Animals, grouping, and dose administration

Wistar rats, Rattus norvegicus (~ 175 g) were purchased from Central Animal Facility, IISc, Bangalore. All the animals were housed in polypropylene cages and kept in controlled lighting of 12 h light/dark cycle throughout the experiment. Standard pellet diet (Agro Corporation Pvt. Ltd. India) and water ad libitum have been provided to the animals. Maintenance and handling of the animals were according to the guidelines of the animal ethical committee. The experimental design and methods followed institutional guidelines and were approved by Institutional Animal Ethics Committee of Indian Institute of Science, Bangalore (Ethical committee approval No: CAF/Ethics/551/2017 and CAF/Ethics/744/2020).

All the studies were designed according to the guidelines of the Central Drug Standard Control Organization (Schedule-Y-CDSCO, Appendix III), India (https://cdsco.gov.in/opencms/opencms/en/Drugs/New-Drugs/). Single-dose toxicity analysis was performed in male Wistar rats. Animals were segregated in groups of five, and Disarib was administered using oral gavage (100, 1000, 2000 mg/kg b. wt.). Carboxymethyl cellulose, used for preparing Disarib, served as vehicle control to feed control animals. Mortality, general appearance, and behavior were initially observed for 24 h. Animals were further observed for 14 days and were further subjected to toxicity analysis.