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Veterinary research communications2022; 47(2); 473-485; doi: 10.1007/s11259-022-09964-x

In vitro and in vivo evaluation of kinase and protease inhibitors against Trypanosoma evansi.

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 IC 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 IC 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 IC 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.
© 2022. The Author(s), under exclusive licence to Springer Nature B.V.
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Publication Date: 2022-06-25 PubMed ID: 35751782PubMed Central: 3789323DOI: 10.1007/s11259-022-09964-xGoogle Scholar: Lookup
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Summary

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This study investigates the effectiveness of various kinase and protease inhibitors in combating Trypanosoma evansi, a harmful parasite causing fatal disease in livestock and wild animals. The findings suggest that some of these inhibitors could potentially serve as therapeutic agents against the parasite.

Background

  • Trypanosoma evansi is a parasite causing surra, a fatal disease in animals that can lead to chronic wasting.
  • Kinase and protease inhibitors are often used in the treatment of cancer and other diseases. This study explores if they can also be effective against T. evansi.

Experiment and Results

  • The researchers examined different inhibitors (Barasertib-HQPA – BAR, Palbociclib isethionate – PAL, Z-pro-prolinal – Z-PRO, and Leupeptin hemisulphate – LEU) for their efficacy against T. evansi within a specific growth medium.
  • BAR, PAL, and Z-PRO inhibited the growth of the parasite to varying degrees, while LEU showed no significant inhibitory effect.
  • The researchers analyzed the quantities of specific T. evansi genes (oligopeptidase B and aurora kinase) to determine how the drugs affected them. Significant upregulation of these genes was observed in populations exposed to BAR and PAL.

Cytotoxicity Assay

  • An assay was performed to determine the cytotoxic effect of the inhibitors on equine PBMC’s and Vero cell lines.
  • BAR showed 52% and 41% cytotoxicity at 50 µM concentration (about five folds the IC value) on equine PBMC’s and Vero cell line, respectively.
  • The cytotoxicity of 25% and 24% was recorded at 10 µM concentration (about ten folds to the IC value) of PAL in equine PBMC’s and Vero cell line, respectively.

In Vivo Trials

  • BAR and PAL, noted as effective in the in-vitro trials were test on mice. The results showed these compounds increased the lifespan of infected mice at higher doses.

Conclusion

  • The study reveals that kinase inhibitors, in particular, have a promising potential as therapeutic agents against the parasite T. evansi.

Cite This Article

APA
Bhutia WD, Gupta S, Rani R, Batra K, Sethi K, Kumar S, Kumar R. (2022). In vitro and in vivo evaluation of kinase and protease inhibitors against Trypanosoma evansi. Vet Res Commun, 47(2), 473-485. https://doi.org/10.1007/s11259-022-09964-x

Publication

Veterinary research communications
ISSN: 1573-7446
NlmUniqueID: 8100520
Country: Switzerland
Language: English
Volume: 47
Issue: 2
Pages: 473-485

Researcher Affiliations

Bhutia, Wangchuk Dorjee
  • Parasitology Lab, ICAR-National Research Centre on Equines, Hisar, Haryana, 125001, India.
Gupta, Snehil
  • Department of Veterinary Parasitology, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana, 125001, India.
Rani, Ruma
  • Parasitology Lab, ICAR-National Research Centre on Equines, Hisar, Haryana, 125001, India.
Batra, Kanisht
  • Department of Animal Biotechnology, Lala Lajpat Rai University of Veterinary and Animal Sciences, Hisar, Haryana, 125001, India.
Sethi, Khushboo
  • Parasitology Lab, ICAR-National Research Centre on Equines, Hisar, Haryana, 125001, India.
Kumar, Sanjay
  • Parasitology Lab, ICAR-National Research Centre on Equines, Hisar, Haryana, 125001, India.
Kumar, Rajender
  • Parasitology Lab, ICAR-National Research Centre on Equines, Hisar, Haryana, 125001, India. rkg.nrce@gmail.com.

MeSH Terms

  • Animals
  • Horses
  • Mice
  • Protease Inhibitors
  • Leukocytes, Mononuclear
  • Trypanosoma
  • Animals, Wild
  • Aurora Kinases

Grant Funding

  • F. No. F No. 3(1)/2020-Budget- NRCE sub-scheme / Indian Council of Agriculture Research

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