Skip to main content
NCBI home page
Antimicrobial Agents and Chemotherapy logoLink to Antimicrobial Agents and Chemotherapy
. 1997 Feb;41(2):419–426. doi: 10.1128/aac.41.2.419

Inhibition of multiple phases of human immunodeficiency virus type 1 replication by a dithiane compound that attacks the conserved zinc fingers of retroviral nucleocapsid proteins.

W G Rice 1, D C Baker 1, C A Schaeffer 1, L Graham 1, M Bu 1, S Terpening 1, D Clanton 1, R Schultz 1, J P Bader 1, R W Buckheit Jr 1, L Field 1, P K Singh 1, J A Turpin 1
PMCID: PMC163723  PMID: 9021201

Abstract

The human immunodeficiency virus type 1 (HIV-1) nucleocapsid p7 protein contains two retrovirus-type zinc finger domains that are required for multiple phases of viral replication. Chelating residues (three Cys residues and one His residue) of the domains are absolutely conserved among all strains of HIV-1 and other retroviruses, and mutations in these residues in noninfectious virions. These properties establish the zinc finger domains as logical targets for antiviral chemotherapy. Selected dithiobis benzamide (R-SS-R) compounds were previously found to inhibit HIV-1 replication by mediating an electrophilic attack on the zinc fingers. Unfortunately, reaction of these disulfide-based benzamides with reducing agents yields two monomeric structures (two R-SH structures) that can dissociated and no longer react with the zinc fingers, suggesting that in vivo reduction would inactivate the compounds. Through an extensive drug discovery program of the National Cancer Institute, a nondissociable tethered dithiane compound (1,2-dithiane-4,5-diol, 1,1-dioxide, cis; NSC 624151) has been identified. This compound specifically attacks the retroviral zinc fingers, but not other antiviral targets. The lead compound demonstrated broad antiretroviral activity, ranging from field isolates and drug-resistant strains of HIV-1 to HIV-2 and simian immunodeficiency virus. The compound directly inactivated HIV-1 virions and blocked production of infectious virus from cells harboring integrated proviral DNA. NSC 624151 provides a scaffold from which medicinal chemists can develop novel compounds for the therapeutic treatment of HIV infection.

Full Text

The Full Text of this article is available as a PDF (956.1 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aboulker J. P., Swart A. M. Preliminary analysis of the Concorde trial. Concorde Coordinating Committee. Lancet. 1993 Apr 3;341(8849):889–890. doi: 10.1016/0140-6736(93)93096-j. [DOI] [PubMed] [Google Scholar]
  2. Aldovini A., Young R. A. Mutations of RNA and protein sequences involved in human immunodeficiency virus type 1 packaging result in production of noninfectious virus. J Virol. 1990 May;64(5):1920–1926. doi: 10.1128/jvi.64.5.1920-1926.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bader J. P., McMahon J. B., Schultz R. J., Narayanan V. L., Pierce J. B., Harrison W. A., Weislow O. S., Midelfort C. F., Stinson S. F., Boyd M. R. Oxathiin carboxanilide, a potent inhibitor of human immunodeficiency virus reproduction. Proc Natl Acad Sci U S A. 1991 Aug 1;88(15):6740–6744. doi: 10.1073/pnas.88.15.6740. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Berg J. M. Potential metal-binding domains in nucleic acid binding proteins. Science. 1986 Apr 25;232(4749):485–487. doi: 10.1126/science.2421409. [DOI] [PubMed] [Google Scholar]
