Please do not adjust margins
ChemComm
Page 4 of 4
ARTICLE
Journal Name
decrease in MIC of imipenem (Table S3 and S4). Also, Cu(DTC)2 and
its analogues exhibited potential in restoring imipenem efficacy,
with a 8-64-fold reduction in MICs against E. coli-MβLs (Table. S6).
Such excellent synergistic antibacterial efficacy is also verified by
gradually elevated cellular uptake of Cu(II) and time kill curves (Fig.
S20 and S21). The MIC of imipenem remained unchanged in the
presence of DSF and Cu(DTC)2 at a dose of 16 μg/mL against E. coli-
NDM-1-C208A and E. coli-L1 (Table S7), also implying that the
Cys208 is essential for inhibition of NDM-1 by DSF and Cu(DTC)2.
Importantly, Cu(DTC)2 is able to circumvent the reduction by the
intracellular thiols and retain its ability to oxidize Zn(II) thiolate site
of NDM-1 to specifically kill carbapenem-resistant bacteria (Fig.
S22a and Table S8).
Jaskolski, Drug Resistance Updates, 2018, 40, 1-12.
DOI: 10.1039/C9CC09074F
4. C. T. Lohans, E. I. Freeman, E. V. Groesen, C. L. Tooke, P. Hinchliffe, J.
Spencer, J. Brem and C. J. Schofield, Sci. Rep., 2019, 9, 13608-13617.
5. J. Brem, R. Cain, S. Cahill, M. A. McDonough, I. J. Clifton, J.
JiménezCastellanos, M. B. Avison, J. Spencer, C. W. G. Fishwick and C.
J. Schofield, Nat Commun, 2016, 7, 12406-12417.
6. B. Ma, C. Fang, L. Lu, M. Wang, X. Xue, Y. Zhou, M. Li, Y. Hu, X. Luo and
Z. Hou, Nat Commun, 2019, 10, 3517-3528.
7. L. Ejim, M. A. Farha, S. B. Falconer, J. Wildenhain, B. K. Coombes, M.
Tyers, E. D. Brown and G. D. Wright, Nat. Chem. Biol., 2011, 7, 348-
351.
8. H. Feng, J. Ding, D. Zhu, X. Liu, X. Xu, Y. Zhang, S. Zang, D. C. Wang and
W. Liu, J. Am. Chem. Soc., 2014, 136, 14694-14697.
9. A. M. King, S. A. Reid-Yu, W. Wang, D. T. King, G. De Pascale, N. C.
Strynadka, T. R. Walsh, B. K. Coombes and G. D. Wright, Nature, 2014,
510, 503-506.
10. J. Chiou, S. Wan, K. F. Chan, P. K. So, D. He, E. W. Chan, T. H. Chan, K. Y.
Wong, J. Tao and S. Chen, Chem. Commun. (Camb.), 2015, 51, 9543-
9546.
11. X. Y. Ouyang, Y. N. Chang, K. W. Yang, W. M. Wang, J. J. Bai, J. W. Wang,
Y. J. Zhang, S. Y. Wang, B. B. Xie and L. L. Wang, Chem. Commun.
(Camb.), 2017, 53, 8878-8881.
Just as it was observed in the case of inhibitor-resistant TEM-1
and KPC-2,24, 25 owing to the rapid evolution of variant MβLs, most
organic molecule-based inhibitors may easily encounter resistance.
The combination of Cu(DTC)2 with imipenem significantly
suppressed the development of high-level resistance in NDM-1 12. M. M. Wang, W. C. Chu, Y. Yang, Q. Q. Yang, S. S. Qin, E. Zhang, Bioorg.
Med. Chem. Lett., 2018, 28, 3436-3440.
13. Z. Skrott, M. Mistrik, K. K. Andersen, S. Friis, D. Majera, J. Gursky, T.
producers, compared to that of original strain treated by imipenem
alone (Fig. S22b), which may be due to the reduced protein levels of
Ozdian, J. Bartkova, Z. Turi, P. Moudry, M. Kraus, M. Michalova, J.
Vaclavkova, P. Dzubak, I. Vrobel, P. Pouckova, J. Sedlacek, A.
Miklovicova, A. Kutt, J. Li, J. Mattova, C. Driessen, Q. P. Dou, J. Olsen,
M. Hajduch, B. Cvek, R. J. Deshaies and J. Bartek, Nature, 2017, 552,
194-199.
NDM-1 in the combination therapy.
