Reversal of P-Glycoprotein-Mediated Drug Efflux by Eudesmin from Haplophyllum perforatum and Cytotoxicity Pattern versus Diphyllin, Podophyllotoxin and Etoposide

Abstract

The present study focuses on eudesmin (bicyclic lignan, 0.15 % of dry leaves) and diphyllin (arylnaphthalene lignan, 0.1 % of dry roots), both isolated from H. perforatum Kar. et Kir, a Rutaceae species endemic to Uzbekistan. We first compared their specificity for cancer cells with those of etoposide and podophyllotoxin by screening their cytotoxicity on 3 healthy cell-lines and 7 sensitive or resistant human solid cancer lines. We then tested their capacity to reverse P-glycoprotein-mediated multidrug resistance (MDR) by assaying dye and drug uptake in MDR1-transfected Madin-Darby canine kidney (MDCK-MDR1) and doxorubicine-resistant human breast carcinoma cells (MCF7/Dox). Eudesmin displays IC₅₀ values > 100 μM on all tested lines. Our data provide the first demonstration that this non-toxic lignan reverses Pgp-mediated drug efflux and supports the hypothesis that it may inhibit resistance mediated by MDR1 and MRP proteins. Even if its reversal activity is insufficient for clinical application, its capacity to accumulate [³ H]-vinblastine in MDCK/MDR1 and MCF7/Dox cells suggests that eudesmin may positively affect the bioavailability and, thereby, the therapeutic potency of anticancer drugs in Pgp-overexpressing cells. Diphyllin exhibits IC₅₀ values ranging from 10 ^{- 6} to 10 ^{- 4} M. It is markedly less toxic than podophyllotoxin (IC₅₀ : 13 - 61 nM), but exhibits tumoricidal effects close to those of etoposide. Unfortunatly, it is 65-fold more toxic than etoposide on human primary fibroblasts. Consequently, it has no value as an anticancer drug. Its value as raw material for the hemisynthesis of anticancer drugs is discussed.

Key words

Haplophyllum perforatum - Rutaceae - lignans - diphyllin - eudesmin - P-glycoprotein - cytotoxicity

References

• 1 Ibn Sina AA ( Avicenna). The handbook of medical sciences. Selected parts. Tashkent; FAN 1985: 264-5.
• 2 Razzakova D M, Bessonova I A, Yunusov S Yu. Eudesmin, a lignan from Haplophyllum acutifolium and H. perforatum .  Chem Nat Compounds. 1972;  8 646-7.
• 3 Akhmedkhodzhaeva Kh S, Kurmukov A G. Some pharmacological properties of eudesmin.  Reports Uzb Acad Sci. 1975;  1 34-5.
• 4 Cho J Y, Yoo E S, Baik K U, Park M H. Eudesmin inhibits tumor necrosis factor-alpha production and T cell proliferation.  Arch Pharm Res. 1999;  22 348-53.
• 5 Nesmelova E F, Razakova D M, Akhmedjanova V I, Bessonova I A. Diphyllin from Haplophyllum alberti-regelii, H. bucharicum, and H. perforatum .  Chem Nat Compounds. 1983;  19 608-9.
• 6 Al-Abed Y, Abu-Zarga M, Sabri S, Atta-Ur-Rahman , Voelter W. A arylnaphthalene lignan from Haplophyllum buxbaumii .  Phytochemistry. 1998;  49 1779-81.
• 7 Okigawa M, Maeda T, Kawano N. The isolation and structure of three new lignans from Justica procumbens Linn. var. leucantha Honda.  Tetrahedron. 1970;  26 4301-5.
• 8 Fukamiya N, Lee K H. Antitumor agents, 81. Justicidin-A and diphyllin, two cytotoxic principles from Justicia procumbens .  J Nat Prod. 1986;  49 348-50.
• 9 Susplugas S, Hung N V, Bignon J, Thoison O, Kruczynski A, Sevenet T. et al . Cytotoxic arylnaphthalene lignans from a Vietnamese acanthaceae, Justicia patentiflora .  J Nat Prod. 2005;  68 734-8.
• 10 Di Giorgio C, Delmas F, Akhmedjanova V, Ollivier E, Bessonova I, Riad E. et al . In vitro antileishmanial activity of diphyllin isolated from Haplophyllum bucharicum .  Planta Med. 2005;  71 366-9.
• 11 Leonard G D, Fojo T, Bates S E. The role of ABC transporters in clinical practice.  Oncologist. 2003;  8 11-24.
• 12 Barthomeuf C, Debiton E, Mshvildadze V, Kemertelidze E, Balansard G. In vitro activity of hederacolchiside A1 compared with other saponins from Hedera colchica against the proliferation of human carcinoma and melanoma cells.  Planta Med. 2002;  68 672-5.
• 13 Govindachari T R, Sathe S S, Viswanathan N, Pai B R, Srinivasan M. Revised structures of diphyllin and justicidin A.  Tetrahedron Lett. 1967;  36 3517-9.
• 14 Barthomeuf C, Grassi J, Demeule M, Fournier C, Boivin D, Beliveau R. Inhibition of P-glycoprotein transport function and, reversion of MDR1 multidrug resistance by cnidiadin.  Cancer Chemother Pharmacol. 2005;  56 173-81.
• 15 Barthomeuf C, Demeule M, Grassi M, Saidkhodjaev A, Beliveau R. Conferone from Ferula schtschurowskiana enhances vinblastine cytotoxicity in MDCK-MDR1 cells by competively inhibiting P-glycoprotein transport function.  Planta Med. 2006;  72 634-9.
• 16 Pradheepkumar C P, Shanmugam G. Anticancer potential of cleistanthin A isolated from the tropical plant Cleistanthus collinus .  Oncol Res. 1999;  11 225-32.
• 17 Meenakshi J, Shanmugam G. Inhibition of matrix metalloproteinase-9 (MMP-9) activity by cleistanthin A, a diphyllin glycoside from Cleistanthus collinus .  Drug Dev Res. 2000;  50 93-4.
• 18 Day S H, Lin Y C, Tsai M L, Tsao L T, Ko H H, Chung M I. et al . Potent cytotoxic lignans from Justicia procumbens and their effects on nitric oxide and tumor necrosis factor-alpha production in mouse macrophages.  J Nat Prod. 2002;  65 379-81.
• 19 Prieto J M, Giner R M, Recio M C, Schinella G, Manez S, Rios J L. Diphyllin acetylapioside, a 5-lipoxygenase inhibitor from Haplophyllum hispanicum .  Planta Med. 2002;  68 359-60.
• 20 Innocenti G, Puricelli L, Piacente S, Caniato R, Filippini R, Cappelletti E M. Patavine, a new arylnaphthalene lignan glycoside from shoot cultures of Haplophyllum patavinum .  Chem Pharm Bull. 2002;  50 844-6.