Abstract
Concerns on increasing trends of multidrug resistance (MDR) around the world have triggered the need to investigate and develop new therapeutic strategies and potent antibacterial drugs. Polymyxins, a class of polycationic antimicrobial peptides, have been regarded as the last-line therapy against Gram-negative bacteria due to limited new antibiotics and phenylalanine-arginine β-naphthylamide (PAβN), a peptidomimetic compound has been characterised as an efflux pump inhibitor (EPI) that have been investigated to overcome efflux-mediated multidrug resistance. In this work, the antibacterial activity of two Schiff base ligands derived from the condensation of S-benzyl dithiocarbazate with 4-carboxybenzaldehye (SB4CB) and 4-formyl-3-hydroxybenzoic acid (SBFH) their copper(II) complexes (Cu(SB4CB)2 and Cu(SBFH)2) were tested individually, and the most promising compound was tested in combination with polymyxin B (POLY) and PAβN against different bacteria, such as antibiotic-susceptible strains: Acinetobacter baumannii ATCC 19606, Escherichia coli ATCC 25922, Pseudomonas aeruginosa ATCC 27853, and Staphylococcus aureus ATCC 35923; multidrug-resistant strains: A. baumannii ATCC BAA-1797, E. coli BAA-196, P. aeruginosa ATCC BAA-2108 and S. aureus ATCC 43300. Initial minimum inhibition concentration (MIC) results showed the Cu(II) complexes Cu(SB4CB)2 and Cu(SBFH)2, demonstrated obvious antibacterial activity as compared to the ligand alone. Fractional inhibitory concentration (FIC) index showed improved MIC values with additivity and synergistic effect for Cu(SBFH)2 in combination with POLY and PAβN. From the in silico molecular docking investigation, Cu(SBFH)2 was shown to engage in hydrophobic interactions via its phenyl rings with surrounding hydrophobic residues in the binding pocket of S. aureus NorA, E. coli AcrB, P. aeruginosa MexB and A. baumannii AdeB efflux pumps. The phenyl rings of the ligand could also form π-π stacking with adjacent residues in the binding site of A. baumannii AdeB. Besides, hydrogen bonding and π-cation interactions were also observed via the carboxyl group, hydroxyl group and phenyl ring of SBFH moiety, respectively with nearby residues in the E. coli AcrB binding pocket. This study indicates that the combination strategy of Cu(SBFH)2 with POLY and PAβN enhances therapeutic potential and sheds light on the binding pockets inside bacteria efflux pumps and the binding interactions of ligand in the binding site.
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References
Adly OMI, El-shafiy HF (2019) New metal complexes derived from S-benzyldithiocarbazate (SBDTC) and chromone-3-carboxaldehyde: synthesis, characterization, antimicrobial, antitumor activity and DFT calculations. J Coord Chem 72(2):1–16. https://doi.org/10.1080/00958972.2018.1564912
Akbar Ali M, Huq mirza A, Ai Fong G (2004) Synthesis, characterization and X-ray crystal structures of the bis-ligand zinc(II) and cadmium(II) complexes of the methylpyruvate Schiff base of S-methyldithiocarbazate. Transit Met Chem 29:613–619. https://doi.org/10.1007/s11243-004-2511-7
Alav I, Sutton JM, Rahman KM (2018) Role of bacterial efflux pumps in biofilm formation. J Antimicrob Chemother 73(8):2003–2020. https://doi.org/10.1093/jac/dky042
Aparna V, Dineshkumar K, Mohanalakshmi N, Velmurugan D, Hopper W (2014) 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 9(7):e101840. https://doi.org/10.1371/journal.pone.0101840