In conclusion, our results validated that the disulfiram is a
promising candidate for the development of NDM-1 inhibitor,
which covalently bind to NDM-1 by forming an S-S bond with the
Cys208 residue at the active site. Cu(DTC)2, a copper-containing
metabolite in vivo of DSF, also inactivated NDM-1 through a unique
mechanism with oxidizing Zn(II) thiolate in the active site of
enzyme, thereby exhibiting dual functional inhibitory mechanisms
against both B1 and B2 subclasses MβLs. Also, DSF and Cu(DTC)2
effectively restored imipenem efficacy against clinical isolates E.
coli, K. pneumoniae and P. aeruginosa producing-NDM-1.
Importantly, Cu(DTC)2 could avoid the reduction by the intracellular
thiols to specifically kill CRE and slow down the development of
higher-level resistance in bacteria producing NDM-1. Although the
disulfiram and Cu(DTC)2 are already used in clinic or in clinical phase
II, their toxicity is a concern. Therefore, future strategy should focus
on enhancing their specificity to carbapenem-resistant Gram-
negative pathogens mediated by MβLs and reducing their toxicity.
14. Y. J. Zhang, W. M. Wang, P. Oelschlaeger, C. Chen, J. E. Lei, M. Lv and K.
W. Yang, ACS Infect Dis, 2018, 4, 1671-1678.
15. Y. Xiang, C. Chen, W. M. Wang, L. W. Xu, K. W. Yang, P. Oelschlaeger
and Y. He, ACS Med. Chem. Lett., 2018, 9, 359-364.
16. Z. Skrott, D. Majera, J. Gursky, T. Buchtova, M. Hajduch, M. Mistrik
and J. Bartek, Oncogene, 2019, DOI: 10.1038/s41388-019-0915-2.
17. Y. N. Chang, Y. Xiang, Y. J. Zhang, W. M. Wang, C. Chen, P. Oelschlaeger
and K. W. Yang, ACS Med. Chem. Lett., 2017, 8, 527-532.
18. A. Y. Chen, W. T. Pei, A. C. Stewart, A. Bergstrom, Z. Cheng, C. Miller, C.
R. Bethel, S. H. Marshall, C. V. Credille and C. L. Riley, J. Med. Chem.,
2017, 60, 7267-7283.
19. R. Wang, T. P. Lai, P. Gao, H. Zhang, P. L. Ho, P. C. Woo, G. Ma, R. Y.
Kao, H. Li and H. Sun, Nat Commun, 2018, 9, 439-450.
20. C. Chen, Y. Xiang, Y. Liu, X. D. Hu and K. W. Yang. Med. Chem. Comm,
2018, 9, 1172-1178.
21. H. Huang, H. Li,, A.J. Wang., S.X. Zhong, K.M. Fang and J.J. Feng,
Analyst, 2014, 139, 6536-6541.
22. S. Poulston, P. M. Parlett, P. Stone and M. Bowker, Surface & Interface
Analysis, 1996, 24, 811-820.
23. Y. L. Wang, S. Liu, Z. J. Yu, Y. Lei, M. Y. Huang, Y. H. Yan, Q. Ma, Y.
Zheng, H. Deng, Y. Sun, C. Wu, Y. Yu, Q. Chen, Z. Wang, Y. Wu and G. B.
Li, J. Med. Chem., 2019, 62, 7160-7184.
24. R. Canton, M. I. Morosini, O. M. de la Maza, E. G. de la Pedrosa, Clin.
Microbiol. Infect. 2008, 14, 53–62.
Conflicts of interest
There are no conflicts to declare
.
25. K. M. Papp-Wallace, M. L. Winkler, M. A. Taracila, R. A. Bonomo,
Antimicrob. Agents Chemother. 2015, 59, 3710–3717.
Acknowledgements
This work was supported by Grants (21572179 and 2019KW-068 to
K.W.Y) from National Natural Science Foundation of China and the
Shaanxi Province International Cooperation Project.
Notes and references
1. Y. Zong, F. Fang, K. J. Meyer, L. Wang, Z. Ni, H. Gao, K. Lewis, J. Zhang
and Y. Rao, Nat Commun, 2019, 10, 3268-3280.
2. Y. Cai, C. Chen, M. Zhao, X. G. Yu, K. Lan, K. Liao, P. H. Guo, W. Z. Zhang,
X. Y. Ma, Y. T. He, J. M. Zeng, L. Chen, W. Jia, Y. W. Tang and B. Huang,
Front Microbiol, 2019, 10, 1610-1625.
4 | J. Name., 2012, 00, 1-3
This journal is © The Royal Society of Chemistry 20xx
Please do not adjust margins