The influenza RNA-dependent RNA polymerase harbours an endonuclease subunit characterized by a catalytic site housing two divalent metal ions. By effectively chelating both Mg2+ and Mn2+ ions, a small-molecule inhibitor with a metal-binding pharmacophore can halt endonuclease activity. Herein, two 3'-dehydroxypurpurogallin-4-carboxamide series, namely twelve C-4' unsubstituted and twelve C-4' phenyl substituted congeners were designed and prepared to be tested as inhibitors of the metal-dependent viral enzyme. These inhibitors were accessed through the chemoenzymatic reaction of gallic acid with either pyrocatechol or phenylpyrocatechol moderated by laccase, followed by amidation. Experimental IC50 values were determined using AlphaScreen technology, with the most potent inhibitors exhibiting IC50 values around 0.35 μM. Using X-ray crystallography, we analyzed structure of the endonuclease in complex with one potent 3'-dehydroxypurpurogallin-carboxamide at 2.0 Å resolution, revealing the coordination of the compound's triad of oxygen atoms with the two metal ions in the influenza A endonuclease active site.

1st Faculty of Medicine Charles University Prague Kateřinská 1660 32 Prague 2 121 08 Czech Republic

Department of Organic Chemistry Faculty of Science Charles University Prague Hlavova 2030 8 Prague 2 128 43 Czech Republic

Institute of Microbiology of the Czech Academy of Sciences Vídeňská 1083 Prague 4 140 00 Czech Republic

Institute of Organic Chemistry and Biochemistry of the Czech Academy of Sciences Flemingovo náměstí 542 2 Prague 6 160 00 Czech Republic

Zobrazit více v PubMed

A. Dastan, H. Kilic, N. Saracoglu, Beilstein J. Org. Chem. 2018, 14, 1120–1180;

S. N. Momm, R. Brückner, J. Org. Chem. 2022, 87, 15415–15420;

P. Koblischek, R. Brückner, Eur. J. Org. Chem. 2022, 2022, e202200686.

J. A. Barltrop, J. S. Nicholson, J. Chem. Soc. 1948, 116–120;

L. Horner, S. Gowecke, W. Durckheimer, Chem. Ber.-Recl. 1961, 94, 1276–1291.

T. W. Wu, L. H. Zeng, J. Wu, K. P. Fung, R. D. Weisel, A. Hempel, N. Camerman, Biochem. Pharmacol. 1996, 52, 1073–1080.

S. Kitada, M. Leone, S. Sareth, D. Y. Zhai, J. C. Reed, M. Pellecchia, J. Med. Chem. 2003, 46, 4259–4264.

S. M. Sang, J. D. Lambert, S. Y. Tian, J. L. Hong, Z. Hou, J. H. Ryu, R. E. Stark, R. T. Rosen, M. T. Huang, C. S. Yang, C. T. Ho, Bioorg. Med. Chem. 2004, 12, 459–467;

T. H. Kim, S. K. Ku, I. C. Lee, J. S. Bae, BMB Rep. 2012, 45, 200–205.

V. Zima, K. Radilová, M. Kozísek, C. B. Albiñana, E. Karlukova, J. Brynda, J. Fanfrlík, M. Flieger, J. Hodek, J. Weber, P. Majer, J. Konvalinka, A. Machara, Eur. J. Med. Chem. 2020, 208, 112754.

A. Dias, D. Bouvier, T. Crepin, A. A. McCarthy, D. J. Hart, F. Baudin, S. Cusack, R. W. Ruigrok, Nature 2009, 458, 914–918.

K. Cheng, X. H. Wang, S. T. Zhang, H. Yin, Angew. Chem. Int. Ed. 2012, 51, 12246–12249.

D. Mesa-Siverio, A. Estévez-Braun, A. G. Ravelo, J. R. Murguia, A. Rodríguez-Afonso, Eur. J. Org. Chem. 2003, 2003, 4243–4247.