Auda IG, Ali Salman IM, Odah JG (2020) Efflux pumps of Gram-negative bacteria in brief. Gene Reports 20:100666. https://doi.org/10.1016/j.genrep.2020.100666
Bolla JM, Alibert Franco S, Handzlink J (2011) Strategies for bypassing the membrane barrier in multidrug resistant gram-negative bacteria. Febsletters 585(11):1682–2690. https://doi.org/10.1016/j.febslet.2011.04.054
Breijyeh Z, Jubeh B, Karaman R (2020) Resistance of Gram-negative bacteria to current antibacterial agents and approaches to resolve it. Molecules 25(6):1340. https://doi.org/10.3390/molecules25061340
Brown OC, Torres JB, Holt KB, Blower PJ, Went MJ (2017) Copper complexes with dissymmetrically substituted bis(thiosemicarbazone) ligands as a basis for PET radiopharmaceuticals: control of redox potential and lipophilicity. Dalton Trans 46(42):14612–14630. https://doi.org/10.1039/C7DT02008B
Charkhi P, Haghshenas MR, Mirzaei B, Davoodi L, Bazgir ZN, Goli HR (2020) Comparison of the effect of phenylalanine arginine beta naphthylamide (PAΒN) and curcumin on minimum inhibitory concentration of aminoglycosides on Pseudomonas aeruginosa clinical isolates. J Adv Med Biomed Res 28(127):105–110. https://doi.org/10.30699/jambs.28.127.105
Choo XY, Liddell JR, Huuskonen MT, Grubman A, Moujalled D, Roberts J, Kysenius K, Patten L, Quek H, Oikari LE, Duncan C, James SA, McInnes LE, Hayne DJ, Donnelly PS, Pollari E, Vähätalo S, Lejavová K, Kettunen MI, Malm T, Koistinaho J, White AR, Kanninen KM (2018) CuII (atsm) attenuates neuroinflammation. Front Neurosci 12:668. https://doi.org/10.3389/fnins.2018.00668
Chung PY, Khoo REY, Liew HS, Low ML (2021) Antimicrobial and antibiofilm activities of Cu (II) Schiff base complexes against methicillin-susceptible and resistant Staphylococcus aureus. Ann Clin Microbiol Antimicrob 20(1):67. https://doi.org/10.1186/s12941-021-00473-4
Claudel M, Schwarte JV, Fromm KM (2020) New antimicrobial strategies based on metal complexes. Chemistry 2(4):849–899. https://doi.org/10.3390/chemistry2040056
Crouse KA, Chew K-B, Tarafder MT, Kasbollah A, Ali A, Yamin B, Fun HK (2004) Synthesis, characterization and bio-activity of S-2-picolyldithiocarbazate (S2PDTC), some of its schiff bases and their Ni(II) complexes and X-ray structure of S-2-picolyl-β-N-(2-acetylpyrrole)dithiocarbazate. Polyhedron 23(1):161–168. https://doi.org/10.1016/J.POLY.2003.09.025
Dai C, Wan Y, Sharma G, Shen J, Velkov T, Xiao X (2020) Polymyxins–curcumin combination antimicrobial therapy: safety implications and efficacy for infection treatment. Antioxidants 9(6):506. https://doi.org/10.3390/antiox9060506
Damit NSHH, Hamid MHSA, Rahman NSRHA, Ilias SNHH, Keasberry NA (2021) Synthesis, structural characterisation and antibacterial activities of lead(II) and some transition metal complexes derived from quinoline-2-carboxaldehyde 4-methyl-3-thiosemicarbazone. Inorg Chim Acta 527(14):120557. https://doi.org/10.1016/j.ica.2021.120557
Deng J, Yu P, Zhang Z, Wang J, Cai J, Wu N, Sun H, Liang H, Yang F (2018) Designing anticancer copper (II) complexes by optimizing 2-pyridine-thiosemicarbazone ligands. Eur J Med Chem 158:442–452. https://doi.org/10.1016/j.ejmech.2018.09.020
