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The Pharmaceutical Society of Korea (대한약학회)
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Synthesis, Characterization and In Vitro Evaluation of Triptolide-lysozyme Conjugate for Renal Targeting Delivery of Triptolide

  • Zheng, Qiang (Key Laboratory of Drug Targeting and Drug Delivery, Ministry of Education, West China School of Pharmacy, Sichuan University) ;
  • Gong, Tao (Key Laboratory of Drug Targeting and Drug Delivery, Ministry of Education, West China School of Pharmacy, Sichuan University) ;
  • Sun, Xun (Key Laboratory of Drug Targeting and Drug Delivery, Ministry of Education, West China School of Pharmacy, Sichuan University) ;
  • Zhang, Zhi-Rong (Key Laboratory of Drug Targeting and Drug Delivery, Ministry of Education, West China School of Pharmacy, Sichuan University)
  • Published : 2006.12.31
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Abstract

A triptolide-lysozyme (TP-LZM) conjugate was synthesized to achieve renal specific delivery and to reduce the side effects of triptolide. Triptolide was coupled to lysozyme through succinic via an ester bond with an average coupling degree of 1 mol triptolide per 1 mol lysozyme. The lysozyme can specifically accumulate in the proximal tubular cells of the kidney, making it a potential carrier for targeting drugs to the kidney. The structure of triptolide succinate (TPS) was confirmed by IR, $^{1}H-NMR$, MS and UV. The concentrations of triptolide in various samples were determined by reversed-phase high-performance liquid chromatography (HPLC). In this study, the physicochemical and stability profiles of TP-LZM under various conditions were investgated the stability and releasing profiles of triptolide-lysozyme (TP-LZM) under various conditions. In vitro release trails showed triptolide-lysozyme was relatively stable in plasma (less than 30% of free triptolide released) and could release triptolide quickly in lysosome (more than 80% of free triptolide released) at $37^{\circ}C$ for 24 h. In addition, the biological activities of the conjugate on normal rat kidney proximal tubular cells (NRK52E) were also tested. The conjugate can effectively reduce NO production in the medium of NRK52E induced by lipopolysaccharide (LPS) but with much lower toxicity. These studies suggest the possibility to promote curative effect and reduce its extra-renal toxicity of triptolide by TP-LZM conjugate.

