Objective: The aim of this study was to evaluate the 99mTc-HYNIC-D4 peptide as a new radiotracer for
tumor targeting in non small cell lung tumor that overexpresses epidermal growth factor receptor (EGFR).
Method: D4 small peptide (Leu-Ala-Arg-Leu-Leu-Thr) was selected as a candidate for specific targeting on EGFR. It
was conjugated with HYNIC on N-terminus and labeled with 99mTc using tricine as a co-ligand. The cellular and
animal studies were evaluated for specific binding of this labeled peptide.
Result: Radiochemical purity was determined to be 98% by instant thin layer chromatography and reverse-phase high
performance liquid chromatography. Stability of radiolabeled peptide was more than 90% up to 24 h incubation in
solution at 37 oC. Specific binding of the radiolabeled peptide to the EGFR showed dissociation constant of 181 ± 41
nM. Animal biodistribution in normal and A-549 xenografted nude mice showed rapid clearance from blood and other
non target organs. Tumor uptake values as %ID/g (percentage of injection dose per gram of tissue) were 8.07% at 1 h
and 3.82% at 4 h after injection that was blocked by presaturation of EGFR using an excess of cold peptide.
Conclusion: This study showed that 99mTc-HYNIC-D4 is a promising tumor targeting radiotracer on the non small cell