  5. Berg J. M., Shi Y. The galvanization of biology: a growing appreciation for the roles of zinc. Science. 1996 Feb 23;271(5252):1081–1085. doi: 10.1126/science.271.5252.1081. [DOI] [PubMed] [Google Scholar]
  6. Bowles N. E., Damay P., Spahr P. F. Effect of rearrangements and duplications of the Cys-His motifs of Rous sarcoma virus nucleocapsid protein. J Virol. 1993 Feb;67(2):623–631. doi: 10.1128/jvi.67.2.623-631.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Buckheit R. W., Jr, Kinjerski T. L., Fliakas-Boltz V., Russell J. D., Stup T. L., Pallansch L. A., Brouwer W. G., Dao D. C., Harrison W. A., Schultz R. J. Structure-activity and cross-resistance evaluations of a series of human immunodeficiency virus type-1-specific compounds related to oxathiin carboxanilide. Antimicrob Agents Chemother. 1995 Dec;39(12):2718–2727. doi: 10.1128/aac.39.12.2718. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Buckheit R. W., Jr, Roberson J. L., Lackman-Smith C., Wyatt J. R., Vickers T. A., Ecker D. J. 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. doi: 10.1089/aid.1994.10.1497. [DOI] [PubMed] [Google Scholar]
  9. Chance M. R., Sagi I., Wirt M. D., Frisbie S. M., Scheuring E., Chen E., Bess J. W., Jr, Henderson L. E., Arthur L. O., South T. L. Extended x-ray absorption fine structure studies of a retrovirus: equine infectious anemia virus cysteine arrays are coordinated to zinc. Proc Natl Acad Sci U S A. 1992 Nov 1;89(21):10041–10045. doi: 10.1073/pnas.89.21.10041. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Clanton D. J., Buckheit R. W., Jr, Terpening S. J., Kiser R., Mongelli N., Borgia A. L., Schultz R., Narayanan V., Bader J. P., Rice W. G. Novel sulfonated and phosphonated analogs of distamycin which inhibit the replication of HIV. Antiviral Res. 1995 Aug;27(4):335–354. doi: 10.1016/0166-3542(95)00017-g. [DOI] [PubMed] [Google Scholar]
  11. Clanton D. J., Moran R. A., McMahon J. B., Weislow O. S., Buckheit R. W., Jr, Hollingshead M. G., Ciminale V., Felber B. K., Pavlakis G. N., Bader J. P. Sulfonic acid dyes: inhibition of the human immunodeficiency virus and mechanism of action. J Acquir Immune Defic Syndr. 1992;5(8):771–781. [PubMed] [Google Scholar]
  12. Coffin J. M. HIV population dynamics in vivo: implications for genetic variation, pathogenesis, and therapy. Science. 1995 Jan 27;267(5197):483–489. doi: 10.1126/science.7824947. [DOI] [PubMed] [Google Scholar]
  13. Concorde: MRC/ANRS randomised double-blind controlled trial of immediate and deferred zidovudine in symptom-free HIV infection. Concorde Coordinating Committee. Lancet. 1994 Apr 9;343(8902):871–881. [PubMed] [Google Scholar]
  14. Cushman M., Golebiewski W. M., McMahon J. B., Buckheit R. W., Jr, Clanton D. J., Weislow O., Haugwitz R. D., Bader J. P., Graham L., Rice W. G. Design, synthesis, and biological evaluation of cosalane, a novel anti-HIV agent which inhibits multiple features of virus reproduction. J Med Chem. 1994 Sep 16;37(19):3040–3050. doi: 10.1021/jm00045a008. [DOI] [PubMed] [Google Scholar]
  15. Darlix J. L., Vincent A., Gabus C., de Rocquigny H., Roques B. Trans-activation of the 5' to 3' viral DNA strand transfer by nucleocapsid protein during reverse transcription of HIV1 RNA. C R Acad Sci III. 1993 Aug;316(8):763–771. [PubMed] [Google Scholar]
