Combination of carbonic anhydrase isoform IX inhibitors and gefitinib suppresses on the invasive potential of non-small cell lung cancer cells

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Abstract

Human carbonic anhydrase isoform IX (CA IX) plays a key role in maintaining the pH homeostasis of malignant neoplasms, creating a favorable microenvironment for the growth, invasion and metastasis of tumor cells. Recent studies have established that inhibition of the activity of CA IX, expressed on the surface of tumor cells, significantly increases the effectiveness of classical chemotherapeutic agents and makes it possible to suppress the resistance of tumor cells to chemotherapy, as well as increase their sensitivity to the drugs used (including reducing the required dose of cytostatics). In this work, we studied the ability of new CA IX inhibitors based on substituted 1,2,4-oxadiazole-containing primary aromatic sulfonamides to potentiate the cytostatic effect of gefitinib (a selective inhibitor of the tyrosine kinase domain of the epidermal growth factor receptor) under hypoxic conditions. In this work, we studied the combined effect of gefitinib and CA IX inhibitors – 4-(3-phenyl-1,2,4-oxadiazol-5-yl)thiophene-2-sulfonamide (1), 4-(5-(thiophene-3-yl)-1,2,4-oxadiazol-3-yl)benzenesulfonamide (2), 4-(3-(pyridin-2-yl)-1,2,4-oxadiazol-5-yl)thiophene-2-sulfonamide (3) and 4-(5-methyl-1,2,4-oxadiazol-3-yl)benzenesulfonamide (4) on cytotoxicity, proliferation, activation of caspases 3/7 and cell cycle using the example of human lung adenocarcinoma cell line A549 under conditions of hypoxia. It was found that combination of compounds 1 and 2 with gefitinib inhibits the invasive potential of A549 cells, of which inhibitor 1 had the greatest effect and can be considered as a promising candidate for further research.

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About the authors

A. S. Bunev

Togliatti State University

Email: sharoyko@gmail.com

Medicinal Chemistry Center

Russian Federation, 445020 Togliatti

A. A. Shetnev

Institute of Biophysics of the Future

Email: sharoyko@gmail.com
Russian Federation, 141701 Dolgoprudny, Moscow Region

O. S. Shemchuk

Pavlov First St. Petersburg State Medical University

Email: sharoyko@gmail.com
Russian Federation, 197022 St. Petersburg

P. K. Kozhukhov

Pavlov First St. Petersburg State Medical University

Email: sharoyko@gmail.com
Russian Federation, 197022 St. Petersburg

T. V. Sharonova

St. Petersburg State University

Email: sharoyko@gmail.com
Russian Federation, 199034 St. Petersburg

I. I. Tyuryaeva

St. Petersburg State University; Institute of Cytology, Russian Academy of Sciences

Email: sharoyko@gmail.com
Russian Federation, 199034 St. Petersburg; 194064 St. Petersburg

M. G. Khotin

Institute of Cytology, Russian Academy of Sciences

Email: sharoyko@gmail.com
Russian Federation, 194064 St. Petersburg

S. V. Ageev

Pavlov First St. Petersburg State Medical University; St. Petersburg State University

Email: sharoyko@gmail.com
Russian Federation, 197022 St. Petersburg; 199034 St. Petersburg

D. K. Kholmurodova

Samarkand State Medical University

Email: sharoyko@gmail.com

Scientific and Practice Center for Immunology, Allergology and Human Genomics

Uzbekistan, 100400 Samarkand

J. A. Rizaev

Samarkand State Medical University

Email: sharoyko@gmail.com

Scientific and Practice Center for Immunology, Allergology and Human Genomics

Uzbekistan, 100400 Samarkand

K. N. Semenov

Pavlov First St. Petersburg State Medical University; St. Petersburg State University; Samarkand State Medical University

Email: sharoyko@gmail.com

Scientific and Practice Center for Immunology, Allergology and Human Genomics, Samarkand State Medical University

Russian Federation, 197022 St. Petersburg; 199034 St. Petersburg; 100400 Samarkand, Uzbekistan

V. V. Sharoyko

Togliatti State University; Pavlov First St. Petersburg State Medical University; St. Petersburg State University; Samarkand State Medical University

