Characterization of the PEGylated Functional Upstream Domain Peptide (PEG-FUD): a Potent Fibronectin Assembly Inhibitor with Potential as an Anti-Fibrotic Therapeutic
| Autor: | Donna M. Peters, Bianca R. Tomasini-Johansson, Pawel Zbyszynski, Glen S. Kwon |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Male Biodistribution Cell Survival Pharmaceutical Science Peptide Article Polyethylene Glycols 03 medical and health sciences 0302 clinical medicine PEG ratio Humans Pharmacology (medical) IC50 Pharmacology chemistry.chemical_classification PUR4 Dose-Response Relationship Drug Chemistry Organic Chemistry technology industry and agriculture Fibroblasts Antifibrinolytic Agents Peptide Fragments Fibronectins 030104 developmental biology Fibronectin binding Biochemistry 030220 oncology & carcinogenesis Spectrometry Mass Matrix-Assisted Laser Desorption-Ionization PEGylation Molecular Medicine Biotechnology Conjugate |
| Zdroj: | Pharmaceutical research. 35(7) |
| ISSN: | 1573-904X |
| Popis: | To develop PEGylated variants of pUR4/FUD (FUD), a fibronectin assembly inhibitor, using 10 kDa, 20 kDa, and 40 kDa PEGs to evaluate their binding affinity and inhibitory potency. The FUD peptide was recombinantly expressed, purified, and PEGylated at the N-terminus using 10 kDa, 20 kDa, and 40 kDa methoxy-PEG aldehyde. The PEGylates were purified and fractionated using ion-exchange chromatography. The molecular weight and degree of PEGylation of each conjugate was verified using MALDI-TOF. The binding affinity of each PEG-FUD conjugate was studied using isothermal titration colorimetry (ITC) and their inhibitory potency was characterized by a cell-based matrix assembly in vitro assay. The 10 kDa, 20 kDa, and 40 kDa PEG-FUD conjugates were synthesized and isolated in good purity as determined by HPLC analysis. Their molecular weight was consistent with attachment of a single PEG molecule to one FUD peptide. The binding affinity (Kd) and the fibronectin fibrillogenesis inhibitory potency (IC50) of all PEG-FUD conjugates remained nanomolar and unaffected by the addition of PEG. Retention of FUD fibronectin binding activity following PEGylation with three different PEG sizes suggest that PEG-FUD holds promise as an effective anti-fibrotic with therapeutic potential and a candidate for further pharmacokinetic and biodistribution studies. |
| Databáze: | OpenAIRE |
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