4-aminobenzoic acid (PABA), an essential nutrient for many human pathogens, but dispensable for humans, and its derivatives have exhibited various biological activities. In this study, we combined two pharmacophores using a molecular hybridization approach: this vitamin-like molecule and various aromatic aldehydes, including salicylaldehydes and 5-nitrofurfural, via imine bond in one-step reaction. Resulting Schiff bases were screened as potential antimicrobial and cytotoxic agents. The simple chemical modification of non-toxic PABA resulted in constitution of antibacterial activity including inhibition of methicillin-resistant Staphylococcus aureus (minimum inhibitory concentrations, MIC, from 15.62 µM), moderate antimycobacterial activity (MIC ≥ 62.5 µM) and potent broad-spectrum antifungal properties (MIC of ≥ 7.81 µM). Some of the Schiff bases also exhibited notable cytotoxicity for cancer HepG2 cell line (IC50 ≥ 15.0 µM). Regarding aldehyde used for the derivatization of PABA, it is possible to tune up the particular activities and obtain derivatives with promising bioactivities.

Department of Biological and Medical Sciences Faculty of Pharmacy in Hradec Králové Charles University Akademika Heyrovského 1203 500 05 Hradec Králové Czech Republic

Department of Organic and Bioorganic Chemistry Faculty of Pharmacy in Hradec Králové Charles University Akademika Heyrovského 1203 500 05 Hradec Králové Czech Republic

Department of Pharmacology and Toxicology Faculty of Pharmacy in Hradec Králové Charles University Akademika Heyrovského 1203 500 05 Hradec Králové Czech Republic

Laboratory for Mycobacterial Diagnostics and Tuberculosis Regional Institute of Public Health in Ostrava Partyzánské námĕstí 7 702 00 Ostrava Czech Republic

Unipetrol Centre of Research and Education 436 70 Litvínov Záluží 1 Czech Republic

Zobrazit více v PubMed

Patel H.M., Bhardwaj V., Sharma P., Noolvi M.N., Lohan S., Bansal S., Sharma A. Quinoxaline-PABA bipartite hybrid derivatization approach: Design and search for antimicrobial agents. J. Mol. Struct. 2019;1184:562–568. doi: 10.1016/j.molstruc.2019.02.074. DOI

Akberova S.I. New Biological Properties of p-Aminobenzoic Acid. Biol. Bull. 2002;29:390–393. doi: 10.1023/A:1016871219882. PubMed DOI

Kadhum W.R., Oshizaka T., Ichiro H., Todo H., Sugibayashi K. Usefulness of liquid-crystal oral formulations to enhance the bioavailability and skin tissue targeting of p-amino benzoic acid as a model compound. Eur. J. Pharm. Sci. 2016;88:282–290. doi: 10.1016/j.ejps.2016.04.003. PubMed DOI

Crisan M.E., Bourosh P., Maffei M.E., Forni A., Pieraccini S., Sironi M., Chumakov Y.M. Synthesis, Crystal Structure and Biological Activity of 2-Hydroxyethylammonium Salt of p-Aminobenzoic Acid. PLoS One. 2014;9:e101892. doi: 10.1371/journal.pone.0101892. PubMed DOI PMC

Sowinska M., Morawiak M., Bochyńska-Czyż M., Lipkowski A.W., Ziemińska E., Zabłocka B., Urbanczyk-Lipkowska Z. Molecular Antioxidant Properties and In Vitro Cell Toxicity of the p-Aminobenzoic Acid (PABA) Functionalized Peptide Dendrimers. Biomolecules. 2019;9:89. doi: 10.3390/biom9030089. PubMed DOI PMC

Richards R.M., Xing D.K., King T.P. Activity of p-aminobenzoic acid compared with other organic acids against selected bacteria. J. Appl. Bacteriol. 1995;78:209–215. doi: 10.1111/j.1365-2672.1995.tb05018.x. PubMed DOI

Markitantova Y.V., Akberova S.I., Ryabtseva A.A., Stroeva O.G. Effect of para-Aminobenzoic Acid on Apoptosis Processes in the Adult Rat Conjunctiva and Corneal Epithelium in vivo after Hypobaric Hypoxia. Biol. Bull. Russ. Acad. Sci. 2018;45:226–234. doi: 10.1134/S1062359018020061. DOI

