Methotrexate α-tert-butyl ester is primarily utilized in research and development settings as a prodrug or intermediate in the synthesis of methotrexate derivatives. Its modified structure allows for improved stability and controlled release properties, making it valuable in the study of drug delivery systems. Researchers explore its potential in targeted cancer therapies, where it can be engineered to release methotrexate selectively in tumor tissues, minimizing systemic toxicity. Additionally, it serves as a tool in biochemical studies to investigate the mechanisms of action of methotrexate and its analogs. The ester form is also investigated for its potential to enhance bioavailability and optimize pharmacokinetic profiles in therapeutic applications.