Dhahagani K, Kesavan MP, Kumar GGV, Ravi L, Rajagopal G, Rajesh J (2018) Crystal structure, optical properties, DFT analysis of new morpholine based Schiff base ligands and their copper(II) complexes: DNA, protein docking analyses, antibacterial study and anticancer evaluation. Mater Sci Eng C 90:119–130. https://doi.org/10.1016/j.msec.2018.04.032
Djoko KY, Goytia MM, Donnelly PS, Schembri MA, Shafer WM, McEwan AG (2015) Copper(II)-bis(thiosemicarbazonato) complexes as antibacterial agents: insights into their mode of action and potential as therapeutics. Antimicrob Agents Chemother 59(10):6444–6453. https://doi.org/10.1128/aac.01289-15
Fazly Bazzaz BS, Seyedi S, Hoseini Goki N, Khameneh B (2021) Human antimicrobial peptides: spectrum, mode of action and resistance mechanisms. Int J Pept Res Ther 27:801–816. https://doi.org/10.1007/s10989-020-10127-2
Ferrer-Espada R, Shahrour H, Pitts B, Stewart PS, Sánchez-Gómez S, Martínez-de-Tejada G (2019) A permeability-increasing drug synergizes with bacterial efflux pump inhibitors and restores susceptibility to antibiotics in multi-drug resistant Pseudomonas aeruginosa strains. Sci Rep. https://doi.org/10.1038/s41598-019-39659-4
Figueiredo J, Serrano JL, Soares M, Ferreira S, Domingues FC, Almeida P, Silvestre S (2019) 5-Hydrazinylethylidenepyrimidines effective against multidrug-resistant Acinetobacter baumannii: synthesis and in vitro biological evaluation of antibacterial, radical scavenging and cytotoxic activities. Eur J Pharm Sci 137:104964. https://doi.org/10.1016/j.ejps.2019.104964
Foo JB, Ng LS, Lim JH, Tan PX, Lor YZ, Loo JSE, Low ML, Chan LC, Beh CY, Leong SW, Yazan LS, Tor YS, How CW (2019) Induction of cell cycle arrest and apoptosis by copper complex Cu (SBCM) 2 towards oestrogen-receptor positive MCF-7 breast cancer cells. RSC Adv 9(32):18359–18370. https://doi.org/10.1039/C9RA03130H
Glišić BĐ, Aleksic I, Comba P, Wadepohl H, Ilic-Tomic T, Nikodinovic-Runic J, Djuran MI (2016) Copper (II) complexes with aromatic nitrogen-containing heterocycles as effective inhibitors of quorum sensing activity in Pseudomonas aeruginosa. RSC Adv 6(89):86695–86709. https://doi.org/10.1039/C6RA19902J
Grace JL, Huang JX, Cheah SE, Truong NP, Cooper MA, Li J, Davis TP, Quinn JF, Velkov T, Whittaker MR (2016) Antibacterial low molecular weight cationic polymers: dissecting the contribution of hydrophobicity, chain length and charge to activity. RSC Adv 6(19):15469–15477. https://doi.org/10.1039/C5RA24361K
Guelfi KC, Tognim MCB, Cardoso CL, Gales AC, Carrara-Marrone FE, Garcia LB (2008) In vitro evaluation of the antimicrobial activity of meropenem in combination with polymyxin B and gatifloxacin against Pseudomonas aeruginosa and Acinetobacter baumannii. J Chemother 20(2):180–185. https://doi.org/10.1179/joc.2008.20.2.180
Hejchman E, Kruszewska H, Maciejewska D, Sowirka-Taciak B, Tomczyk M, Sztokfisz-Ignasiak A, Jankowski J, Młynarczuk-Biały I (2019) Design, synthesis, and biological activity of Schiff bases bearing salicyl and 7-hydroxycoumarinyl moieties. Monatsh Chem 150(17):255–266. https://doi.org/10.1007/s00706-018-2325-5
How FN-F, Crouse KA, Tahir MIM, Tarafder MTH, Cowley AR (2008) Synthesis, characterization and biological studies of S-benzyl-β-N-(benzoyl) dithiocarbazate and its metal complexes. Polyhedron 27(15):3325–3329. https://doi.org/10.1016/j.poly.2008.07.022
Hsieh MH, Yu CM, Yu VL, Chow JW (1993) Synergy assessed by checkerboard a critical analysis. Diagn Microbiol Infect Dis 16(4):343–349. https://doi.org/10.1016/0732-8893(93)90087-N
