Objectives/Goals: The overarching objective of this study is to inform clinicians, patients, and other stakeholders of the level of evidence and the real-world risk-to-benefit profiles associated with older antidepressant drugs that are frequently used off-label. Methods/Study Population: A PubMed literature review was performed to identify clinical trials conducted in the USA between 2013 and 2023 for trazodone and 2000 and 2023 for escitalopram and citalopram. These studies were examined for robustness, due to sample size, study design, and generalizability. Findings were compared with information provided on UpToDate® LexiDrug™, a primary database used by clinicians to inform prescribing practice. To explore risks associated with off-label use, the FDA adverse event reporting system was probed to identify adverse events reported for each drug; results were systematically categorized by reason for use. To compare the volume of on-label to off-label prescriptions, data will be extracted from electronic health records from University of Southern California-affiliated hospitals. Results/Anticipated Results: Studies conducted on off-label prescriptions of these drugs show primarily small sample sizes, pointing to a limitation in generalizability. For citalopram (N = 77) and trazodone (N = 42), over half of their off-label studies had samples of 50 participants or less. These two drugs also showed low evidence rating for off-label prescription on LexiDrug due to limited power studies. Multiple health agencies recommend against off-label prescriptions for trazodone due to insufficient evidence. There is limited data in the US regarding the volume of off-label prescriptions; however, trazodone’s FAERS analysis indicated a large proportion of adverse event reports (1099/7239) come from cases where trazodone was used for insomnia, an off-label indication, compared to depression, the on-label indication (464/7239). Discussion/Significance of Impact: With 1 in 6 Americans taking antidepressants and 40%–80% of these psychiatric prescriptions being employed off-label, there is a serious and present risk for patients regarding the safety and efficacy of these medications. Awareness must be brought to clinicians to protect patients and encourage evidence-based practice.