Antifungal Activity and Cytotoxicity of Imidazole- and Morpholine-Based Lysosomotropic Detergents
DOI:
https://doi.org/10.20535/ibb.2025.9.1.315523Keywords:
lysosomotropic detergents, 1-alkylimidazole, N-alkylmorpholine, anti-Candida activity, cytotoxicity, molecular docking, QSARAbstract
Background. The spread of infectious diseases caused by drug-resistant bacteria and fungi, especially nosocomial strains, is currently considered a serious medical problem. It is known that the fungus Candida albicans is the most common causative agent of candidal infection, including the severe type. The emergence and rapid formation of drug resistance, as a risk factor in the treatment of oncological diseases burdened by candidal infection, requires new therapeutic approaches, including the study of synthetic bioregulators with antifungal and anticancer efficacy.
Objective. To synthesize and to determine the antifungal activity and cytotoxicity of imidazole- and morpholine-based lysosomotropic detergents (LDs) comprising both dodecyl radicals and ester-functionalized long alkyl chains.
Methods. To develop the QSAR models by the OCHEM platform, machine learning methods such as Transformer Convolutional Neural Network (Trans-CNN), Transformer Convolutional Neural Fingerprint (Trans-CNF), and Random Forest (RF) were used. Imidazole- and morpholine-based lysosomotropic detergents comprising both dodecyl radicals and ester-functionalized long alkyl chains were synthesized and characterized by 1H Nuclear Magnetic Resonance spectroscopy. The antifungal properties of studied compounds were estimated by the disc diffusion method against the C. albicans ATCC 10231, C. albicans, C. glabrata and C. krusei clinical isolates. The in vitro cytotoxic activity of LDs was evaluated by IC50 value against the throat cancer HEp-2 cell lines. AutoDock Vina software was used for the evaluation of the synthesis compounds as ligands of several antifungal targets.
Results. The identified and synthesized imidazole- and morpholine-based LDs showed high antifungal potential against all Candida spp., specifically against the fluconazole-resistant C. albicans, C. glabrata, and C. krusei clinical isolates. Imidazole-based LD 1 (IM-C12) and morpholine-based LD 4 (Mor-C12) were the most active against tested fungal strains. Molecular docking results suggest that the antifungal mechanisms of the compounds may be related to the inhibition of fungal lanosterol 14a-demethylase. The cytotoxic results of the synthesized compounds against the HEp-2 cell line demonstrated that morpholine-based LDs are less cytotoxic compared to imidazole-based
Conclusions. It was found that LD IM-C12 (1) is the most promising cytostatic and antifungal agent based on the obtained results. LDs IM-CH2COOC10 (2) and Mor-C12 (4), which, under conditions of chemical modification, including through the carbon chain, may also be interesting for developing potential anticancer agents.