Jamshidi S, Sutton JM, Rahman KM (2017) Computational study reveals the molecular mechanism of the interaction between the efflux inhibitor PAβN and the AdeB transporter from Acinetobacter baumannii. ACS Omega 2(6):3002–3016. https://doi.org/10.1021/acsomega.7b00131
Kremer E, Facchin G, Estévez E, Alborés P, Baran EJ, Ellena J, Torre MH (2006) Copper complexes with heterocyclic sulfonamides: synthesis, spectroscopic characterization, microbiological and SOD-like activities: crystal structure of [Cu(sulfisoxazole)2(H2O)4]·2H2O. J Inorg Biochem 100(7):1167–1175. https://doi.org/10.1016/j.jinorgbio.2006.01.042
Kuete V, Ngameni B, Tangmouo JG, Bolla JM, Alibert-Franco S, Ngadjui BT, Pagès JM (2010) Efflux pumps are involved in the defense of Gram-negative bacteria against the natural products isobavachalcone and diospyrone. Antimicrob Agents Chemother 54(5):1749–1752. https://doi.org/10.1128/AAC.01533-09
Lacmata ST, Kuete V, Dzoyem JP, Tankeo SB, Teke GN, Kuiate JR, Pages JM (2012) Antibacterial activities of selected Cameroonian plants and their synergistic effects with antibiotics against bacteria expressing MDR phenotypes. Evid Based Complem Alternat Med 2012:623723. https://doi.org/10.1155/2012/623723
Lamers RP, Cavallari JF, Burrows LL (2013) The efflux inhibitor phenylalanine-arginine beta-naphthylamide (PAβN) permeabilizes the outer membrane of gram-negative bacteria. PLoS ONE 8(3):e60666. https://doi.org/10.1371/journal.pone.0060666
Latheef L, Kurup MRP (2008) Spectral and structural studies of copper(II) complexes of thiosemicarbazones derived from salicylaldehyde and containing ring incorporated at N(4)-position. Spectrochim Acta Part A 70(1):86–93. https://doi.org/10.1016/j.saa.2007.07.015
Leite A, Bessa LJ, Silva AMG, Gameiro P, de Castro B, Rangel M (2019) Antibacterial activity of naphthyl derived bis-(3-hydroxy-4-pyridinonate) copper (II) complexes against multidrug-resistant bacteria. J Inorg Biochem 197:110704. https://doi.org/10.1016/j.jinorgbio.2019.110704
Lenhard JR, Nation RL, Tsuji BT (2016) Synergistic combinations of polymyxins. Int J Antimicrob Agents 48(6):607–613. https://doi.org/10.1016/j.ijantimicag.2016.09.014
Li XZ, Nikaido H (2009) Efflux-mediated drug resistance in bacteria: an update. Drugs 69(12):1555–1623. https://doi.org/10.2165/11317030-000000000-00000
Lima FC, Silva TS, Martins CHG, Gatto CC (2018) Synthesis, crystal structures and antimicrobial activity of dimeric copper (II) complexes with 2-hydroxyphenyl-ethylidene-dithiocarbazates. Inorg Chim Acta. https://doi.org/10.1016/j.ica.2018.08.032
Lomovskaya O, Warren MS, Lee A, Galazzo J, Fronko R, Lee M, Blais J, Cho D, Chamberland S, Renau T, Leger R, Hecker S, Watkins W, Hoshino K, Ishida H, Lee VJ (2001) Identification and characterization of inhibitors of multidrug resistance efflux pumps in Pseudomonas aeruginosa: novel agents for combination therapy. Antimicrob Agents Chemother 45(1):105–116. https://doi.org/10.1128/AAC.45.1.105-116.2001
Low ML, Maigre L, Tahir MIM, Tiekink ERT, Dorlet P, Guillot R, Ravoof TB, Rosli R, Pagès J-M, Policar C, Delsuc N, Crouse KA (2016) New insight into the structural, electrochemical and biological aspects of macroacyclic Cu(II) complexes derived from S-substituted dithiocarbazate schiff bases. Eur J Med Chem 14(120):1–12. https://doi.org/10.1016/j.ejmech.2016.04.027