B. Kim, S. Jo, S. B. Park, C. H. Chae, K. Lee, B. Koh, I. Shin, Bioorg. Med. Chem. Lett. 2020, 30, 126756.

J. A. Souto, F. Sarno, A. Nebbioso, C. Papulino, R. Alvarez, J. Lombino, U. Perricone, A. Padova, L. Altucci, A. R. de Lera, Front. Chem. 2020, 8, 312;

L. Zhang, Y. Chen, Z. J. Li, C. C. Lin, T. T. Zhang, G. Wang, Drug Discov. Today 2023, 28, 103519.

A. Y. Chen, R. N. Adamek, B. L. Dick, C. V. Credille, C. N. Morrison, S. M. Cohen, Chem. Rev. 2019, 119, 1323–1455.

J. C. Hastings, H. Selnick, B. Wolanski, J. E. Tomassini, Antimicrob. Agents Chemother. 1996, 40, 1304–1307;

A. Stevaert, R. Dallocchio, A. Dessì, N. Pala, D. Rogolino, M. Sechi, L. Naesens, J. Virol. 2013, 87, 10524–10538.

T. Kuzuhara, Y. Iwai, H. Takahashi, D. Hatakeyama, N. Echigo, PLoS Curr. 2009, 1, RRN1052;

E. Kowalinski, C. Zubieta, A. Wolkerstorfer, O. H. J. Szolar, R. W. H. Ruigrok, S. Cusack, PloS Pathog. 2012, 8, e1002831.

S. B. Singh, J. E. Tomassini, J. Org. Chem. 2001, 66, 5504–5516.

R. Reiberger, K. Radilová, M. Král, V. Zima, P. Majer, J. Brynda, M. Dracínsky, J. Konvalinka, M. Kozísek, A. Machara, Int. J. Mol. Sci. 2021, 22, 7735.

J. C. Jones, B. M. Marathe, C. Lerner, L. Kreis, R. Gasser, P. N. Q. Pascua, I. Najera, E. A. Govorkova, Antimicrob. Agents Chemother. 2016, 60, 5504–5514.

H. Y. Sagong, J. D. Bauman, D. Patel, K. Das, E. Arnold, E. J. LaVoie, J. Med. Chem. 2014, 57, 8086–8098.

A. K. Parhi, A. Xiang, J. D. Bauman, D. Patel, R. S. K. Vijayan, K. Das, E. Arnold, E. J. LaVoie, Bioorg. Med. Chem. 2013, 21, 6435–6446;

H. Y. Sagong, J. D. Bauman, A. Nogales, L. Martínez-Sobrido, E. Arnold, E. J. LaVoie, ChemMedChem 2019, 14, 1204–1223;

C. V. Credille, C. N. Morrison, R. W. Stokes, B. L. Dick, Y. F. Feng, J. X. Sun, Y. Chen, S. M. Cohen, J. Med. Chem. 2019, 62, 9438–9449;

R. W. Stokes, A. J. Kohlbrand, H. Seo, B. Sankaran, J. Karges, S. M. Cohen, ACS Med. Chem. Lett. 2023, 14, 75–82.

Y. X. Liao, Y. L. Ye, S. M. Li, Y. L. Zhuang, L. Y. Chen, J. X. Chen, Z. N. Cui, L. J. Huo, S. W. Liu, G. P. Song, Eur. J. Med. Chem. 2020, 189, 112048.

D. Rogolino, L. Naesens, J. Bartoli, M. Carcelli, L. De Luca, G. Pelosi, R. W. Stokes, R. Van Berwaer, S. Vittorio, A. Stevaert, S. M. Cohen, Bioorg. Chem. 2021, 116, 105388.

C. V. Credille, Y. Chen, S. M. Cohen, J. Med. Chem. 2017, 60, 9912–9912;

H. Ju, J. Zhang, B. S. Huang, D. W. Kang, B. Huang, X. Y. Liu, P. Zhan, J. Med. Chem. 2017, 60, 3533–3551;

C. V. Credille, B. L. Dick, C. N. Morrison, R. W. Stokes, R. N. Adamek, N. C. Wu, I. A. Wilson, S. M. Cohen, J. Med. Chem. 2018, 61, 10206–10217.

