PREPARATION OF CHEMICAL AND PHYSICAL CONJUGATES OF SELF-ASSEMBLING NANOPARTICLES WITH CELL-PENETRATING PEPTIDE AND DOXORUBICIN

Zhadyra Sagykyzy Shagyrova, Yerzhan Yersaiynuly Zhienbay, Mikhail Seregeyevich Voikov, Alexander Vyacheslavovich Shustov

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.


Keywords


virus-like particles, drug carrier, targeting, conjugate, transportan, Doxorubicin

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DOI: http://dx.doi.org/10.12955/cbup.v3.630

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