Low ML, Maigre L, Dorlet P, Guillot R, Pagès J-M, Crouse KA, Policar C, Delsuc N (2014) Conjugation of a new series of dithiocarbazate Schiff base copper(II) complexes with vectors selected to enhance antibacterial activity. Bioconjug Chem 25(12):2269–2284. https://doi.org/10.1021/bc5004907
Manikal VM, Landman D, Saurina G, Oydna E, Lal H, Quale J (2000) Endemic carbapenem-resistant Acinetobacter species in Brooklyn, New York: citywide prevalence, interinstitutional spread, and relation to antibiotic usage. Clin Infect Dis 31(1):101–106. https://doi.org/10.1086/313902
Mbaveng AT, Ignat AG, Ngameni B, Zaharia V, Ngadjui BT, Kuete V (2016) In vitro antibacterial activities of p-toluenesulfonyl-hydrazinothiazoles and hydrazinoselenazoles against multi-drug resistant Gram-negative phenotypes. BMC Pharmacol Toxicol. https://doi.org/10.1186/s40360-016-0046-0
Meer Ahmad AM (2019) Antibiotic resistance in Malaysia, and its public health implications. J Drug Deliv Therap 9(2):534–541. https://doi.org/10.22270/jddt.v9i2.2427
Morgan CE, Glaza P, Leus I, Trinh A, Su C-C, Cui M, Zgurskaya HI, Yu EW (2021) Cyroelectron microscopy structures of AdeB illuminate mechanisms of simultaneous binding and exporting of substrates. J Clin Microbiol. https://doi.org/10.1128/mBio.03690-20
Nair MS, Arish D, Joseyphus RS (2012) Synthesis, characterization, antifungal, antibacterial and DNA cleavage studies of some heterocyclic Schiff base metal complexes. J Saudi Chem Soc 16(1):83–88. https://doi.org/10.1016/j.jscs.2010.11.002
Nang SC, Azad MAK, Velkov T, Zhou Q, Li J (2021) Rescuing the last-line polymyxins: achievements and challenges. Pharmacol Rev 73(2):679–728. https://doi.org/10.1124/pharmrev.120.000020
Nikaido H, Pagès JM (2012) Broad-specificity efflux pumps and their role in multidrug resistance of Gram-negative bacteria. FEMS Microbiol Rev 36(2):340–363. https://doi.org/10.1111/j.1574-6976.2011.00290.x
Oliveira AA, Oliveira APA, Franco LL, Ferencs MO, Ferreira JFG, Bachi SMPS, Speziali NL, Farias LM, Magalhães PP, Beraldo H (2018) 5-Nitroimidazole-derived Schiff bases and their copper (II) complexes exhibit potent antimicrobial activity against pathogenic anaerobic bacteria. Biometals 31(4):571–584. https://doi.org/10.1007/s10534-018-0106-6
Rampioni G, Pillai CR, Longo F, Bondì R, Baldelli V, Messina M, Imperi F, Visca P, Leoni L (2017) Effect of efflux pump inhibition on Pseudomonas aeruginosa transcriptome and virulence. Sci Rep. https://doi.org/10.1038/s41598-017-11892-9
Rebolledo AP, Vieites M, Gambino D, Piro OE, Castellano EE, Zani CL, Souza-Fagundes EM, Teixeira LR, Batista AA, Beraldo H (2005) Palladium(II) complexes of 2-benzoylpyridine-derived thiosemicarbazones: spectral characterization, structural studies and cytotoxic activity. J Inorg Biochem 99(3):698–706. https://doi.org/10.1016/j.jinorgbio.2004.11.022
Said MA, Al-Harbi WS, Shanmugam M, Aljohani FS, Bouqellah NA, Al-Kaff NS (2020) Synthesis, XRD, HAS, in silico molecular docking studies and biological assessment of novel Schiff base compounds as anti-cancer and antimicrobial agents. J Taibah Univ Sci 14(1):1590–1603. https://doi.org/10.1080/16583655.2020.1849492
Samal S, Mishra SB, Patra SK, Rath A, Dash A, Nayak B, Mohanty D (2021) Polymyxin monotherapy vs. combination therapy for the treatment of multidrug-resistant infections: a systematic review and meta-analysis. Indian J Crit Care Med 25(2):199–206. https://doi.org/10.5005/jp-journals-10071-23720