References
Firestone RA, Pisano JM, Garrity GM, Fromtling RA, Zimmerman SB. Lysosomotropic agents. Broad-spectrum antifungal activity of lysosomotropic detergents. Journal of Medicinal Chemistry. 1987;30(8):1519-21. DOI: 10.1021/jm00391a043
Hu M, Chen J, Liu S, Xu H. The acid gate in the lysosome. Autophagy. 2023;19(4):1368-70. DOI: 10.1080/15548627.2022.2125629
Tancini B, Buratta S, Delo F, Sagini K, Chiaradia E, Pellegrino RM, et al. Lysosomal exocytosis: The extracellular role of an intracellular organelle. Membranes. 2020;10(12):406. DOI: 10.3390/membranes10120406
Hussain M, Leibowitz MJ, Lenard J. Killing of Saccharomyces cereviciae by the lysosomotropic detergent N-dodecylimidazole. Antimicrobial Agents and Chemotherapy. 1987;31(4):512-17. DOI: 10.1128/AAC.31.4.512
Miller DK, Griffiths E, Lenard J, Firestone RA. Cell killing by lysosomotropic detergents. Journal of Cell Biology. 1983;97(6):1841-51. DOI: 10.1083/jcb.97.6.1841
Wilson PD, Firestone RA, Lenard J. The role of lysosomal enzymes in killing of mammalian cells by the lysosomotropic detergent N-dodecylimidazole. Journal of Cell Biology. 1987;104(5):1223-9. DOI: 10.1083/jcb.104.5.1223
Eriksson I, Öllinger K. Lysosomes in cancer – At the crossroad of good and evil. Cells. 2024;13(5):459. DOI: 10.3390/cells13050459
Trybus W, Trybus E, Król T. Lysosomes as a target of anticancer therapy. International Journal of Molecular Sciences. 2023;24(3):2176. DOI: 10.3390/ijms24032176
Hao G, Xu ZP, Li L. Manipulating extracellular tumour pH: an effective target for cancer therapy. RSC Advances. 2018;8(39):22182-92. DOI: 10.1039/c8ra02095g
Boyer MJ, Horn I, Firestone RA, Steele-Norwood D, Tannock IF. pH dependent cytotoxicity of N-dodecylimidazole: a compound that acquires detergent properties under acidic conditions. British Journal of Cancer. 1993;67(1):81-7. DOI: 10.1038/bjc.1993.13
Wilson PD, Hreniuk D, Lenard J. Reduced cytotoxicity of the lysosomotropic detergent N-dodecylimidazole after differentiation of HL60 promyelocytes. Cancer Research. 1989;49(3):507-10.
Dubowchik GM, Gawlak SL, Firestone RA. The in vitro effects of three lysosomotropic detergents against three human tumor cell lines. Bioorganic & Medicinal Chemistry Letters. 1995;5(8):893-8. DOI: 10.1016/0960-894X(95)00136-H
Gryniukova A, Borysko P, Myziuk I, Alieksieieva D, Hodyna D, Semenyuta I, et al. Anticancer activity features of imidazole-based ionic liquids and lysosomotropic detergents: in silico and in vitro studies. Molecular Diversity. 2024;28(6):3817-33. DOI: 10.1007/s11030-023-10779-4
Pappas PG, Lionakis MS, Arendrup MС, Ostrovsky-Zeichner L, Kullberg BJ. Invasive candidiasis. Nature Reviews Disease Primers. 2018;4:18026. DOI: 10.1038/nrdp.2018.26
Ruijters VJ, Oosterom N, van der Perk MEM, Wolfs TFW, Meijer AJM, van den Heuvel-Eibrink MM, van Grotel M. Clinical characteristics of invasive fungal infections in pediatric oncology patients with solid tumors. Journal of Pediatric Hematology/Oncology. 2021;43(3):e408-e413. DOI: 10.1097/MPH.0000000000001761
De Rosa FG, Busca A, Capparella MR, Yan JL, Aram JA. Invasive candidiasis in patients with solid tumors treated with anidulafungin: a post hoc analysis of efficacy and safety of six pooled studies. Clinical Drug Investigation. 2021;41(6):539-48. DOI: 10.1007/s40261-021-01024-7
Bai X, Luo J. Invasive Candidiasis in Patients with Solid Tumors: A Single-Center Retrospective Study. International Journal of General Medicine. 2023;16:2419-26. DOI: 10.2147/IJGM.S411006
Pokharel M, Konarzewska P, Roberge JY, Han G-S, Wang Y, Carman GM, Xue C. The anticancer drug bleomycin shows potent antifungal activity by altering phospholipid biosynthesis. Microbiology Spectrum. 2022;10(5):e0086222. DOI: 10.1128/spectrum.00862-22
Hamill RJ. Amphotericin B Formulations: A Comparative Review of Efficacy and Toxicity. Drugs. 2013;73(9):919-34. DOI: 10.1007/s40265-013-0069-4
Dixon DM, Walsh TJ. Antifungal Agents. In: Medical Microbiology. 4th ed. Galveston, TX: University of Texas Medical Branch at Galveston, 1996.
