Preparation of boron-containing s-nitrosothiol based on homocysteinylamides of human serum albumin for combined no-chemical and boron-neutron-capture therapy

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Abstract

The strategic aim of this work is to create a fluorophore-labelled, clinically relevant exogenous NO donor carrying a boron-containing compound residue on the basis of human serum albumin (HSA) for the implementation of combined NO-chemotherapy and boron-neutron-capture therapy. By selective modification of the Cys34 residue of albumin with a maleimide derivative of a fluorescent dye and subsequent N-homocysteinylation with a thiolactone derivative of homocysteine containing a clozo-dodecaborate residue, a nanoconstruct for boron-neutron-capture therapy was obtained. An analogue based on the natural modifier, boron-containing homocysteine thiolactone, was synthesised by alkylation of the amino group of thiolactone with a dioxonium derivative of clozo-dodecaborate. Post-synthetic modification of the lysine residues of the protein using the boron thiolactone of homocysteine provided the introduction of SH groups into the protein and the possibility of subsequent trans-S-nitrosylation of the protein using S-nitrosoglutathione. It was found that 2 mol of NO was conjugated to 1 mol of boron-containing HSA. Boron-containing S-nitrosothiol based on albumin homocysteinylamide, without epithermal neutron irradiation, was demonstrated to be more cytotoxic against human glioblastoma cell lines than the boron-containing albumin conjugate. Thus, the approach used allows obtaining a boron-enriched structure based on a biocompatible tumor-specific protein, containing a fluorescent label and an increased number of S-nitroso groups. It is necessary for the manifestation of a chemotherapeutic effect of the construct. The practical significance of this structure lies in the possibility of a cancer treating, combining chemo- and boron-neutron capture therapy.

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About the authors

T. V. Popova

Institute of Chemical Biology and Fundamental Medicine SB RAS; Novosibirsk National Research State University

Author for correspondence.
Email: v.silnikov@mail.ru
Russian Federation, Novosibirsk; Novosibirsk

M. Van

Novosibirsk National Research State University

Email: v.silnikov@mail.ru
Russian Federation, Novosibirsk

T. N. Kurochkin

Novosibirsk National Research State University

Email: v.silnikov@mail.ru
Russian Federation, Novosibirsk

S. A. Tsyrempilov

Novosibirsk National Research State University

Email: v.silnikov@mail.ru
Russian Federation, Novosibirsk

O. D. Zakharova

Institute of Chemical Biology and Fundamental Medicine SB RAS

Email: v.silnikov@mail.ru
Russian Federation, Novosibirsk

V. N. Silnikov

Institute of Chemical Biology and Fundamental Medicine SB RAS

Email: v.silnikov@mail.ru
Russian Federation, Novosibirsk

T. S. Godovikova

Institute of Chemical Biology and Fundamental Medicine SB RAS; Novosibirsk National Research State University

