A translatable evaluation tool to study the biodistribution of clinically-available doxorubicin liposomes: PET imaging of Zr-Talidox

INTRODUCTIONDoxil/Caelyx is a PEGylated liposomal formulation of the chemotherapeutic doxorubicin used in the clinic for Kaposis sarcoma, advanced ovarian cancer, progressive multiple myeloma and metastatic breast cancer. Talidox(R), a smaller doxorubicin PEGylated liposome is undergoing clinical trials and has been proposed as an improvement on previous liposomal formulations for the treatment of advanced solid tumors. We aimed to validate an easily translatable radiolabeling method using zirconium-89 (89Zr) that enables quantitative whole-body PET imaging of these formulations to study their biodistribution and pharmacokinetics. METHODS[89Zr][Zr(oxinate)4] was produced using a kit-based approach followed by use as a direct radiolabeling agent of the liposomal formulations. DFT studies were performed to elucidate the mechanism behind the radiolabeling stability observed within the liposomes. Purified 89Zr-labelled Doxil/Talidox(R) liposomes (5 mg/kg doxorubicin dose) were administered in female BALB/c mice bearing 4T1 tumors. PET/CT imaging was acquired at 20 min, 24 h, 48 h, and 72 h, followed by post-mortem biodistribution at 72 h. RESULTS and DISCUSSIONBoth formulations were radiolabeled efficiently with high stability in serum in vitro for 72 h. In vivo, both formulations showed high tumor uptake at 72 h (18.5 {+/-} 2.4 % IA/g for Doxil and 20.2 {+/-} 2.3 % IA/g for Talidox). In general, ex vivo biodistribution showed similar uptake values for both formulations with high spleen/liver uptake and low bone uptake, confirming stability. Talidox(R) showed significantly lower spleen uptake and higher uptake in bone than Doxil. DFT studies confirmed that doxorubicin can form complexes with 89Zr that are more stable than [89Zr][Zr(oxinate)4], explaining the radiolabeling mechanism and stability results in vitro and in vivo. CONCLUSIONSClinically available PEGylated liposomes containing doxorubicin can be efficiently radiolabelled with 89Zr for PET imaging studies, using a clinically translatable radiolabelling method. HighlightsO_LIDoxorubicin-containing liposomes can be labeled with the positron-emitting radionuclide 89Zr with no impact on their original physicochemical properties. C_LIO_LIRadiolabeling is stable in vivo and enables imaging and biodistribution studies of the liposomes using positron emission tomography (PET). C_LIO_LIThe radiolabeling method is clinically translatable and would allow early assessment of existing and novel doxorubicin liposome biodistribution in humans or personalized medicine (nanotheranostic) approaches. C_LI