Kanafani ZA, Perfect JR. Resistance to antifungal agents: mechanism and clinical impact. Clinical Infectious Diseases. 2008;46(1):120-8. DOI: 10.1086/524071
Draskau MK, Svingen T. Azole fungicides and their endocrine disrupting properties: perspectives on sex hormone-dependent reproductive development. Frontiers in Toxicology. 2022;4:883254. DOI: 10.3389/ftox.2022.883254
Munkboel CH, Rasmussen TB, Elgaard C, Olesen M-LK, Kretschmann AC, Styrishave B. The classic azole antifungal drugs are highly potent endocrine disruptors in vitro inhibiting steroidogenic CYP enzymes at concentrations lower than therapeutic Cmax. Toxicology. 2019;425:152247. DOI: 10.1016/j.tox.2019.152247
Nishimoto AT, Sharma C, Rogers PD. Molecular and genetic basis of azole antifungal resistance in the opportunistic pathogenic fungus Candida albicans. Journal of Antimicrobial Chemotherapy. 2020;75(2):257-70. DOI: 10.1093/jac/dkz400
Sucher AJ, Chahine EB, Balcer HE. Echinocandins: the newest class of antifungals. Annals of Pharmacotherapy. 2009;43(10):1647-57. DOI: 10.1345/aph.1M237
Zhu P, Li Y, Guo T, Liu S, Tancer RJ, Hu C, et al. New antifungal strategies: drug combination and co-delivery. Advanced Drug Delivery Reviews. 2023;198:114874. DOI: 10.1016/j.addr.2023.114874
Perlin DS. Echinocandin resistance in Candida. Clinical Infectious Diseases. 2015;61(suppl_6):S612-S617. DOI: 10.1093/cid/civ791
Pristov KE, Ghannoum MA. Resistance of Candida to azoles and echinocandins worldwide. Clinical Microbiology and Infection. 2019;25(7):792-8. DOI: 10.1016/j.cmi.2019.03.028
Weng N, Zhang Z, Tan Y, Zhang X, Wei X, Zhu Q. Repurposing antifungal drugs in cancer therapy. Journal of Advanced Research. 2023;48:259-73. DOI: 10.1016/j.jare.2022.08.018
Seo I, Jha BK, Lim J-G, Suh S-I, Suh M-H, Baek W-K. Identification of lysosomotropic compounds based on the distribution and size of lysosomes. Biochemical and Biophysical Research Communications. 2014;450(1):189-94. DOI: 10.1016/j.bbrc.2014.05.091
Trush M, Metelytsia L, Semenyuta I, Kalashnikova L, Papeykin O, Venger I, et al. Reduced ecotoxicity and improved biodegradability of cationic biocides based on ester-functionalized pyridinium ionic liquids. Environmental Science and Pollution Research. 2019;26(5):4878-89. DOI: 10.1007/s11356-018-3924-8
Hughes JA, Aronsohn AI, Avrutskaya AV, Juliano RL. Evaluation of adjuvants that enhance the effectiveness of antisense oligodeoxynucleotides. Pharmaceutical Research. 1996;13(3):404-10. DOI: 10.1023/a:1016044609972
Chen F-J, Asokan A, Cho MJ. Cytosolic delivery of macromolecules: I. Synthesis and characterization of pH-sensitive acyloxyalkylimidazoles. Biochimica et Biophysica Acta (BBA) - Biomembranes. 2003;1611(1-2):140-50. DOI: 10.1016/s0005-2736(03)00049-x
Mendez D, Gaulton A, Bento AP, Chambers J, De Veij M, Félix E, et al. ChEMBL: towards direct deposition of bioassay data. Nucleic Acids Research. 2019;47(D1):D930-D940. DOI: 10.1093/nar/gky1075
