Virus-Derived Peptides for Hepatic Enzyme Delivery
| Autor: | Stephan Urban, Anna Pratsinis, Philipp Uhl, Dominik Witzigmann, Walter Mier, Susanne H. Schenk, Jörg Huwyler, Patrick Hauswirth, Jan Stephan Bolten |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
Embryo
Nonmammalian Enzyme Therapy Organic Anion Transporters Sodium-Dependent Pharmaceutical Science 02 engineering and technology digestive system 030226 pharmacology & pharmacy Horseradish peroxidase Lipopeptides Mice 03 medical and health sciences chemistry.chemical_compound 0302 clinical medicine Non-competitive inhibition In vivo Cell Line Tumor Drug Discovery Animals Humans Prodrugs Tissue Distribution Receptor Zebrafish Drug Carriers Symporters biology Chemistry Calcium-Binding Proteins Lipopeptide Zebrafish Proteins Prodrug 021001 nanoscience & nanotechnology In vitro Enzymes HEK293 Cells Liver Biochemistry Models Animal Hepatocytes biology.protein Molecular Medicine 0210 nano-technology Ex vivo |
| Zdroj: | Molecular Pharmaceutics. 18:2004-2014 |
| ISSN: | 1543-8392 1543-8384 |
| DOI: | 10.1021/acs.molpharmaceut.0c01222 |
| Popis: | Recently, a lipopeptide derived from the hepatitis B virus (HBV) large surface protein has been developed as an HBV entry inhibitor. This lipopeptide, called MyrcludexB (MyrB), selectively binds to the sodium taurocholate cotransporting polypeptide (NTCP) on the basolateral membrane of hepatocytes. Here, the feasibility of coupling therapeutic enzymes to MyrB was investigated for the development of enzyme delivery strategies. Hepatotropic targeting shall enable enzyme prodrug therapies and detoxification procedures. Here, horseradish peroxidase (HRP) was conjugated to MyrB via maleimide chemistry, and coupling was validated by SDS-PAGE and reversed-phase HPLC. The specificity of the target recognition of HRP-MyrB could be shown in an NTCP-overexpressing liver parenchymal cell line, as demonstrated by competitive inhibition with an excess of free MyrB and displayed a strong linear dependency on the applied HRP-MyrB concentration. In vivo studies in zebrafish embryos revealed a dominating interaction of HRP-MyrB with scavenger endothelial cells vs xenografted NTCP expressing mammalian cells. In mice, radiolabeled 125I-HRP-MyrBy, as well as the non-NTCP targeted control HRP-peptide-construct (125I-HRP-alaMyrBy) demonstrated a strong liver accumulation confirming the nonspecific interaction with scavenger cells. Still, MyrB conjugation to HRP resulted in an increased and NTCP-mediated hepatotropism, as revealed by competitive inhibition. In conclusion, the model enzyme HRP was successfully conjugated to MyrB to achieve NTCP-specific targeting in vitro with the potential for ex vivo diagnostic applications. In vivo, target specificity was reduced by non-NTCP-mediated interactions. Nonetheless, tissue distribution experiments in zebrafish embryos provide mechanistic insight into underlying scavenging processes indicating partial involvement of stabilin receptors. |
| Databáze: | OpenAIRE |
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