The cell surface receptor claudin-4 (Cld-4) is upregulated in various tumours and represents an important emerging target for both diagnosis and treatment of solid tumours of epithelial origin. The C-terminal fragment of the Clostridium perfringens enterotoxin cCPE290-319 appears as a suitable ligand for targeting Cld-4. The synthesis of this 30mer peptide was attempted via several approaches, which has revealed sequential SPPS using three pseudoproline dipeptide building blocks to be the most efficient one. Labelling with fluorine-18 was achieved on solid phase using N-succinimidyl 4-[18F]fluorobenzoate ([18F]SFB) and 4-[18F]fluorobenzoyl chloride as 18F-acylating agents, which was the most advantageous when [18F]SFB was reacted with the resin-bound 30mer containing an N-terminal 6-aminohexanoic spacer. Binding to Cld-4 was demonstrated via surface plasmon resonance using a protein construct containing both extracellular loops of Cld-4. In addition, cell binding experiments were performed for 18F-labelled cCPE290-319 with the Cld-4 expressing tumour cell lines HT-29 and A431 that were complemented by fluorescence microscopy studies using the corresponding fluorescein isothiocyanate-conjugated peptide. The 30mer peptide proved to be sufficiently stable in blood plasma. Studying the in vivo behaviour of 18F-labelled cCPE290-319 in healthy mice and rats by dynamic PET imaging and radiometabolite analyses has revealed that the peptide is subject to substantial liver uptake and rapid metabolic degradation in vivo, which limits its suitability as imaging probe for tumour-associated Cld-4.
- MeSH
- buňky HT-29 MeSH
- cílená molekulární terapie MeSH
- claudin-4 antagonisté a inhibitory chemie metabolismus MeSH
- enterotoxiny chemická syntéza chemie farmakokinetika farmakologie MeSH
- izotopové značení MeSH
- krysa rodu rattus MeSH
- lidé MeSH
- ligandy MeSH
- molekulární mimikry fyziologie MeSH
- molekulární zobrazování MeSH
- myši nahé MeSH
- myši MeSH
- nádory farmakoterapie MeSH
- potkani Wistar MeSH
- pozitronová emisní tomografie MeSH
- radioizotopy fluoru chemie MeSH
- techniky syntézy na pevné fázi MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- lidé MeSH
- mužské pohlaví MeSH
- myši MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- MeSH
- Bacteria MeSH
- bakteriologie trendy MeSH
- enterotoxiny * farmakologie imunologie izolace a purifikace MeSH
- imunoterapie * metody trendy využití MeSH
- infekce spojené se zdravotní péčí diagnóza komplikace terapie MeSH
- kontaminace potravin analýza prevence a kontrola MeSH
- lidé MeSH
- osteomyelitida diagnóza etiologie terapie MeSH
- stafylokokové infekce kůže diagnóza imunologie terapie MeSH
- stafylokokové infekce * diagnóza etiologie imunologie MeSH
- virové nemoci diagnóza komplikace terapie MeSH
- Check Tag
- lidé MeSH