Chang T.-Y., Hu M.-L. Concentrations and lipid peroxidation in tissues and toxicity of para-aminobenzoic acid fed to rats in drinking water. J. Nutr. Biochem. 1996;7:408–413. doi: 10.1016/S0955-2863(96)00065-4. DOI

Nortje C., Jansen van Rensburg P., Cooke C., Erasmus E. The simultaneous detection and quantification of p-aminobenzoic acid and its phase 2 biotransformation metabolites in human urine using LC–MS/MS. Bioanalysis. 2015;7:1211–1224. doi: 10.4155/bio.15.54. PubMed DOI

Švarcová M., Krátký M., Vinšová J. Investigation of potential inhibitors of chorismate-utilizing enzymes. Curr. Med. Chem. 2015;22:1383–1399. doi: 10.2174/0929867322666150209152446. PubMed DOI

Yun M.-K., Wu Y., Li Z., Zhao Y., Waddell M.B., Ferreira A.M., Lee R.E., Bashford D., White S.W. Catalysis and Sulfa Drug Resistance in Dihydropteroate Synthase: Crystal structures reveal the catalytic mechanism of DHPS and the structural basis of sulfa drug action and resistance. Science. 2012;335:1110–1114. doi: 10.1126/science.1214641. PubMed DOI PMC

Kluczyk A., Popek T., Kiyota T., de Macedo P., Stefanowicz P., Lazar C., Konishi Y. Drug evolution: p-aminobenzoic acid as a building block. Curr. Med. Chem. 2002;9:1871–1892. doi: 10.2174/0929867023368872. PubMed DOI

Beran J., Šalapová E., Špajdel M. Inosine pranobex is safe and effective for the treatment of subjects with confirmed acute respiratory viral infections: Analysis and subgroup analysis from a Phase 4, randomised, placebo-controlled, double-blind study. BMC Infect. Dis. 2016;16:648. doi: 10.1186/s12879-016-1965-5. PubMed DOI PMC

Zheng J., Rubin E.J., Bifani P., Mathys V., Lim V., Au M., Jang J., Nam J., Dick T., Walker J.R., et al. para-Aminosalicylic acid is a prodrug targeting dihydrofolate reductase in Mycobacterium tuberculosis. J. Biol. Chem. 2013;288:23447–23456. doi: 10.1074/jbc.M113.475798. PubMed DOI PMC

Marc G., Oniga S., Pirnau A., Duma M., Vlase L., Oniga O. Rational Synthesis of Some New para-Aminobenzoic Acid Hybrids with Thiazolidin-2,4-diones with Antimicrobial Properties. ADMET and molecular docking evaluation. Rev. Chim. (Buchar.) 2019;70:769–775.

Scherman M.S., North E.J., Jones V., Hess T.N., Grzegorzewicz A.E., Kasagami T., Kim I.H., Merzlikin O., Lenaerts A.J., Lee R.E., et al. Screening a library of 1600 adamantyl ureas for anti-Mycobacterium tuberculosis activity in vitro and for better physical chemical properties for bioavailability. Bioorg. Med. Chem. 2012;20:3255–3262. doi: 10.1016/j.bmc.2012.03.058. PubMed DOI PMC

Mureşan-Pop M., Kacsó I., Martin F., Simon S., Ştefan R., Bratu I. Ambazone salt with p-aminobenzoic acid. J. Therm. Anal. Calorim. 2015;120:905–912. doi: 10.1007/s10973-015-4452-0. DOI

Farooq M., Al Marhoon Z.M., Taha N.A., Baabbad A.A., Al-Wadaan M.A., El-Faham A. Synthesis of Novel Class of N-Alkyl-isatin-3-iminobenzoic Acid Derivatives and Their Biological Activity in Zebrafish Embryos and Human Cancer Cell Lines. Biol. Pharm. Bull. 2018;41:350–359. doi: 10.1248/bpb.b17-00674. PubMed DOI

Krátký M., Vinšová J., Volková M., Buchta V., Trejtnar F., Stolaříková J. Antimicrobial activity of sulfonamides containing 5-chloro-2-hydroxybenzaldehyde and 5-chloro-2-hydroxybenzoic acid scaffold. Eur. J. Med. Chem. 2012;50:433–440. doi: 10.1016/j.ejmech.2012.01.060. PubMed DOI