  16. Dorfman T., Luban J., Goff S. P., Haseltine W. A., Göttlinger H. G. Mapping of functionally important residues of a cysteine-histidine box in the human immunodeficiency virus type 1 nucleocapsid protein. J Virol. 1993 Oct;67(10):6159–6169. doi: 10.1128/jvi.67.10.6159-6169.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Dreyer G. B., Metcalf B. W., Tomaszek T. A., Jr, Carr T. J., Chandler A. C., 3rd, Hyland L., Fakhoury S. A., Magaard V. W., Moore M. L., Strickler J. E. Inhibition of human immunodeficiency virus 1 protease in vitro: rational design of substrate analogue inhibitors. Proc Natl Acad Sci U S A. 1989 Dec;86(24):9752–9756. doi: 10.1073/pnas.86.24.9752. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Déméné H., Dong C. Z., Ottmann M., Rouyez M. C., Jullian N., Morellet N., Mely Y., Darlix J. L., Fournié-Zaluski M. C., Saragosti S. 1H NMR structure and biological studies of the His23-->Cys mutant nucleocapsid protein of HIV-1 indicate that the conformation of the first zinc finger is critical for virus infectivity. Biochemistry. 1994 Oct 4;33(39):11707–11716. doi: 10.1021/bi00205a006. [DOI] [PubMed] [Google Scholar]
  19. Frederickson C. J., Kasarskis E. J., Ringo D., Frederickson R. E. A quinoline fluorescence method for visualizing and assaying the histochemically reactive zinc (bouton zinc) in the brain. J Neurosci Methods. 1987 Jun;20(2):91–103. doi: 10.1016/0165-0270(87)90042-2. [DOI] [PubMed] [Google Scholar]
  20. Gorelick R. J., Chabot D. J., Ott D. E., Gagliardi T. D., Rein A., Henderson L. E., Arthur L. O. Genetic analysis of the zinc finger in the Moloney murine leukemia virus nucleocapsid domain: replacement of zinc-coordinating residues with other zinc-coordinating residues yields noninfectious particles containing genomic RNA. J Virol. 1996 Apr;70(4):2593–2597. doi: 10.1128/jvi.70.4.2593-2597.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Gorelick R. J., Chabot D. J., Rein A., Henderson L. E., Arthur L. O. The two zinc fingers in the human immunodeficiency virus type 1 nucleocapsid protein are not functionally equivalent. J Virol. 1993 Jul;67(7):4027–4036. doi: 10.1128/jvi.67.7.4027-4036.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Gorelick R. J., Henderson L. E., Hanser J. P., Rein A. Point mutants of Moloney murine leukemia virus that fail to package viral RNA: evidence for specific RNA recognition by a "zinc finger-like" protein sequence. Proc Natl Acad Sci U S A. 1988 Nov;85(22):8420–8424. doi: 10.1073/pnas.85.22.8420. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Graham N. M., Zeger S. L., Park L. P., Vermund S. H., Detels R., Rinaldo C. R., Phair J. P. The effects on survival of early treatment of human immunodeficiency virus infection. N Engl J Med. 1992 Apr 16;326(16):1037–1042. doi: 10.1056/NEJM199204163261601. [DOI] [PubMed] [Google Scholar]
  24. Henderson L. E., Bowers M. A., Sowder R. C., 2nd, Serabyn S. A., Johnson D. G., Bess J. W., Jr, Arthur L. O., Bryant D. K., Fenselau C. Gag proteins of the highly replicative MN strain of human immunodeficiency virus type 1: posttranslational modifications, proteolytic processings, and complete amino acid sequences. J Virol. 1992 Apr;66(4):1856–1865. doi: 10.1128/jvi.66.4.1856-1865.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Henderson L. E., Copeland T. D., Sowder R. C., Smythers G. W., Oroszlan S. Primary structure of the low molecular weight nucleic acid-binding proteins of murine leukemia viruses. J Biol Chem. 1981 Aug 25;256(16):8400–8406. [PubMed] [Google Scholar]
