New Chalcone–Triazole Hybrids with Promising Antimicrobial Activity in Multidrug Resistance Strains
Abstract
:1. Introduction
2. Results and Discussion
2.1. Synthesis and Structure Elucidation
2.2. Antimicrobial Activity and Potentiation of Antimicrobial Activity
2.3. Efflux Pump Inhibition Assay
2.4. Inhibition of Biofilm Formation and Quorum-Sensing
2.5. In Silico Prediction of Druglikeness and Toxicity
3. Materials and Methods
3.1. Chemistry
3.1.1. Synthesis and Structure Elucidation of Chalcones
Synthesis of Chalcones 7 and 8
Synthesis of Chalcones 9 and 10
3.2. Docking Studies
3.3. Biological Activity
3.3.1. Culture Media and Chemicals
3.3.2. Bacterial and Fungal Strains
3.3.3. Antibacterial Assay
3.3.4. Antifungal Assay
3.3.5. Combination with Antibiotics
3.3.6. Efflux Pump Inhibition Assays
3.3.7. Inhibition of Biofilm Formation
3.3.8. Quorum-Sensing Assay
3.3.9. Cytotoxicity in Mouse Embryonic Fibroblasts
4. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
- Tanwar, J.; Das, S.; Fatima, Z.; Hameed, S. Multidrug Resistance: An Emerging Crisis. Interdiscip. Perspect. Infect. Dis. 2014, 2014, 541340. [Google Scholar] [CrossRef] [Green Version]
- Blanco, P.; Hernando-Amado, S.; Reales-Calderon, J.A.; Corona, F.; Lira, F.; Alcalde-Rico, M.; Bernardini, A.; Sanchez, M.B.; Martinez, J.L. Bacterial Multidrug Efflux Pumps: Much More Than Antibiotic Resistance Determinants. Microorganisms 2016, 4, 14. [Google Scholar] [CrossRef] [Green Version]
- Alav, I.; Sutton, J.M.; Rahman, K.M. Role of bacterial efflux pumps in biofilm formation. J. Antimicrob. Chemother. 2018, 73, 2003–2020. [Google Scholar] [CrossRef] [Green Version]
- Durães, F.; Resende, D.I.S.P.; Palmeira, A.; Szemerédi, N.; Pinto, M.M.M.; Spengler, G.; Sousa, E. Xanthones Active against Multidrug Resistance and Virulence Mechanisms of Bacteria. Antibiotics 2021, 10, 600. [Google Scholar] [CrossRef]
- Durães, F.; Pinto, M.; Sousa, E. Medicinal Chemistry Updates on Bacterial Efflux Pump Modulators. Curr. Med. Chem. 2018, 25, 6030–6069. [Google Scholar] [CrossRef]
- Durães, F.; Cravo, S.; Freitas-Silva, J.; Szemerédi, N.; Martins-da-Costa, P.; Pinto, E.; Tiritan, M.E.; Spengler, G.; Fernandes, C.; Sousa, E.; et al. Enantioselectivity of Chiral Derivatives of Xanthones in Virulence Effects of Resistant Bacteria. Pharmaceuticals 2021, 14, 1141. [Google Scholar] [CrossRef]
- Durães, F.; Palmeira, A.; Cruz, B.; Freitas-Silva, J.; Szemerédi, N.; Gales, L.; da Costa, P.M.; Remião, F.; Silva, R.; Pinto, M.; et al. Antimicrobial Activity of a Library of Thioxanthones and Their Potential as Efflux Pump Inhibitors. Pharmaceuticals 2021, 14, 572. [Google Scholar] [CrossRef]
- Durães, F.; Szemerédi, N.; Kumla, D.; Pinto, M.; Kijjoa, A.; Spengler, G.; Sousa, E. Metabolites from Marine-Derived Fungi as Potential Antimicrobial Adjuvants. Mar. Drugs 2021, 19, 475. [Google Scholar] [CrossRef]
- Pinheiro, P.G.; Santiago, G.M.P.; da Silva, F.E.F.; de Araújo, A.C.J.; de Oliveira, C.R.T.; Freitas, P.R.; Rocha, J.E.; Neto, J.B.d.A.; da Silva, M.M.C.; Tintino, S.R.; et al. Ferulic acid derivatives inhibiting Staphylococcus aureus tetK and MsrA efflux pumps. Biotechnol. Rep. 2022, 34, e00717. [Google Scholar] [CrossRef]
- Samreen; Qais, F.A.; Ahmad, I. In silico screening and in vitro validation of phytocompounds as multidrug efflux pump inhibitor against E. coli. J. Biomol. Struct. Dyn. 2022, 1–13. [Google Scholar] [CrossRef]
