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Antimicrob Agents Chemother. 1995 November; 39(11): 2426–2435.
PMCID: PMC162960

T30177, an oligonucleotide stabilized by an intramolecular guanosine octet, is a potent inhibitor of laboratory strains and clinical isolates of human immunodeficiency virus type 1.


T30177, an oligonucleotide composed of only deoxyguanosine and thymidine, is 17 nucleotides in length and contains single phosphorothioate internucleoside linkages at its 5' and 3' ends for stability. This oligonucleotide does not share significant primary sequence homology with or possess any complementary (antisense) sequence motifs to the human immunodeficiency virus type 1 (HIV-1) genome. T30177 inhibited replication of multiple laboratory strains of HIV-1 in human T-cell lines, peripheral blood lymphocytes, and macrophages. T30177 was also found to be capable of inhibiting multiple clinical isolates of HIV-1 and preventing the cytopathic effect of HIV-1 in primary CD4+ T lymphocytes. In assays with human peripheral blood lymphocytes there was no observable toxicity associated with T30177 at the highest concentration tested (100 microM), while the median inhibitory concentration was determined to be in the range of 0.1 to 1.0 microM for the clinical isolates tested, resulting in a high therapeutic index for this drug. In temporal studies, the kinetics of addition of T30177 to infected cell cultures indicated that, like the known viral adsorption blocking agents dextran sulfate and Chicago sky blue, T30177 needed to be added to cells during or very soon after viral infection. However, analysis of nucleic acids extracted at 12 h postinfection from cells treated with T30177 at the time of virus infection established the presence of unintegrated viral cDNA, including circular proviral DNA, in the treated cells. In vitro analysis of viral enzymes revealed that T30177 was a potent inhibitor of HIV-1 integrase, reducing enzymatic activity by 50% at concentrations in the range of 0.050 to 0.09 microM. T30177 was also able to inhibit viral reverse transcriptase activity; however, the 50% inhibitory value obtained was in the range of 1 to 10 microM, depending on the template used in the enzymatic assay. No observable inhibition of viral protease was detected at the highest concentration of T30177 used (10 microM). In experiments in which T30177 was removed from infected cell cultures at 4 days post-HIV-1 infection, total suppression of virus production was observed for more than 27 days. PCR analysis of DNA extracted from cells treated in this fashion was unable to detect the presence of viral DNA 11 days after removal of the drug from the infected cell cultures. The ability of T30177 to inhibit both laboratory and clinical isolates of HIV-1 and the experimental data which suggest that T30177 represents a novel class of integrase inhibitors indicate that this compound is a viable candidate for evaluation as a therapeutic agent against HIV-1 in humans.

