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Preclinical evaluation of bozepinib in bladder cancer cell lines: modulation of the NPP1 enzyme

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Abstract

Bladder cancer (BC) is the most common cancer of the urinary tract. Bozepinib (BZP), a purine-derived molecule, is a potential compound for the treatment of cancer. Purinergic signaling consists of the activity of nucleosides and nucleotides present in the extracellular environment, modulating a variety of biological actions. In cancer, this signaling is mainly controlled by the enzymatic cascade involving the NTPDase/E-NPP family and ecto-5’-nucleotidase/CD73, which hydrolyze extracellular adenosine triphosphate (ATP) to adenosine (ADO). The aim of this work is to evaluate the activity of BZP in the purinergic system in BC cell lines and to compare its in vitro antitumor activity with cisplatin, a chemotherapeutic drug widely used in the treatment of BC. In this study, two different BC cell lines, grade 1 RT4 and the more aggressive grade 3 T24, were used along with a human fibroblast cell line MRC-5, a cell used to predict the selectivity index (SI). BZP shows strong antitumor activity, with notable IC50 values (8.7 ± 0.9 µM for RT4; 6.7 ± 0.7 µM for T24), far from the SI for cisplatin (SI for BZP: 19.7 and 25.7 for RT4 and T24, respectively; SI for cisplatin: 1.7 for T24). BZP arrests T24 cells in the G2/M phase of the cell cycle, inducing early apoptosis. Moreover, BZP increases ATP and ADP hydrolysis and gene/protein expression of the NPP1 enzyme in the T24 cell line. In conclusion, BZP shows superior activity compared to cisplatin against BC cell lines in vitro.

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Acknowledgements

This study was supported by Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq/PQ n°302879/2017-0, Battastini AM and CNPq n°406035/2021-0, CNPq/PQ n° 311580/2021-1, Figueiró F). J.S. received support from the Natural Sciences and Engineering Research Council of Canada (NSERC; RGPIN-2023-05498).

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Authors and Affiliations

  1. Programa de Pós-Graduação em Ciências Biológicas: Bioquímica, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul, Rua Ramiro Barcelos, 2600 – anexo, Porto Alegre, CEP 90035-003, RS, Brazil

    Álisson Coldebella da Silva, Juliete Nathali Scholl, Amanda de Fraga Dias, Augusto Ferreira Weber, Fabrício Figueiró & Ana Maria Oliveira Battastini

  2. Laboratório de Farmacologia Aplicada, Escola de Ciências da Saúde e da Vida, Pontifícia Universidade Católica do Rio Grande do Sul, Porto Alegre, RS, Brazil

    Fernanda Bueno Morrone

  3. Departamento de Química Farmacéutica y Orgánica, Facultad de Farmacia, c/ Campus de Cartuja s/n, Granada, Spain

    Olga Cruz-López, Ana Conejo-García & Joaquín María Campos

  4. Instituto de Investigación Biosanitaria de Granada (ibs.GRANADA), Granada, Spain

    Olga Cruz-López, Ana Conejo-García & Joaquín María Campos

  5. Département de microbiologie-infectiologie et d’immunologie, Faculté de Médecine, Université Laval, Québec, QC, Canada

    Jean Sévigny

  6. Axe maladies infectieuses et immunitaires, Centre de recherche du CHU de Québec - Université Laval, Quebec city, QC, Canada

    Jean Sévigny

  7. Departamento de Bioquímica, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil

    Fabrício Figueiró

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  1. Álisson Coldebella da Silva
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  2. Juliete Nathali Scholl
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  3. Amanda de Fraga Dias
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  8. Joaquín María Campos
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  11. Ana Maria Oliveira Battastini
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Contributions

Álisson C. da Silva: Methodology, investigation, writing-original draft preparation. Juliete N. Scholl, Amanda F. Dias and Augusto F. Weber: Investigation, data curation. Fernanda B. Morrone, Olga Cruz-López, Ana Conejo-García, and Jean Sévigny: Investigation. Joaquín María Campos: Investigation and writing-reviewing. Fabrício Figueiró and Ana M. O. Battastini: Supervision, writing-reviewing and editing, funding acquisition.

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da Silva, Á.C., Scholl, J.N., de Fraga Dias, A. et al. Preclinical evaluation of bozepinib in bladder cancer cell lines: modulation of the NPP1 enzyme . Purinergic Signalling (2023). https://doi.org/10.1007/s11302-023-09975-6

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