Synthesis and Preliminary Evaluation of a New ^99mTc Labeled Substance P Analogue as a Potential Tumor Imaging Agent

Abstract

Neurokinin 1 receptors (NK ₁R) are overexpressed on several types of important human cancer cells. Substance P (SP) is the most specific endogenous ligand known for NK ₁Rs. Accordingly,a new SP analogue was synthesized and evaluated for detection of NK ₁R positive tumors.[6-hydrazinopyridine-3-carboxylic acid (HYNIC)-Tyr ⁸-Met(O) ¹¹-SP] was synthesized and radiolabeled with ^99mTc using ethylenediamine-N,N'-diacetic acid (EDDA)and Tricine as coligands. Common physicochemical properties of radioconjugate were studied and in-vitro cell line biological tests were accomplished to determine the receptor mediated characteristics. In-vivo biodistribution in normal and tumor bearingnude mice was also assessed. The cold peptide was prepared in high purity (>99%) and radiolabeled with ^99mTc at high specific activities (84-112GBq/µmol) with an acceptable labeling yield (>95%). The radioconjugate was stable in-vitro in the presence of human serum and showed 44% protein binding to human serumalbumin. In-vitro cell line studies on U373MG cells showed an acceptable uptake up to 4.91 ± 0.22% with the ratio of 60.21 ± 1.19% for its specific fraction and increasing specific internalization during 4 h. Receptor binding assays on U373MG cells indicated a mean Kd of 2.46 ± 0.43 nM and Bmax of 128925 ± 8145 sites/cell. In-vivo investigations determined the specific tumor uptake in 3.36 percent of injected dose per gram (%ID/g) for U373MG cells and noticeable accumulations of activity in the intestines and lung. Predominant renal excretion pathway was demonstrated.

Therefore, this new radiolabeled peptide could be a promising radiotracer for detection of NK ₁R positive primary or secondary tumors.