Author for correspondence.
Email: sharoyko@gmail.com

Medicinal Chemistry Center, Togliatti State University; Scientific and Practice Center for Immunology, Allergology and Human Genomics, Samarkand State Medical University

Russian Federation, 445020 Togliatti; 197022 St. Petersburg; 199034 St. Petersburg; 100400 Samarkand, Uzbekistan

References

  1. Jemal, A., Bray, F., Center, M. M., Ferlay, J., Ward, E., and Forman, D. (2011) Global cancer statistics, Cancer J. Clinic., 61, 69-90, https://doi.org/10.3322/CAAC.20107.
  2. Lynch, T. J., Bell, D. W., Sordella, R., Gurubhagavatula, S., Okimoto, R. A., Brannigan, B. W., Harris, P. L., Haserlat, S. L., Supko, J. G., Haluska, F. G., Louis, D. C., Christiani, M. D.,Settleman, J., and Haber, D. A. (2004) Activating mutations in the epidermal growth factor receptor underlying responsiveness of non-small-cell lung cancer to gefitinib, New Eng. J. Med., 350, 2129-2139, https://doi.org/10.1056/NEJMOA040938.
  3. Pao, W., and Chmielecki, J. (2010) Rational, biologically based treatment of EGFR-mutant non-small-cell lung cancer, Nat. Rev. Cancer, 10, 760, https://doi.org/10.1038/NRC2947.
  4. Sim, E. H. A., Yang, I. A., Wood-Baker, R., Bowman, R. V., and Fong, K. M. (2018) Gefitinib for advanced non-small cell lung cancer, Cochrane Database Syst. Rev., 1, https://doi.org/10.1002/14651858.CD006847.PUB2.
  5. Sowa, T., Menju, T., Chen-Yoshikawa, T. F., Takahashi, K., Nishikawa, S., Nakanishi, T., Shikuma, K., Motoyama, H., Hijiya, K., Aoyama, A., Sato, T., Sonobe, M., Harada, H., and Date, H. (2017) Hypoxia-inducible factor 1 promotes chemoresistance of lung cancer by inducing carbonic anhydrase IX expression, Cancer Med., 6, 288-297, https://doi.org/10.1002/CAM4.991.
  6. Lie, M., Mazure, N. M., Hofman, V., Ammadi, R. E., Ortholan, C., Bonnetaud, C., Havet, K., Venissac, N., Mograbi, B., Mouroux, J., Pouysségur, J., and Hofman, P. (2010) High levels of carbonic anhydrase IX in tumour tissue and plasma are biomarkers of poor prognostic in patients with non-small cell lung cancer, Br. J. Cancer, 102, 1627, https://doi.org/10.1038/SJ.BJC.6605690.
  7. Becker, H. M. (2020) Carbonic anhydrase IX and acid transport in cancer, Br. J. Cancer, 122, 157, https://doi.org/ 10.1038/S41416-019-0642-Z.
  8. Lee, S. H., McIntyre, D., Honess, D., Hulikova, A., Pacheco-Torres, J., Cerdán, S., Swietach, P., Harris, A. L., and Griffiths, J. R. (2018) Carbonic anhydrase IX is a pH-stat that sets an acidic tumour extracellular pH in vivo, Br. J. Cancer, 119, 622, https://doi.org/10.1038/S41416-018-0216-5.
  9. Debreova, M., Csaderova, L., Burikova, M., Lukacikova, L., Kajanova, I., Sedlakova, O., Kery, M., Kopacek, J., Zatovicova, M., Bizik, J., Pastorekova, S., and Svastova E. (2019) CAIX regulates invadopodia formation through both a pH-dependent mechanism and interplay with actin regulatory proteins, Int. J. Mol. Sci., 20, 2745, https://doi.org/10.3390/IJMS20112745.
  10. Succoio, M., Amiranda, S., Sasso, E., Marciano, C., Finizio, A., De Simone, G., Garbi, C., and Zambrano, N. (2023) Carbonic anhydrase IX subcellular localization in normoxic and hypoxic SH-SY5Y neuroblastoma cells is assisted by its C-terminal protein interaction domain, Heliyon, 9, e18885, https://doi.org/10.1016/J.HELIYON. 2023.E18885.
  11. Peppicelli, S., Andreucci, E., Ruzzolini, J., Bianchini, F., Nediani, C., Supuran, C. T., and Calorini, L. (2020) The carbonic anhydrase IX inhibitor SLC-0111 as emerging agent against the mesenchymal stem cell-derived pro-survival effects on melanoma cells, J. Enzyme Inhibit. Med. Chem., 35, 1185-1193, https://doi.org/10.1080/14756366.2020.1764549.
  12. Yap, T. A., Omlin, A., and De Bono, J. S. (2013) Development of therapeutic combinations targeting major cancer signaling pathways, J. Clin. Oncol., 31, 1592-1605, https://doi.org/10.1200/JCO.2011.37.6418.
