Characterization of Neuraminidase Inhibitors in Korean Papaver rhoeas Bee Pollen Contributing to Anti-Influenza Activities In Vitro

Abstract

The active constituents of Korean Papaver rhoeas bee pollen conferring neuraminidase inhibitory activities (H1N1, H3N2, and H5N1) were investigated. Six flavonoids and one alkaloid were isolated and characterized by nuclear magnetic resonance and mass spectrometry data. These included kaempferol-3-sophoroside (1), kaempferol-3-neohesperidoside (2), kaempferol-3-sambubioside (3), kaempferol-3-glucoside (4), quercetin-3-sophoroside (5), luteolin (6), and chelianthifoline (7). All compounds showed neuraminidase inhibitory activities with IC₅₀ values ranging from 10.7 to 151.1 µM. The most potent neuraminidase inhibitor was luteolin, which was the dominant content in the ethyl acetate fraction. All tested compounds displayed noncompetitive inhibition of H3N2 neuraminidase. Furthermore, compounds 1–7 all reduced the severity of virally induced cytopathic effects as determined by the Madin-Darby canine kidney cell-based assay showing antiviral activity with IC₅₀ values ranging from 10.7 to 33.4 µM (zanamivir: 58.3 µM). The active compounds were quantified by high-performance liquid chromatography, and the total amount of compounds 1–7 made up about 0.592 g/100 g bee pollen, contributing a rich resource of a natural antiviral product.

Key words

Papaver rhoeas - Papaveraceae - bee pollen - Apis mellifera - neuraminidase inhibitors - anti-influenza activity - flavonoid glycosides

^* In-Kyoung Lee and Byung Soon Hwang contributed equally to this work.

• References

• 1 Graikou K, Kapeta S, Aligiannis N, Sotiroudis G, Chondrogianni N, Gonos E, Chinou I. Chemical analysis of Greek pollen – antioxidant, antimicrobial and proteasome activation properties. Chem Cent J 2011; 5: 33-41
• 2 Morais M, Moreira L, Feás X, Estevinho LM. Honeybee-collected pollen from five Portuguese natural parks: palynological origin, phenolic content, antioxidant properties and antimicrobial activity. Food Chem Toxicol 2011; 49: 1096-1101
• 3 Maruyama H, Sakamoto T, Araki Y, Hara H. Anti-inflammatory effect of bee pollen ethanol extract from Cistus sp. of Spanish on carrageenan-induced rat hind paw edema. BMC Complement Altern Med 2010; 10: 30-41
• 4 Medeiros KC, Figueiredo CA, Figueredo TB, Freire KR, Santos FA, Alcantara-Neves NM, Silva TM, Piuvezam MR. Anti-allergic effect of bee pollen phenolic extract and myricetin in ovalbumin-sensitized mice. J Ethnopharmacol 2008; 119: 41-46
• 5 Ozcan M. Inhibition of Aspergillus parasiticus NRRL 2999 by pollen and propolis extracts. J Med Food 2004; 7: 114-116
• 6 Furusawa E, Chou SC, Hirazumi A, Melera A. Antitumour potential of pollen extract on lewis lung carcinoma implanted intraperitoneally in syngeneic mice. Phytother Res 1995; 9: 255-259
• 7 Pinto B, Cagiagli F, Riccio E, Reali D, Saric A, Balog T, Likic S, Scarpato R. Antiestrogenic and antigenotoxic activity of bee pollen from Cystus incanus and Salix alba as evaluated by the yeast estrogen screen and the micronucleus assay in human lymphocytes. Eur J Med Chem 2010; 45: 4122-4128
• 8 Pascoal A, Rodrigues S, Teixeira A, Feas X, Estevinho LM. Biological activities of commercial bee pollens: antimicrobial, antimutagenic, antioxidant and anti-inflammatory. Food Chem Toxicol 2014; 63: 233-239
• 9 Negri G, Teixeira EW, Alves ML, Moreti AC, Otsuk IP, Borguini RG, Salatino A. Hydroxycinnamic acid amide derivatives, phenolic compounds and antioxidant activities of extracts of pollen samples from Southeast Brazil. J Agric Food Chem 2011; 59: 5516-5522
• 10 Grienke U, Schmidtke M, Grafenstein SV, Kirchmair J, Liedl KR, Rollinger JM. Influenza neuraminidase: a druggable target for natural products. Nat Prod Rep 2012; 29: 11-36
• 11 Hillenbrand M, Zapp J, Becker H. Depsides from the petals of Papaver rhoeas . Planta Med 2004; 70: 380-382
• 12 Niu XF, Xu HB, Liu X, Fan T, Qi L. Isoquinoline alkaloids from Corydalis impatiens . Chem Nat Compd 2013; 49: 187-189
• 13 Wynne PM, Vine JH, Amiet RG. Protopine alkaloids in horse urine. J Chromatogr B Analyt Technol Biomed Life Sci 2004; 811: 85-91
• 14 Ryu YB, Kim JH, Park SJ, Chang JS, Rho MC, Bae KH, Park KH, Lee WS. Inhibition of neuraminidase activity by polyphenol compounds isolated from the roots of Glycyrrhiza uralensis . Bioorg Med Chem Lett 2010; 20: 971-974
• 15 Choi HJ, Song JH, Park KS, Kwon DH. Inhibitory effects of quercetin 3-rhamnoside on influenza A virus replication. Eur J Pharm Sci 2009; 37: 329-333