Effects of triterpenoids and flavonoids isolated from alnus firma on HIV-1 viral enzymes

  • Young -Beob Yu
  • Hirotsugu Miyashiro
  • Norio Nakamura
  • Masao Hattori
  • Jong Cheol ParkEmail author
Article Drug discovery


Triterpenoids and flavonoids isolated from Alnus firma S. Z. were found to inhibit HIV-1 virus replication and controlled its essential enzymes. In this study, the inhibition of HIV-1 viral replication and its essential enzymes, such as reverse transcriptase, protease and α-glucosidase, were observed using 18 Korean plant extracts. Among the extracts, the methanol extract of Alnus firma leaves showed potent inhibition against the HIV-1 induced cytopathic effect (CPE) in MT-4 cells on microscopic observation (the minimum concentration for complete inhibition of HIV-1 induced CPE, IC=50 μg/mL). Thus, 14 compounds were isolated and identified from the methanol extract of Alnus firma leaves. Of these compounds, the alnustic acid methyl ester exhibited inhibition against HIV-1 protease, with an IC50 of 15.8 μM, and quercetin, quercitrin and myricetin 3-O-β-D-galactopyranoside displayed inhibition against HIV-1 reverse transcriptase, all with IC50 values of 60 μM. Based on these results, the viral replication inhibition of the methanol extract of Alnus firma leaves was adjudged to be acutely related to the protease inhibition activation of the alnustic acid methyl ester as well as the reverse transcriptase inhibition activation of flavonoids.

Key words

Alnus firma HIV-1 Reverse transcriptase Protease α-Glucosidase Alnustic acid methyl ester 