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Selected References

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  • Agrawal S, Tang JY. GEM 91--an antisense oligonucleotide phosphorothioate as a therapeutic agent for AIDS. Antisense Res Dev. 1992 Winter;2(4):261–266. [PubMed]
  • Andrei G, De Clercq E. Inhibitory effect of selected antiviral compounds on arenavirus replication in vitro. Antiviral Res. 1990 Oct-Nov;14(4-5):287–299. [PubMed]
  • Bader JP, McMahon JB, Schultz RJ, Narayanan VL, Pierce JB, Harrison WA, Weislow OS, Midelfort CF, Stinson SF, Boyd MR. Oxathiin carboxanilide, a potent inhibitor of human immunodeficiency virus reproduction. Proc Natl Acad Sci U S A. 1991 Aug 1;88(15):6740–6744. [PubMed]
  • Bock LC, Griffin LC, Latham JA, Vermaas EH, Toole JJ. Selection of single-stranded DNA molecules that bind and inhibit human thrombin. Nature. 1992 Feb 6;355(6360):564–566. [PubMed]
  • Buckheit RW, Jr, Roberson JL, Lackman-Smith C, Wyatt JR, Vickers TA, Ecker DJ. Potent and specific inhibition of HIV envelope-mediated cell fusion and virus binding by G quartet-forming oligonucleotide (ISIS 5320). AIDS Res Hum Retroviruses. 1994 Nov;10(11):1497–1506. [PubMed]
  • Buckheit RW, Jr, Swanstrom R. Characterization of an HIV-1 isolate displaying an apparent absence of virion-associated reverse transcriptase activity. AIDS Res Hum Retroviruses. 1991 Mar;7(3):295–302. [PubMed]
  • Carteau S, Mouscadet JF, Goulaouic H, Subra F, Auclair C. Inhibitory effect of the polyanionic drug suramin on the in vitro HIV DNA integration reaction. Arch Biochem Biophys. 1993 Sep;305(2):606–610. [PubMed]
  • Ciminale V, Felber BK, Campbell M, Pavlakis GN. A bioassay for HIV-1 based on Env-CD4 interaction. AIDS Res Hum Retroviruses. 1990 Nov;6(11):1281–1287. [PubMed]
  • Clanton DJ, Moran RA, McMahon JB, Weislow OS, Buckheit RW, Jr, Hollingshead MG, Ciminale V, Felber BK, Pavlakis GN, Bader JP. Sulfonic acid dyes: inhibition of the human immunodeficiency virus and mechanism of action. J Acquir Immune Defic Syndr. 1992;5(8):771–781. [PubMed]
  • Crowe S, Mills J, McGrath MS. Quantitative immunocytofluorographic analysis of CD4 surface antigen expression and HIV infection of human peripheral blood monocyte/macrophages. AIDS Res Hum Retroviruses. 1987 Summer;3(2):135–145. [PubMed]
  • De Clercq E. Antiviral and antimetabolic activities of neplanocins. Antimicrob Agents Chemother. 1985 Jul;28(1):84–89. [PMC free article] [PubMed]
  • De Clercq E, Yamamoto N, Pauwels R, Balzarini J, Witvrouw M, De Vreese K, Debyser Z, Rosenwirth B, Peichl P, Datema R, et al. Highly potent and selective inhibition of human immunodeficiency virus by the bicyclam derivative JM3100. Antimicrob Agents Chemother. 1994 Apr;38(4):668–674. [PMC free article] [PubMed]
  • Fesen MR, Kohn KW, Leteurtre F, Pommier Y. Inhibitors of human immunodeficiency virus integrase. Proc Natl Acad Sci U S A. 1993 Mar 15;90(6):2399–2403. [PubMed]
  • Gao WY, Stein CA, Cohen JS, Dutschman GE, Cheng YC. Effect of phosphorothioate homo-oligodeoxynucleotides on herpes simplex virus type 2-induced DNA polymerase. J Biol Chem. 1989 Jul 5;264(19):11521–11526. [PubMed]
  • Garry RF. Potential mechanisms for the cytopathic properties of HIV. AIDS. 1989 Nov;3(11):683–694. [PubMed]
  • Gowda SD, Stein BS, Mohagheghpour N, Benike CJ, Engleman EG. Evidence that T cell activation is required for HIV-1 entry in CD4+ lymphocytes. J Immunol. 1989 Feb 1;142(3):773–780. [PubMed]
  • Hirt B. Selective extraction of polyoma DNA from infected mouse cell cultures. J Mol Biol. 1967 Jun 14;26(2):365–369. [PubMed]
  • Ho DD, Neumann AU, Perelson AS, Chen W, Leonard JM, Markowitz M. Rapid turnover of plasma virions and CD4 lymphocytes in HIV-1 infection. Nature. 1995 Jan 12;373(6510):123–126. [PubMed]
  • Kimpton J, Emerman M. Detection of replication-competent and pseudotyped human immunodeficiency virus with a sensitive cell line on the basis of activation of an integrated beta-galactosidase gene. J Virol. 1992 Apr;66(4):2232–2239. [PMC free article] [PubMed]
  • Letsinger RL, Zhang GR, Sun DK, Ikeuchi T, Sarin PS. Cholesteryl-conjugated oligonucleotides: synthesis, properties, and activity as inhibitors of replication of human immunodeficiency virus in cell culture. Proc Natl Acad Sci U S A. 1989 Sep;86(17):6553–6556. [PubMed]
  • Lewis AF, Drach JC, Fennewald SM, Huffman JH, Ptak RG, Sommadossi JP, Revankar GR, Rando RF. Inhibition of human cytomegalovirus in culture by alkenyl guanine analogs of the thiazolo[4,5-d]pyrimidine ring system. Antimicrob Agents Chemother. 1994 Dec;38(12):2889–2895. [PMC free article] [PubMed]
