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. 1996 Jan;40(1):133–138. doi: 10.1128/aac.40.1.133

Characterization of siamycin I, a human immunodeficiency virus fusion inhibitor.

P F Lin 1, H Samanta 1, C M Bechtold 1, C A Deminie 1, A K Patick 1, M Alam 1, K Riccardi 1, R E Rose 1, R J White 1, R J Colonno 1
PMCID: PMC163071  PMID: 8787894

Abstract

The human immunodeficiency virus (HIV) fusion inhibitor siamycin I, a 21-residue tricyclic peptide, was identified from a Streptomyces culture by using a cell fusion assay involving cocultivation of HeLa-CD4+ cells and monkey kidney (BSC-1) cells expressing the HIV envelope gp160. Siamycin I is effective against acute HIV type 1 (HIV-1) and HIV-2 infections, with 50% effective doses ranging from 0.05 to 5.7 microM, and the concentration resulting in a 50% decrease in cell viability in the absence of viral infection is 150 microM in CEM-SS cells. Siamycin I inhibits fusion between C8166 cells and CEM-SS cells chronically infected with HIV (50% effective dose of 0.08 microM) but has no effect on Sendai virus-induced fusion or murine myoblast fusion. Siamycin I does not inhibit gp120 binding to CD4 in either gp120- or CD4-based capture enzyme-linked immunosorbent assays. Inhibition of HIV-induced fusion by this compound is reversible, suggesting that siamycin I binds noncovalently. An HIV-1 resistant variant was selected by in vitro passage of virus in the presence of increasing concentrations of siamycin I. Drug susceptibility studies on a chimeric virus containing the envelope gene from the siamycin I-resistant variant indicate that resistance maps to the gp160 gene. Envelope-deficient HIV complemented with gp160 from siamycin I-resistant HIV also displayed a resistant phenotype upon infection of HeLa-CD4-LTR-beta-gal cells. A comparison of the DNA sequences of the envelope genes from the resistant and parent viruses revealed a total of six amino acid changes. Together these results indicate that siamycin I interacts with the HIV envelope protein.