  13. Partridge, A. H., Burstein, H. J., and Winer, E. P. (2001) Side effects of chemotherapy and combined chemohormonal therapy in women with early-stage breast cancer, J. Natl. Cancer Inst. Monogr., 2001, 135-142, https:// doi.org/10.1093/oxfordjournals.jncimonographs.a003451.
  14. Blagosklonny, M. V. (2005) Overcoming limitations of natural anticancer drugs by combining with artificial agents, Trends Pharmacol. Sci., 26, 77-81, https://doi.org/10.1016/J.TIPS.2004.12.002.
  15. Lu, Y., Liu, Y., Oeck, S., and Glazer, P. M. (2018) Hypoxia promotes resistance to EGFR inhibition in NSCLC cells via the histone demethylases, LSD1 and PLU-1, Mol. Cancer Res., 16, 1458-1469, https://doi.org/10.1158/1541-7786.MCR-17-0637.
  16. Ma, J. S. Y., Kim, J. Y., Kazane, S. A., Choi, S., Yun, H. Y., Kim, M. S., Rodgers, D. T., Pugh H. M., Singer, O., Sun, S. B., Fonslow, B. R., Kochenderfer, J. N., Wright, T. M., Schultz, P. G., Young, T. S., Kim, C. H., and Cao, Y. (2016) Versatile strategy for controlling the specificity and activity of engineered T cells, Proc. Natl. Acad. Sci. USA, 113, E450-E458, https://doi.org/10.1073/PNAS.1524193113.
  17. Swinson, D. E. B., and O’Byrne, K. J. (2006) Interactions between hypoxia and epidermal growth factor receptor in non-small-cell lung cancer, Clin. Lung Cancer, 7, 250-256, https://doi.org/10.3816/CLC.2006.N.002.
  18. Ono, M., Hirata, A., Kometani, T., Miyagawa, M., Ueda, S., Kinoshita, H., Fujii, T., and Kuwano M. (2004) Sensitivity to gefitinib (Iressa, ZD1839) in non-small cell lung cancer cell lines correlates with dependence on the epidermal growth factor (EGF) receptor/extracellular signal-regulated kinase 1/2 and EGF receptor/Akt pathway for proliferation, Mol. Cancer Ther., 3, 465-472, https://doi.org/10.1158/1535-7163.465.3.4.
  19. Tracy, S., Mukohara, T., Hansen, M., Meyerson, M., Johnson, B. E., and Jänne, P. A. (2004) Gefitinib induces apoptosis in the EGFRL858R non-small-cell lung cancer cell line H3255, Cancer Res., 64, 7241-7244, https://doi.org/ 10.1158/0008-5472.CAN-04-1905.
  20. Krasavin, M., Shetnev, A., Sharonova, T., Baykov, S., Kalinin, S., Nocentini, A., Sharoyko, V., Poli, G., Tuccinardi, T., Presnukhina, S., Tennikova, T. B., and Supuran, C. T. (2019) Continued exploration of 1,2,4-oxadiazole periphery for carbonic anhydrase-targeting primary arene sulfonamides: Discovery of subnanomolar inhibitors of membrane-bound hCA IX isoform that selectively kill cancer cells in hypoxic environment, Eur. J. Med. Chem., 164, 92-105, https://doi.org/10.1016/J.EJMECH.2018.12.049.
  21. Wu, D., and Yotnda, P. (2011) Induction and testing of hypoxia in cell culture, J. Visual. Exp., 54, 2899, https://doi.org/10.3791/2899.
  22. Paneerselvam, C., and Ganapasam, S. (2020) β-Escin alleviates cobalt chloride-induced hypoxia-mediated apoptotic resistance and invasion via ROS-dependent HIF-1α/TGF-β/MMPs in A549 cells, Toxicol. Res., 9, 191-201, https://doi.org/10.1093/TOXRES/TFAA019.
  23. Bahadori, M. B., Vandghanooni, S., Dinparast, L., Eskandani, M., Ayatollahi, S. A., Ata, A., and Nazemiyeh, H. (2019) Triterpenoid corosolic acid attenuates HIF-1 stabilization upon cobalt (II) chloride-induced hypoxia in A549 human lung epithelial cancer cells, Fitoterapia, 134, 493-500, https://doi.org/10.1016/J.FITOTE.2019.03.013.
  24. Pastorekova, S., and Gillies, R. J. (2019) The role of carbonic anhydrase IX in cancer development: links to hypoxia, acidosis, and beyond, Cancer Metastasis Rev., 38, 65-77, https://doi.org/10.1007/S10555-019-09799-0.
  25. Hedlund, E. M. E., McDonald, P. C., Nemirovsky, O., Awrey, S., Jensen, L. D. E., and Dedhar, S. (2019) Harnessing induced essentiality: targeting carbonic anhydrase IX and angiogenesis reduces lung metastasis of triple negative breast cancer xenografts, Cancers, 11, 1002, https://doi.org/10.3390/CANCERS11071002.