- Shyam, M.; Verma, H.; Bhattacharje, G.; Mukherjee, P.; Singh, S.; Kamilya, S.; Jalani, P.; Das, S.; Dasgupta, A.; Mondal, A.; et al. Mycobactin Analogues with Excellent Pharmacokinetic Profile Demonstrate Potent Antitubercular Specific Activity and Exceptional Efflux Pump Inhibition. J. Med. Chem. 2022, 65, 234–256. [Google Scholar] [CrossRef] [PubMed]
- Cushnie, T.P.T.; Lamb, A.J. Antimicrobial activity of flavonoids. Int. J. Antimicrob. Agents 2005, 26, 343–356. [Google Scholar] [CrossRef]
- Gatadi, S.; Gour, J.; Nanduri, S. Natural product derived promising anti-MRSA drug leads: A review. Bioorg. Med. Chem. 2019, 27, 3760–3774. [Google Scholar] [CrossRef]
- Górniak, I.; Bartoszewski, R.; Króliczewski, J. Comprehensive review of antimicrobial activities of plant flavonoids. Phytochem. Rev. 2019, 18, 241–272. [Google Scholar] [CrossRef] [Green Version]
- Aparna, V.; Dineshkumar, K.; Mohanalakshmi, N.; Velmurugan, D.; Hopper, W. Identification of Natural Compound Inhibitors for Multidrug Efflux Pumps of Escherichia coli and Pseudomonas aeruginosa Using In Silico High-Throughput Virtual Screening and In Vitro Validation. PLoS ONE 2014, 9, e101840. [Google Scholar] [CrossRef] [Green Version]
- Bame, J.R.; Graf, T.N.; Junio, H.A.; Bussey, R.O.; Jarmusch, S.A.; El-Elimat, T.; Falkinham, J.O.; Oberlies, N.H.; Cech, R.A.; Cech, N.B. Sarothrin from Alkanna orientalis Is an Antimicrobial Agent and Efflux Pump Inhibitor. Planta Med. 2013, 79, 327–329. [Google Scholar] [CrossRef] [Green Version]
- Chan, B.C.L.; Ip, M.; Gong, H.; Lui, S.L.; See, R.H.; Jolivalt, C.; Fung, K.P.; Leung, P.C.; Reiner, N.E.; Lau, C.B.S. Synergistic effects of diosmetin with erythromycin against ABC transporter over-expressed methicillin-resistant Staphylococcus aureus (MRSA) RN4220/pUL5054 and inhibition of MRSA pyruvate kinase. Phytomedicine 2013, 20, 611–614. [Google Scholar] [CrossRef]
- Chan, B.C.L.; Ip, M.; Lau, C.B.S.; Lui, S.L.; Jolivalt, C.; Ganem-Elbaz, C.; Litaudon, M.; Reiner, N.E.; Gong, H.; See, R.H.; et al. Synergistic effects of baicalein with ciprofloxacin against NorA over-expressed methicillin-resistant Staphylococcus aureus (MRSA) and inhibition of MRSA pyruvate kinase. J. Ethnopharmacol. 2011, 137, 767–773. [Google Scholar] [CrossRef]
- Falcão-Silva, V.S.; Silva, D.A.; Souza, M.d.F.V.; Siqueira-Junior, J.P. Modulation of drug resistance in staphylococcus aureus by a kaempferol glycoside from herissantia tiubae (malvaceae). Phytother. Res. 2009, 23, 1367–1370. [Google Scholar] [CrossRef]
- Holler, J.G.; Christensen, S.B.; Slotved, H.-C.; Rasmussen, H.B.; Gúzman, A.; Olsen, C.-E.; Petersen, B.; Mølgaard, P. Novel inhibitory activity of the Staphylococcus aureus NorA efflux pump by a kaempferol rhamnoside isolated from Persea lingue Nees. J. Antimicrob. Chemother. 2012, 67, 1138–1144. [Google Scholar] [CrossRef]
- Maia, G.L.d.A.; Falcão-Silva, V.d.S.; Aquino, P.G.V.; Araújo-Júnior, J.X.d.; Tavares, J.F.; Silva, M.S.d.; Rodrigues, L.C.; Siqueira-Júnior, J.P.d.; Barbosa-Filho, J.M. Flavonoids from Praxelis clematidea R.M. King and Robinson Modulate Bacterial Drug Resistance. Molecules 2011, 16, 4828–4835. [Google Scholar] [CrossRef]
- Holler, J.G.; Slotved, H.-C.; Mølgaard, P.; Olsen, C.E.; Christensen, S.B. Chalcone inhibitors of the NorA efflux pump in Staphylococcus aureus whole cells and enriched everted membrane vesicles. Bioorg. Med. Chem. 2012, 20, 4514–4521. [Google Scholar] [CrossRef]
