154
Compositions and Methods for the Inhibition of Fibrosis
Wisconsin Alumni Research Foundation
Peptide
Fibrosis
Hit To Lead or Lead Optimization
• A PEGylated functional upstream domain peptide (PEG-FUD), which tightly binds to fibronectin and inhibits the development of the fine fibrils that precede collagen deposits, preventing the formation of scar tissue.
• 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 ionexchange chromatography. The molecular weight and degree of PEGylation of each conjugate was verified using MALDITOF.
• 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.
US10828372B2
Characterization of the PEGylated Functional Upstream Domain Peptide (PEG-FUD): a Potent Fibronectin Assembly Inhibitor with Potential as an Anti-Fibrotic Therapeutic. Pharmaceutical Research, (2018)