Krátký M., Dzurková M., Janoušek J., Konečná K., Trejtnar F., Stolaříková J., Vinšová J. Sulfadiazine Salicylaldehyde-Based Schiff Bases: Synthesis, Antimicrobial Activity and Cytotoxicity. Molecules. 2017;22:1573. doi: 10.3390/molecules22091573. PubMed DOI PMC

Krátký M., Bősze S., Baranyai Z., Stolaříková J., Vinšová J. Synthesis and biological evolution of hydrazones derived from 4-(trifluoromethyl)benzohydrazide. Bioorg. Med. Chem. Lett. 2017;27:5185–5189. doi: 10.1016/j.bmcl.2017.10.050. PubMed DOI

Krátký M., Vinšová J., Stolaříková J. Antimicrobial activity of rhodanine-3-acetic acid derivatives. Bioorg. Med. Chem. 2017;25:1839–1845. doi: 10.1016/j.bmc.2017.01.045. PubMed DOI

Shrivastava S.K., Srivastava P., Upendra T.V.R., Tripathi P.N., Sinha S.K. Design, synthesis and evaluation of some N-methylenebenzenamine derivatives as selective acetylcholinesterase (AChE) inhibitor and antioxidant to enhance learning and memory. Bioorg. Med. Chem. 2017;25:1471–1480. doi: 10.1016/j.bmc.2017.01.010. PubMed DOI

Shetty M.M., Sadanandam Y.S., Sattur P.B. Synthese und Pharmakologie von substituierten N-Benzyliden-Derivaten. Arch. Pharm. 1979;312:721–726. doi: 10.1002/ardp.19793120902. PubMed DOI

Brewster C.M., Millam L.H. Phototropic and Thermotropic Anils from 5-Bromosalicylaldehyde. J. Am. Chem. Soc. 1933;55:763–766. doi: 10.1021/ja01329a049. DOI

Mayadeo M.S., Dhakappa V.P. Determination of stability constants of N-[2-hydroxy-5-methylbenzylidene]-4-carboxyaniline complexes with Y3+, La3+, Pr3+, Nd3+, Sm3+, Gd3+ and Dy3+ J. Indian Chem. Soc. 1981;58:1010.

Nasr G., Cristian A., Barboiu M., Vullo D., Winum J.-Y., Supuran C.T. Carbonic anhydrase inhibitors. Inhibition of human cytosolic isoforms I and II with (reduced) Schiff’s bases incorporating sulfonamide, carboxylate and carboxymethyl moieties. Bioorg. Med. Chem. 2014;22:2867–2874. doi: 10.1016/j.bmc.2014.03.041. PubMed DOI

UNIVERSITÀ DEGLI STUDI DI SALERNO. Longo P., Saturnino C., Arra C., Palma G., Mariconda A., Sinicropi M.S., Puoci F., Iacopetta D. Hydroxybenzene derivatives having a N-aryl substitute imino group and use thereof in the treatment of solid tumours. WO2018/116158. Patent. 2018 Jun 28; A1.

Cronenberger L., Gaige T., Pacheco H., Pillon D. Nouvelles bases de Schiff dérivées des aldehydes dihalogéno-3,5 salicyliques et possédant des propriétés antifongiques et antibactériennes. Chim. Ther. 1968;3:87–99. PubMed

Ujiie T. Experimental Anticancer Studies. XXVIII. Anticancer Activity of Some Nitrofuran Derivatives. Chem. Pharm. Bull. 1966;14:461–466. doi: 10.1248/cpb.14.461. PubMed DOI

Silku P., Özkinali S., Öztürk Z., Asan A., Köse D.A. Synthesis of novel Schiff Bases containing acryloyl moiety and the investigation of spectroscopic and electrochemical properties. J. Mol. Struct. 2016;1116:72–83. doi: 10.1016/j.molstruc.2016.03.028. DOI

Bhunia M., Hota P.K., Vijaykumar G., Adhikari D., Mandal S.K. A Highly Efficient Base-Metal Catalyst: Chemoselective Reduction of Imines to Amines Using An Abnormal-NHC–Fe(0) Complex. Organometallics. 2016;35:2930–2937. doi: 10.1021/acs.organomet.6b00478. DOI