Email: v.silnikov@mail.ru
Russian Federation, Novosibirsk; Novosibirsk

References

  1. Lancaster J.R. // Biochem. Pharmacol. 2020. V. 176. P. 113793. https://doi.org/10.1016/j.bcp.2020.113793
  2. Lundberg J.O., Weitzberg E. // Cell. 2022. V. 185. P. 2853–2878. https://doi.org/10.1016/j.cell.2022.06.010
  3. Kannan M.S., Guiang S., Johnson D.E. // Indian J. Pediatr. 1998. V. 65. P. 333–345. https://doi.org/10.1007/BF02761123
  4. Moncada S., Palmer R.M., Higgs E.A. // Pharmacol. Rev. 1991. V. 43. P. 109–142.
  5. Thomas D.D., Liu X., Kantrow S.P., Lancaster J.R. // Proc. Natl. Acad. Sci. USA. 2001. V. 98. P. 355–360. https://doi.org/10.1073/pnas.011379598
  6. Осипов А.Н., Борисенко Г.Г., Владимиров Ю.А. // Усп. биол. химии. 2007. Т. 47. С. 259–292.
  7. Ванин А.Ф. // Мол. биология. 2023. Т. 57. С. 925–937.
  8. Sharma V., Fernando V.R., Letson J., Walia Y., Zheng X., Fackelman D., Furuta S. // Int. J. Mol. Sci. 2021. V. 22. P. 4600. https://doi.org/10.3390/ijms22094600
  9. Soni Y., Softness K., Arora H., Ramasamy R. // Am. J. Mens. Health. 2020. V. 14. P. 1557988320903191. https://doi.org/10.1177/1557988320903191
  10. Kamm A., Przychodzen P., Kuban-Jankowska A., Jacewicz D., Dabrowska A.M., Nussberger S., Wozniak M., Gorska-Ponikowska M. // Nitric Oxide. 2019. V. 93. P. 102–114. https://doi.org/10.1016/j.niox.2019.09.005
  11. Mintz J., Vedenko A., Rosete O., Shah K., Goldstein G., Hare J.M., Ramasamy R., Arora H. // Vaccines (Basel). 2021. V. 9. P. 94. https://doi.org/10.3390/vaccines9020094
  12. Andrabi S.M., Sharma N.S., Karan A., Shahriar S.M.S., Cordon B., Ma B., Xie J. // Adv. Sci. (Weinh). 2023. V. 10. P. e2303259. https://doi.org/10.1002/advs.202303259
  13. Gao D., Asghar S., Hu R., Chen S., Niu R., Liu J., Chen Z., Xiao Y. // Acta. Pharm. Sin. B. 2023. V. 13. P. 1498–1521. https://doi.org/10.1016/j.apsb.2022.11.016
  14. Huang W., Zhang J., Luo L., Yu Y., Sun T. // ACS Biomater. Sci. Eng. 2023. V. 9. P. 139–152. https://doi.org/10.1021/acsbiomaterials.2c01247
  15. Alimoradi H., Greish K., Barzegar-Fallah A., Alshaibani L., Pittalà V. // Int. J. Nanomedicine. 2018. V. 20. P. 7771–7787. https://doi.org/10.2147/IJN.S187089
  16. Ishima Y., Kragh-Hansen U., Maruyama T., Otagiri M. // Biomed. Res. Int. 2013. V. 2013. P. 353892. https://doi.org/10.1155/2013/353892
  17. Sinha B.K. // Int. J. Mol. Sci. 2023. V. 24. P. 13611. https://doi.org/10.3390/ijms241713611
  18. Zhao Z., Shan X., Zhang H., Shi X., Huang P., Sun J., He Z., Luo C., Zhang S. // J. Controll. Releas. 2023. V. 362. P. 151–169.
  19. Kim J., Thomas S.N. // Pharmacol. Rev. 2022. V. 74. P. 1146–1175. https://doi.org/10.1124/pharmrev.121.000500
  20. Stamler J.S., Jaraki O., Osborne J., Simon D.I., Keaney J., Vita J., Singel D., Valeri C.R., Loscalzo J. // Proc. Natl. Acad. Sci. USA. 1992. V. 89. P. 7674–7677. https://doi.org/10.1073/pnas.89.16.7674
  21. Tsiountsioura M., Cvirn G., Schlagenhauf A., Haidl H., Zischmeier K., Janschitz N., Koestenberger M., Wonisch W., Paar M., Wagner T., Weiss E.C., Hallström S. // Biomedicines. 2022. V. 10. P. 649. https://doi.org/10.3390/biomedicines10030649
  22. Rungatscher A., Hallstrom S., Linardi D., Milani E., Gasser H., Podesser B.K., Scarabelli T.M., Luciani G.B., Faggian G. // J. Heart Lung Transplant. 2015. V. 34. P. 479–488.
  23. Hallstrom S., Franz M., Gasser H., Vodrazka M., Semsroth S., Losert U.M., Haisjackl M., Podesser B.K., Malinski T. // Cardiovasc. Res. 2008. V. 77. P. 506–514.
  24. Hallstrom S., Gasser H., Neumayer C., Fugl A., Nanobashvili J., Jakubowski A., Huk I., Schlag G., Malinski T. // Circulation. 2002. V. 105. P. 3032–3038.
  25. Enayati M., Schneider K.H., Almeria C., Grasl C., Kaun C., Messner B., Rohringer S., Walter I., Wojta J., Budinsky L., Walpoth B.H., Schima H., Kager G., Hallström S., Podesser B.K., Bergmeister H. // Acta Biomaterialia. 2021. V. 134. P. 276–288. https://doi.org/10.1016/j.actbio.2021.07.048
  26. Schaefer A.-K., Kiss A., Oszwald A., Nagel F., Acar E., Aliabadi-Zuckermann A., Hackl M., Zuckermann A., Kain R., Jakubowski A., Ferdinandy P., Hallström S. // Transpl. Int. 2022. V. 35. P. 10057. https://doi.org/10.3389/ti.2022.10057
  27. Ishima Y., Maruyama T., Otagiri M., Chuang V.T.G., Ishida T. // Chem. Pharm. Bull. (Tokyo). 2022. V. 70. P. 330–333. https://doi.org/10.1248/cpb.c21-01024
  28. Ishima Y., Maruyama T., Otagiri M., Ishida T. // Chem. Pharm. Bull. 2020. V. 68. P. 583–588. https://doi.org/10.1248/cpb.c20-00026
  29. Bihari S., Bannard-Smith J., Bellomo R. // Crit. Care Resusc. 2020. V. 22. P. 257–265. https://doi.org/10.1016/S1441-2772(23)00394-0
  30. Ishima Y. // Biol. Pharm. Bull. 2017. V. 40. P. 128–134. https://doi.org/10.1248/bpb.b16-00867
  31. Maeda H. // J. Pers. Med. 2021. V. 11. P. 229.
  32. Fang J. // J. Pers. Med. 2022. V. 12. P. 95.