Keywords

References

  1. Banu, N. and Meyers, C. M., IFN-gamma and LPS differentially modulate class II MHC and B7-1 expression on murine renal tubular epithelial cells. Kidney Int., 55, 2250-63 (1999) https://doi.org/10.1046/j.1523-1755.1999.00495.x
  2. Fischer, D., Bieber, T., Li, Y., Elsasser, H. P., and Kissel, T., A novel non-viral vector for DNA delivery based on low molecular weight, branched polyethylenimine: effect of molecular weight on transfection efficiency and cytotoxicity. Pharm. Res., 16, 1273-1279 (1999) https://doi.org/10.1023/A:1014861900478
  3. Eric, J. F. F., Moolenaar, F., de Zeeuw, D., and Meijer, D. K. F., Low molecular weight proteins as carriers for renal drug targeting: Naproxen coupled to lysozyme via the spacer Llactic acid. Pharmacol. Res., 10, 963-969 (1993) https://doi.org/10.1023/A:1018946219057
  4. Gu, W. Z., Brandwein, S. R., and Banerjee, S., Inhibition of type II collagen induced arthritis in mice by an immunosuppressive extract of Tripterygium wilfordii Hook F. J . Rheumatol., 19, 682-8 (1992)
  5. Gu, W. Z., Chen, R., Brandwein, S., McAlpine, J., and Burres, N., Isolation, purification, and characterization of immunosuppressive compounds from Tripter ygium: triptolide andtripdiolide. Int. J. Immunopharmacol., 17, 241-51(1995)
  6. Haas, M., de Zeeuw, D., Zanten, A. V., and Meijer, D. K. F., Quantification of renal low-molecular-weight protein handling in the intact rat. Kidney Int., 43, 949-954 (1993) https://doi.org/10.1038/ki.1993.133
  7. Woods, J. S., Ellis, M. E., Dieguez-Acun, F. J., and Corral, J., Activation of NF-kappa in normal rat kidney epithelial (NRK52E) cells is mediated via a redox-insensitive, calciumdependent pathway. Toxicology and Applied Pharmacology, 154, 219-227 (1999) https://doi.org/10.1006/taap.1998.8583
  8. Jevnikar, A. M., Wuthrich, R. P., Takei, F., Xu, H. W., Brennan, D. C., and Glimcher, L. H., Differing regulation and function of ICAM-1 and clas s II antigens on renal tubular cells. Kidney Int., 38, 417-25 (1990) https://doi.org/10.1038/ki.1990.221
  9. Kok, R. J., Grijpstra, F., Walthuis, R. B., Moolenaar, F., de Zeeuw, D., and Meijer, K. F., Specific delivery of captopril to the kidney with the prodrug captopril-lysozyme. J. Pharmacol. Exp. Ther., 288, 281-285 (1999)
  10. Li, H., Liu, Z. H., Dai, C. S., Liu, D., and Li, L. S., Triptolide inhibits proinflammatory factor-induced over-expression of class II MHC and B7 molecules in renal tubular epithelial cells. Acta Pharmacol Sin., 23, 75-81 (2002)
  11. Maack, T., Johnson, V., Kau, S. T., Figueiredo, J., and Sigulem, D., Renal filtration, transport, and metabolism of lowmolecular- weight proteins: A review. Kidney Int.,16, 251-270 (1979) https://doi.org/10.1038/ki.1979.128
  12. Mehvar, R., Dann, R. O., and Hoganson, D. A., Kinetics of hydrolysis of dextran-methylprednisolone succinate,a macromolecular prodrug of methylprednisolone, in rat blood and liver lysosomes. Journal of Controlled Release, 68, 53-61 (2000) https://doi.org/10.1016/S0168-3659(00)00234-0
  13. Folgert, R., Haverdings, G., and Haas, M., Renal targeting of catopril selectively enhances the intrarenal over the systematic effects of ACE inhibition in rats. British Journal of Pharmacology, 136, 1107-1116 ( 2002) https://doi.org/10.1038/sj.bjp.0704814
  14. Tao, X. L., Davis, L. S., and Lipsky, P. E., Effect of the Chinese herbal remedy Tripterygium wilfordii Hook F on human immune responsiveness. Arthritis Rheum., 34, 1274-81 (1991) https://doi.org/10.1002/art.1780341011
  15. Yang, Y, Liu, Z. H., Tolosa, E., Yang, J. W., and Li, L. S., Triptolide induces apoptotic death of T lymphocyte. Immunopharmacology, 40,139-49 (1998) https://doi.org/10.1016/S0162-3109(98)00036-8
  16. Kim, Y. H., Lee, S. H., Lee, J. Y., Choi, S. W., Parka, J. W., and Kwon, T. K., Triptolide inhibits murine-inducible nitric oxide synthase expression by down-regulating lipopolysaccharideinduced activity of nuclear factor-kB and c-Jun NH2-terminal kinase. Eur. J. Pharmacol., 494, 1-9 (2004) https://doi.org/10.1016/j.ejphar.2004.04.040
  17. Mei, Z., Chen, H., and Weng, T., Solid lipid nanoparticle and microemulsion for topical delivery of triptolide. Eur. J. Pharmaceutics and Biopharmaceutics, 56, 189-196 (2003) https://doi.org/10.1016/S0939-6411(03)00067-5
  18. Zheng, Q. and Zhang Z. R., Progress in renal drug targeting. Acta Phamacutica Sinica, 40, 199-203 (2005)
  19. van Kooten, C., Daha, M. R., and van Es, L. A., Tubular epithelial cells: A critical cell type in the regulation of renal inflammatory processes. Exp. Nephrol., 7, 429-37 (1999) https://doi.org/10.1159/000020622