- Jesus, A.; Durães, F.; Szemerédi, N.; Freitas-Silva, J.; da Costa, P.M.; Pinto, E.; Pinto, M.; Spengler, G.; Sousa, E.; Cidade, H. BDDE-Inspired Chalcone Derivatives to Fight Bacterial and Fungal Infections. Mar. Drugs 2022, 20, 315. [Google Scholar] [CrossRef]
- Moreira, J.; Durães, F.; Freitas-Silva, J.; Szemerédi, N.; Resende, D.I.S.P.; Pinto, E.; da Costa, P.M.; Pinto, M.; Spengler, G.; Cidade, H.; et al. New diarylpentanoids and chalcones as potential antimicrobial adjuvants. Bioorg. Med. Chem. Lett. 2022, 67, 128743. [Google Scholar] [CrossRef]
- Lal, K.; Yadav, P.; Kumar, A.; Kumar, A.; Paul, A.K. Design, synthesis, characterization, antimicrobial evaluation and molecular modeling studies of some dehydroacetic acid-chalcone-1,2,3-triazole hybrids. Bioorg. Chem. 2018, 77, 236–244. [Google Scholar] [CrossRef]
- Yadav, P.; Lal, K.; Kumar, L.; Kumar, A.; Kumar, A.; Paul, A.K.; Kumar, R. Synthesis, crystal structure and antimicrobial potential of some fluorinated chalcone-1,2,3-triazole conjugates. Eur. J. Med. Chem. 2018, 155, 263–274. [Google Scholar] [CrossRef]
- Pereira, D.; Pinto, M.; Correia-da-Silva, M.; Cidade, H. Recent Advances in Bioactive Flavonoid Hybrids Linked by 1,2,3-Triazole Ring Obtained by Click Chemistry. Molecules 2022, 27, 230. [Google Scholar] [CrossRef]
- Pereira, D.; Gonçalves, C.; Martins, B.T.; Palmeira, A.; Vasconcelos, V.; Pinto, M.; Almeida, J.R.; Correia-da-Silva, M.; Cidade, H. Flavonoid Glycosides with a Triazole Moiety for Marine Antifouling Applications: Synthesis and Biological Activity Evaluation. Mar. Drugs 2021, 19, 5. [Google Scholar] [CrossRef]
- Tacconelli, E.; Carrara, E.; Savoldi, A.; Harbarth, S.; Mendelson, M.; Monnet, D.L.; Pulcini, C.; Kahlmeter, G.; Kluytmans, J.; Carmeli, Y.; et al. Discovery, research, and development of new antibiotics: The WHO priority list of antibiotic-resistant bacteria and tuberculosis. Lancet Infect. Dis. 2018, 18, 318–327. [Google Scholar] [CrossRef]
- Bessa, L.J.; Barbosa-Vasconcelos, A.; Mendes, Â.; Vaz-Pires, P.; Martins da Costa, P. High prevalence of multidrug-resistant Escherichia coli and Enterococcus spp. in river water, upstream and downstream of a wastewater treatment plant. J. Water Health 2014, 12, 426–435. [Google Scholar] [CrossRef]
- Shi, X.; Chen, M.; Yu, Z.; Bell, J.M.; Wang, H.; Forrester, I.; Villarreal, H.; Jakana, J.; Du, D.; Luisi, B.F.; et al. In situ structure and assembly of the multidrug efflux pump AcrAB-TolC. Nat. Commun. 2019, 10, 2635. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Aron, Z.; Opperman, T.J. The hydrophobic trap—The Achilles heel of RND efflux pumps. Res. Microbiol. 2018, 169, 393–400. [Google Scholar] [CrossRef] [PubMed]
- Parai, D.; Banerjee, M.; Dey, P.; Mukherjee, S.K. Reserpine attenuates biofilm formation and virulence of Staphylococcus aureus. Microb. Pathog. 2020, 138, 103790. [Google Scholar] [CrossRef]
- Zimmermann, S.; Klinger-Strobel, M.; Bohnert, J.A.; Wendler, S.; Rödel, J.; Pletz, M.W.; Löffler, B.; Tuchscherr, L. Clinically Approved Drugs Inhibit the Staphylococcus aureus Multidrug NorA Efflux Pump and Reduce Biofilm Formation. Front. Microbiol. 2019, 10, 2762. [Google Scholar] [CrossRef] [Green Version]
- Gajdács, M.; Spengler, G. Standard operating procedure (SOP) for disk diffusion-based quorum sensing inhibition assays. Acta Pharm. Hung. 2020, 89, 117–125. [Google Scholar] [CrossRef] [Green Version]
- Benomar, S.; Evans Kara, C.; Unckless Robert, L.; Chandler Josephine, R.; Parales Rebecca, E. Efflux Pumps in Chromobacterium Species Increase Antibiotic Resistance and Promote Survival in a Coculture Competition Model. Appl. Environ. Microbiol. 2019, 85, e00908–e00919. [Google Scholar] [CrossRef] [Green Version]