  33. Zi Y., Yang K., He J., Wu Z., Liu J., Zhang W. // Adv. Drug Deliv. Rev. 2022. V. 188. P. 114449.
  34. Kim J., Cho H., Lim D.K., Joo M.K., Kim K. // Int. J. Mol. Sci. 2023. V. 24. P. 10082. https://doi.org/10.3390/ijms241210082
  35. Subhan M.A., Parveen F., Filipczak N., Yalamarty S.S.K., Torchilin V.P. // J. Pers. Med. 2023. V. 13. P. 389. https://doi.org/10.3390/jpm13030389
  36. Xu Y., Ren. H., Liu J., Wang Y., Meng Z., He Z., Miao W., Chen G., Li X. // Nanoscale. 2019. V. 11. P. 5474–5488.
  37. Zhao Y., Ouyang X., Peng Y., Peng S. // Pharmaceutics. 2021. V. 13. P. 1917. https://doi.org/10.3390/pharmaceutics1311191
  38. Ji P., Yang K., Xu Q., Qin G., Zhu Q., Qian Y., Yao W. // Pharmaceuticals (Basel). 2023. V. 16. P. 1394. https://doi.org/10.3390/ph16101394
  39. Fan W., Bu W., Zhang Z., Shen B., Zhang H., He Q., Ni D., Cui Z., Zhao K., Bu J., Du J., Liu J., Shi J. // Angew. Chem. Int. Ed. Engl. 2015. V. 54. P. 14026– 14030. https://doi.org/10.1002/anie.201504536
  40. Barth R.F., Zhang Z., Liu T. // Cancer Commun. 2018. V. 38. P. 36.
  41. Иванов А.А., Смирнов А.Н., Таскаев С.Ю., Баянов Б.Ф., Бельченко Ю.И., Давыденко В.И., Дунаевский А., Емелев И.С., Касатов Д.А., Макаров А.Н., Микенс М., Куксанов Н.К., Попов С.С., Салимов Р.А., Санин А.Л., Сорокин И.Н., Сычева Т.В., Щудло И.М., Воробьев Д.С., Черепков В.Г., Фадеев С.Н. // Усп. физ. химии. 2022. Т. 192. С. 894– 912.
  42. Popova T. V., Pyshnaya I.A., Zakharova O.D., Akulov A.E., Shevelev O.B., Poletaeva J., Zavjalov E.L., Silnikov V.N., Ryabchikova E.I., Godovikova T.S. // Biomedicines. 2021. V. 9. P. 1–15.
  43. Peters T.J. // Adv. Protein Chem. 1985. V. 37. P. 161– 245.
  44. Ishima Y., Hiroyama S., Kragh-Hansen U., Maruyama T., Sawa T., Akaike T., Kai T., Otagiri M. // Nitric Oxide. 2010. V. 23. P. 121–127.
  45. Era H., Terada S., Minami T., Takahashi T., Arikawa T. // Heterogenity of Commercially Available Human Serum Albumin Products: Thiol Oxidation and Protein Carbonylation. 37th Congress of IUPS. Birmingham, UK, 2013.
  46. Miyamura S., Imafuku T., Anraku M., Taguchi K., Yamasaki K., Tominaga Y., Maeda H., Ishima Y., Watanabe H., Otagiri M., Maruyama T. // J. Pharm. Sci. 2016. V. 105. P. 1043–1049.
  47. Xu Y., Liu J., Liu Z., Chen G., Li X., Ren H. // Int. J. Nanomedicine. 2021. V. 16. P. 2597–2613. https://doi.org/10.2147/IJN.S295445
  48. Ikeda M., Ishima Y., Chuang V.T.G., Ikeda T., Kinoshita R., Watanabe H., Ishida T., Otagiri M., Maruyama T. // Nitric Oxide. 2017. V. 69. P. 28–34. https://doi.org/10.1016/j.niox.2017.04.005
  49. Lisitskiy V.A., Khan H., Popova T.V., Chubarov A.S., Zakharova O.D., Akulov A.E., Shevelev O.B., Zavjalov E.L., Koptyug I.V., Moshkin M.P., Silnikov V.N., Ahmad S., Godovikova T.S. // Bioorg. Med. Chem. Lett. 2017. V. 27. P. 3925–3930.
  50. Popova T.V., Krumkacheva O.A., Burmakova A.S., Spitsyna A.S., Zakharova O.D., Lisitskiy V.A., Kirilyuk I.A., Silnikov V.N., Bowman M.K., Bagryanskaya E.G., Godovikova T.S. // RSC Medicinal Chemistry. 2020. V. 11. P. 1314–1325.
  51. Raskolupova V.I., Wang M., Dymova M.A., Petrov G.O., Shchudlo I.M., Taskaev S.Y., Abramova T.V., Godovikova T.S., Silnikov V.N., Popova T.V. // Molecules. 2023. V. 28. P. 2672–2689.
  52. Popova T.V., Dymova M.A., Koroleva L.S., Zakharova O.D., Lisitskiy V.A., Raskolupova V.I., Sycheva T.V., Taskaev S.Yu., Silnikov V.N., Godovikova T.S. // Molecules. 2021. V. 26. P. 6537–6553.
  53. Popova T.V., Khan H., Chubarov A.S., Lisitskiy V.A., Antonova N.M., Akulov A.E., Shevelev O.B., Zavjalov E.L., Silnikov V.N., Ahmad S., Godovikova T.S. // Bioorg. Med. Chem. Lett. 2018. V. 28. P. 260–264. https://doi.org/10.1016/j.bmcl.2017.12.061
  54. Kashiwagi H., Kawabata S., Yoshimura K., Fukuo Y., Kanemitsu T., Takeuchi K., Shiba H., Hiramatsu R., Nishimura K., Kawai K., Takata T., Tanaka H., Watanabe T., Suzuki M., Miyatake S.I., Nakamura H., Wanibuchi M. // Investigat. New Drugs. 2022. V. 40. P. 255–264.
  55. Sivaev I.B., Kulikova N.Y., Nizhnik E.A., Vichuzhanin M.V., Starikova Z.A., Semioshkin A.A., Bregadze V.I. // J. Organomet. Chem. 2008. V. 693. P. 519– 525.
  56. Semioshkin A., Nizhnik E., Godovikov I., Starikiva Z., Bregadze V. // J. Organomet. Chem. 2007. V. 692. P. 4020–4028.
  57. Kikuchi S., Kanoh D., Sato S., Sakurai Y., Suzuki M., Nakamura H. // J. Control. Release. 2016. V. 237. P. 160–166.
  58. Mosmann T. // J. Immunol. Methods. 1983. V. 65. P. 55–63.
  59. Peters R.A. // In: Advances in Enzymology / Eds. Nord F.F. Interscience Publishers Inc., Geneva, Switzerland, 1957. P. 113–159.
  60. Laemmli U.K. // Nature. 1970. V. 227. P. 680–685.
  61. Terance W. // Tetrahedron Letters. 1985. V. 26. P. 2013– 2016.
  62. Chubarov A.S., Zakharova O.D., Koval O.A., Romaschenko A.V., Akulov A.E., Zavjalov E.L., Razumov I.A., Koptyug I.V., Knorre D.G., Godovikova T.S. // Bioorg. Med. Chem. 2015. V. 23. P. 6943–6954.