F. G. Hayden, N. Sugaya, N. Hirotsu, N. Lee, M. D. de Jong, A. C. Hurt, T. Ishida, H. Sekino, K. Yamada, S. Portsmouth, K. Kawaguchi, T. Shishido, M. Arai, K. Tsuchiya, T. Uehara, A. Watanabe, B. M. Investigators, N. Engl. J. Med. 2018, 379, 913–923.

J. C. Jones, G. Kumar, S. Barman, I. Najera, S. W. White, R. J. Webby, E. A. Govorkova, MBio 2018, 9, e00430-18;

M. Nakauchi, E. Takashita, S. Fujisaki, M. Shirakura, R. Ogawa, H. Morita, H. Miura, S. Saito, S. Watanabe, T. Odagiri, T. Kageyama, Influenza Other Resp. 2020, 14, 436–443.

B. Wagner, R. Öhrlein, B. Herzog, K. Eichin, G. Baisch, S. Portmann, Patent US9707417B2, 2017.

J. A. Souto, ChemistrySelect 2021, 6, 10717–10721.

M. Tanc, M. Cleenewerck, A. Kurdi, R. Roelandt, W. Declercq, G. De Meyer, K. Augustyns, W. Martinet, P. Van der Veken, Bioorg. Chem. 2019, 87, 163–168.

M. Fukuoka, M. Minakuchi, A. Kawaguchi, K. Nagata, Y. O. Kamatari, K. Kuwata, Bba-Gen. Subj. 2012, 1820, 90–95.

T. Crépin, A. Dias, A. Palencia, C. Swale, S. Cusack, R. W. H. Ruigrok, J. Virol. 2010, 84, 9096–9104;

K. Hara, F. I. Schmidt, M. Crow, G. G. Brownlee, J. Virol. 2006, 80, 7789–7798.

G. Kumar, M. Cuypers, R. R. Webby, T. R. Webb, S. W. White, Nucleic Acids Res. 2021, 49, 1609–1618.

R. M. DuBois, P. J. Slavish, B. M. Baughman, M. K. Yun, J. Bao, R. J. Webby, T. R. Webb, S. W. White, PloS Pathog. 2012, 8, e1002830.

U. Mueller, R. Förster, M. Hellmig, F. U. Huschmann, A. Kastner, P. Malecki, S. Pühringer, M. Röwer, K. Sparta, M. Steffien, M. Ühlein, P. Wilk, M. S. Weiss, Eur. Phys. J. Plus 2015, 130, 141.

M. Krug, M. S. Weiss, U. Heinemann, U. Mueller, J. Appl. Crystallogr. 2012, 45, 568–572;

W. Kabsch, Acta Crystallogr. Sect. D 2010, 66, 125–132.

A. Vagin, A. Teplyakov, J. Appl. Crystallogr. 1997, 30, 1022–1025.

M. D. Winn, C. C. Ballard, K. D. Cowtan, E. J. Dodson, P. Emsley, P. R. Evans, R. M. Keegan, E. B. Krissinel, A. G. W. Leslie, A. McCoy, S. J. McNicholas, G. N. Murshudov, N. S. Pannu, E. A. Potterton, H. R. Powell, R. J. Read, A. Vagin, K. S. Wilson, Acta Crystallogr. D 2011, 67, 235–242.

P. Emsley, K. Cowtan, Acta Crystallogr. D 2004, 60, 2126–2132.

G. N. Murshudov, P. Skubák, A. A. Lebedev, N. S. Pannu, R. A. Steiner, R. A. Nicholls, M. D. Winn, F. Long, A. A. Vagin, Acta Crystallogr. D 2011, 67, 355–367.

V. B. Chen, W. B. Arendall, J. J. Headd, D. A. Keedy, R. M. Immormino, G. J. Kapral, L. W. Murray, J. S. Richardson, D. C. Richardson, Acta Crystallogr. D 2010, 66, 12–21.

Beyond natural flavonoids: exploring bioisosterism in design and synthesis of influenza endonuclease inhibitors