The investigation aimed to reveal in-vivo therapeutic efficacy of the D. trifoliata's leaves ethyl acetate extract (EACE) in mice model in addition to the isolation and detection of phyto-molecules.

Phyto-molecules were isolated through chromatographic techniques and detected via nuclear magnetic resonance (NMR) approaches. Tail immersion, and acetic acid-induced writhing method were used for central and peripheral analgesic, respectively, whereas hypoglycemic and CNS depression efficacy were evaluated by oral glucose tolerance test, and thiopental-induced test, respectively. The 200, 400, and 600 mg/kg b.w. doses of EACE were used for all in vivo experiments.

Four different compounds (4-hydroxy benzoic acid A, kaempferol B, Baicalein C, stigmasterol D) were detected via NMR technique, where A and C were firstly isolated molecules from this species. The 600 mg/kg b.w. significantly prolonged reaction times (14.07 ± 0.44 s at 90 min; p < 0.001), confirming strong central analgesic activity. In the peripheral analgesic model, it produced a prominent dose-dependent efficacy with 34.91%, 58.49%, and 65.03% inhibition at 200, 400, and 600 mg/kg, respectively. The EACE also exerted a significant antidiarrheal activity (82.20% inhibition at 600 mg/kg b.w.). In the hypoglycemic assay, the extract elicited mild glucose-lowering effects, the highest reduction observed at 600 mg/kg (4.90 ± 0.42 mmol/L at 90 min). Additionally, CNS depressant activity was at moderate level, as the extract reduced sleep onset (33.75 ± 4.56 min) and extended sleep duration (182.5 ± 10.50 min; p < 0.01) in mice.

EACE showed some specific in vivo efficacy in dose dependent manner such as analgesic and diarrhea. Further extensive investigation, is needed to identify the responsible lead compounds.