Anti-adherent effects of bark and leaf extracts and computational prediction of the effects of its compound on β-tubulin interaction in genotype 4.

BACKGROUND AND AIM

, an opportunistic protozoan, exists widely in natural sources and can cause infections in humans and animals. The absence of effective monotherapy after the initial infection leads to chronic disease and recurrence. Tubulin protein is a vital target for design-targeted drug discovery. Anti-tubulin drugs are also used to treat infections, although resistance to these drugs has been observed. Therefore, it is necessary to identify a new targeted drug for infections. Therefore, this study aimed to assess the activity of ethanol extracts of extracts (RAE) against spp. and to predict its chemical compound on β-tubulin interaction.

MATERIALS AND METHODS

In this study, anti- activity with minimal inhibitory concentration (MIC) and minimal parasiticidal concentration (MPC) determination of ethanolic RAE from leaves, blossoms, buds, branches, and barks was tested on four trophozoites and cysts: WU 19001, American Type Culture Collection (ATCC) 30461, ATCC 50739, and ATCC 30010. The inhibitory effect on adherence was determined by the ability of adherence on 96-well plates, and its adhesive acanthopodia structure was evaluated using scanning electron microscopy analysis. In addition, the minimum cytotoxic concentrations (MCC) of leaf extract (RALE) and bark extract (RABE) were evaluated on Vero and HaCaT cell lines using the MTT assay. Phytochemical compounds from RALE and RABE were also analyzed by gas chromatography-mass spectrometry (GC-MS). Molecular docking and molecular dynamic analysis predicted the binding sites of chemicals in extracts and β-tubulin protein.

RESULTS

The results revealed that . and trophozoites had the highest inhibition at 90% at a MIC of 8 mg/mL after treatment with RALE and RABE, respectively, at 24 h. Those MPC values were exhibited at 16 mg/mL against . trophozoites. In addition, both extracts inhibited the adhesive properties of all approximately 80%-90% at 4 mg/mL, as well as adherent structural acanthopodia loss. MCC was 0.25 mg/mL, provided to be harmless to mammalian cells. GC-MS analysis supported that 8 and 11 major phytochemicals were from RABE and RALE, respectively. Molecular docking and molecular dynamics demonstrated that -β-tubulin exhibited potent root-mean-square deviation, root mean square fluctuation, and binding free energy values with clionasterol (from RABE and RALE) and stigmasterol (from RALE). Based on our results, ethanolic RABE and RALE exhibited anti- activity in reducing adhesion. showed that promising clionasterol and stigmasterol interacted with a targeting β-tubulin.

CONCLUSION

The RABE and RALE exhibited a potential anti-adherent effect on , low toxicity, and the clionasterol and stigmasterol in RABE and RALE predicted to interact the targeted β-tubulin. These agents may be used as alternative therapeutic agents in the management of disease using a sustainable one-heath approach.