Beilstein J. Org. Chem. 2013, 9, 1501–1507.
4. Vandeputte, P.; Ferrari, S.; Coste, A. T. Int. J. Microbiol. 2012,
efforts to determine the efficacy of ML212 against diverse
mechanisms of fluconazole resistance, including biofilm forma-
tion, drug-target mutations, and efflux-pump amplification.
5. Pfaller, M. A.; Diekema, D. J. Clin. Microbiol. Rev. 2007, 20, 133–163.
6. Sanglard, D.; Odds, F. C. Lancet Infect. Dis. 2002, 2, 73–85.
Conclusion
7. Lavigne, J.-P.; Brunel, J.-M.; Chevalier, J.; Pagès, J.-M.
8. Gallo, S.; Chevalier, J.; Mahamoud, A.; Eyraud, A.; Pagès, J.-M.;
Barbe, J. Int. J. Antimicrob. Agents 2003, 22, 270–273.
High-throughput screening of 300,000 compounds from the
NIH’s MLSMR collection identified several substances that
potentiate the effect of fluconazole in fluconazole-resistant
Candida albicans clinical isolates. Among the numerous hits,
3-phenylindazole 1 was selected for chemical optimization,
resulting in the identification of 3-(3-anisoyl)indazole 36 as new
small-molecule probe (ML212) to facilitate investigation of the
various mechanisms used by C. albicans to withstand flucona-
zole. Elucidation of ML212’s mechanism of action may afford
new targets to exploit in the continuing efforts to develop novel
antimycotics and combat increasingly prevalent drug-resistance.
Samples of ML212 are available free of charge, on request.
9. Kim, J.; Campbell, B.; Mahoney, N.; Chan, K.; Molyneux, R.; May, G.
Biochem. Biophys. Res. Commun. 2008, 372, 266–271.
10.Cernicka, J.; Kozovska, Z.; Hnatova, M.; Valachovic, M.; Hapala, I.;
Riedl, Z.; Hajós, G.; Subik, J. Int. J. Antimicrob. Agents 2007, 29,
11.The complete results of the HTS assay can be viewed online free of
charge on PubChem.
12.Redding, S.; Smith, J.; Farinacci, G.; Rinaldi, M.; Fothergill, A.;
Rhine-Chalberg, J.; Pfaller, M. Clin. Infect. Dis. 1994, 18, 240–242.
Supporting Information
13.DiGirolamo, J. A.; Li, X.-C.; Jacob, M. R.; Clark, A. M.; Ferreira, D.
14.Gamarra, S.; Rocha, E. M. F.; Zhang, Y.-Q.; Park, S.; Rao, R.;
Perlin, D. S. Antimicrob. Agents Chemother. 2010, 54, 1753–1761.
Detailed experimental protocols for cellular assays and for
the preparation of representative compounds 25 and 36 are
provided. Proton NMR spectra for all prepared compounds
are also available.
15.Guo, X.-L.; Leng, P.; Yang, Y.; Yu, L.-G.; Lou, H.-X. J. Appl. Microbiol.
16.Mai, A.; Rotili, D.; Massa, S.; Brosch, G.; Simonetti, G.; Passariello, C.;
Palamara, A. T. Bioorg. Med. Chem. Lett. 2007, 17, 1221–1225.
Supporting Information File 1
Detailed assay protocols and compound synthesis.
17.Additional information about the MLSMR can be found online at
Supporting Information File 2
18.The Present Communication details a portion of the work previously
described in official Probe Report submitted to the NIH upon project
completion. The official Probe Report has been made available online
by the NIH, free of charge: Hartland, C. L.; Youngsaye, W.; Morgan, B.;
Ting, A.; Nag, P.; Burhlage, S.; Johnston, S.; Bittker, J.; Vincent, B.;
Whitesell, L.; Dandapani, S.; MacPherson, L.; Munoz, B.; Palmer, M.;
Lindquist, S.; Schreiber, S. L. “Identification of small molecules that
selectively inhibit fluconazole-resistant Candida albicans in the
presence of fluconazole but not in its absence - Probe 2”. In: Probe
Reports from the NIH Molecular Libraries Program [Internet]. Bethesda
(MD): National Center for Biotechnology Information (US); 2010-.
19.Youngsaye, W.; Vincent, B.; Hartland, C. L.; Morgan, B. J.;
Buhrlage, S. J.; Johnston, S.; Bittker, J. A.; MacPherson, L.;
Dandapani, S.; Palmer, M.; Whitesell, L.; Lindquist, S.; Schreiber, S. L.;
Munoz, B. Bioorg. Med. Chem. Lett. 2011, 21, 5502–5505.
NMR spectra of reported compounds.
Acknowledgements
This work was funded by the NIH’s MLPCN program (1 U54
HG005032-1 awarded to S.L.S.). BV, LW, and SL are grateful
for funding from the NIH (1 R03 MH086456-01). The authors
are grateful to Dr. Spencer Redding (University of Texas Health
Science Center San Antonio) for graciously providing samples
of C. albicans CaCi-2 and CaCi-8.
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