Supplementary files

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2. Fig. 1. The effect of inhibitor 1 and gefitinib on A549 cell migration depending on the concentration of inhibitor 1 and the culturing time. a – Incubation with gefitinib (10 μM) and inhibitor 1 (25 μM) added to the cells before the start of the cell index (CI) measurement; b – incubation with gefitinib (10 μM) and inhibitor 1 (50 μM) added to the cells before the start of the CI measurement. The results of one representative measurement from three different experiments are shown.

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3. Fig. 2. The effect of inhibitor 2 and gefitinib on A549 cell migration depending on the concentration of inhibitor 2 and the culturing time. a – Incubation with gefitinib (10 μM) and inhibitor 2 (25 μM) added to the cells before the start of the cell index (CI) measurement; b – incubation with gefitinib (10 μM) and inhibitor 2 (50 μM) added to the cells before the start of the CI measurement. The results of one representative measurement from three different experiments are shown.

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4. Fig. 3. The effect of inhibitor 3 and gefitinib on the migration of A549 cells depending on the concentration of inhibitor 3 and the culturing time. a – Incubation with gefitinib (10 μM) and inhibitor 3 (25 μM) added to the cells before the start of the cell index (CI) measurement; b – incubation with gefitinib (10 μM) and inhibitor 3 (50 μM) added to the cells before the start of the CI measurement. The results of one representative measurement from three different experiments are shown.

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5. Fig. 4. The effect of inhibitor 4 and gefitinib on the migration of A549 cells depending on the concentration of inhibitor 4 and the culturing time. a – Incubation with gefitinib (10 μM) and inhibitor 4 (25 μM) added to the cells before the start of the cell index (CI) measurement; b – incubation with gefitinib (10 μM) and inhibitor 4 (50 μM) added to the cells before the start of the CI measurement. The results of one representative measurement from three different experiments are shown.

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6. Fig. 5. Distribution of A549 cells by cell cycle phases in the presence of QAC IX inhibitors and gefitinib (24 h and 48 h) under physiological hypoxia conditions (1% O2)

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7. Fig. 6. caspase IX inhibitors in combination with gefitinib induce caspase 3/7-dependent apoptosis in A549 cells. a – Incubation with gefitinib (10 μM) for 24 h and 48 h; b – incubation with gefitinib (10 μM) and inhibitor 1 (25 and 50 μM) for 24 h and 48 h; c – incubation with gefitinib (10 μM) and inhibitor 2 (25 and 50 μM) for 24 h and 48 h. Cell types: D – dead cells; N – late apoptotic cells; L – living cells; A – early apoptotic cells

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8. Tab. 1, fig. 1

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9. Tab. 1, fig. 2

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10. Tab. 1, fig. 3

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11. Tab. 1, fig. 4

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12. Appendix
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