Full Papers
doi.org/10.1002/cmdc.202100042
ChemMedChem
°
[9] D. Yong, M. A. Toleman, C. G. Giske, H. S. Cho, K. Sundman, K. Lee, T. R.
inoculated into tryptic soy broth (TSB) and incubated at 37 C. Once
the bacteria cells grew to an OD600 of 0.5, the suspension was
diluted with MHB (final concentration 106 CFU/mL) and then added
to the microplates containing the test compounds (50 μL to each
1
2
3
4
5
6
7
8
9
[10] T. Li, Q. Wang, F. Chen, X. Li, S. Luo, H. Fang, D. Wang, Z. Li, X. Hou, H.
Wang, PLoS One 2013, 8, 1–5.
[11] N. Laraki, N. Franceschini, G. M. Rossolini, P. Santucci, C. Meunier, E.
[12] L. C. Ju, Z. Cheng, W. Fast, R. A. Bonomo, M. W. Crowder, Trends
[13] A. M. King, S. A. Reid-Yu, W. Wang, D. T. King, G. De Pascale, N. C.
[15] A. J. Turner in The Protective Arm of the Renin Angiotensin System (RAS),
(Eds.: Thomas Unger, Ulrike M. Steckelings, Robson A. S. dos Santos),
Academic Press, 2015, pp. 185–189, Chapter 25 – ACE2 Cell Biology,
Regulation, and Physiological Functions, ISBN 9780128013649, https://
doi.org/10.1016/B978-0-12-801364-9.00025-0.
[17] B. M. R. Liénard, G. Garau, L. Horsfall, A. I. Karsisiotis, C. Damblon, P.
Lassaux, C. Papamicael, G. C. K. Roberts, M. Galleni, O. Dideberg, J. M.
[19] T. Yuan, Captopril. In Handbook of Metabolic Pathways of Xenobiotics
(eds P. W. Lee, H. Aizawa, L. L. Gan, C. Prakash and D. Zhong), 2014.
[20] A. M. Somboro, D. Tiwari, L. A. Bester, R. Parboosing, L. Chonco, H. G.
Kruger, P. I. Arvidsson, T. Govender, T. Naicker, S. Y. Essack, J. Antimicrob.
Chemother. 2014, 70, 1594–1596.
[21] R. Azumah, J. Dutta, A. M. Somboro, M. Ramtahal, L. Chonco, R.
Parboosing, L. A. Bester, H. G. Kruger, T. Naicker, S. Y. Essack, T.
[22] Q. Wang, Y. He, R. Lu, W. M. Wang, K. W. Yang, H. M. Fan, Y. Jin, G.
Michael Blackburn, Biosci. Rep. 2018, 38, 1–11.
[23] L. E. Horsfall, G. Garau, B. M. R. Liénard, O. Dideberg, C. J. Schofield, J. M.
[24] F. M. Klingler, T. A. Wichelhaus, D. Frank, J. Cuesta-Bernal, J. El-Delik,
H. F. Müller, H. Sjuts, S. Göttig, A. Koenigs, K. M. Pos, D. Pogoryelov, E.
[25] C. Mollard, C. Moali, C. Papamicael, C. Damblon, S. Vessilier, G.
Amicosante, C. J. Schofield, M. Galleni, J. M. Frère, G. C. K. Roberts, J.
[26] N. Li, Y. Xu, Q. Xia, C. Bai, T. Wang, L. Wang, D. He, N. Xie, L. Li, J. Wang,
H. G. Zhou, F. Xu, C. Yang, Q. Zhang, Z. Yin, Y. Guo, Y. Chen, Bioorg. Med.
[27] W. S. Shin, A. Bergstrom, J. Xie, R. A. Bonomo, W. Michael, R. Muthyala,
[28] “Inhibitors of Metallo-Beta-Lactamases”, B. Jurgen, A. M. Rydzik, M. A.
McDonough, C. J. Schofield, A. Morrison, J. Hewitt, A. Pannifer, P. Jones,
WO/2017/093727, 2017.
°
well, final volume: 100 μL). After incubation at 37 C for 16–20 h,
the microplates were inspected for growth inhibition. MIC values
were defined as the lowest concentration of the compound that
prevented visible growth of bacteria.
