Optimization and Validation of Polyherbal Formulation by Applying Box- Behnken Design for the Treatment of Inflammatory Bowel Disease in Experimental Animals

Background: The use of herbal medicine in inflammatory bowel disease (IBD) has increased significantly. Allopathic treatment of IBD leads to several side effects; therefore, the use of the herbal formulation is promising. Aegle Marmelos, Bombax malabericum, and Hollarrhena antidysentrica plants have been used to treat IBD. Objective: This study aimed to evaluate a designed polyherbal formulation in experimentally induced inflammatory bowel disease in rats and validate a mathematical model derived by Box-Behnken experimental design for optimized polyherbal formulation for the treatment of IBD in experimental rats by checkpoint analysis. Method: Three-level Box-Behnken design was selected to optimize the dose. Polyherbal formulation that consisted of plant extracts of Aegle Marmelos (X1), Bombax malabericum (X2), and Hollarrhena antidysentrica (X3) in different ratios was selected as an independent variable. Polynomial equations were established based on analysis of variance (ANOVA). To validate the chosen polynomial equation, checkpoint analyses were performed. The percentage of predictive error is presented. Results: ANOVA reveals that X2 plant does not have any significant impact on the response surface. The checkpoint batch showed the experimental values of CMDI and Disease activity index (DAI) as 1.33 and 0.66, respectively. It is worthwhile to note that the observed values were quite close to the calculated values of CMDI. A little difference in the value of DAI may be attributed to the inherent variation observed in animal studies. Conclusion: From this study, it was concluded that a dose of 100 mg/kg of Aegle marmelos, a dose of 300 mg/kg of Bombax malabericum, and a dose of 200 mg/kg of Holarrhena antidysentrica will be effective in IBD patients.