Identification and characterization of a novel peptide ligand of epidermal growth factor receptor for targeted delivery of therapeutics

Abstract

Epidermal growth factor receptor (ErbB1, EGFR) is overexpressed in a variety of human cancer cells. It has been considered as a rational target for drug delivery. To identify novel ligands with specific binding capabilities to EGFR, we screened a phage display peptide library and found an enriched phage clone encoding the amino acid sequence YHWYGYTPQNVI (designated as GE11). Competitive binding assay and Scatchard analysis revealed that GE11 peptide bound specifically and efficiently to EGFR with a dissociation constant of approximately 22 nM, but with much lower mitogenic activity than with EGF. We showed that the peptides were internalized preferentially into EGFR highly expressing cells, and they accumulated in EGFR overexpressing tumor xenografts after i.v. delivery in vivo. In gene delivery studies, GE11-conjugated polyethylenimine (PEI) vectors were less mitogenic, but still quite efficient at transfecting genes into EGFR highly expressing cells and tumor xenografts. Taken together, GE11 is a potentially safe and efficient targeting moiety for selective drug delivery systems mediated through EGFR.

Disconnecting the Yin and Yang Relation of Epidermal Growth Factor Receptor (EGFR)-Mediated Delivery: A Fully Synthetic, EGFR-Targeted Gene Transfer System Avoiding Receptor Activation

“…This is in line with observations made by Li and colleagues, where GE11 did not promote the mitosis of hepatoma cells (Li et al, 2005). For this purpose, we further evaluated the GE11 peptide on other tumor types, that is, hepatoma and prostate carcinoma.…”

“…The absorption of the thiopyridone group released was measured at 343 nm (e = 8080 M -1 ), and the LPEI/OPSS ratio was determined as 1:1.5 (M/M). To a peptide sequence derived from the middle loop of human EGF (MYIEALD KYA; Komoriya et al, 1984) a terminal cysteine was added, either at the N terminus (CMYIEALDKYA, termed CMY) or the C terminus (MYIEALDKYAC, termed MYI); the peptide sequence GE11 (YHWYGYFPQNVI; Li et al, 2005) was synthesized with an N-terminal cysteine (CYHWY GYTPQNVI, termed GE11). Cysteine or cysteine-modified peptides were added to LPEI-PEG-OPSS and the reaction of peptides or cysteine with the OPSS group was quantitative as measured by thiopyridone release at 343 nm.…”

“…Although GE11 peptide has a 10-fold lower affinity for EGFR (K d = 20 nM; Li et al, 2005), GE11 polyplexes were able to induce similar or even higher transgene expression levels when compared with EGF polyplexes (Figs. 1 and 3).…”

Population pharmacokinetic data analysis of three phase I studies of matuzumab, a humanised anti-EGFR monoclonal antibody in clinical cancer development

“…colon, mamma and bronchial carcinoma) and is often associated with a high metastatic rate, poor prognosis and advanced disease progression (Dassonville et al, 1993;Rusch et al, 1993;Salomon et al, 1995). Epidermal growth factor receptor can be used as a target for therapies based on blockade of receptor -ligand interactions and inhibition of downstream signalling pathways such as cell proliferation, angiogenesis and invasion as well as increase of apoptosis (Ritter and Arteaga, 2003;Li et al, 2005). Epidermal growth factor receptor and other members of this receptor family have already been successful targets for cancer therapy (Wells, 1999).…”

Potential of phage-displayed peptide library technology to identify functional targeting peptides

“…phage-displayed peptide library against recombinant human epidermal growth factor receptor (hEGFR) in vitro and isolated the phage displaying YHWYGYTPQNVI peptide (GE11) [81]. Synthetic free GE11 peptide and epidermal growth factor (EGF) inhibited phage binding to human EGF receptor (hEGFR) as well as the EGFR-positive human hepatocarcinoma cell line SMMC-7721.…”