Singh A, Barman P (2021) Recent advances in Schiff base ruthenium metal complexes: synthesis and applications. Top Curr Chem 379(4):29. https://doi.org/10.1007/s41061-021-00342-w
Singh NK, Kumbhar AA, Pokharel YR, Yadav PN (2020) Anticancer potency of copper (II) complexes of thiosemicarbazones. J Inorg Biochem 210:111134. https://doi.org/10.1016/j.jinorgbio.2020.111134
Singh S, Kalia N, Joshi P, Kumar A, Sharma P, Kumar A, Bharate S, Inshad K (2017) Boeravinone B, A novel dual inhibitor of NorA Bacterial efflux pump of Staphylococcus aureus and human P-glycoprotein, reduces the biofilm formation and intracellular invasion of bacteria. Front Microbiol 8:1868. https://doi.org/10.3389/fmicb.2017.01868
Soto SM (2013) Role of efflux pumps in the antibiotic resistance of bacteria embedded in a biofilm. Virulence 4(3):223–229. https://doi.org/10.4161/viru.23724
Tidwell TT (2008) Hugo (Ugo) Schiff, Schiff bases, and a century of beta-lactam synthesis. Angew Chem Int Ed 47(6):1016–1020. https://doi.org/10.1002/anie.200702965
Vergalli J, Atzori A, Pajovic J, Dumont E, Malloci G, Masi M, Vargiu AV, Winterhalter M, Réfrégiers M, Ruggerone P, Pagès JM (2020) The challenge of intracellular antibiotic accumulation, a function of fluoroquinolone influx versus bacterial efflux. Commun Biol. https://doi.org/10.1038/s42003-020-0929-x
Walsh TJ, Peter J, McGough DA, Fothergill AW, Rinaldi MG, Pizzo PA (1995) Activities of amphotericin B and antifungal azoles alone and in combination against Pseudallescheria boydii. Antimicrob Agents Chemother 39(6):1361–1364. https://doi.org/10.1128/AAC.39.6.1361
World Health Organisation (2014) Antimicrobial resistance: global report on surveillance. https://apps.who.int/iris/handle/10665/112642
Yoon J, Urban C, Terzian C, Mariano N, Rahal JJ (2004) In vitro double and triple synergistic activities of polymyxin B, imipenem, and rifampin against multidrug-resistant Acinetobacter baumannii. Antimicrob Agents Chemother 48(3):753–757. https://doi.org/10.1128/AAC.48.3.753-757.2004
Zharkova MS, Orlov DS, Golubeva OY, Chakchir OB, Eliseev IE, Grinchuk TM, Shamova OV (2019) Application of antimicrobial peptides of the innate immune system in combination with conventional antibiotics—a novel way to combat antibiotic resistance? Front Cell Infect Microbiol 9:128. https://doi.org/10.3389/fcimb.2019.00128
Zusman O, Altunin S, Koppel F, Dishon Benattar Y, Gedik H, Paul M (2017) Polymyxin monotherapy or in combination against carbapenem-resistant bacteria: systematic review and meta-analysis. J Antimicrob Chemother 72(1):29–39. https://doi.org/10.1093/jac/dkw377
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The authors would like to thank the School of Postgraduate Studies and the Institute for Research, Development and Innovation (IRDI) of the International Medical University for the support and funding (Project ID No: MAPC I/2019(2)-IR118) provided.
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Gan, W.K., Liew, H.S., Pua, L.J.W. et al. Novel Cu(II) Schiff Base Complex Combination with Polymyxin B/Phenylalanine-Arginine β-Naphthylamide Against Various Bacterial Strains. Int J Pept Res Ther 28, 60 (2022). https://doi.org/10.1007/s10989-021-10358-x
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DOI: https://doi.org/10.1007/s10989-021-10358-x