  1. Agrawal, P. K., Carbon-13 NMR of flavonoids. Elsevier, Amsterdam, pp. 128–228, (1989).Google Scholar
  2. Aoki T., Ohta S., Aratani S., Hirata T., and Suga T., The structures of four Novel C31-Secodammarane-type triterpenoid saponins from the female flowers ofAlnus serrulatioides.J. Chem. Soc. Perkin Trans., 1,1398–1403 (1982).Google Scholar
  3. Peumans, W., and Declercq, E., The mannose-specific plants lectins fromCymvidium hybrid andEpipactis helleborine and the (N-acetylglucosamine)n-specific plant lectin fromUrtica dioica are potent and selective inhibitors of human immunodeficiency virus and cytomegalovirus replication in vitro.Antiviral Res., 18, 191–207(1992).PubMedCrossRefGoogle Scholar
  4. Balzarini, J. and Van Damme L, Microbicide drug candidates to prevent HIV infection.Lancet, 369, 787–97 (2007)PubMedCrossRefGoogle Scholar
  5. Batinic, D. and Robey, R. A., The V3 region of the envelope glycoprotein of human immunodeficiency virus type 1 binds sulfated polysaccharides and CD4-derived synthetic peptides.J. Biol. Chem., 267,6664–6671 (1992).PubMedGoogle Scholar
  6. Borges de Melo, E., Silveira Gomes, A. D., and Carvalho, I., α- and β-Glucosidase inhibitors: chemical structure and biological activity.Tetrahedron, 62,10277–10302 (2006).CrossRefGoogle Scholar
  7. Camarasa, M. J., Velázquez, S., San-Félix, A., Pérez-Pérez, M. J., and Gago, F., Dimerization inhibitors of HIV-1 reverse transcriptase, protease and integrase: A single mode of inhibition for the three HIV enzymes?.Antiviral Res., 71, 260–267 (2006).PubMedCrossRefGoogle Scholar
  8. Choi, S. Z., Choi, S. U., and Lee, K. R., Phytochemical constituents of the aerial parts fromSolidago virgaaurea var.gigantean. Arch. Pharm. Res., 27,164–168 (2004).CrossRefGoogle Scholar
  9. Fauci, A. S., The human immunodeficiency virus: Infectivity and mechanism of pathogenesis.Science, 239, 617–622 (1988).PubMedCrossRefGoogle Scholar
  10. Greene, W. C., The molecular biology of human immunodeficiency virus type 1 infection.N. Engl. J. Med., 324, 308–317 (1991).PubMedGoogle Scholar
  11. Hattori, M., Kusumoto, I. T, Soga, M., and Namba, T., Screening of various Ayurvedic medicines for their inhibitory activities on reverse transcriptase and identification of arecatannins and embelin as major inhibitory substances fromAreca catechu andEmbelia rives.J. Med. and Pharm. Soc. for Wakan-Yaku, 10,141–148 (1993).Google Scholar
  12. Kusumoto, I. T, Hattori, M., Ishii H., and Namba, T., A comparative study on the inhibitory effects of flavonoid and alkaloids on reverse transcriptase of different retroviruses.Shoyakugaku Zasshi, 47, 291–294 (1993).Google Scholar
  13. Lee, Y A. and Lee, M. W., Tannins fromRubus coreanum. Kor. J. Pharmacogn., 26, 27–30 (1995).Google Scholar
  14. Lipinski, C. A., Lombardo, F., Dominy, B. W., and Feeney, P. J., Experimental and computational approaches to estimate solubility and permeability in drug discovery and development settings.Adv. Drug Deliv. Rev., 23, 3–25 (1997).CrossRefGoogle Scholar
  15. Makoto, I., Takashi, T, Gen-Ichiro, N., and Itsuo, N., Alnusnins A and B from the leavesof Alnus sieboldiana.Phytochemistry, 28,3179–3184(1989).CrossRefGoogle Scholar
  16. Matsui, T, Yoshimoto, C., Osajima K., Oki, T, and Osajima, Y., In vitro survey of alpha-glucosidase inhibitory food components.Biosci. Biotechnol. Biochem., 60,2019–2022 (1996).PubMedGoogle Scholar
  17. Mayaux, J. F., Bousseau, A., Pauwels, R., Huet, T, Henin, Y, Dereu, N., Evers, M., Soler, F, Poujade, C, De Clercq, E. and Le Pecq, J. B., Triterpene derivatives that block entry of human immunodeficiency virus type 1 into cells.Proc. Natl. Acad. Sci. U.S.A., 91, 3564–3568 (1994).PubMedCrossRefGoogle Scholar
  18. Park, J. C, Hur, J. M., Park, J. G, Hatano, T, Yoshida, T, Miyashiro, H., Min, B. S., and Hattori, M., Inhibitory effects of Korean medicinal plants and camelliatannin H fromCamellia japonica on human immunodeficiency virus type 1 protease.Phytother. Res., 16,422–426 (2002).PubMedCrossRefGoogle Scholar
  19. Park, J. C, Kim, S. C, Choi, M. R., Song, S. H., Yoo E. J., Kim S. H., Miyashiro H., and Hattori M., Anti-HIV protease activity from rosa family plant extracts and rosamultin fromRosa rugosa.J. Med. Food, 8,107–109 (2005).PubMedCrossRefGoogle Scholar
  20. Tan, G T., Pezzuto, J. M., and Kinghorn, A. D., Evaluation of natural products as inhibitors of human immunodeficiency virus type 1 (HIV-1) reverse transcriptase.J. Nat. Prod., 54, 143–153(1991).PubMedCrossRefGoogle Scholar
  21. Yasimina, A. L, Ma, C. M., Kusumoto, I. T., Miyashiro, H., Hattori, M., Mahabir, P. G, and Mireye, C, HIV-1 reverse transcriptase inhibitory principles fromChamaesyce hyssopifolia.Phytother. Res., 11, 22–27 (1997).CrossRefGoogle Scholar
  22. Yoshinori, A., Chemical constituents ofAlnus sieboldiana (Betulaceae). II. The isolation and structure of flavonoids and stilbenes.Bull. Chem. Soc. Jpn., 44, 2761–2766 (1971).CrossRefGoogle Scholar
  23. Yu, Y B., Park, J. C, Lee, J. H., Kim, G E., Cho, S. K., Byun, M. W., Miyashiro, H., and Hattori, M., Screening of some plant extracts for inhibitory effects on HIV-1 and Its essential enzymes.Kor. J. Pharmacogn., 29, 338–346 (1998).Google Scholar

Copyright information

© The Pharmaceutical Society of Korea 2007

Authors and Affiliations

  • Young -Beob Yu
    • 1
  • Hirotsugu Miyashiro
    • 2
  • Norio Nakamura
    • 2
  • Masao Hattori
    • 2
  • Jong Cheol Park
    • 3
    Email author
  1. 1.Department of Herbal Pharmaceutical DevelopmentKorea Institute of Oriental MedicineDaejeonKorea
  2. 2.Institute of Natural MedicineToyama Medical and Pharmaceutical UniversityToyamaJapan
  3. 3.Deparement of Oriental Medicine ResourcesSunchon National UniversitySuncheon, JeonnamKorea

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