Enhancement of Solubility and Bioavailability of β-Lapachone Using Cyclodextrin Inclusion Complexes
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Purpose. To explore the use of cyclodextrins (CD) to form inclusion complexes with β-lapachone (β-lap) to overcome solubility and bioavailability problems previously noted with this drug.
Methods. Inclusion complexes between β-lap and four cyclodextrins (α-, β-, γ-, and HPβ-CD) in aqueous solution were investigated by phase solubility studies, fluorescence, and 1H-NMR spectroscopy. Biologic activity and bioavailability of β-lap inclusion complexes were investigated by in vitro cytotoxicity studies with MCF-7 cells and by in vivo lethality studies with C57Blk/6 mice (18-20 g).
Results. Phase solubility studies showed that β-lap solubility increased in a linear fashion as a function of α-, β-, or HPβ-CD concentrations but not γ-CD. Maximum solubility of β-lap was achieved at 16.0 mg/ml or 66.0 mM with HPβ-CD. Fluorescence and 1H-NMR spectroscopy proved the formation of 1:1 inclusion complexes between β-CD and HPβ-CD with β-lap. Cytotoxicity assays with MCF-7 cells showed similar biologic activities of β-lap in β-CD or HPβ-CD inclusion complexes (TD50 = 2.1 μM). Animal studies in mice showed that the LD50 value of β-lap in an HPβ-CD inclusion complex is between 50 and 60 mg/kg.
Conclusions. Complexation of β-lap with HPβ-CD offers a major improvement in drug solubility and bioavailability.
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- 1.A. Marin, A. Lopex de Cerain, E. Hamilton, A. D. Lewis, J. M. Martinez-Penuela, M. A. Idoate, and J. Bello. DT-diaphorase and cytochrome B5 reductase in human lung and breast tumors. Br. J. Cancer 76:923-929 (1997).Google Scholar
- 2.A. M. Malkinson, D. Siegel, G. L. Forrest, A. F. Gazdar, H. K. Oie, D. C. Chan, P. A. Bunn, M. Mabry, D. J. Dykes, S. D. Harrison, and D. Ross. Elevated DT-diaphorase activity and messenger RNA content in human non-small cell lung carcinoma: relationship to the response of lung tumor xenografts to mitomycin Cl. Cancer Res. 52:4752-4757 (1992).Google Scholar
- 3.A. M. Malkinson. Molecular comparison of human and mouse pulmonary adenocarcinomas. Exp. Lung Res. 24:541-555 (1998).Google Scholar
- 4.M. Belinsky and A. K. Jaiswal. NAD(P)H:quinone oxidoreductase1 (DT-diaphorase) expression in normal and tumor tissues. Cancer Metastasis Rev. 12(2):103-117 (1993).Google Scholar
- 5.V. J. Stella and R. A. Rajewski. Cyclodextrins: Their future drug formulation and delivery. Pharm. Res. 14:556-567 (1997).Google Scholar
- 6.J. Szejtli. Introduction and general overview of cyclodextrin chemistry. Chem. Rev. 98:1743-1753 (1998).Google Scholar
- 7.T. Loftsson and M. E. Brewster. Pharmaceutical applications of cyclodextrins. 1. Drug solubilization and stabilization, J Pharm Sci. 85:1017-1025 (1996).Google Scholar
- 8.R. A. Rajewski and V. J. Stella. Pharmaceutical applications of cyclodextrins. 2. In vivo drug delivery. J. Pharm. Sci. 85:1142-1169 (1996).Google Scholar
- 9.T. Irie and K. Uekama. Pharmaceutical applications of cyclodextrins. 3. Toxicological issues and safety evaluation. J. Pharm. Sci. 86:147-162 (1997).Google Scholar
- 10.M. V. Rekharsky and Y. Inoue. Complexation thermodynamics of cyclodextrin. Chem. Rev. 98:1875-1917 (1998).Google Scholar
- 11.J. L. Lach and T. F. Chin. Interaction of pharmaceuticals with Schardinger dextrins III. J. Pharm. Sci. 53:69-73 (1964).Google Scholar
- 12.S. M. Planchon, S. Wuerzberger, B. Frydman, D. T. Witiak, P. Hutson, D. R. Church, G. Wilding, and D. A. Boothman. Beta-lapachone-mediated apoptosis in human promyelocytic leukemia (HL-60) and human prostate cancer cells: a p53-independent response. Cancer Res. 55:3706-3711 (1995).Google Scholar
- 13.J. J. Pink, S. M. Planshon, C. Tagliarino, S. M. Wuerzberger-Davis, M. E. Varnes, D. Siegel, and D. A. Boothman. NAD(P)H:quinone oxidoreductase (NQO1) activity is the principal determinant of beta-lapachone cytoxicity. J. Biol. Chem. 275:5416-5424 (2000).Google Scholar
- 14.T. Higuchi and K. A. Connors. Phase solubility techniques. Adv. Anal. Chem. Instrum. 4:117-212 (1965).Google Scholar
- 15.A. Botsi, K. Yannakopoulou, B. Perly, and E. Hadjoudis. Positives or adverse effects of methylation on the inclusion behavior of cyclodextrins. A comparative NMR study using pheromone constituents of the olive fruit fly. J. Org. Chem. 60:4017-4023 (1995).Google Scholar
- 16.S. M. Wuerzberger, J. J. Pink, S. M. Planchon, K. L. Byers, W. G. Bornmann, and D. A. Boothman. Induction of apoptosis in MCF-7:WS8 breast cancer cells by beta-lapachone. Cancer Res. 58:1876-1885 (1998).Google Scholar
- 17.C. Labarca and K. Paigen. A simple, rapid, and sensitive DNA assay procedure. Anal. Biochem. 102:344-352 (1980).Google Scholar
- 18.K. A. Connors. The stability of cyclodextrin complexes in solution. Chem. Rev. 97:1325-1357 (1997).Google Scholar
- 19.P. Adell, T. Parella, F. Sanchez-Ferrando, and A. Virgili. Clean selective spin-locking spectra using pulsed field gradients. J. Magn. Reson. 108:77-80 (1995).Google Scholar
- 20.Y. Ikeda, S. Motoune, T. Matsuoka, H. Arima, F. Hirayama, and K. Uekama. Inclusion complex formation of captopril with α-and β-cyclodextrins in aqueous solution: NMR spectroscopic and molecular dynamic studies. J. Pharm. Sci. 91:2390-2398 (2002).Google Scholar
- 21.V. Ramamurthy and D. F. Eaton. Photochemistry and photophysics within cyclodextrin cavities. Acc. Chem. Res. 21:300-306 (1988).Google Scholar
- 22.E. E. Sideris, G. N. Valsami, M. A. Koupparis, and P. E. Macheras. Determination of association constants in cyclodextrin/drug complexation using the Scatchard plot: application to β-cyclodextrin anilinonaphthalenesulfonates. Pharm. Res. 9:1568-1574 (1992).Google Scholar
- 23.C. Tagliarino, J. J. Pink, G. R. Dubyak, A. L. Nieminen, and D. A. Boothman. Calcium is a key signaling molecule in beta-lapachone-mediated cell death. J. Biol. Chem. 276:19150-19159 (2001).Google Scholar
- 24.C. J. Li, Y. Z. Li, A. V. Pinto, and A. B. Pardee. Potent inhibition of tumor survival in vivo by beta-lapachone plus taxol: combining drugs imposes different artificial checkpoints. Proc. Natl. Acad. Sci. USA 96:13369-13374 (1999).Google Scholar