PREPARATION OF CHEMICAL AND PHYSICAL CONJUGATES OF SELF-ASSEMBLING NANOPARTICLES WITH CELL-PENETRATING PEPTIDE AND DOXORUBICIN
Abstract
Abstract: Nano-sized carriers can help to reduce toxicity and improve clinical efficacy of drugs. Virus-like particles (VLPs) are biocompatible and biodegradable self-assembling nanoparticles, which show great promise as carriers for substances for targeted delivery and controlled release. Either chemical conjugation of physical incorporation without formation of covalent bonds is possible to load substances of interest into VLPs.
Objectives: To produce VLPs from recombinant viral capsid protein (HBcAg) and test feasibility of methods of formation of chemical and physical conjugates of VLPs with substances of pharmacological interest.
Methods: Virus-like particles composed from recombinant hepatitis B core antigen (HBcAg) were produced by recombinant expression in E.coli and purified by successive centrifugation through sucrose gradients. Peptide transportan 10 was synthesized and used for carbodiimide (EDC)-mediated conjugation to VLPs. Doxorubicin (DOX) was loaded into the nucleic acid-containing VLPs to form physical conjugate.
Results: VLPs with chemically attached moieties of cell-penetrating peptide transportan 10 were produced. The conjugate was examined in SDS-PAGE to confirm presence of conjugation products. Conjugation efficiency (molar ration peptide/protein in the conjugate) reaches 0.5:1 (i.e. 50% of protein chains have one attached peptide moiety). The nucleic acid-containing VLPs can be loaded with the DOX forming stable non-covalent physical conjugate.
Conclusion: Recombinantly expressed VLPs allow easy attaching of small molecules making them a convenient platform to develop drug carriers.
References
Bally, M. B., Nayar, R., Masin, D., Cullis, P. R., & Mayer, L. D. (1990). Studies on the myelosuppressive activity of doxorubicin entrapped in liposomes. Cancer Chemotherapy and Pharmacology, 27, 13-19, doi: 10.1007/BF00689270
Cai, B., Lin, Y., Xue, X. H., Fang, L., Wang, N., & Wu, Z. Y. (2011). TAT-mediated delivery of neuroglobin protects against focal cerebral ischemia in mice. Experimental Neurology, 227, 224-31. doi: 10.1016/j.expneurol.2010.11.009
Fonseca, S. B., Pereira, M. P., & Kelley, S. O. (2009). Recent advances in the use of cell-penetrating peptides for medical and biological applications. Advanced Drug Delivery Reviews, 61, 953-64. doi: 10.1016/j.addr.2009.06.001
Gabathuler, R. (2010). Development of new peptide vectors for the transport of therapeutic across the blood-brain barrier. Therapeutic Delivery, 1, 571-586. doi:10.4155/tde.10.35
Gallo, G. (2003). Making proteins into drugs: assisted delivery of proteins and peptides into living neurons. Methods in Cell Biology, 71, 325-338. doi: 10.1016/S0091-679X(03)01015-X
Heitz, F., Morris, M. C., & Divita, G. (2009). Twenty years of cell-penetrating peptides: from molecular mechanisms to therapeutics. British Journal of Pharmacology, 157, 195-206. doi:10.1111/j.1476-5381.2008.00057.x
Hwang do, W., Son, S., Jang, J., Youn, H., Lee, S., Lee, D., Lee, Y. S., Jeong, J. M., Kim, W. J., & Lee, D. S. (2011). A brain-targeted rabies virus glycoprotein-disulfide linked PEI nanocarrier for delivery of neurogenic microRNA. Biomaterials, 32, 4968-75. doi: 10.1016/j.biomaterials.2011.03.047
Kaplan, L. D., Du Mond, C., Mamelok, R. D., & Henry, D. H. (1998). Pegylated-liposomal doxorubicin versus doxorubicin, bleomycin, and vincristine in the treatment of AIDS-related Kaposi's sarcoma: results of a randomized phase III clinical trial. Journal of Clinical Oncology, 16, 2445-2451.
