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Radiochemical synthesis and pre-clinical evaluation of [68Ga]Ga-FAPI-4 formulated using in-house developed FAPI-4 kit

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Abstract

The present work describes the formulation of a lyophilized Fibroblast Activation Protein Inhibitor-4 (FAPI-4) kit for preparation of [68Ga]Ga-FAPI-4 in high radiochemical yield (RCY) using either generator or cyclotron produced 68GaCl3, suitable for Positron Emission Tomography (PET) based diagnosis of cancers over-expressing Fibroblast Activation Protein (FAP). Experimental parameters such as concentration of FAPI-4, buffer content, storage conditions and shelf-life were optimized to ensure > 95% RCY to utilize the kit for multi-patient dose formulations. A lyophilized FAPI-4 kit containing 50 µg of FAPI-4 ligand could be successfully optimized which consistently yielded [68Ga]Ga-FAPI-4 in > 95% RCY on labeling with ~ 1.11 GBq of 68GaCl3. [68Ga]Ga-FAPI-4 retained > 95% radiochemical purity (RCP) even after 2 h post labeling when stored at ambient temperature. The kits stored below − 20  °C reproduced the labeling profile up to a period of ~ 3 months (period of study). In the in vitro cell binding experiments performed in HT29 cells, [68Ga]Ga-FAPI-4 exhibited specific binding to FAP (16.1 ± 1.8%ID in 106 cells). In the biodistribution studies performed in female SCID mice bearing HT29 colorectal adenocarcinoma, [68Ga]Ga-FAPI-4 exhibited tumor uptake of 3.8 ± 0.3% ID/g at 1 h post-injection. The above results demonstrate the utility of developed FAPI-4 kits for formulation of [68Ga]Ga-FAPI-4 for PET imaging of FAP over-expressing cancers.

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Acknowledgements

The authors thank Chief Executive, Board of Radiation and Isotope Technology for the constant support and encouragement. The support of Head, Radiation Medicine Centre, Bhabha Atomic Research Centre is gratefully acknowledged.

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Authors and Affiliations

  1. Radiopharmaceutical Laboratory, Board of Radiation and Isotope Technology, Navi Mumbai, 400 703, India

    Navin Sakhare, Arpit Mitra, Laxman Ram, Sanjeev Kumar, Anupam Mathur & Usha Pandey

  2. Department of Chemistry, Mangalore University, Mangalagangotri, Mangaluru, India

    Navin Sakhare & Anupam Mathur

  3. Radiation Medicine Centre, Bhabha Atomic Research Centre, Mumbai, India

    Avik Chakraborty, Sudeep Sahu & Sutapa Rakshit

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  1. Navin Sakhare
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  2. Arpit Mitra
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Correspondence to Usha Pandey.

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Sakhare, N., Mitra, A., Chakraborty, A. et al. Radiochemical synthesis and pre-clinical evaluation of [68Ga]Ga-FAPI-4 formulated using in-house developed FAPI-4 kit. J Radioanal Nucl Chem (2024). https://doi.org/10.1007/s10967-024-09476-9

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