  • Lisziewicz J, Sun D, Metelev V, Zamecnik P, Gallo RC, Agrawal S. Long-term treatment of human immunodeficiency virus-infected cells with antisense oligonucleotide phosphorothioates. Proc Natl Acad Sci U S A. 1993 May 1;90(9):3860–3864. [PubMed]
  • Marshall WS, Beaton G, Stein CA, Matsukura M, Caruthers MH. Inhibition of human immunodeficiency virus activity by phosphorodithioate oligodeoxycytidine. Proc Natl Acad Sci U S A. 1992 Jul 15;89(14):6265–6269. [PubMed]
  • Mazumder A, Cooney D, Agbaria R, Gupta M, Pommier Y. Inhibition of human immunodeficiency virus type 1 integrase by 3'-azido-3'-deoxythymidylate. Proc Natl Acad Sci U S A. 1994 Jun 21;91(13):5771–5775. [PubMed]
  • McGrath MS, Hwang KM, Caldwell SE, Gaston I, Luk KC, Wu P, Ng VL, Crowe S, Daniels J, Marsh J, et al. GLQ223: an inhibitor of human immunodeficiency virus replication in acutely and chronically infected cells of lymphocyte and mononuclear phagocyte lineage. Proc Natl Acad Sci U S A. 1989 Apr;86(8):2844–2848. [PubMed]
  • Milligan JF, Matteucci MD, Martin JC. Current concepts in antisense drug design. J Med Chem. 1993 Jul 9;36(14):1923–1937. [PubMed]
  • Nara PL, Fischinger PJ. Quantitative infectivity assay for HIV-1 and-2. Nature. 1988 Mar 31;332(6163):469–470. [PubMed]
  • Ojwang J, Elbaggari A, Marshall HB, Jayaraman K, McGrath MS, Rando RF. Inhibition of human immunodeficiency virus type 1 activity in vitro by oligonucleotides composed entirely of guanosine and thymidine. J Acquir Immune Defic Syndr. 1994 Jun;7(6):560–570. [PubMed]
  • Parker WB, White EL, Shaddix SC, Ross LJ, Buckheit RW, Jr, Germany JM, Secrist JA, 3rd, Vince R, Shannon WM. Mechanism of inhibition of human immunodeficiency virus type 1 reverse transcriptase and human DNA polymerases alpha, beta, and gamma by the 5'-triphosphates of carbovir, 3'-azido-3'-deoxythymidine, 2',3'-dideoxyguanosine and 3'-deoxythymidine. A novel RNA template for the evaluation of antiretroviral drugs. J Biol Chem. 1991 Jan 25;266(3):1754–1762. [PubMed]
  • Rando RF, Ojwang J, Elbaggari A, Reyes GR, Tinder R, McGrath MS, Hogan ME. Suppression of human immunodeficiency virus type 1 activity in vitro by oligonucleotides which form intramolecular tetrads. J Biol Chem. 1995 Jan 27;270(4):1754–1760. [PubMed]
  • Schultze P, Macaya RF, Feigon J. Three-dimensional solution structure of the thrombin-binding DNA aptamer d(GGTTGGTGTGGTTGG). J Mol Biol. 1994 Feb 4;235(5):1532–1547. [PubMed]
  • Stein CA, Cleary AM, Yakubov L, Lederman S. Phosphorothioate oligodeoxynucleotides bind to the third variable loop domain (v3) of human immunodeficiency virus type 1 gp120. Antisense Res Dev. 1993 Spring;3(1):19–31. [PubMed]
  • Steinkasserer A, Harrison R, Billich A, Hammerschmid F, Werner G, Wolff B, Peichl P, Palfi G, Schnitzel W, Mlynar E, et al. Mode of action of SDZ NIM 811, a nonimmunosuppressive cyclosporin A analog with activity against human immunodeficiency virus type 1 (HIV-1): interference with early and late events in HIV-1 replication. J Virol. 1995 Feb;69(2):814–824. [PMC free article] [PubMed]
  • Wang KY, McCurdy S, Shea RG, Swaminathan S, Bolton PH. A DNA aptamer which binds to and inhibits thrombin exhibits a new structural motif for DNA. Biochemistry. 1993 Mar 2;32(8):1899–1904. [PubMed]
  • Wei X, Ghosh SK, Taylor ME, Johnson VA, Emini EA, Deutsch P, Lifson JD, Bonhoeffer S, Nowak MA, Hahn BH, et al. Viral dynamics in human immunodeficiency virus type 1 infection. Nature. 1995 Jan 12;373(6510):117–122. [PubMed]
  • White EL, Buckheit RW, Jr, Ross LJ, Germany JM, Andries K, Pauwels R, Janssen PA, Shannon WM, Chirigos MA. A TIBO derivative, R82913, is a potent inhibitor of HIV-1 reverse transcriptase with heteropolymer templates. Antiviral Res. 1991 Oct;16(3):257–266. [PubMed]
  • Wyatt JR, Vickers TA, Roberson JL, Buckheit RW, Jr, Klimkait T, DeBaets E, Davis PW, Rayner B, Imbach JL, Ecker DJ. Combinatorially selected guanosine-quartet structure is a potent inhibitor of human immunodeficiency virus envelope-mediated cell fusion. Proc Natl Acad Sci U S A. 1994 Feb 15;91(4):1356–1360. [PubMed]
  • Zack JA, Arrigo SJ, Weitsman SR, Go AS, Haislip A, Chen IS. HIV-1 entry into quiescent primary lymphocytes: molecular analysis reveals a labile, latent viral structure. Cell. 1990 Apr 20;61(2):213–222. [PubMed]

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