- Daina, A.; Michielin, O.; Zoete, V. SwissADME: A free web tool to evaluate pharmacokinetics, drug-likeness and medicinal chemistry friendliness of small molecules. Sci. Rep. 2017, 7, 42717. [Google Scholar] [CrossRef] [Green Version]
- Patonay, T.; Juhász-Tóth, É.; Bényei, A. Base-Induced Coupling of α-Azido Ketones with Aldehydes—An Easy and Efficient Route to Trifunctionalized Synthons 2-Azido-3-hydroxy Ketones, 2-Acylaziridines, and 2-Acylspiroaziridines. Eur. J. Org. Chem. 2002, 2002, 285–295. [Google Scholar] [CrossRef]
- Eicher, T.; Cha, H.-J.; Seeger, M.A.; Brandstätter, L.; El-Delik, J.; Bohnert, J.A.; Kern, W.V.; Verrey, F.; Grütter, M.G.; Diederichs, K.; et al. Transport of drugs by the multidrug transporter AcrB involves an access and a deep binding pocket that are separated by a switch-loop. Proc. Natl. Acad. Sci. USA 2012, 109, 5687. [Google Scholar] [CrossRef] [Green Version]
- Mikolosko, J.; Bobyk, K.; Zgurskaya, H.I.; Ghosh, P. Conformational Flexibility in the Multidrug Efflux System Protein AcrA. Structure 2006, 14, 577–587. [Google Scholar] [CrossRef]
- Koronakis, V.; Sharff, A.; Koronakis, E.; Luisi, B.; Hughes, C. Crystal structure of the bacterial membrane protein TolC central to multidrug efflux and protein export. Nature 2000, 405, 914–919. [Google Scholar] [CrossRef] [PubMed]
- Sussman, J.L.; Lin, D.; Jiang, J.; Manning, N.O.; Prilusky, J.; Ritter, O.; Abola, E.E. Protein Data Bank (PDB): Database of three-dimensional structural information of biological macromolecules. Acta Cryst. D Biol. Cryst. 1998, 54, 1078–1084. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Waterhouse, A.; Bertoni, M.; Bienert, S.; Studer, G.; Tauriello, G.; Gumienny, R.; Heer, F.T.; de Beer, T.A.P.; Rempfer, C.; Bordoli, L.; et al. SWISS-MODEL: Homology modelling of protein structures and complexes. Nucleic Acids Res. 2018, 46, W296–W303. [Google Scholar] [CrossRef] [PubMed] [Green Version]
- Apweiler, R.; Bairoch, A.; Wu, C.H.; Barker, W.C.; Boeckmann, B.; Ferro, S.; Gasteiger, E.; Huang, H.; Lopez, R.; Magrane, M.; et al. UniProt: The universal protein knowledgebase. Nucleic Acids Res. 2017, 45, D158–D169. [Google Scholar] [CrossRef] [PubMed]
- Zárate, S.G.; Morales, P.; Świderek, K.; Bolanos-Garcia, V.M.; Bastida, A. A Molecular Modeling Approach to Identify Novel Inhibitors of the Major Facilitator Superfamily of Efflux Pump Transporters. Antibiotics 2019, 8, 25. [Google Scholar] [CrossRef] [Green Version]
- Trott, O.; Olson, A.J. AutoDock Vina: Improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading. J. Comput. Chem. 2010, 31, 455–461. [Google Scholar] [CrossRef] [Green Version]
- Simões, R.R.; Aires-de-Sousa, M.; Conceição, T.; Antunes, F.; da Costa, P.M.; de Lencastre, H. High Prevalence of EMRSA-15 in Portuguese Public Buses: A Worrisome Finding. PLoS ONE 2011, 6, e17630. [Google Scholar] [CrossRef] [Green Version]
- CLSI. CLSI Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria that Grow Aerobically, 11th ed.; Clinical and Laboratory Standards Institute: Wayne, PA, USA, 2018. [Google Scholar]
- CLSI Document M27-A3; CLSI Clinical and Laboratory Standards Institute: Reference Method for Broth Dilution Antifungal Susceptibility Testing of Yeasts; Approved Standard. Clinical and Laboratory Standards Institute: Wayne, PA, USA, 2008; Volume 28.
