In-XYIMSR-01 is a promising single-photon emission computed tomography (SPECT) imaging agent for identification of tumors that overexpress carbonic anhydrase IX. To translate [ 111 In]In-XYIMSR-01 to phase I trials, we performed animal toxicity and dosimetry studies, determined the maximum dose for human use, and completed the chemistry, manufacturing, and controls component of a standard regulatory application. The production process, quality control testing, stability studies, and specifications for sterile drug product release were based on United States Pharmacopeia chapters <823> and <825>, FDA 21 CFR Part 212. Toxicity was evaluated by using nonradioactive [ 113/115 In]In-XYIMSR-01 according to 21 CFR Part 58 guidelines. Organ Level INternal Dose Assessment/EXponential Modeling (OLINDA/EXM) was used to calculate the maximum single dose for human studies. Three process validation runs at starting radioactivities of 800 MBq were completed with a minimum concentration of 407 MBq/ml and radiochemical purity of >99% at the end of synthesis. A single intravenous dose of 55 μg/ml of [ 113/115 In]In-XYIMSR-01 was well tolerated in male and female Sprague-Dawley rats. The calculated maximum single dose for human injection from dosimetry studies was 390.35 MBq of [ 111 In]In-XYIMSR-01. We have completed toxicity and dosimetry studies as well as validated a manufacturing process to test [ 111 In]In-XYIMSR-01 in a phase I clinical trial. K E Y W O R D S carbonic anhydrase IX, clear cell renal cell carcinoma, clinical trials, molecular imaging, SPECT 1 | INTRODUCTION A total of 73,750 new cases of kidney cancer, representing 5% of all diagnosed malignancies, are estimated to be diagnosed in the United States in 2020. 1 Additionally, 14,830 deaths are estimated from this disease. The clear cell subtype of renal cell carcinoma (ccRCC) accounts for up to 70% of all kidney cancers. 2 Although computed tomography (CT), magnetic resonance imaging (MRI), and ultrasound are useful noninvasive tools for detecting