VPAC1 Targeted 64Cu-TP3805 kit preparation and its evaluation

Introduction: Previously, our laboratory has shown that Cu-64-TP3805 can specifically target VPAC1 receptors and be used for positron emission tomography (PET) imaging of breast (BC) and prostate cancer (PC) in humans. Present work is aimed at the formulation of a freeze-dried diaminedithiol-peptide (N2S2-TP3805) kit and it's evaluation for the preparation of Cu-64 labeled TP3805. Parameters such as pH, temperature and incubation time were examined that influenced the radiolabeling efficiency and stability of the product. Methods: Kits were prepared under different conditions and radiolabeling efficiency of TP3805 kit was evaluated for a range of pH 3.5-8.5, after addition of Cu-64 in 30 mu l, 0.1 M HCI. Incubation temperature (37-90 degrees C) and time (30-120 min.) were also investigated. Kits were stored at -10 degrees C and their long term stability was determined as a function of their radiolabeling efficiency. Further, stability of Cu-64-TP3805 complex was evaluated in presence of fetal bovine serum and bovine serum albumin by using SDS polyacrylamide gel electrophoresis. Kits were then used for PET imaging of BC and PC following eIND (101550) and institutional approvals. Specificity of (64)CuTP3805 for VPAC1 was examined with digital autoradiography (DAR) of prostate tissues obtained after prostatectomy, benign prostatic hyperplasia (BPH) tissue, and benign and malignant lymph nodes. Results were compared with corresponding tissue histology. Results: Radiolabeling efficiency was >= 95% at final pH similar to 7.2 when incubated at 50 degrees C for 90 min. Kits were stable up to 18 months when stored at -10 degrees C, and Cu-64-TP3805 complex exhibited excellent stability for up to 4 h at room temperature. Cu-64-TP3805 complex did not show any transchelation even after 2 h incubation at 37 degrees C in 10% FBS as well as in BSA as determined by SDS PAGE analysis. DAR identified >= 95% of malignant lesions 11 new PC lesions, 20 high grade prostatic intraepithelial neoplasia, 2/2 ejaculatory ducts and 5/5 urethra verumontanum not previously identified The malignant lymph nodes were correctly identified by DAR and for 3/3 BPH patients, and 5/5 cysts, DAR was negative. In human BC (n = 19) and PC (n = 26) were imaged with 100% sensitivity. Conclusion: Availability of ready to use N2S2-peptide kits for Cu-64 labeling is convenient and eliminates possible day to day variation during its routine preparation for clinical use. (C) 2017 Elsevier Inc. All rights reserved.