Inhibition of Porcine Pancreatic Elastase (PPE) by Korean Mistletoe (Viscum album var.coloratum) Fractions

  • Lyu, Su-Yun (School of Pharmaceutical Sciences, Boots Science Building, University of Nottingham) ;
  • Moon, You-Sun (College of Natural Sciences, Seoul Womes University) ;
  • Kwon, Young-Ju (College of Natural Sciences, Seoul Womes University) ;
  • Joo, Hye-Jin (College of Natural Sciences, Seoul Womes University) ;
  • Park, Won-Bong (College of Natural Sciences, Seoul Womes University)
  • Published : 2003.12.01


The serine proteases such as human leukocyte elastase (HLE) and porcine pancreatic elastase (PPE) are classified in the chymotrypsin family, and possibly the most destructive enzymes having the ability to degrade virtually all of the connective components in the body. In the present study, the extracts of water and methanol of Korean mistletoe (Viscum album var. coloratum) inhibited significantly the PPE activity. The fractions eluated on Amberlite XAD-2 from methanol extract were further purified on the repeated $SiO_2$ column chromatography and the fractions A, B and C were eluated. The fractions A, B and C at 3 mg/ml inhibited significantly the PPE activity up to 66%, 95% and 85%, respectively. In conclusion, the fraction A assumed as lignans or phenylpropanes, and fraction B and C assumed as triterpenoids showed the PPE inhibitory effects on the PPE and that these compounds in mistletoe may be used for treatment of pathological processes such as age-dependent tissue loss or inflammation.