Kopecek, J., Rejmanova, P., Duncan, R., & Lloyd, J. B. (1985). Controlled release of drug model from N-[2-hydroxy-propyl] methacrylamide copolymers. Annals of the New York Academy of Sciences, 446, 93-104. doi: 10.1111/j.1749-6632.1985.tb18393.x
Kumar, P., Wu, H., McBride, J. L., Jung, K. E., Kim, M. H., Davidson, B. L., Lee, S. K., Shankar, P., & Manjunath, N. (2007). Transvascular delivery of small interfering RNA to the central nrvous system. Nature, 448, 39-43. doi:10.1038/nature05901
Levi-Schaffer, F., Bernstein, A., Meshorer, A., & Arnon, R. (1982). Reduced toxicity of daunorubicin by conjugation to dextran. Cancer Treatment Reports, 66, 107-114
Lohmann, A. E., Speers, C. H., & Chia, S. K. (2013). Evaluation of the clinical benefits of nanoparticle albumin-bound paclitaxel in women with metastatic breast cancer in British Columbia. Current Oncology, 20, 97-103. doi: 10.3747/co.20.1256.
Maeda, H. (2010). Tumor-selective delivery of macromolecular drugs via the EPR effect: background and future prospects. Bioconjugate Chemistry, 21, 797-802. doi: 10.1021/bc100070g
Northfelt, D. W., Dezube, B. J., Thommes, J. A., Miller, B. J., Fischl, M. A., Friedman-Kien, A., Plummer, E. M., & Manchester, M. (2011). Viral nanoparticles and virus-like particles: platforms for contemporary vaccine design. Wiley Interdisciplinary Reviews. Nanomedicine and Nanobiotechnology, 3, 174-196. doi: 10.1002/wnan.119
Qin, Y., Chen, H., Yuan, W., Kuai, R., Zhang, Q., Xie, F., Zhang, L., Zhang, Z., Liu, J., & He, Q. (2011). Liposome formulated with TAT-modified cholesterol for enhancing the brain delivery. International Journal of Pharmacy, 419, 85-95. doi: 10.1016/j.ijpharm.2011.07.021
Rahman, A., Joher, A., & Neefe, J. R. (1986) Immunotoxicity of multiple dosing regimens of free doxorubicin and doxorubicin entrapped in cardiolipin liposomes. British Journal of Cancer, 54, 401-408. doi:10.1038/bjc.1986.190
Whitacre, D. C., Lee, B. O., & Milich, D. R. (2009). Use of hepadnavirus core proteins as vaccine platforms. Expert Review of Vaccines, 8, 1565-1573. doi: 10.1586/erv.09.121
Xia, H., Gao, X., Gu, G., Liu, Z., Hu, Q., Tu, Y., Song, Q., Yao, L., Pang, Z., Jiang, X., Chen, J. & Chen, H. (2012). Penetratin-functionalized PEG-PLA nanoparticles for brain drug delivery. Int J Pharm., 436, 840-50. doi: 10.1016/j.ijpharm.2012.07.029
Zunino, F., Pratesi, G., & Pezzoni, G. (1987). Increased therapeutic efficacy and reduced toxicity of doxorubicin linked to pyran copolymer via the side chain of the drug. Cancer Treatment Reports, 71, 367-373
Copyright information
- Authors retain copyright and grant the journal right of first publication with the work simultaneously licensed under a Creative Commons Attribution License (Creative Commons Attribution License 3.0 - CC BY 3.0) that allows others to share the work with an acknowledgement of the work's authorship and initial publication in this journal.
- Authors are able to enter into separate, additional contractual arrangements for the non-exclusive distribution of the journal's published version of the work (e.g., post it to an institutional repository or publish it in a book), with an acknowledgement of its initial publication in this journal.
- Authors are permitted and encouraged to post their work online (e.g., in institutional repositories or on their website) prior to and during the submission process, as it can lead to productive exchanges, as well as earlier and greater citation of published work (See The Effect of Open Access).
info@iseic.cz, www.iseic.cz, ojs.journals.cz