  26. Kageyama S., Mimoto T., Murakawa Y., Nomizu M., Ford H., Jr, Shirasaka T., Gulnik S., Erickson J., Takada K., Hayashi H. In vitro anti-human immunodeficiency virus (HIV) activities of transition state mimetic HIV protease inhibitors containing allophenylnorstatine. Antimicrob Agents Chemother. 1993 Apr;37(4):810–817. doi: 10.1128/aac.37.4.810. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Kaplan A. H., Swanstrom R. Human immunodeficiency virus type 1 Gag proteins are processed in two cellular compartments. Proc Natl Acad Sci U S A. 1991 May 15;88(10):4528–4532. doi: 10.1073/pnas.88.10.4528. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. McQuade T. J., Tomasselli A. G., Liu L., Karacostas V., Moss B., Sawyer T. K., Heinrikson R. L., Tarpley W. G. A synthetic HIV-1 protease inhibitor with antiviral activity arrests HIV-like particle maturation. Science. 1990 Jan 26;247(4941):454–456. doi: 10.1126/science.2405486. [DOI] [PubMed] [Google Scholar]
  29. Meek T. D., Lambert D. M., Dreyer G. B., Carr T. J., Tomaszek T. A., Jr, Moore M. L., Strickler J. E., Debouck C., Hyland L. J., Matthews T. J. Inhibition of HIV-1 protease in infected T-lymphocytes by synthetic peptide analogues. Nature. 1990 Jan 4;343(6253):90–92. doi: 10.1038/343090a0. [DOI] [PubMed] [Google Scholar]
  30. Parker W. B., White E. L., Shaddix S. C., Ross L. J., Buckheit R. W., Jr, Germany J. M., Secrist J. A., 3rd, Vince R., Shannon W. M. 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] [Google Scholar]
  31. Prichard M. N., Shipman C., Jr A three-dimensional model to analyze drug-drug interactions. Antiviral Res. 1990 Oct-Nov;14(4-5):181–205. doi: 10.1016/0166-3542(90)90001-n. [DOI] [PubMed] [Google Scholar]
  32. Reardon J. E., Miller W. H. Human immunodeficiency virus reverse transcriptase. Substrate and inhibitor kinetics with thymidine 5'-triphosphate and 3'-azido-3'-deoxythymidine 5'-triphosphate. J Biol Chem. 1990 Nov 25;265(33):20302–20307. [PubMed] [Google Scholar]
  33. Rein A., Ott D. E., Mirro J., Arthur L. O., Rice W., Henderson L. E. Inactivation of murine leukemia virus by compounds that react with the zinc finger in the viral nucleocapsid protein. J Virol. 1996 Aug;70(8):4966–4972. doi: 10.1128/jvi.70.8.4966-4972.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Rice W. G., Bader J. P. Discovery and in vitro development of AIDS antiviral drugs as biopharmaceuticals. Adv Pharmacol. 1995;33:389–438. doi: 10.1016/s1054-3589(08)60675-4. [DOI] [PubMed] [Google Scholar]
  35. Rice W. G., Schaeffer C. A., Graham L., Bu M., McDougal J. S., Orloff S. L., Villinger F., Young M., Oroszlan S., Fesen M. R. The site of antiviral action of 3-nitrosobenzamide on the infectivity process of human immunodeficiency virus in human lymphocytes. Proc Natl Acad Sci U S A. 1993 Oct 15;90(20):9721–9724. doi: 10.1073/pnas.90.20.9721. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Rice W. G., Schaeffer C. A., Harten B., Villinger F., South T. L., Summers M. F., Henderson L. E., Bess J. W., Jr, Arthur L. O., McDougal J. S. Inhibition of HIV-1 infectivity by zinc-ejecting aromatic C-nitroso compounds. Nature. 1993 Feb 4;361(6411):473–475. doi: 10.1038/361473a0. [DOI] [PubMed] [Google Scholar]
  37. Rice W. G., Supko J. G., Malspeis L., Buckheit R. W., Jr, Clanton D., Bu M., Graham L., Schaeffer C. A., Turpin J. A., Domagala J. Inhibitors of HIV nucleocapsid protein zinc fingers as candidates for the treatment of AIDS. Science. 1995 Nov 17;270(5239):1194–1197. doi: 10.1126/science.270.5239.1194. [DOI] [PubMed] [Google Scholar]