  1. Ahn W. Y, Analysis of chemical constituents of saccharides and triterpenoids in the Korean native mistletoes. Mokchae Konghak; 24,27-33 (1995)
  2. Bussing A., Biological and pharmacological properties of viscum album L.; From tissue flask to man. In A. Bussing (eds.), Mistletoe, genus Viscum & other genera, Medicinal and aromatic plants-Industrial profiles. Harwood Academic Publishers, Netherland, pp 45-60 (2000)
  3. Cabaret D., Gonzales M. G., Wakselman M., Adediran S. A., and Pratt R. F, Synthesis, hydrolysis, and evaluation of 3-acylamino3,4- dihydro-2-oxo-2H-l,3-benzoxazinecarboxylic acids and linear azadepsipeptides as potential substrates/inhibitors of $\beta$-lactamrecognizing enzymes. Eur. J. Org. Chem., 1, 141-150 (2001)
  4. Cowan K. N., Heilbut A., Humpl T.,Lam C; Ito S., and Rabinovitch M., Complete reversal of fatal pulmonary hypertension in rats by a serine elastase inhibitor. Nat. Med., 6, 698702 (2000)
  5. Cregge R. J., Durham S. L., Farr R. A., Gallion S. L., Hare C .M., Hoffman R. V; Janusz M. J., Kim H. O., Koehl J. R, Mehdi S., Metz W. A., Peet N. P, Pelton J. P, Schreuder H. A., Sunder S., and Tardif C, Inhibition of human neutrophil elastase. 4. Design, synthesis, X-ray crystallographic analysis, and structure-activity relationships for a series of P2-modified, orally active peptidyl pentafluoroethyl ketones. J. Med. Chem., 41,2461-2480 (1998)
  6. Edwards P. D. and Bemstein P.R, Synthetic inhibitors of elastase. Med. Res. Rev., 14, 127194 (1994)
  7. Fukunaga T., Kajikawa I., Nishiya K., Takeya K. and Itokawa H., Studies on the constituents of the Japanase misteltoe Viscum album L. var. coloratum Ohwi grown on different trees and their antimicrobial and hypotensive properties. Chem. Pharm. Bull., 37, 1543-1546 (1989)
  8. Gerard S., Nollet G., Put J. V.. and Marchand-Brynaert J., 1Alkoxycarbonyl- 3-halogenoazetidin-2-ones as elastase (PPE) inhibitors, Bioorg. Med. Chem., 10, 3955-3964 (2002a)
  9. Gerard S., Dive G., Clamot B., Touillaux R, and Marchand-Brynaert J., Synthesis, hydrolysis, biochemical and theoretical evaluation of 1,4-bis(alkoxycarbonyl)azetidin-2-ones as potential elastase inhibitors, Tetrahedron, 58, 2423-2433 (2002b)
  10. Grinell F. and Zhu M., Identification of neutrophil elastase as the proteinase in bum wound fluid responsible for degradation of fibronectin. J. Invest. Dermatol. 103, 155161 (1994)
  11. Homsy R., Pelletier-Lebon P., Robert L, and Homebeck W., Fibroblast elastase(s). In: L Robert and W Homebeck (eds.), Elastin and elastases Vol. 2, CRC Press, pp. 5762 (1989)
  12. Kerrigan J. E., Walters M. C, Forrester K. J., Crowder J. B.and Christopher L. J, 6-Acylamino-2-[(alkylsulfonyl)oxy]-1H-isoindole1,3- dione mechanism-based inhibitors of human leukocyte elastase. Bioorg. Med. Chem. Lett., 10, 27-30 (2000)
  13. Khorramizadeh M. and Tredget E., Aging differentially modulates the expression of collagen and collagenase in dermal fibroblasts. Mol Cell Biochem,194, 99108 (1999)
  14. Kong D. Y, Li H. T.,and Luo S. Q., Chemical constituents of Viscum coloratum VII. Isolation and structure of viscumneoside VII. Yaoxue Xuebao, 258, 608-611 (1990)
  15. Kuang S. H. R, Venkataraman R, Tu J., Truong T. M., Chan H. K., and Groutas W. C, Potent inhibition of serine proteases by heterocyclic sulfide derivatives of 1,2,5-thiadiazolidin-3-one 1,1 dioxide. Bioorg. Med. Chem., 8, 1713-1717 (2000)
  16. Lee J. K., Zaidi S. H., Liu P., Dawood F, Cheah A. Y, Wen W. H., Saiki Y., and Rabinovitch M., A serine elastase inhibitor reduces inflammation and fibrosis and preserves cardiac function after experimentally-induced murine myocarditis. Nat. Med., 4, 13831391 (1998)
  17. Lyu, S. Y., Park, S. M., Choung, B. Y., and Park, W. B., Comparative study of Korean (Viscum album, var. coloratum) and European mistletoes (Viscum album). Arch. Pharm. Res., 23, 592-598 (2000)
  18. Lyu, S. Y., Park, W. B., Choi, K. H., and Kim, W. H., Involvement of caspase-3 in apoptosis induced by VlScum album var. coloratum agglutinin in HL-60 cells. Biosci. Biotechnol. Biochem., 65, 534-541 (2001)
  19. Lyu, S. Y., Choi S. H., and Park W. B., Korean mistletoe lectin-induced apoptosis in hepatocarcinoma cells is associated with inhibition of telomerase via mitochondrial controlled pathway independent of p53. Arch. Pharm. Res., 25, 93-101 (2002)
  20. Pochet L., Doucet C, Dive G., Wouters J., Masereel B., Reboux Ravaux M., and Pirotte B., Coumarinic derivatives as mechanismbased inhibitors of $\alpha$ -chymotrypsin and human leukocyte elastase. Bioorg. Med. Chem., 8, 1489-1501 (2000)
  21. Rai R and Katzenellenbogen J.A., Effect of conformational mobility and hydrogen-bonding interactions on the selectivity of some guanidinoaryl-substituted mechanism-based inhibitors of trypsinlike serine proteases J. Med. Chem. 35,4297-4305 (1992)
  22. Rinderknecht H.. Pancreatic secretory enzymes. In: Go V. L. W., Dimagno E. P., Gardner J. D., Lebenthal E., and Reber H. A., Scheele GA (eds.) The pancreas biology, pathobiology and disease. 2nd ed. New York, Raven Press, 225-7 (1993)
  23. Robert L. and Robert A. M., Recent advances in elastase inhibitors. Curr. Opin. Ther. Pat. 2,573581 (1992)
  24. Scheele G., Bartelt D., and Bieger w., Characterization of human exocrine pancreatic proteins by two-dimensional isoelectric focusing/sodium dodecyl sulfate gel electrophoresis. Gastroenterology, 80,461-73 (1981)
  25. Shapiro S., Matrix metalloproteinase degradation of extracellular matrix: biological consequences. Curr. Opin. Cell Biol., 10, 602-608 (1998)
  26. Siedle B., Cisielski S., Murillo R, Loser B., Castro V, Klaas C. A., Hucke O., Labahn A., Melzig M. F., and Merfort I., Sesquiterpene lactones as inhibitors of human neutrophil elastase, Bioorg. Med. Chem., 10, 2855-2861 (2002)
  27. Tiefenbacher C. P., Ebert M., Niroomand F., Batkai S., Tillmanns H., Zimmermann R., and Kubler W., Inhibition of elastase improves myocardial function after repetitive ischaemia and myocardial infarction in the rat heart. Pflugers Arch., 433, 563570 (1997)
  28. Travis J., Mechanism for alteration of the proteinaseproteinase inhibitor balance in tissue. In: Katunuma N., Suzuki K., Travis J., and Fritz H. (eds), Biological functions of proteases and inhibitors, Japan Scientific Societies Press, Tokyo, pp. 221229 (1994)
  29. Vogelmeier C., Hubbard R. C., Fells G. A, Schnebli H. P., Thompson R. C., Fritz H., and Crystal R.G., Anti-neutrophil elastase defense of the normal human respiratory epithelial surface provided by the secretory leukoprotease inhibitor. J. Clin. Invest., 87,482488 (1991)
  30. Yager D .R, Chen S. M., Ward S. I., Olutoye O. O., Diegelmann R. F, and Cohen I. K., Ability of chronic wound fluids to degrade peptide growth factors is associated with increased levels of elastase activity and diminished levels of proteinase inhibitors. Wound Repair and Regeneration 5, 2332 (1997)
  31. Yamamoto H., Koizumi T., Kaneki T., Hanaoka M., and .Kubo K., Effects of lecithinized superoxide dismutase and a neutrophil elastase inhibitor (ONO-5046) on hyperoxic lung injury in rat. Eur. J. Pharmacol., 409, 179183 (2000)
  32. Wagner H, Feil B, Seligmann O, Petricic J, and Kalogjera Z, Phenylpropanones and lignans of Viscum album cardioactive drugs. V. Planta Medica, 2, 102-104 (1986)
  33. Wiedow O., Wiese F., Streit V.. Kalm C, and Christophers E., Lesional elastase activity in psoriasis contact dermatitis and atopic dermatitis. J. Invest. Dermatol.. 99, 306-309 (1992)
  34. Zhang H., Patel S. A., Kandil E., Mueller C. M., Lin Y. Y., and Zenilman M. E., Pancreatic elastase is proven to be a mannosebinding proteinimplications for the systemic response to pancreatitis, Surgery, 133, 678-688 (2003)