Sushko I, Novotarskyi S, Körner R, Pandey AK, Rupp M, Teetz W, et al. Online chemical modeling environment (OCHEM): web platform for data storage, model development and publishing of chemical information. Journal of Computer-Aided Molecular Design. 2011;25(6):533-54. DOI: 10.1007/s10822-011-9440-2
Online chemical database with modeling environment. Ochem.eu. Available from: https://ochem.eu/
Karpov P, Godin G, Tetko IV. Transformer-CNN: Swiss knife for QSAR modeling and interpretation. Journal of Cheminformatics. 2020;12(1):17. DOI: 10.1186/s13321-020-00423-w
Tetko IV, Karpov P, Bruno E, Kimber TB, Godin G. Augmentation is what you need! In: Artificial neural networks and machine learning – ICANN 2019: Workshop and special sessions, vol. 11731. Cham: Springer International Publishing, 2019, pp. 831-5. DOI: 10.1007/978-3-030-30493-5_79
Breiman L. Random forests. Machine Learning. 2001;45(1):5-32. DOI: 10.1023/A:1010933404324
Hall LH, Kier LB. Electrotopological state indexes for atom types: A novel combination of electronic, topological, and valence state information. Journal of Chemical Information and Computer Sciences. 1995;35(6):1039-45. DOI: 10.1021/ci00028a014
Tetko IV, Tanchuk VYu. Application of associative neural networks for prediction of lipophilicity in ALOGPS 2.1 program. Journal of Chemical Information and Computer Sciences. 2002;42(5):1136-45. DOI: 10.1021/ci025515j
Willighagen EL, Mayfield JW, Alvarsson J, Berg A, Carlsson L, Jeliazkova N, et al. The Chemistry Development Kit (CDK) v2.0: atom typing, depiction, molecular formulas, and substructure searching. Journal of Cheminformatics. 2017;9(1):33. DOI: 10.1186/s13321-017-0220-4
Tetko IV, Sushko I, Pandey AK, Zhu H, Tropsha A, Papa E, et al. Critical assessment of QSAR models of environmental toxicity against Tetrahymena pyriformis: focusing on applicability domain and overfitting by variable selection. Journal of Chemical Information and Modeling. 2008;48(9):1733-46. DOI: 10.1021/ci800151m
Johnson NL, Leone FC. Statistic and experimental design in engineering and the physical sciences. New York: John Wiley & Sons, 1977, 1082 p.
Sushko I, Novotarskyi S, Körner R, Pandey AK, Kovalishyn VV, Prokopenko VV, Tetko IV. Applicability domain for in silico models to achieve accuracy of experimental measurements. Journal of Chemometrics. 2010;24(3-4):202-8. DOI: 10.1002/cem.1296
OCHEM user's manual - OCHEM Introduction. Ochem.eu. Available from: https://docs.ochem.eu/x/AwBa.html
Bauer AW, Kirby WMM, Sherris JC, Turck M. Antibiotic susceptibility testing by a standardized single disk method. American Journal of Clinical Pathology. 1966;45(4_ts):493-6. DOI: 10.1093/ajcp/45.4_ts.493
Brown A, Kumar S, Tchounwou PB. Cisplatin-based chemotherapy of human cancers. Journal of Cancer Science & Therapy. 2019;11(4):97.