EUCAST DISCUSSION DOCUMENT. E.Dis 5.1. Determination of minimum inhibitory concentrations (MICs) of antibacterial agents by broth dilution. Clin. Microbiol. Infect. 2003;9:1–7. PubMed

Krátký M., Vinšová J., Novotná E., Mandíková J., Trejtnar F., Stolaříková J. Antibacterial Activity of Salicylanilide 4-(Trifluoromethyl)benzoates. Molecules. 2013;18:3674–3688. doi: 10.3390/molecules18043674. PubMed DOI PMC

EUCAST DEFINITIVE DOCUMENT EDEF 7.3.1. Method for the Determination of Broth Dilution Minimum Inhibitory Concentrations of Antifungal Agents for Yeasts. [(accessed on 20 October 2019)];2017 :1–21. Available online: http://www.eucast.org/fileadmin/src/media/PDFs/EUCAST_files/AFST/Files/EUCAST_E_Def_7_3_1_Yeast_testing__definitive.pdf.

EUCAST DEFINITIVE DOCUMENT E.DEF 9.3.1. Method for the Determination of Broth Dilution Minimum Inhibitory Concentrations of Antifungal Agents for Conidia Forming Moulds. [(accessed on 20 October 2019)];2017 :1–23. Available online: http://www.eucast.org/fileadmin/src/media/PDFs/EUCAST_files/AFST/Files/EUCAST_E_Def_9_3_1_Mould_testing__definitive.pdf.

Ordonez A.A., Weinstein E.A., Bambarger L.E., Saini V., Chang Y.S., DeMarco V.P., Klunk M.H., Urbanowski M.E., Moulton K.L., Murawski A.M., et al. A Systematic Approach for Developing Bacteria-Specific Imaging Tracers. J. Nucl. Med. 2017;58:144–150. doi: 10.2967/jnumed.116.181792. PubMed DOI PMC

Elsaman T., Mohamed M.S., Mohamed M.A. Current development of 5-nitrofuran-2-yl derivatives as antitubercular agents. Bioorg. Chem. 2019;88:102969. doi: 10.1016/j.bioorg.2019.102969. PubMed DOI

Laborda P., Zhao Y., Ling J., Hou R., Liu F. Production of Antifungal p-Aminobenzoic Acid in Lysobacter antibioticus OH13. J. Agric. Food Chem. 2018;66:630–636. doi: 10.1021/acs.jafc.7b05084. PubMed DOI

Meir Z., Osherov N. Vitamin Biosynthesis as an Antifungal Target. J. Fungi. 2018;4:72. doi: 10.3390/jof4020072. PubMed DOI PMC

Iliev I., Kontrec D., Detcheva R., Georgieva M., Balacheva A., Galić N., Pajpanova T. Cancer cell growth inhibition by aroylhydrazone derivatives. Biotechnol. Biotec. Eq. 2019;33:756–763. doi: 10.1080/13102818.2019.1608302. DOI

Gerets H.H.J., Tilmant K., Gerin B., Chanteux H., Depelchin B.O., Dhalluin S., Atienzar F.A. Characterization of primary human hepatocytes, HepG2 cells, and HepaRG cells at the mRNA level and CYP activity in response to inducers and their predictivity for the detection of human hepatotoxins. Cell Biol. Toxicol. 2012;28:69–87. doi: 10.1007/s10565-011-9208-4. PubMed DOI PMC

Kamalian L., Chadwick A.E., Bayliss M., French N.S., Monshouwer M., Snoeys J., Park B.K. The utility of HepG2 cells to identify direct mitochondrial dysfunction in the absence of cell death. Toxicol. In Vitro. 2015;29:732–740. doi: 10.1016/j.tiv.2015.02.011. PubMed DOI

Hradilová L., Poláková M., Dvořáková B., Hajdúch M., Petruš L. Synthesis and cytotoxicity of some d-mannose click conjugates with aminobenzoic acid derivatives. Carbohydr. Res. 2012;361:1–6. doi: 10.1016/j.carres.2012.08.001. PubMed DOI

Insight into the Antibacterial Action of Iodinated Imine, an Analogue of Rafoxanide: a Comprehensive Study of Its Antistaphylococcal Activity

Novel Aminoguanidine Hydrazone Analogues: From Potential Antimicrobial Agents to Potent Cholinesterase Inhibitors