A tyrosine kinase inhibitor, vandetanib (Van), is an anticancer drug affecting the signaling of VEGFR, EGFR and RET protooncogenes. Van is primarily used for the treatment of advanced or metastatic medullary thyroid cancer; however, its usage is significantly limited by side effects, particularly cardiotoxicity. One approach to minimize them is the encapsulation or binding of Van in- or onto a suitable carrier, allowing targeted delivery to tumor tissue. Herein, we constructed a nanocarrier based on apoferritin associated with Van (ApoVan). Based on the characteristics obtained by analyzing the average size, the surface ζ-potential and the polydispersive index, ApoVan nanoparticles exhibit long-term stability and maintain their morphology. Experiments have shown that ApoVan complex is relatively stable during storage. It was found that Van is gradually released from its ApoVan form into the neutral environment (pH 7.4) as well as into the acidic environment (pH 6.5). The effect of free Van and ApoVan on neuroblastoma and medullary thyroid carcinoma cell lines revealed that both forms were toxic in both used cell lines, and minimal differences between ApoVan and Van were observed. Thus, we assume that Van might not be encapsulated into the cavity of apoferritin, but instead only binds to its surface.

• Klíčová slova
• apoferritin, cancer targeting, medullary thyroid cancer, neuroblastoma, vandetanib,
• MeSH
• apoferritiny chemie   farmakokinetika   MeSH
• chinazoliny chemie   farmakokinetika   MeSH
• koncentrace vodíkových iontů MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• nanočástice chemie   MeSH
• piperidiny chemie   farmakokinetika   MeSH
• stabilita léku MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• apoferritiny MeSH
• chinazoliny MeSH
• piperidiny MeSH
• vandetanib MeSH Prohlížeč

INTRODUCTION: The present study reports on examination of the effects of encapsulating the tyrosine kinase inhibitors (TKIs) vandetanib and lenvatinib into a biomacromolecular ferritin-based delivery system. METHODS: The encapsulation of TKIs was performed via two strategies: i) using an active reversible pH-dependent reassembly of ferritin´s quaternary structure and ii) passive loading of hydrophobic TKIs through the hydrophobic channels at the junctions of ferritin subunits. After encapsulation, ferritins were surface-functionalized with folic acid promoting active-targeting capabilities. RESULTS: The physico-chemical and nanomechanical analyses revealed that despite the comparable encapsulation efficiencies of both protocols, the active loading affects stability and rigidity of ferritins, plausibly due to their imperfect reassembly. Biological experiments with hormone-responsive breast cancer cells (T47-D and MCF-7) confirmed the cytotoxicity of encapsulated and folate-targeted TKIs to folate-receptor positive cancer cells, but only limited cytotoxic effects to healthy breast epithelium. Importantly, the long-term cytotoxic experiments revealed that compared to the pH-dependent encapsulation, the passively-loaded TKIs exert markedly higher anticancer activity, most likely due to undesired influence of harsh acidic environment used for the pH-dependent encapsulation on the TKIs' structural and functional properties. CONCLUSION: Since the passive loading does not require a reassembly step for which acids are needed, the presented investigation serves as a solid basis for future studies focused on encapsulation of small hydrophobic molecules.

• Klíčová slova
• drug delivery, lenvatinib, nanomedicine, vandetanib,
• MeSH
• biokompatibilní materiály chemie   MeSH
• buněčná smrt účinky léků   MeSH
• buněčné klony MeSH
• buněčné linie MeSH
• chinazoliny chemie   farmakologie   MeSH
• chinoliny chemie   farmakologie   MeSH
• difuze MeSH
• fenylmočovinové sloučeniny chemie   farmakologie   MeSH
• ferritiny chemie   MeSH
• inhibitory proteinkinas farmakologie   MeSH
• koncentrace vodíkových iontů MeSH
• koně MeSH
• kyselina listová chemie   MeSH
• lékové transportní systémy * MeSH
• lidé MeSH
• nosiče léků chemie   MeSH
• piperidiny chemie   farmakologie   MeSH
• pohyb buněk účinky léků   MeSH
• povrchové vlastnosti MeSH
• protinádorové látky farmakologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• biokompatibilní materiály MeSH
• chinazoliny MeSH
• chinoliny MeSH
• fenylmočovinové sloučeniny MeSH
• ferritiny MeSH
• inhibitory proteinkinas MeSH
• kyselina listová MeSH
• lenvatinib MeSH Prohlížeč
• nosiče léků MeSH
• piperidiny MeSH
• protinádorové látky MeSH
• vandetanib MeSH Prohlížeč

BACKGROUND: Currently, the diagnosis and treatment of neuroblastomas-the most frequent solid tumors in children-exploit the norepinephrine transporter (hNET) via radiolabeled norepinephrine analogs. We aim to develop a nanomedicine-based strategy towards precision therapy by targeting hNET cell-surface protein with hNET-derived homing peptides. RESULTS: The peptides (seq. GASNGINAYL and SLWERLAYGI) were shown to bind high-resolution homology models of hNET in silico. In particular, one unique binding site has marked the sequence and structural similarities of both peptides, while most of the contribution to the interaction was attributed to the electrostatic energy of Asn and Arg (< - 228 kJ/mol). The peptides were comprehensively characterized by computational and spectroscopic methods showing ~ 21% β-sheets/aggregation for GASNGINAYL and ~ 27% α-helix for SLWERLAYGI. After decorating 12-nm ferritin-based nanovehicles with cysteinated peptides, both peptides exhibited high potential for use in actively targeted neuroblastoma nanotherapy with exceptional in vitro biocompatibility and stability, showing minor yet distinct influences of the peptides on the global expression profiles. Upon binding to hNET with fast binding kinetics, GASNGINAYLC peptides enabled rapid endocytosis of ferritins into neuroblastoma cells, leading to apoptosis due to increased selective cytotoxicity of transported payload ellipticine. Peptide-coated nanovehicles significantly showed higher levels of early apoptosis after 6 h than non-coated nanovehicles (11% and 7.3%, respectively). Furthermore, targeting with the GASNGINAYLC peptide led to significantly higher degree of late apoptosis compared to the SLWERLAYGIC peptide (9.3% and 4.4%, respectively). These findings were supported by increased formation of reactive oxygen species, down-regulation of survivin and Bcl-2 and up-regulated p53. CONCLUSION: This novel homing nanovehicle employing GASNGINAYLC peptide was shown to induce rapid endocytosis of ellipticine-loaded ferritins into neuroblastoma cells in selective fashion and with successful payload. Future homing peptide development via lead optimization and functional analysis can pave the way towards efficient peptide-based active delivery of nanomedicines to neuroblastoma cells.

• Klíčová slova
• Ferritin, Homing peptide, Neuroblastoma, Norepinephrine transporter, Targeted therapy,
• MeSH
• endocytóza genetika   MeSH
• ferritiny chemie   MeSH
• lékové transportní systémy metody   MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• nanomedicína MeSH
• nanostruktury chemie   MeSH
• neuroblastom metabolismus   MeSH
• peptidy chemie   genetika   metabolismus   MeSH
• proteiny přenášející noradrenalin přes plazmatickou membránu * chemie   genetika   metabolismus   MeSH
• protinádorové látky chemie   farmakokinetika   farmakologie   MeSH
• viabilita buněk účinky léků   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• Názvy látek
• ferritiny MeSH
• peptidy MeSH
• proteiny přenášející noradrenalin přes plazmatickou membránu * MeSH
• protinádorové látky MeSH