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Selected References

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  1. Acosta E. P., Fletcher C. V. Agents for treating human immunodeficiency virus infection. Am J Hosp Pharm. 1994 Sep 15;51(18):2251–2287. [PubMed] [Google Scholar]
  2. Alam M., Bechtold C. M., Patick A. K., Skoog M. T., Gant T. G., Colonno R. J., Meyers A. I., Li H., Trimble J., Lin P. F. Substituted naphthalenones as a new structural class of HIV-1 reverse transcriptase inhibitors. Antiviral Res. 1993 Oct;22(2-3):131–141. doi: 10.1016/0166-3542(93)90091-v. [DOI] [PubMed] [Google Scholar]
  3. Balzarini J., Neyts J., Schols D., Hosoya M., Van Damme E., Peumans W., De Clercq E. The mannose-specific plant lectins from Cymbidium hybrid and Epipactis helleborine and the (N-acetylglucosamine)n-specific plant lectin from Urtica dioica are potent and selective inhibitors of human immunodeficiency virus and cytomegalovirus replication in vitro. Antiviral Res. 1992 Jun;18(2):191–207. doi: 10.1016/0166-3542(92)90038-7. [DOI] [PubMed] [Google Scholar]
  4. Barnes W. M. PCR amplification of up to 35-kb DNA with high fidelity and high yield from lambda bacteriophage templates. Proc Natl Acad Sci U S A. 1994 Mar 15;91(6):2216–2220. doi: 10.1073/pnas.91.6.2216. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Barrish J. C., Gordon E., Alam M., Lin P. F., Bisacchi G. S., Chen P., Cheng P. T., Fritz A. W., Greytok J. A., Hermsmeier M. A. Aminodiol HIV protease inhibitors. 1. Design, synthesis, and preliminary SAR. J Med Chem. 1994 Jun 10;37(12):1758–1768. doi: 10.1021/jm00038a005. [DOI] [PubMed] [Google Scholar]
  6. Bechtold C. M., Patick A. K., Alam M., Greytok J., Tino J. A., Chen P., Gordon E., Ahmad S., Barrish J. C., Zahler R. Antiviral properties of aminodiol inhibitors against human immunodeficiency virus and protease. Antimicrob Agents Chemother. 1995 Feb;39(2):374–379. doi: 10.1128/aac.39.2.374. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cao J., Bergeron L., Helseth E., Thali M., Repke H., Sodroski J. Effects of amino acid changes in the extracellular domain of the human immunodeficiency virus type 1 gp41 envelope glycoprotein. J Virol. 1993 May;67(5):2747–2755. doi: 10.1128/jvi.67.5.2747-2755.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Constantine K. L., Friedrichs M. S., Detlefsen D., Nishio M., Tsunakawa M., Furumai T., Ohkuma H., Oki T., Hill S., Bruccoleri R. E. High-resolution solution structure of siamycin II: novel amphipathic character of a 21-residue peptide that inhibits HIV fusion. J Biomol NMR. 1995 Apr;5(3):271–286. doi: 10.1007/BF00211754. [DOI] [PubMed] [Google Scholar]
  9. Earl P. L., Moss B. Mutational analysis of the assembly domain of the HIV-1 envelope glycoprotein. AIDS Res Hum Retroviruses. 1993 Jul;9(7):589–594. doi: 10.1089/aid.1993.9.589. [DOI] [PubMed] [Google Scholar]
  10. Fisher R. A., Bertonis J. M., Meier W., Johnson V. A., Costopoulos D. S., Liu T., Tizard R., Walker B. D., Hirsch M. S., Schooley R. T. HIV infection is blocked in vitro by recombinant soluble CD4. Nature. 1988 Jan 7;331(6151):76–78. doi: 10.1038/331076a0. [DOI] [PubMed] [Google Scholar]
  11. Fréchet D., Guitton J. D., Herman F., Faucher D., Helynck G., Monegier du Sorbier B., Ridoux J. P., James-Surcouf E., Vuilhorgne M. Solution structure of RP 71955, a new 21 amino acid tricyclic peptide active against HIV-1 virus. Biochemistry. 1994 Jan 11;33(1):42–50. doi: 10.1021/bi00167a006. [DOI] [PubMed] [Google Scholar]
  12. Helynck G., Dubertret C., Mayaux J. F., Leboul J. Isolation of RP 71955, a new anti-HIV-1 peptide secondary metabolite. J Antibiot (Tokyo) 1993 Nov;46(11):1756–1757. doi: 10.7164/antibiotics.46.1756. [DOI] [PubMed] [Google Scholar]
  13. Hu S. L., Kosowski S. G., Dalrymple J. M. Expression of AIDS virus envelope gene in recombinant vaccinia viruses. Nature. 1986 Apr 10;320(6062):537–540. doi: 10.1038/320537a0. [DOI] [PubMed] [Google Scholar]
  14. Jansen R. W., Molema G., Pauwels R., Schols D., De Clercq E., Meijer D. K. Potent in vitro anti-human immunodeficiency virus-1 activity of modified human serum albumins. Mol Pharmacol. 1991 Jun;39(6):818–823. [PubMed] [Google Scholar]
  15. Jansen R. W., Schols D., Pauwels R., De Clercq E., Meijer D. K. Novel, negatively charged, human serum albumins display potent and selective in vitro anti-human immunodeficiency virus type 1 activity. Mol Pharmacol. 1993 Nov;44(5):1003–1007. [PubMed] [Google Scholar]
  16. Japour A. J., Mayers D. L., Johnson V. A., Kuritzkes D. R., Beckett L. A., Arduino J. M., Lane J., Black R. J., Reichelderfer P. S., D'Aquila R. T. Standardized peripheral blood mononuclear cell culture assay for determination of drug susceptibilities of clinical human immunodeficiency virus type 1 isolates. The RV-43 Study Group, the AIDS Clinical Trials Group Virology Committee Resistance Working Group. Antimicrob Agents Chemother. 1993 May;37(5):1095–1101. doi: 10.1128/aac.37.5.1095. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Jiang S., Lin K., Strick N., Neurath A. R. Inhibition of HIV-1 infection by a fusion domain binding peptide from the HIV-1 envelope glycoprotein GP41. Biochem Biophys Res Commun. 1993 Sep 15;195(2):533–538. doi: 10.1006/bbrc.1993.2078. [DOI] [PubMed] [Google Scholar]