Supplementary files

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2. Fig. 1. Approaches to enhancing the effectiveness of NO therapy by exposure to exogenous (light, ultrasound, X-rays) or endogenous (glutathione, acid, glucose) stimuli [37].

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3. Fig. 2. Schematic diagram of BNCT [40].

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4. Fig. 3. Scheme of synthesis of boron-containing analogue of homocysteine ​​thiolactone (HTL-B12H11).

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5. Fig. 4. Scheme for the preparation of S-nitrosylated boron-containing homocysteinylamide albumin containing a fluorescent dye (paths a, b and c) and S-nitrosylated albumin (path d).

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6. Fig. 5. Characteristics of multifunctional conjugates of human serum albumin. (a) – Electronic absorption spectra of HSA and its homocystamides in PBS buffer, pH 7.4; (b) – MALDI-TOF mass spectra of HSA and its homocystamides; (c) – SDS-PAGE of homocystamide conjugates of HSA under Laemmli conditions followed by staining with Coomassie blue.

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7. Fig. 6. Viability of T98G cells treated with HSA, HSA-Cy5-HcyB12H11, and HSA-Cy5-HSA-Cy5-HcyB12H11-NO. The dose of conjugates was 0.02–40 μM protein equivalent. All data are presented as mean ± SD (n = 3). Two-way ANOVA was used to compare more than two data sets. **** p ≤ 0.0001.

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