- CLSI Document M38-A2; CLSI Reference Method for Broth Dilution Antifungal Susceptibility Testing of Filamentous Fungi; Approved Standard—Second Edition. Clinical and Laboratory Standards Institute: Wayne, PA, USA, 2008.
- Franklin, R.; Cockerill, M.D., III. Performance Standards for Antimicrobial Susceptibility Testing, Twenty-First Informational Supplement M100-S21; Clinical and Laboratory Standards Institute (CLSI): Wayne, PA, USA, 2011. [Google Scholar]
- Spengler, G.; Takács, D.; Horváth, A.; Szabó, A.M.; Riedl, Z.; Hajós, G.; Molnár, J.; Burián, K. Efflux pump inhibiting properties of racemic phenothiazine derivatives and their enantiomers on the bacterial AcrAB-TolC system. In Vivo 2014, 28, 1071–1075. [Google Scholar]
- Kincses, A.; Szabó, S.; Rácz, B.; Szemerédi, N.; Watanabe, G.; Saijo, R.; Sekiya, H.; Tamai, E.; Molnár, J.; Kawase, M.; et al. Benzoxazole-Based Metal Complexes to Reverse Multidrug Resistance in Bacteria. Antibiotics 2020, 9, 649. [Google Scholar] [CrossRef]
- Nové, M.; Kincses, A.; Szalontai, B.; Rácz, B.; Blair, J.M.A.; González-Prádena, A.; Benito-Lama, M.; Domínguez-Álvarez, E.; Spengler, G. Biofilm Eradication by Symmetrical Selenoesters for Food-Borne Pathogens. Microorganisms 2020, 8, 566. [Google Scholar] [CrossRef] [Green Version]
- Gajdács, M.; Spengler, G. The Role of Drug Repurposing in the Development of Novel Antimicrobial Drugs: Non-Antibiotic Pharmacological Agents as Quorum Sensing-Inhibitors. Antibiotics 2019, 8, 270. [Google Scholar] [CrossRef] [PubMed]
- Ferreira, R.J.; Kincses, A.; Gajdács, M.; Spengler, G.; dos Santos, D.J.V.A.; Molnár, J.; Ferreira, M.-J.U. Terpenoids from Euphorbia pedroi as Multidrug-Resistance Reversers. J. Nat. Prod. 2018, 81, 2032–2040. [Google Scholar] [CrossRef] [PubMed]
Compound | E. coli SA/2 | E. faecalis B3/101 |
---|---|---|
MIC CTX = 256 µg/mL (562 µM) | MIC VAN = 1024 µg/mL (707 µM) | |
MIC Reduction | ||
1 | None | 2-fold |
2 | None | 2-fold |
3 | None | None |
4 | 2-fold | None |
5 | 2-fold | 2-fold |
6 | None | None |
7 | None | 8-fold |
8 | None | None |
9 | None | 4-fold |
10 | None | None |
Compound | RFI ± SD | |
---|---|---|
S. aureus 272123 | S. Typhimurium SL1344 | |
1 | −0.06 ± 0.11 | 0.01 ± 0.02 |
2 | −0.04 ± 0.20 | 0.02 ± 0.03 |
3 | 0.06 ± 0.05 | 0.45 ± 0.02 |
4 | −0.09 ± 0.05 | 0.15 ± 0.05 |
5 | 0.20 ± 0.08 | 0.40 ± 0.06 |
6 | 0.33 ± 0.04 | 0.52 ± 0.07 |
7 | 0.06 ± 0.11 | 0.85 ± 0.02 |
8 | 0.05 ± 0.02 | 0.59 ± 0.12 |
9 | 0.10 ± 0.09 | −0.01 ± 0.01 |
