Subchronic Toxicity Evaluation of Ssanghwa-tang Following 13-Week Repeated Oral Administration in SD Rats and Clinical Dose Extrapolation

Objectives: The present study evaluated the potential subchronic toxicity of Ssanghwa-tang following repeated oral administration to Sprague-Dawley rats for 13 weeks. And this study is to investigate the correlation between nonclinical and clinical dosages of Ssanghwa-tang.Methods: Ssanghwa-tang was administered orally once daily for approximately 13 weeks to 10 males and 10 females per group at doses of 0, 1,000, 2,000, and 5,000 mg/kg/day.Results: During the study period, no mortalities were observed in any dose group of either sex. No treatment-related changes were observed in body weight, food consumption, ophthalmic findings, gross necropsy, or organ weights for both male and female rats. Although intermittent salivation was noted during clinical observations, it was not considered toxicologically significant. Hematological analysis revealed a decrease in red blood cell count, accompanied by increases in reticulocyte count, reticulocyte ratio, mean corpuscular volume, and mean corpuscular hemoglobin. Furthermore, biochemical analysis showed an elevation in total bilirubin, while urinalysis indicated the presence of bilirubin and urobilinogen. Histopathological assessment revealed increased extramedullary hematopoiesis and pigmentation in the spleen. Consequently, no observed adverse effect level of Ssanghwa-tang was determined to be 5,000 mg/kg therefore human equivalent dose is 41.4 g and 48.6 g, and maximum recommended starting doses for clinical trials are 4.1 g and 4.8 g.Conclusions: Administration of Ssanghwa-tang to Sprague-Dawley rats at doses up to 5,000 mg/kg/day resulted in no significant toxicological concerns.