Synergy assay (CMIC/4). The test compounds were serially diluted with
MHB starting from a maximum concentration of 1000 μM (25 μL in
each well). Meropenem (25 μL) was then added to the wells to
achieve a final concentration of 16 μg/mL. E. coli RC0089 (NDM-1)
was cultured and added to the microplates as described above
(50 μL to each well, final volume: 100 μL). The CMIC/4 value was
defined as the lowest concentration of the inhibitor that prevented
10
11
12
13
14
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
42
43
44
45
46
47
48
49
50
51
52
53
54
55
56
57
the visible growth of the bacteria when combined with meropenem
1
at
=
of its MIC. FICI values were established by applying the
4
following formula where an FICI<0.5 indicates synergy:
in combination
MICMeropenem alone
MICInhibitor in combination
+
MICInhibitor alone
FICI=MIC
Meropenem
OD600 checkerboard assay. The test compounds were serially diluted
with MHB starting from a maximum concentration of 125 μM (25 μL
in each well). Meropenem was serially diluted to 4x the final
concentration before being added to the test compounds (25 μl). E.
coli RC0089 was cultured and added to the microplates as
described above (50 μL to each well, final volume: 100 μL). The
microplates were then incubated at 37 C with shaking. After 18 h,
the optical density of each well was measured using a Tecan Spark
plate reader at 600 nm.
°
Acknowledgements
We thank Robbert Q. Kim and Angeliki Moutsiopoulou of the
LUMC Protein Facility for assistance with protein expression and
purification. We thank Ioli Kotsogianni for her guidance with the
ITC experiments. Financial support was provided by the European
Research Council (ERC consolidator grant to NIM, grant agreement
no. 725523).
Conflict of Interest
[29] “Antibacterial Thiazolecarboxylic Acids”, M. Lemonnier, D. Davies, D.
Pallin, WO/2014/198849, 2014.
The authors declare no conflict of interest.
[30] S. Leiris, A. Coelho, J. Castandet, M. Bayet, C. Lozano, J. Bougnon, J.
Bousquet, M. Everett, M. Lemonnier, N. Sprynski, M. Zalacain, T. D. Pallin,
M. C. Cramp, N. Jennings, G. Raphy, M. W. Jones, R. Pattipati, B. Shankar,
R. Sivasubrahmanyam, A. K. Soodhagani, R. R. Juventhala, N. Pottabathi-
ni, S. Pothukanuri, M. Benvenuti, C. Pozzi, S. Mangani, F. De Luca, G.
[31] C. A. Thomas, Z. Cheng, K. Yang, E. Hellwarth, C. J. Yurkiewicz, F. M.
Baxter, S. A. Fullington, S. A. Klinsky, J. L. Otto, A. Y. Chen, S. M. Cohen,
M. W. Crowder, J. Inorg. Biochem. 2020, 210, 111–123.
[32] S. S. Van Berkel, J. Brem, A. M. Rydzik, R. Salimraj, R. Cain, A. Verma, R. J.
[33] K. H. M. E. Tehrani, H. Fu, N. C. Brüchle, V. Mashayekhi, A. Prats Luján,
Keywords: antibiotic resistance · MBL inhibitors · metallo-beta-
lactamases · synergy · zinc binding
[1] T. P. Van Boeckel, S. Gandra, A. Ashok, Q. Caudron, B. T. Grenfell, S. A.
[3] R. P. Ambler, Philos. Trans. R. Soc. London Ser. B 1980, 289, 321–331.
[4] D. de A Viana Marques, S. E. F. Machado, V. Carvalho Santos Ebinuma, C.
de A L Duarte, A. Converti, A. L. F. Porto, Antibiotics 2018, 7, 1–26.
[5] H. Feng, J. Ding, D. Zhu, X. Liu, X. Xu, Y. Zhang, S. Zang, D. C. Wang, W.
[6] V. L. Green, A. Verma, R. J. Owens, S. E. V. Phillips, S. B. Carr, Acta
[7] N. O. Concha, C. A. Janson, P. Rowling, S. Pearson, C. A. Cheever, B. P.
Clarke, C. Lewis, M. Galleni, J. M. Frère, D. J. Payne, J. H. Bateson, S. S.
[34] S. Minisola, J. Pepe, S. Piemonte, C. Cipriani, BMJ 2015, 350, 1–9.
[35] J. Soar, G. D. Perkins, G. Abbas, A. Alfonzo, A. Barelli, J. J. L. M. Bierens, H.
Brugger, C. D. Deakin, J. Dunning, M. Georgiou, A. J. Handley, D. J.
Lockey, P. Paal, C. Sandroni, K. C. Thies, D. A. Zideman, J. P. Nolan,
[36] A. Krężel, W. Maret, Arch. Biochem. Biophys. 2016, 611, 3–19.
ChemMedChem 2021, 16, 1–10
8
© 2021 The Authors. ChemMedChem published by Wiley-VCH GmbH
��
These are not the final page numbers!