10 | 0.17 ± 0.06 | −0.06 ± 0.03 |
Reserpine | 0.48 ± 0.04 | - |
CCCP | - | 0.40 ± 0.06 |
Compound | Biofilm Inhibition (%) ± SD | |
---|---|---|
S. aureus ATCC 25923 | S. aureus 272123 | |
1 | 0.21 ± 1.82 | 94.58 ± 1.15 |
2 | 41.10 ± 2.12 | 32.77 ± 3.65 |
3 | 0 | 85.76 ± 2.61 |
4 | 57.40 ± 1.64 | 61.23 ± 4.63 |
5 | 0 | 36.89 ± 1.37 |
6 | 1.29 ± 0.42 | 59.56 ± 2.51 |
7 | 0.55 ± 2.00 | 89.36 ± 0.56 |
8 | 0 | 72.33 ± 0.89 |
9 | 0 | 26.47 ± 0.29 |
10 | 0.08 ± 0.34 | 83.51 ± 1.02 |
Thioridazine | 97.07 ± 1.01 | ND |
Reserpine | 0.53 ± 1.05 | 81.11 ± 1.18 |
Structure | Site | Position | Dimension | ||||
---|---|---|---|---|---|---|---|
X | Y | Z | X | Y | Z | ||
AcrA (2F1M) | HH | 27.4205 | 14.1758 | 175.9638 | 15.9628 | 12.8742 | 17.6756 |
LD | 26.8634 | −2.5985 | 207.5824 | 15.9628 | 11.6319 | 27.9448 | |
AcrB (4DX5) | SBS | 24.3266 | −32.1670 | −7.0000 | 18.4129 | 26.7613 | 20.1435 |
HT | 20.8792 | 17.7378 | −7.0708 | 14.5855 | 17.7378 | 15.3042 | |
TolC (1EK9) | −7.8482 | 84.1409 | 63.4236 | 39.5596 | 29.8075 | 15.9794 | |
NorA | BCR | −4.3807 | −19.3774 | 20.8856 | 14.5855 | 17.2122 | 20.5459 |
CS | −9.2889 | −27.7277 | 42.4691 | 14.5855 | 17.2122 | 17.3139 |
Publisher’s Note: MDPI stays neutral with regard to jurisdictional claims in published maps and institutional affiliations. |
© 2022 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
Share and Cite
Pereira, D.; Durães, F.; Szemerédi, N.; Freitas-da-Silva, J.; Pinto, E.; Martins-da-Costa, P.; Pinto, M.; Correia-da-Silva, M.; Spengler, G.; Sousa, E.; et al. New Chalcone–Triazole Hybrids with Promising Antimicrobial Activity in Multidrug Resistance Strains. Int. J. Mol. Sci. 2022, 23, 14291. https://doi.org/10.3390/ijms232214291
Pereira D, Durães F, Szemerédi N, Freitas-da-Silva J, Pinto E, Martins-da-Costa P, Pinto M, Correia-da-Silva M, Spengler G, Sousa E, et al. New Chalcone–Triazole Hybrids with Promising Antimicrobial Activity in Multidrug Resistance Strains. International Journal of Molecular Sciences. 2022; 23(22):14291. https://doi.org/10.3390/ijms232214291
Chicago/Turabian StylePereira, Daniela, Fernando Durães, Nikoletta Szemerédi, Joana Freitas-da-Silva, Eugénia Pinto, Paulo Martins-da-Costa, Madalena Pinto, Marta Correia-da-Silva, Gabriella Spengler, Emília Sousa, and et al. 2022. "New Chalcone–Triazole Hybrids with Promising Antimicrobial Activity in Multidrug Resistance Strains" International Journal of Molecular Sciences 23, no. 22: 14291. https://doi.org/10.3390/ijms232214291