Plant Phenolics Inhibit Neutrophil Elastase

Abstract

Human neutrophil elastase (HNE) is a serine protease, which is present in its active form in inflamed tissue as well as in psoriatic lesions. In extension of our research on natural compounds as inhibitors of HNE or of its release, several phenolics of different size were tested. The ellagitannins agrimoniin and pedunculagin were the most potent direct HNE inhibitors (IC₅₀ = 0.9 and 2.8 μM, respectively). Ligand docking calculations provided evidence that inhibition may occur in an unspecific manner. Agrimoniin also showed anti-proliferative effects in the ATP assay (IC₅₀ = 3.2 μM), suggesting that this type of tannin could have beneficial effects in the treatment of diseases such as psoriasis. Tests with other phenolics combined with ligand docking experiments revealed that, besides the presence of ortho-dihydroxy groups, a specific lipophilic shape is necessary for an inhibitory activity. The phenolic genistein deserves special interest as an inhibitor of elastase release because its effect was remarkably potent (IC₅₀ = 0.6 μM).

Key words

Human neutrophil elastase - proliferation - phenolics - genistein - tannins - agrimoniin - resveratrol - ligand docking

References

• 1 Wiedow O, Wiese F, Christophers E. Lesional elastase activity in psoriasis - Diagnostic and prognostic-significance.  Arch Dermat Res. 1995;  287 632-5
• 2 Ohbayashi H. Current synthetic inhibitors of human neutrophil elastase.  Exp Opin Invest Drugs. 2002;  12 65-84
• 3 Meyer-Hoffert U, Wingertszahn J, Wiedow O. Human leukocyte elastase induces keratinocyte proliferation by epidermal growth factor receptor activation.  J Investig Dermatol. 2004;  123 338-45
• 4 Wiedow O, Meyer-Hoffert U. Neutrophil serine proteases: potential key regulators of cell signalling during inflammation.  J Int Med. 2005;  257 319-28
• 5 Bos M A, Vennat B, Meunier M T, Pouget M P, Pourrat A, Fialip J. Procyanidins from tormentil: Antioxidant properties towards lipoperoxidation and anti-elastase activity.  Biol Pharm Bull. 1996;  19 146-8
• 6 Shimizu M, Deguchi A, Joe A K, McKoy J F, Moriwaki H, Weinstein I B. EGCG inhibits activation of HER3 and expression of cyclooxygenase-2 in human colon cancer cells.  J Exp Ther Oncol. 2005;  5 69-78
• 7 Siedle B, Murillo R, Hucke O, Labahn A, Merfort I. Structure activity relationship studies of cinnamic acid derivatives as inhibitors of human neutrophil elastase revealed by ligand docking calculations.  Pharmazie. 2003;  58 337-9
• 8 Geiss F, Heinrich M, Hunkler D, Rimpler H. Proanthocyanidins with (+)-epicatechin units from Byrsonima crassifolia bark.  Phytochemistry. 1995;  39 635-43
• 9 Geiger C, Rimpler H. Ellagitannins from Tormentillae rhizoma and Alchemillae herba.  Planta Med. 1990;  56 585-6
• 10 Schorr K, Rott A, Da Costa F B, Merfort I. Optimisation of a human neutrophil elastase assay and investigation of the effect of sesquiterpene lactones.  Biologicals. 2005;  33 175-84
• 11 Boukamp P, Petrussevska R T, Breitkreutz D, Hornung J, Markham A, Fusenig N E. Normal keratinization in a spontaneously immortalized aneuploid human keratinocyte cell line.  J Cell Biol. 1988;  106 761-71
• 12 Kramer B, Rarey M, Lengauer T. Evaluation of the FlexX incremental construction algorithm for protein-ligand dockings.  Proteins. 1999;  37 228-41
• 13 Rarey M, Kramer B, Lengauer T, Klebe G. A fast flexible docking method using an incremental construction algorithm.  J Mol Biol. 1996;  261 470-89
• 14 Navia M A, McKeever B M, Springer J P, Lin T Y, Williams H R, Fluder E M. et al . Structure of human neutrophil elastase in complex with a peptide chloromethyl ketone inhibitor at 1.84-Å resolution.  Proc Natl Acad Sci USA. 1989;  86 7-11
• 15 Vriend G. WHAT IF: a molecular modeling and drug design program.  J Mol Graph. 1990;  8 52-6
• 16 Siedle B, Cisielski S, Murillo R, Löser B, Castro V, Klaas C A. et al . Sesquiterpene lactones as inhibitors of human neutrophil elastase.  Bioorg Med Chem. 2002;  10 2855-61
• 17 Scholz E. Pflanzliche Gerbstoffe.  Dtsch Apoth Ztg. 1994;  134 17-29
• 18 Sartor L, Pezzato E, Dell’Aica I, Caniato R, Biggin S, Garbisa S. Inhibition of matrix-proteases by polyphenols: chemical insights for anti-inflammatory and anti-invasion drug design.  Biochem Pharmacol. 2002;  64 229-37
• 19 Rotondo S, Rajtar G, Manarini S, Celardo A, Rotilio D, de Gaetano G. et al . Effect of trans-resveratrol, a natural polyphenolic compound, on human polymorphonuclear leukocyte function.  Brit J Pharmacol. 1998;  123 1691-9
• 20 Tou J S. Differential regulation of phosphatidic acid (PA) formation and degranulation by polyphenolic antioxidants in stimulated human neutrophils.  Faseb J. 2002;  16 A538-9
• 21 Tuluc F, Garcia A, Bredetean O, Meshki J, Kunapuli S P. Primary granule release from human neutrophils is potentiated by soluble fibrinogen through a mechanism depending on multiple intracellular signaling pathways.  Am J Physiol Cell Physiol. 2004;  287 C1264-72
• 22 Melzig M F, Löser B, Cisielski S. Inhibition of neutrophil elastase activity by phenolic compounds from plants.  Pharmazie. 2001;  56 967-70
• 23 Melzig M F, Löser B, Lobitz G O, Tamayo-Castillo G, Merfort I. Inhibition of granulocyte elastase activity by caffeic acid derivatives.  Pharmazie. 1999;  54 712
• 24 Steinbrecher T, Case D A, Labahn A. A multistep approach to structure-based drug design: studying ligand binding at the human neutrophil elastase.  J Med Chem. 2006;  49 1837-44
• 25 Shen G X, Jeong W S, Hu R, Kong A NT. Regulation of Nrf2, NF-kappaB, and AP-1 signaling pathways by chemopreventive agents.  Antioxid Redox Signal. 2005;  7 1648-63
• 26 Sottong P R, Rosebrock J A, Britz J A, Kramer T R. Measurement of T-lymphocyte responses in whole-blood cultures using newly synthesized DNA and ATP.  Clin Diagn Lab Immunol. 2000;  7 307-11
• 27 Crouch S PM, Kozlowski R, Slater K J, Fletcher J. The use of ATP bioluminescence as a measure of cell proliferation and cytotoxicity.  J Immunol Methods. 1993;  160 81-8
• 28 Dvorakova K, Dorr R T, Valcic S, Timmermann B, Alberts D S. Pharmacokinetics of the green tea derivative, EGCG, by the topical route of administration in mouse and human skin.  Cancer Chemother Pharmacol. 1999;  43 331-5