  18. 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. doi: 10.1128/jvi.66.4.2232-2239.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Kowalski M., Bergeron L., Dorfman T., Haseltine W., Sodroski J. Attenuation of human immunodeficiency virus type 1 cytopathic effect by a mutation affecting the transmembrane envelope glycoprotein. J Virol. 1991 Jan;65(1):281–291. doi: 10.1128/jvi.65.1.281-291.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Kowalski M., Potz J., Basiripour L., Dorfman T., Goh W. C., Terwilliger E., Dayton A., Rosen C., Haseltine W., Sodroski J. Functional regions of the envelope glycoprotein of human immunodeficiency virus type 1. Science. 1987 Sep 11;237(4820):1351–1355. doi: 10.1126/science.3629244. [DOI] [PubMed] [Google Scholar]
  21. Lee P. P., Linial M. L. Efficient particle formation can occur if the matrix domain of human immunodeficiency virus type 1 Gag is substituted by a myristylation signal. J Virol. 1994 Oct;68(10):6644–6654. doi: 10.1128/jvi.68.10.6644-6654.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Lin P. F., Samanta H., Rose R. E., Patick A. K., Trimble J., Bechtold C. M., Revie D. R., Khan N. C., Federici M. E., Li H. Genotypic and phenotypic analysis of human immunodeficiency virus type 1 isolates from patients on prolonged stavudine therapy. J Infect Dis. 1994 Nov;170(5):1157–1164. doi: 10.1093/infdis/170.5.1157. [DOI] [PubMed] [Google Scholar]
  23. Maione R., Fimia G. M., Amati P. Inhibition of in vitro myogenic differentiation by a polyomavirus early function. Oncogene. 1992 Jan;7(1):85–93. [PubMed] [Google Scholar]
  24. Masuda M., Nakashima H., Ueda T., Naba H., Ikoma R., Otaka A., Terakawa Y., Tamamura H., Ibuka T., Murakami T. A novel anti-HIV synthetic peptide, T-22 ([Tyr5,12,Lys7]-polyphemusin II). Biochem Biophys Res Commun. 1992 Dec 15;189(2):845–850. doi: 10.1016/0006-291x(92)92280-b. [DOI] [PubMed] [Google Scholar]
  25. Mayaux J. F., Bousseau A., Pauwels R., Huet T., Hénin Y., Dereu N., Evers M., Soler F., Poujade C., De Clercq E. Triterpene derivatives that block entry of human immunodeficiency virus type 1 into cells. Proc Natl Acad Sci U S A. 1994 Apr 26;91(9):3564–3568. doi: 10.1073/pnas.91.9.3564. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Nakashima H., Masuda M., Murakami T., Koyanagi Y., Matsumoto A., Fujii N., Yamamoto N. Anti-human immunodeficiency virus activity of a novel synthetic peptide, T22 ([Tyr-5,12, Lys-7]polyphemusin II): a possible inhibitor of virus-cell fusion. Antimicrob Agents Chemother. 1992 Jun;36(6):1249–1255. doi: 10.1128/aac.36.6.1249. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Okada Y. Sendai virus-induced cell fusion. Methods Enzymol. 1993;221:18–41. doi: 10.1016/0076-6879(93)21005-s. [DOI] [PubMed] [Google Scholar]
  28. Owens R. J., Burke C., Rose J. K. Mutations in the membrane-spanning domain of the human immunodeficiency virus envelope glycoprotein that affect fusion activity. J Virol. 1994 Jan;68(1):570–574. doi: 10.1128/jvi.68.1.570-574.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Owens R. J., Tanner C. C., Mulligan M. J., Srinivas R. V., Compans R. W. Oligopeptide inhibitors of HIV-induced syncytium formation. AIDS Res Hum Retroviruses. 1990 Nov;6(11):1289–1296. doi: 10.1089/aid.1990.6.1289. [DOI] [PubMed] [Google Scholar]
  30. Patick A. K., Rose R., Greytok J., Bechtold C. M., Hermsmeier M. A., Chen P. T., Barrish J. C., Zahler R., Colonno R. J., Lin P. F. Characterization of a human immunodeficiency virus type 1 variant with reduced sensitivity to an aminodiol protease inhibitor. J Virol. 1995 Apr;69(4):2148–2152. doi: 10.1128/jvi.69.4.2148-2152.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Pleskoff O., Seman M., Alizon M. Amphotericin B derivative blocks human immunodeficiency virus type 1 entry after CD4 binding: effect on virus-cell fusion but not on cell-cell fusion. J Virol. 1995 Jan;69(1):570–574. doi: 10.1128/jvi.69.1.570-574.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Pontecorvo G. Production of mammalian somatic cell hybrids by means of polyethylene glycol treatment. Somatic Cell Genet. 1975 Oct;1(4):397–400. doi: 10.1007/BF01538671. [DOI] [PubMed] [Google Scholar]
  33. Richman D. D. Resistance of clinical isolates of human immunodeficiency virus to antiretroviral agents. Antimicrob Agents Chemother. 1993 Jun;37(6):1207–1213. doi: 10.1128/aac.37.6.1207. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Takeuchi H., Baba M., Shigeta S. An application of tetrazolium (MTT) colorimetric assay for the screening of anti-herpes simplex virus compounds. J Virol Methods. 1991 Jun;33(1-2):61–71. doi: 10.1016/0166-0934(91)90008-n. [DOI] [PubMed] [Google Scholar]
  35. Walker R. E., Parker R. I., Kovacs J. A., Masur H., Lane H. C., Carleton S., Kirk L. E., Gralnick H. R., Fauci A. S. Anemia and erythropoiesis in patients with the acquired immunodeficiency syndrome (AIDS) and Kaposi sarcoma treated with zidovudine. Ann Intern Med. 1988 Mar;108(3):372–376. doi: 10.7326/0003-4819-108-3-372. [DOI] [PubMed] [Google Scholar]
  36. Weislow O. S., Kiser R., Fine D. L., Bader J., Shoemaker R. H., Boyd M. R. New soluble-formazan assay for HIV-1 cytopathic effects: application to high-flux screening of synthetic and natural products for AIDS-antiviral activity. J Natl Cancer Inst. 1989 Apr 19;81(8):577–586. doi: 10.1093/jnci/81.8.577. [DOI] [PubMed] [Google Scholar]
  37. Wild C. T., Shugars D. C., Greenwell T. K., McDanal C. B., Matthews T. J. Peptides corresponding to a predictive alpha-helical domain of human immunodeficiency virus type 1 gp41 are potent inhibitors of virus infection. Proc Natl Acad Sci U S A. 1994 Oct 11;91(21):9770–9774. doi: 10.1073/pnas.91.21.9770. [DOI] [PMC free article] [PubMed] [Google Scholar]

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