Abstract
Trypanosoma evansi is a causative agent of chronic wasting and fatal disease of livestock and wild animals known as surra. In this study, repurposing approach based on drug target was used to investigate the efficacy of kinase inhibitors (Barasertib-HQPA, BAR and Palbociclib isethionate, PAL) and protease inhibitors (Z-pro-prolinal, Z-PRO and Leupeptin hemisulphate, LEU) against T. evansi in HMI-9 medium. BAR, PAL and Z-PRO exhibited IC50 values of 13.52 µM, 0.6375 µM and 63.20 µM against T. evansi in terms of growth inhibition, in the contrary, LEU failed to exhibit a significant growth inhibition at any time interval. Furthermore, oligopeptidase B and aurora kinase genes of T. evansi were targeted to determine the effect of these drugs on quantitative mRNA expression, which showed significant (p < 0.01) up-regulation of both genes in the BAR and PAL-exposed population at 12 h of exposure, whereas, Z-PRO showed only significant (p < 0.05) up-regulation of aurora kinase gene at 12 h interval. In cytotoxicity assay, BAR exhibited 52% and 41% cytotoxicity at 50 μM concentration (about five folds the IC50 value) on equine PBMC's and Vero cell line, respectively. Similarly, the cytotoxicity of 25% and 24% were recorded at 10 μM concentration (about ten folds to the IC50 value) of PAL in equine PBMC's and Vero cell line, respectively. Of these, BAR and PAL, which were found effective under in vitro trials, raised the longevity of mice at higher doses during in vivo trials. Data generated showed that kinase inhibitors have higher potential to explore therapeutic molecules against surra organism.
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Acknowledgements
The authors gratefully thank the Director, ICAR-National Research Centre on Equines, Hisar, India, for providing all the necessary facilities for conducting this study. The authors are thankful to Mr. Raj Kumar Dayal for technical assistance provided during the experiments.
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The authors received funding from Indian Council of Agricultural Research (ICAR) via. financial sanction number (F No. 3(1)/2020-Budget- NRCE sub-scheme) for completion of present research work.
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WDB, SG, RR, KB, and KS conceived the study, performed the main experiments, and wrote the manuscript. SK and RK designed the experiment, supervised the study and provided help in drafting the final document. All authors read and approved the final manuscript.
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All authors certified that the animal experimentations were carried out in the present work after approval of Institute Animal Ethic Committee (IAEC) and as per guidelines set by the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), Animal Welfare Division, Government of India. The ethical approvals for the present study was accorded by the IAEC vide Approval No. NRCE/CPCSEA/2019–20 Dated 15/02/2020.
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Bhutia, W.D., Gupta, S., Rani, R. et al. In vitro and in vivo evaluation of kinase and protease inhibitors against Trypanosoma evansi. Vet Res Commun 47, 473–485 (2023). https://doi.org/10.1007/s11259-022-09964-x
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DOI: https://doi.org/10.1007/s11259-022-09964-x