  38. Rice W. G., Turpin J. A., Schaeffer C. A., Graham L., Clanton D., Buckheit R. W., Jr, Zaharevitz D., Summers M. F., Wallqvist A., Covell D. G. Evaluation of selected chemotypes in coupled cellular and molecular target-based screens identifies novel HIV-1 zinc finger inhibitors. J Med Chem. 1996 Sep 13;39(19):3606–3616. doi: 10.1021/jm960375o. [DOI] [PubMed] [Google Scholar]
  39. Roberts N. A., Martin J. A., Kinchington D., Broadhurst A. V., Craig J. C., Duncan I. B., Galpin S. A., Handa B. K., Kay J., Kröhn A. Rational design of peptide-based HIV proteinase inhibitors. Science. 1990 Apr 20;248(4953):358–361. doi: 10.1126/science.2183354. [DOI] [PubMed] [Google Scholar]
  40. Rodríguez-Rodríguez L., Tsuchihashi Z., Fuentes G. M., Bambara R. A., Fay P. J. Influence of human immunodeficiency virus nucleocapsid protein on synthesis and strand transfer by the reverse transcriptase in vitro. J Biol Chem. 1995 Jun 23;270(25):15005–15011. doi: 10.1074/jbc.270.25.15005. [DOI] [PubMed] [Google Scholar]
  41. South T. L., Blake P. R., Sowder R. C., 3rd, Arthur L. O., Henderson L. E., Summers M. F. The nucleocapsid protein isolated from HIV-1 particles binds zinc and forms retroviral-type zinc fingers. Biochemistry. 1990 Aug 28;29(34):7786–7789. doi: 10.1021/bi00486a002. [DOI] [PubMed] [Google Scholar]
  42. South T. L., Summers M. F. Zinc fingers. Adv Inorg Biochem. 1990;8:199–248. [PubMed] [Google Scholar]
  43. St Clair M. H., Richards C. A., Spector T., Weinhold K. J., Miller W. H., Langlois A. J., Furman P. A. 3'-Azido-3'-deoxythymidine triphosphate as an inhibitor and substrate of purified human immunodeficiency virus reverse transcriptase. Antimicrob Agents Chemother. 1987 Dec;31(12):1972–1977. doi: 10.1128/aac.31.12.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Summers M. F., Henderson L. E., Chance M. R., Bess J. W., Jr, South T. L., Blake P. R., Sagi I., Perez-Alvarado G., Sowder R. C., 3rd, Hare D. R. Nucleocapsid zinc fingers detected in retroviruses: EXAFS studies of intact viruses and the solution-state structure of the nucleocapsid protein from HIV-1. Protein Sci. 1992 May;1(5):563–574. doi: 10.1002/pro.5560010502. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Tanchou V., Gabus C., Rogemond V., Darlix J. L. Formation of stable and functional HIV-1 nucleoprotein complexes in vitro. J Mol Biol. 1995 Oct 6;252(5):563–571. doi: 10.1006/jmbi.1995.0520. [DOI] [PubMed] [Google Scholar]
  46. Turpin J. A., Terpening S. J., Schaeffer C. A., Yu G., Glover C. J., Felsted R. L., Sausville E. A., Rice W. G. Inhibitors of human immunodeficiency virus type 1 zinc fingers prevent normal processing of gag precursors and result in the release of noninfectious virus particles. J Virol. 1996 Sep;70(9):6180–6189. doi: 10.1128/jvi.70.9.6180-6189.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Yarchoan R., Klecker R. W., Weinhold K. J., Markham P. D., Lyerly H. K., Durack D. T., Gelmann E., Lehrman S. N., Blum R. M., Barry D. W. Administration of 3'-azido-3'-deoxythymidine, an inhibitor of HTLV-III/LAV replication, to patients with AIDS or AIDS-related complex. Lancet. 1986 Mar 15;1(8481):575–580. doi: 10.1016/s0140-6736(86)92808-4. [DOI] [PubMed] [Google Scholar]
  48. You J. C., McHenry C. S. Human immunodeficiency virus nucleocapsid protein accelerates strand transfer of the terminally redundant sequences involved in reverse transcription. J Biol Chem. 1994 Dec 16;269(50):31491–31495. [PubMed] [Google Scholar]

Articles from Antimicrobial Agents and Chemotherapy are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES