Abstract
Fungal infection is a significant global health challenge in part due to the emergence of strains exhibiting resistance to nearly all classes of antifungals. This underscores the urgent need for the development of new antifungal agents that can circumvent this burgeoning problem. For the present research, a new selected set of pyrazolo[5,1-c][1,2,4]triazine derivatives 3a–g was prepared in high yield via the reaction of N1-(5-methylpyrazol-3-yl)hydrazonoyl chloride 1 with morpholine, thiomorpholine, 4-phenylpiperidine and N-(substituted)piperazines. The new compounds were evaluated for their in vitro antifungal and antibacterial activities. The screening revealed compounds with specific activity against pathogenic fungi, including Candida albicans, Candida auris, and Cryptococcus. Compound 3d, which incorporated N-phenylpiperazine moiety, exhibited the highest growth inhibition against C. albicans with a minimum inhibitory concentration of 16 µg/mL. The compounds were superior to fluconazole in inhibiting Candida biofilm mass at sub-inhibitory concentration. Furthermore, the MTS assay confirmed that compounds 1 and 3d exhibited an excellent toxicity profile (not toxic, up to 256 μg/mL, for mammalian cells). Collectively, the presented results demonstrate that the synthesized pyrazolo-triazines warrant further exploration for potential use as antifungal agents.
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References
Abadleh MM, El-Abadelah MM, Sabri SS, Mohammed HH, Zihlif MA, Voelter W (2014) Synthesis and antitumor activity of some N2-(Thien-3-yl) amidrazones. Z Naturforsch B 69:811–816. https://doi.org/10.5560/znb.2014-4062
AbdelKhalek A, Ashby CR Jr, Patel BA, Talele TT, Seleem MN (2016) In vitro antibacterial activity of rhodanine derivatives against pathogenic clinical isolates. PLoS One 11:e0164227. https://doi.org/10.1371/journal.pone.0164227
AbdelKhalek A, Abutaleb NS, Mohammad H, Seleem MN (2019) Antibacterial and antivirulence activities of auranofin against Clostridium difficile. Int J Antimicrob Agents 53:54–62. https://doi.org/10.1016/j.ijantimicag.2018.09.018
Al-Adiwish WM, Tahir MIM, Siti-Noor-Adnalizawati A, Hashim SF, Ibrahim N, Yaacob WA (2013) Synthesis, antibacterial activity and cytotoxicity of new fused pyrazolo [1, 5-a] pyrimidine and pyrazolo [5, 1-c][1, 2, 4] triazine derivatives from new 5-aminopyrazoles. Eur J Med Chem 64:464–476. https://doi.org/10.1016/j.ejmech.2013.04.029
Baldwin JE (1976) Rules for ring closure. J Chem Soc Chem Commun 18:734–736. https://doi.org/10.1039/C39760000734
Baldwin JE, Thomas RC, Kruse LI, Silberman L (1977) Rules for ring closure: ring formation by conjugate addition of oxygen nucleophiles. J Org Chem 42:3846–3852. https://doi.org/10.1021/jo00444a011
Barrett GC, El-Abadelah MM, Hargreaves MK (1970) Cleavage of 2-acetyl-2-phenylazopropionanilide and related compounds by boron trifluoride. New Japp–Klingemann reactions. J Chem Soc C Organic 14:1986–1989. https://doi.org/10.1039/J39700001986
Benincori T, Sannicolo F (1988) New access to 2-(arylazo)-, 2-(arylhydrazo)-, and 2-aminoindoles, -benzofurans, and -thianaphthenes. J Org Chem 53:1309–1312. https://doi.org/10.1021/jo00241a035
Butler RN, Scott FL (1970) Versatile reactive intermediates-hydrazidic halides. Chem Ind 38:1216. https://doi.org/10.2174/1874847300902010008
Cao X, Sun Z, Cao Y, Wang R, Cai T, Chu W, Hu Y, Yang Y (2014) Design, synthesis, and structure–activity relationship studies of novel fused heterocycles-linked triazoles with good activity and water solubility. J Med Chem 57:3687–3706. https://doi.org/10.1021/jm4016284
Ciciani G, Coronnello M, Guerrini G, Selleri S, Cantore M, Failli P, Mini E, Costanzo A (2008) Synthesis of new pyrazolo [5, 1-c][1, 2, 4] benzotriazines, pyrazolo [5, 1-c] pyrido [4, 3-e][1, 2, 4] triazines and their open analogues as cytotoxic agents in normoxic and hypoxic conditions. Bioorg Med Chem 16:9409–9419. https://doi.org/10.1016/j.bmc.2008.09.055
CLSI (2008) Reference method for broth dilution antifungal susceptibility testing of yeasts; approved standard-third edition. CLSI document M27-A3. Clinical and Laboratory Standards Institute, Wayne, p 28
Coronnello M, Ciciani G, Mini E, Guerrini G, Caciagli B, Selleri S, Costanzo A, Mazzei T (2005) Cytotoxic activity of 3-nitropyrazolo [5, 1-c][1, 2, 4] benzotriazine derivatives: a new series of anti-proliferative agents. Anticancer Drugs 16:645–651 PMID: 15930893
Costanzo A, Ciciani G, Guerrini G, Bruni F, Selleri S, Donato R, Sacco C, Musiu C, Milia C, Longu S, Minnei C (1999) Antiproliferative and antimicrobial activities of pyrazolo[5,1- c][1,2,4]benzotriazine 5-oxides and their 5-deoxyderivatives. Med Chem Res 9:223–238
Dalla Croce P, Del Buttero P, Licandro E, Maiorana S (1979) Synthesis of arylazomethylenetriphenylphosphoranes from arylhydrazonoyl chlorides (via nitrilimines in situ) and triphenylphosphine. Synthesis 04:299–300. https://doi.org/10.1055/s-1979-28658
Debruyne D (1997) Clinical pharmacokinetics of fluconazole in superficial and systemic mycoses. Clin pharmacokinetics 33:52–77. https://doi.org/10.2165/00003088-199733010-00005
Del Pozo JL, Patel R (2007) The challenge of treating biofilm-associated bacterial infections. Clin Pharmacol Ther 82:204–209. https://doi.org/10.1038/sj.clpt.6100247
Denning DW (2003) Echinocandin antifungal drugs. Lancet 362:1142–1151
Douglas LJ (2002) Medical importance of biofilms in Candida infections. Rev Iberoam Micol 19:139–143. https://doi.org/10.1016/S0140-6736(03)14472-8
Eldesouky HE, Li X, Abutaleb NS, Mohammad H, Seleem MN (2018a) Synergistic interactions of sulfamethoxazole and azole antifungal drugs against emerging multidrug-resistant Candida auris. Int J Antimicrob Agents 52:754–761. https://doi.org/10.1016/j.ijantimicag.2018.08.016
Eldesouky HE, Mayhoub A, Hazbun TR, Seleem MN (2018b) Reversal of azole resistance in Candida albicans by sulfa antibacterial drugs. Antimicrob Agents Chemother 62:e00701–e00717. https://doi.org/10.1128/AAC.00701-17
Elnagdi MH, Zayed EM, Khalifa ME, Ghozlan SA (1981) Reactions with heterocyclic amidines, VII: synthesis of some new pyrazolo [1, 5-c]-1, 2, 4-triazines, pyrazolo [1, 5-a]-1, 3, 5-triazines and pyrazolo [1, 5-a] pyrimidines. Monatsh Chem/Chemical Monthly 112:245–252
Galishev VA, Chistokletov VN, Petrov AA (1975) 1, 3-Dipolar addition to unsaturated-compounds. 34. Reactions of diphenylmethyleneaminodiphenyl-phosphine with nitrilimines. Russ J Gen Chem 45:1695–1697
Groll AH, Lumb J (2012) New developments in invasive fungal disease. Future Microbiol 7(2):179–184. https://doi.org/10.2217/fmb.11.154
Hassaneen HM, Mousa HA, Abed NM, Shawali AS (1988) Chemistry of C-heteroarylhydrazidoyl halides. Synthesis and reactions of N-(p-Nitrophenyl)-C-(2-thienyl)-formohydrazidoyl halides. Heterocycles 27:695–706. https://doi.org/10.3987/COM-87-4381
Heeres J, Meerpoel L, Lewi P (2010) Conazoles. Molecules 15:4129–4188. https://doi.org/10.3390/molecules15064129
Hegarty AF, Aylward JB, Scott FL (1967) Synthesis and rearrangement of hydrazidic azides. J Chem Soc C Organic. https://doi.org/10.3390/molecules15064129
Heubach G (1980) Synthese neuer 2, 5-dihydro-1, 2, 3, 5-thiatriazol-1-oxide und 3, 4-dihydro-2H-1, 2, 4, 3-triazaphosphol-3-oxide. Liebigs Ann Chem 1980:1376–1383. https://doi.org/10.1002/jhet.5570170501
Jain A, Jain S, Rawat S (2010) Emerging fungal infections among children: a review on its clinical manifestations, diagnosis, and prevention. J Pharm Bioallied Sci 2(4):314–320. https://doi.org/10.4103/0975-7406.72131
Kabir MA, Ahmad Z (2012) Candida infections and their prevention. ISRN preventive medicine, 2013. https://doi.org/10.5402/2013/763628
Kanafani ZA, Perfect JR (2008) Resistance to antifungal agents: mechanisms and clinical impact. Clin Infect Dis 46:120–128. https://doi.org/10.5402/2013/763628
Kojic EM, Darouiche RO (2004) Candida infections of medical devices. Clin Microbiol Rev 17:255–267. https://doi.org/10.1128/CMR.17.2.255-267.2004
Kotb A, Abutaleb NS, Seleem MA, Hagras M, Mohammad H, Bayoumi A, Ghiaty A, Seleem MN, Mayhoub AS (2018) Phenylthiazoles with tert-Butyl side chain: metabolically stable with anti-biofilm activity. Eur J Med Chem 151:110–120. https://doi.org/10.1016/j.ejmech.2018.03.044
Larkin E, Hager C, Chandra J, Mukherjee PK, Retuerto M, Salem I, Long L, Isham N, Kovanda L, Borroto-Esoda K, Wring S (2017) The emerging pathogen Candida auris: growth phenotype, virulence factors, activity of antifungals, and effect of SCY-078, a novel glucan synthesis inhibitor, on growth morphology and biofilm formation. Antimicrob Agents Chemother 61:e02396–e02416. https://doi.org/10.1128/AAC.02396-16
Lebeaux D, Ghigo JM, Beloin C (2014) Biofilm-related infections: bridging the gap between clinical management and fundamental aspects of recalcitrance toward antibiotics. Microbiol Mol Biol Rev 78:510–543. https://doi.org/10.1128/MMBR.00013-14
Lee WG, Shin JH, Uh Y, Kang MG, Kim SH, Park KH, Jang HC (2011) First three reported cases of nosocomial fungemia caused by Candida auris. J Clin Microbiol 49:3139–3142. https://doi.org/10.1128/JCM.00319-11
Magill SS, Swoboda SM, Shields CE, Colantuoni EA, Fothergill AW, Merz WG, Lipsett PA, Hendrix CW (2009) The epidemiology of Candida colonization and invasive candidiasis in a surgical intensive care unit where fluconazole prophylaxis is utilized: follow-up to a randomized clinical trial. Ann Surg 249:657–665. https://doi.org/10.1097/SLA.0b013e31819ed914
Marriott DJ, Playford EG, Chen S, Slavin M, Nguyen Q, Ellis D, Sorrell TC (2009) Determinants of mortality in non-neutropenic ICU patients with candidaemia. Crit Care 13:R115. https://doi.org/10.1186/cc7964
Mateus C, Crow SA, Ahearn DG (2004) Adherence of Candida albicans to silicone induces immediate enhanced tolerance to fluconazole. Antimicrob Agents Chemother 48:3358–3366. https://doi.org/10.1128/AAC.48.9.3358-3366.2004
Mohamed MF, Hammac GK, Guptill L, Seleem MN (2014) Antibacterial activity of novel cationic peptides against clinical isolates of multi-drug resistant Staphylococcus pseudintermedius from infected dogs. PLoS One 9:e116259. https://doi.org/10.1371/journal.pone.0116259
Mohamed MF, Brezden A, Mohammad V, Chmielewski V, Seleem MN (2017) Targeting biofilms and persisters of ESKAPE pathogens with P14KanS, a kanamycin peptide conjugate. Biochimica et Biophysica Acta (BBA)-General Subjects 1861:848–859. https://doi.org/10.1016/j.bbagen.2017.01.029
Mohammad H, Mayhoub AS, Ghafoor A, Soofi M, Alajlouni RA, Cushman M, Seleem MN (2014) Discovery and characterization of potent thiazoles versus methicillin-and vancomycin-resistant Staphylococcus aureus. J Med Chem 57:1609–1615. https://doi.org/10.1021/jm401905m
Mohammad H, Cushman M, Seleem MN (2015a) Antibacterial evaluation of synthetic thiazole compounds in vitro and in vivo in a methicillin-resistant Staphylococcus aureus (MRSA) skin infection mouse model. PLoS One 10:e0142321. https://doi.org/10.1371/journal.pone.0142321
Mohammad H, Mayhoub AS, Cushman M, Seleem MN (2015b) Anti-biofilm activity and synergism of novel thiazole compounds with glycopeptide antibiotics against multidrug-resistant Staphylococci. J Antibiot 68:259–266. https://doi.org/10.1038/ja.2014.142
Mohammad H, Elghazawy NH, Eldesouky HE, Hegazy YA, Younis W, Avrimova L, Hazbun T, Arafa RK, Seleem MN (2018) Discovery of a novel dibromoquinoline compound exhibiting potent antifungal and antivirulence activity that targets metal ion homeostasis. ACS Infect Dis 4:403–414. https://doi.org/10.1021/acsinfecdis.7b00215
Moudgal V, Sobel J (2010) Antifungals to treat Candida albicans. Expert Opin Pharmacother 11:2037–2048. https://doi.org/10.1517/14656566.20
Mukherjee PK, Chandra J (2004) Candida biofilm resistance. Drug Resist Updat 7:301–309. https://doi.org/10.1016/j.drup.2004.09.002
Mukherjee PK, Chandra J, Kuhn DM, Ghannoum MA (2003) Mechanism of fluconazole resistance in Candida albicans biofilms: phase-specific role of efflux pumps and membrane sterols. Infect Immun 71:4333–4340. https://doi.org/10.1128/IAI.71.8.4333-4340.2003
Neplyuev VM, Bazavova IM, Lozinskii MO (1989) Japp-Klingemann reaction-nontraditional objects and leaving groups. Russ J Org Chem 25:2225–2236
Pappas PG, Kauffman CA, Andes DR, Clancy CJ, Marr KA, Ostrosky-Zeichner L, Walsh TJ, Zaoutis TE (2015) Clinical practice guideline for the management of candidiasis: 2016 update by the Infectious Diseases Society of America. Clin Infect Dis 62:e1–e50. https://doi.org/10.1093/cid/civ933
Pfaller MA, Pappas PG, Wingard JR (2006) Invasive fungal pathogens: current epidemiological trends. Clin Infect Dis 43(Supplement_1):S3–S14. https://doi.org/10.1086/504490
Ramage G, Saville SP, Wickes BL, López-Ribot JL (2002) Inhibition of Candida albicans biofilm formation by farnesol, a quorum-sensing molecule. Appl Environ Microbiol 68:5459–5463. https://doi.org/10.1128/AEM.68.11.5459-5463.2002
Seneviratne CJ, Wang Y, Jin L, Abiko Y, Samaranayake LP (2008) Candida albicans biofilm formation is associated with increased anti-oxidative capacities. Proteomics 8:2936–2947. https://doi.org/10.1002/pmic.200701097
Shawali AS, Párkanyi C (1980) Hydrazidoyl halides in the synthesis of heterocycles. J Heterocycl Chem 17:833–854. https://doi.org/10.1002/jhet.5570170501
Sheng C, Zhang W, Ji H, Zhang M, Song Y, Xu H, Zhu J, Miao Z, Jiang Q, Yao J, Zhou Y (2006) Structure-based optimization of azole antifungal agents by CoMFA, CoMSIA, and molecular docking. J Med Chem 49:2512–2525. https://doi.org/10.1021/jm051211n
Sorbera LA, Aravamudan J, Rosa E (2011) Therapeutic targets for candidiasis. Drugs Fut 36:627–630
Thangamani S, Mohammad H, Abushahba MF, Sobreira TJ, Seleem MN (2016) Repurposing auranofin for the treatment of cutaneous staphylococcal infections. Int J Antimicrob Agents 47:195–201. https://doi.org/10.1016/j.ijantimicag.2015.12.016
Thangamani S, Eldesouky HE, Mohammad H, Pascuzzi PE, Avramova L, Hazbun TR, Seleem MN (2017a) Ebselen exerts antifungal activity by regulating glutathione (GSH) and reactive oxygen species (ROS) production in fungal cells. Biochim Biophys Acta 1861:3002–3010. https://doi.org/10.1358/dof.2011.36.8.1686467
Thangamani S, Maland M, Mohammad H, Pascuzzi PE, Avramova L, Koehler CM, Hazbun TR, Seleem MN (2017b) Repurposing approach identifies auranofin with broad spectrum antifungal activity that targets Mia40-Erv1 pathway. Front Cell Infect Microbiol 7:4. https://doi.org/10.3389/fcimb.2017.00004
Tobudic S, Kratzer C, Lassnigg A, Presterl E (2012a) Antifungal susceptibility of Candida albicans in biofilms. Mycoses 55:199–204. https://doi.org/10.1111/j.1439-0507.2011.02076.x
Tobudic S, Kratzer C, Presterl E (2012b) Azole-resistant Candida spp.–emerging pathogens? Mycoses 55:24–32. https://doi.org/10.1111/j.1439-0507.2011.02146.x
Ulrich H (1968) Chemistry of imidoyl halides. Springer Science & Business Media, New York
Yao HC, Resnick P (1962) Azo-hydrazone conversion. I. The Japp–Klingemann reaction. J Am Chem Soc 84:3514–3517. https://doi.org/10.1021/ja00877a018
Acknowledgements
We wish to thank the Deanship of Scientific Research at Mutah University, Al-Karak, Jordan for financial support. Also, the authors would like to thank Biodefense and Emerging Infections Research Resources Repository (BEI Resources) and the American Type Culture Collection (ATCC) for providing clinical isolates used in this study.
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Al-Trawneh, S.A., Al-Dawdieh, S.A., Abutaleb, N.S. et al. Synthesis of new pyrazolo[5,1-c][1,2,4]triazines with antifungal and antibiofilm activities. Chem. Pap. 74, 1241–1252 (2020). https://doi.org/10.1007/s11696-019-00974-9
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DOI: https://doi.org/10.1007/s11696-019-00974-9