Cloning, characterization, and inhibition of the novel β-carbonic anhydrase from parasitic blood fluke, Schistosoma mansoni

Abstract

Schistosoma mansoni is an intestinal parasite with one β-class carbonic anhydrase, SmaBCA. We report the sequence enhancing, production, catalytic activity, and inhibition results of the recombinant SmaBCA. It showed significant catalytic activity on CO₂ hydration in vitro with k_cat 1.38 × 10⁵ s⁻¹ and k_cat/K_m 2.33 × 10⁷ M⁻¹ s⁻¹. Several sulphonamide inhibitors, from which many are clinically used, showed submicromolar or nanomolar inhibitory effects on SmaBCA. The most efficient inhibitor with a K_I of 43.8 nM was 4-(2-amino-pyrimidine-4-yl)-benzenesulfonamide. Other effective inhibitors with K_Is in the range of 79.4–95.9 nM were benzolamide, brinzolamide, topiramate, dorzolamide, saccharin, epacadostat, celecoxib, and famotidine. The other tested compounds showed at least micromolar range inhibition against SmaBCA. Our results introduce SmaBCA as a novel target for drug development against schistosomiasis, a highly prevalent parasitic disease.

Keywords:

• Carbonic anhydrase
• anti-parasitic agents
• inhibitor
• sulphonamide
• Schistosoma mansoni

Acknowledgements

We thank Marianne Kuuslahti and Sanna Kavén for their skillful technical assistance; Juha Määttä, PhD, and Niklas Kähkönen for the help with protein purification and Maarit Patrikainen, PhD, for the help with refining the article text. We acknowledge the Tampere Facility of Protein Services (PS) for their service.

Author contributions

SH, AA, SP, and CTS designed the experiments. SH and AA performed the experiments in the laboratory. MT and RZE planned the modifications to the sequence. MT planned and prepared the bioinformatic analyses. SH drafted this manuscript. SP, MT, and CTS reviewed the drafts and modified the manuscript. All authors read and approved the final manuscript.

Disclosure statement

No potential conflict of interest was reported by all authors except Claudiu T. Supuran. C. T. Supuran is Editor-in-Chief of the Journal of Enzyme Inhibition and Medicinal Chemistry. He was not involved in the assessment, peer review, or decision-making process of this paper. The authors have no relevant affiliations of financial involvement with any organisation or entity with a financial interest in or financial conflict with the subject matter or materials discussed in the manuscript. This includes employment, consultancies, honoraria, stock ownership or options, expert testimony, grants or patents received or pending, or royalties.

Data availability statement

The data that support the findings of this study are openly available in MarttiT/S.-mansoni-BCA at https://github.com/MarttiT/S.-mansoni-BCA.

Additional information

Funding

This work was supported by the Finnish Medical Foundation under Grant number 5299; and the Academy of Finland under Grant number 348972. The funders had no role in study design, data collection and analysis, publication decision, or manuscript preparation.