Assessment of phytochemicals from Syzygium aqueum as inhibitors of ATP-dependent 6-phosphofructokinase: in silico and in vitro studies

Helminth infections can be effectively treated with various anthelmintics. However, the potential for drug resistance and the need for repeated doses to ensure successful treatment are some limitations to the treatment. This study aimed to evaluate the ability of phytochemicals from Syzygium aqueum to inhibit ATP-dependent 6-phosphofructokinase (6-PFK) from Ascaris lumbricoides and Ascaris suum using the in silico method (molecular docking, molecular dynamics simulation, and ADMET analysis) and in vitro evaluation using mortality, paralysis, and ovicidal activity testing. Molecular docking outcomes indicated that out of 76 compounds from Syzygium aqueum, 18 molecules exhibited strong inhibition against the target enzyme, based on binding free energy and interaction patterns. Molecular dynamics simulation demonstrated that samarangenin A and butyrospermol complexes remained stable for up to 200 ns within a range below 1 nm. The most promising compounds also exhibited good ADMET properties. In the in vitro egg hatch test, a 10% concentration of Syzygium aqueum extract showed potent ovicidal properties by significantly reducing the number of fertile eggs but was less effective than 0.12% albendazole. However, ferulic acid at various concentrations exhibited minimal effect on egg fertility but showed increased mortality and incidence of paralysis at higher doses against male worms. These findings yielded valuable insights into the underlying mechanisms, providing essential clues for the development of highly efficient inhibitors for parasitic worm infections through structure-based design. Keywords: Syzygium aqueum , Ascaris species, ATP-dependent 6-phosphofructokinase, in silico study, in vitro approach