da Nóbrega Alves D, Monteiro AFM, Andrade PN, Lazarini JG, Abílio GMF, Guerra FQS, et al. Docking prediction, antifungal activity, anti-biofilm effects on Candida spp., and toxicity against human cells of cinnamaldehyde. Molecules. 2020;25(24):5969. DOI: 10.3390/molecules25245969
Li H, Kong Y, Hu W, Zhang S, Wang W, Yang M, Luo Y. Litsea cubeba essential oil: Component analysis, anti-Candida albicans activity and mechanism based on molecular docking. Journal of Oleo Science. 2022;71(8):1221-8. DOI: 10.5650/jos.ess22108
Tonthat NK, Juvvadi PR, Zhang H, Lee SC, Venters R, Spicer L, et al. Structures of pathogenic fungal FKBP12s reveal possible self-catalysis function. mBio. 2016;7(2):e00492-16. DOI: 10.1128/mBio.00492-16
Srivastava V, Singla RK, Dubey AK. Inhibition of biofilm and virulence factors of Candida albicans by partially purified secondary metabolites of Streptomyces chrestomyceticus strain ADP4. Current Topics in Medicinal Chemistry. 2018;18(11):925-45. DOI: 10.2174/1568026618666180711154110
Flamandita D, Sahlan M, Lischer K, Pratami DK. Molecular Docking Analysis of Anti-Candida albicans Biomarkers in Sulawesi Propolis Against Secreted Aspartic Proteinase-5. In: 2019 IEEE 6th International Conference on Engineering Technologies and Applied Sciences (ICETAS), Kuala Lumpur, Malaysia, 2019, pp. 1-5. DOI: 10.1109/ICETAS48360.2019.9117300
Sinha K, Rule GS. The structure of thymidylate kinase from Candida albicans reveals a unique structural element. Biochemistry. 2017;56(33):4360-70. DOI: 10.1021/acs.biochem.7b00498
Berman HM, Westbrook J, Feng Z, Gilliland G, Bhat TN, Weissig H, et al. The Protein Data Bank. Nucleic Acids Research. 2000;28(1):235-42. DOI: 10.1093/nar/28.1.235
Marvin n. n. n. (5.2.4.), 201n. Chemaxon.com, 2009. Available from: http://www.chemaxon.com
Hanwell MD, Curtis DE, Lonie DC, Vandermeersch T, Zurek E, Hutchison GR. Avogadro: an advanced semantic chemical editor, visualization, and analysis platform. Journal of Cheminformatics. 2012;4(1):17. DOI: 10.1186/1758-2946-4-17
Alonso H, Bliznyuk AA, Gready JE. Combining docking and molecular dynamic simulations in drug design. Medicinal Research Reviews. 2006;26(5):531-68. DOI: 10.1002/med.20067
Trott O, Olson AJ. AutoDock Vina: Improving the speed and accuracy of docking with a new scoring function, efficient optimization and multithreading. Journal of Computational Chemistry. 2010;31(2):455-61. DOI: 10.1002/jcc.21334
Trush MM, Kovalishyn V, Ocheretniuk AD, Kobzar OL, Kachaeva MV, Brovarets VS, Metelytsia LO. QSAR Study of Some 1,3-Oxazolylphosphonium Derivatives as New Potent Anti-Candida Agents and Their Toxicity Evaluation. Current Drug Discovery Technologies. 2019;16(2):204-9. DOI: 10.2174/1570163815666180418145422
Hodyna D, Kovalishyn V, Semenyuta I, Rogalsky S, Trokhimenko O, Gryniukova A, Metelytsia L. Ester-Functionalized Imidazolium- and Pyridinium-Based Ionic Liquids: Design, Synthesis and Cytotoxicity Evaluation. Biointerface Research in Applied Chemistry. 2022;12(3):2905-57. DOI: 10.33263/BRIAC123.29052957
Monk BC, Sagatova AA, Hosseini P, Ruma YN, Wilson RK, Keniya MV. Fungal Lanosterol 14α-demethylase: A target for next-generation antifungal design. Biochimica et Biophysica Acta (BBA) - Proteins and Proteomics. 2020;1868(3):140206. DOI: 10.1016/j.bbapap.2019.02.008
