Aylina Glasenapp Jens P. Bankstahl Heike Bähre Silke Glage Marion Bankstahl ^*

• Hannover Medical School, Hanover, Germany

Surgical interventions in mice require appropriate pain relief to ensure animal welfare and to avoid influence of pain on research findings. Carprofen is a non-steroidal anti-inflammatory drug commonly used as an analgesic for interventions inducing mild to moderate pain in laboratory rodents. Despite its frequent use, species-specific data on pharmacokinetics (PK), side effects, and potential impact on behavioral pain indicators are limited. We determined PK and tolerability profiles of carprofen in healthy male and female C57BL/6J mice (n=42), administered at highest recommended doses via single subcutaneous (s.c.) injection (20 mg/kg) and oral self-administration (25 mg/kg/24 h) per drinking water (d.w.) for 5 days. Plasma concentrations were measured at various time points after treatment start (n=6 per time point), and side effects evaluated using a modified Irwin test battery, hematology and histopathology. Additionally, potential interference with cage-side behaviors commonly used for pain assessment, such as the mouse grimace scale, wheel running, burrowing, nesting and grooming activity was investigated. Maximum plasma concentrations of 133.4 ± 11.3 µg/ml were reached 1 h after single s.c. injection with an elimination half-life of 8.52 h. Intake from d.w. resulted in a steady-state within 24 h after treatment start with plasma levels of around 60 µg/ml over 5 days in both sexes. The medicated water was well accepted, and increased d.w. intake was observed in the first 24 h after exposure (p < 0.0001). The Irwin test revealed only minor influence on tested behavior and physiological functions. However, during treatment via d.w., an increase in body temperature (p < 0.0001) was observed, as well as a reduction in voluntary wheel running activity by 49-70% in male mice. Moreover, burrowing, nesting, and grooming behaviors were slightly affected. Hematology and histopathology where without pathological findings that could be attributed to carprofen treatment. High-dose carprofen can be considered safe and of favorable PK for both administration routes assessed in healthy C57BL/6J mice of both sexes. Further efficacy evaluation of carprofen as monoanalgesic or component of multimodal post-surgical regimens is clearly encouraged, however, impact on behavioral markers used for pain assessment should be considered in this context.