyacrylamide gel methods, I100-15 appears to form a structure in which two stacked guanosine tetrads are connected by three two-base long loops. Structure/activity experiments indicated that formation of intramolecular guanosine tetrads was necessary to achieve maximum antiviral activity. The single deoxyguanosine nucleotide present in each loop was found to be extremely important for the overall antiviral activity. The toxicity of I100-15 was determined to be well above the 50% effective dose (ED(50)) in culture which yielded a high therapeutic index (>100). The addition of a cholesterol moiety to the 3' terminus of I100-15 (1100-23) reduced the ED(50) value to less than 50 nM (from 0.12 mu M for I100-15) and increased the duration of viral suppression to greater than 21 days (versus 7-10 days for I100-16) after removal of the drug from infected cell cultures. The favorable therapeutic index of such molecules coupled with the prolonged suppression of HIV-1, suggest that such compounds further warrant investigation as potential therapeutic agents.