In vitro and in vivo anthelmintic evaluation with molecular docking studies of bioactive compounds from Croton joufra...

Croton joufra Roxb. (Euphorbiaceae), locally known as "Gochmahudi," is traditionally used against helminth infections by the Mishing tribe in Assam, India. Although the leaves of C. joufra have shown potential activity as anthelmintic, the bioactive constituents responsible for their activity have not yet been investigated.

The present study aimed to isolate, identify, and evaluate the anthelmintic compounds from C. joufra leaf extract.

Bioactivity-guided fractionation was carried out to isolate the bioactive compounds from C. joufra leaf extract. The structures of isolated compounds were determined using Nuclear Magnetic Resonance spectroscopy and Electrospray Ionization Mass Spectrometry. The isolated compounds were evaluated for their in vitro anthelmintic activity on the basis of their effects on the motility and survival of adult zoonotic parasites, Hymenolepis diminuta and Fasciolopsis buski. In vivo efficacy was assessed using an H. diminuta-infected rat model, where therapeutic outcomes were measured by determining the egg counts per gram of feces and worm burden within the host. The active compounds were subjected to molecular docking using CDOCKER of Biovia Discovery Studios Client against the RCSB PDB sourced protein tubulin-colchicine: stathmin-like domain complex (PDB ID: 1SA0) followed by molecular dynamics simulation using Schrodinger Desmond Software for 200 ns.

Bioactivity-guided fractionation led to isolation and characterization of two compounds: a known compound 1, 2-(N-benzoylamino) acetophenone, and compound 2, with a structurally undescribed sesquiterpene, 5-isopropyl-6,7,8-trimethylnon-1-ene. The results showed that both compounds possessed notable anthelmintic activity under in vitro and in vivo conditions, with effects either comparable to or slightly greater than those of the reference drug praziquantel. The molecular docking revealed favorable binding interactions, with 2-(N-benzoylamino)acetophenone showing a stronger binding affinity comparable to albendazole. Molecular dynamics simulations indicated stable yet dynamic protein-ligand interactions, primarily driven by hydrophobic and van der Waals forces. Among the two compounds, compound 1 demonstrated significantly greater activity than compound 2 and also exhibited stronger binding interactions, as supported by in silico analysis.

This study represents the first report identifying two active compounds with anthelmintic potential in C. joufra leaf extract.Future studies are warranted to investigate detailed mechanism of action and toxicity of these compounds.