Aimanianda V, Simenel C, Garnaud C, Clavaud C, Tada R, Barbin L, et al. The Dual Activity Responsible for the Elongation and Branching of β-(1,3)-Glucan in the Fungal Cell Wall. mBio. 2017;8(3):e00619-17. DOI: 10.1128/mBio.00619-17
Jayasheela K, Al-Wahaibi LH, Periandy S, Hassan HM, Sebastian S, Xavier S, et al. Probing vibrational activities, electronic properties, molecular docking and Hirshfeld surfaces analysis of 4-chlorophenyl ({[(1E)-3-(1H-imidazol-1-yl)-1-phenylpropylidene]amino}oxy)methanone: A promising anti-Candida agent. Journal of Molecular Structure. 2018;1159:83-95. DOI: 10.1016/j.molstruc.2018.01.042
Li W, Yu D, Gao S, Lin J, Chen Z, Zhao W. Role of Candida albicans-Secreted Aspartyl Proteinases (Saps) in Severe Early Childhood Caries. International Journal of Molecular Sciences. 2014;15(6):10766-79. DOI: 10.3390/ijms150610766
Sun S, Yang Q, Sheng Y, Fu Y, Sun C, Deng C. Investigational drugs with dual activity against HBV and HIV (Review). Experimental and Therapeutic Medicine. 2021;21(1):35. DOI: 10.3892/etm.2020.9467
Sun M, Qin J, Kang Y, Zhang Y, Ba M, Yang H, et al. 2-Methoxydiol derivatives as new tubulin and HDAC dual-targeting inhibitors, displaying antitumor and antiangiogenic response. Bioorganic Chemistry. 2022;120:105625. DOI: 10.1016/j.bioorg.2022.105625
Balbuena-Rebolledo I, Rivera-Antonio AM, Sixto-López Y, Correa-Basurto J, Rosales-Hernández MC, Mendieta-Wejebe JE, et al. Dihydropyrazole-carbohydrazide derivatives with dual activity as antioxidant and anti-proliferative drugs on breast cancer targeting the HDAC6. Pharmaceuticals. 2022;15(6):690. DOI: 10.3390/ph15060690
Shaker AMM, Abdelall EKA, Abdellatif KRA, Abdel-Rahman HM. Synthesis and biological evaluation of 2-(4-methylsulfonyl phenyl) indole derivatives: multi-target compounds with dual antimicrobial and anti-inflammatory activities. BMC Chemistry. 2020;14(1):23. DOI:10.1186/s13065-020-00675-5
Jachak GR, Ramesh R, Sant DG, Jorwekar SU, Jadhav MR, Tupe SG, et al. Silicon Incorporated Morpholine Antifungals: Design, Synthesis, and Biological Evaluation. ACS Medicinal Chemistry Letters. 2015;6(11):1111-6. DOI: 10.1021/acsmedchemlett.5b00245
Rani N, Sharma A, Gupta GK, Singh R. Imidazoles as Potential Antifungal Agents: A Review. Mini-Reviews in Medicinal Chemistry. 2013;13(11):1626-55. DOI: 10.2174/13895575113139990069
Schrekker HS, Donato RK, Fuentefria AM, Bergamo V, Oliveira LF, Machado MM. Imidazolium salts as antifungal agents: activity against emerging yeast pathogens, without human leucocyte toxicity. MedChemComm. 2013;4(11):1457-60. DOI: 10.1039/C3MD00222E
Bergamo VZ, Donato RK, Dalla Lana DF, Donato KJZ, Ortega GG, Schrekker HS, Fuentefria AM. Imidazolium salts as antifungal agents: strong antibiofilm activity against multidrug-resistant Candida tropicalis isolates. Letters in Applied Microbiology. 2015;60(1):66-71. DOI: 10.1111/lam.12338
Reddy GKK, Nancharaiah YV. Alkylimidazolium Ionic Liquids as Antifungal Alternatives: Antibiofilm Activity Against Candida albicans and Underlying Mechanism of Action. Frontiers in Microbiology. 2020;11:730. DOI: 10.3389/fmicb.2020.00730
Przestalski S, Sarapuk J, Kleszczyńska H, Gabrielska J, Hładyszowski J, Trela Z, Kuczera J. Influence of amphiphilic compounds on membranes. Acta Biochimica Polonica. 2000;47(3):627-38. DOI: 10.18388/abp.2000_3984
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