Virginia Research Day 2025

Medical Student Research Biomedical

28 A Novel Non-Opioid Analgesic Does Not Alter Mouse Hippocampal Protein Expression

James Lee, Class of 2027; Romina Boortalary, Class of 2027; De’Yana Hines; Nakia Phillip; Seth C. Boehringer; Ramu Anandakrishnan; Blaise M. Costa Corresponding author: rboortalary@vcom.edu

Edward Via College of Osteopathic Medicine - Virginia Campus

Pain affects more people than diabetes, heart disease, and cancer combined, accounting for 20% of physician visits in the USA1. Physicians face limited options for pharmacological pain management, having to choose between opioids—which carry the risk of harmful side effects and addiction—or non opioid alternatives such as NSAIDs, ketamine, and gabapentin. However, these pharmacologic options often leave lasting effects on the brain. Proteomic studies have shown statistically significant differences in protein expression between control groups and drug-treated groups2, highlighting potential long-term impacts. CNS4 is a novel non-opioid analgesic that primarily acts on the NMDA receptor as a glutamate

concentration- dependent allosteric modulator, with a preference for the 2D subtype of the NMDA receptor. In this study, we examined differences in protein expression in the hippocampal brain tissue of male and female mice treated with CNS4 (100 mg/kg) or saline. After treatment via the intraperitoneal route, animals underwent fear conditioning, open field, and sucrose preference tests. Three days after treatment, they were euthanized, and brain tissue was harvested for analysis. Across the four groups (male treated, male control, female treated, female control; n=5 per group), 4,932 proteins were reproducibly quantified by the proteomic analysis. Among these, no proteins showed

significant differences in expression (p < 0.05) between any of the four groups studied. These results are encouraging, suggesting that CNS4—a centrally acting compound that produces potent analgesic effects without altering fear memory—does not induce lasting changes in protein expression in one of the brain’s critical regions for learning and memory. Further testing is necessary to compare CNS4 against pharmacologic therapies with addictive profiles to confirm whether CNS4 truly lacks proteomic changes or if confounding factors may have influenced the observed results.

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