Antifungal Activity and Toxicity of the 3,4,5-Trihydroxybenzoic and 3,4,5-Tris(Acetyloxy)Benzoic Acids
Author(s)
Jéssica Pinto1,
Diego Silva1,
João Honorato2,
Antônio Menezes1,
Eugénia Pinto3,
Plínio Naves1*
Affiliation(s)
1 Unit of Exact and Technological Sciences, Universidade Estadual de Goiás, Anápolis, Brazil. 2 Department of Chemistry, Universidade Federal de Sao Carlos, Sao Carlos, Brazil. 3 Department of Biological Sciences, Faculty of Pharmacy, Universidade do Porto, Porto, Portugal.
1 Unit of Exact and Technological Sciences, Universidade Estadual de Goiás, Anápolis, Brazil. 2 Department of Chemistry, Universidade Federal de Sao Carlos, Sao Carlos, Brazil. 3 Department of Biological Sciences, Faculty of Pharmacy, Universidade do Porto, Porto, Portugal.
ABSTRACT
Invasive fungal infections have been gaining notoriety due to several factors, mainly their increasing incidence in immunocompromised patients. The aim of the present study was to evaluate the antifungal activity and toxicity of the 3,4,5-trihydroxybenzoic acid (3,4,5-THB) and of its derivative, the 3,4,5-tris(acetyloxy)benzoic acid (3,4,5-TAB). The 3,4,5-THB was purchased and its derivative was obtained by purifying and characterizing performed using semisynthesis reactions (esterification), recrystallization, column chromatography and infrared analytical techniques and nuclear magnetic resonance. Minimum inhibitory concentration (MIC) and minimum lethal concentration (MLC) were determined in order to evaluate the antifungal activity of the compounds against four clinical isolates and four standard strains of Candida sp. and five clinical isolates of dermatophytes, following the Clinical and Laboratory Standards Institute protocols. The toxicity of the compounds was evaluated by determining the lethal dosis (LD50) using lethality assay of Artemia salina. The most sensitive yeasts to the 3,4,5-THB were C. albicans ATCC 10231 and C. krusei ATCC 6258, both presenting a MIC of 128 μg·mL-1. For Trichophyton sp. and Epidermophyton floccosum, the MIC was 32 μg·mL-1. The 3,4,5-TAB showed a lower inhibitory activity against Candida and dermatophyte species tested. The LD50 of 3,4,5-THB was 222.60 μg·mL-1 and the 3,4,5-TAB showed 481.69 μg·mL-1 of LD50. In conclusion, the 3,4,5-trihydroxybenzoic acid showed antifungal activity against species of medical importance, mainly dermatophytosis-causing fungi, and the 3,4,5-tris(acetyloxy)benzoic acid showed no increasing antifungal activity and toxicity in relation to the original compound.
Invasive fungal infections have been gaining notoriety due to several factors, mainly their increasing incidence in immunocompromised patients. The aim of the present study was to evaluate the antifungal activity and toxicity of the 3,4,5-trihydroxybenzoic acid (3,4,5-THB) and of its derivative, the 3,4,5-tris(acetyloxy)benzoic acid (3,4,5-TAB). The 3,4,5-THB was purchased and its derivative was obtained by purifying and characterizing performed using semisynthesis reactions (esterification), recrystallization, column chromatography and infrared analytical techniques and nuclear magnetic resonance. Minimum inhibitory concentration (MIC) and minimum lethal concentration (MLC) were determined in order to evaluate the antifungal activity of the compounds against four clinical isolates and four standard strains of Candida sp. and five clinical isolates of dermatophytes, following the Clinical and Laboratory Standards Institute protocols. The toxicity of the compounds was evaluated by determining the lethal dosis (LD50) using lethality assay of Artemia salina. The most sensitive yeasts to the 3,4,5-THB were C. albicans ATCC 10231 and C. krusei ATCC 6258, both presenting a MIC of 128 μg·mL-1. For Trichophyton sp. and Epidermophyton floccosum, the MIC was 32 μg·mL-1. The 3,4,5-TAB showed a lower inhibitory activity against Candida and dermatophyte species tested. The LD50 of 3,4,5-THB was 222.60 μg·mL-1 and the 3,4,5-TAB showed 481.69 μg·mL-1 of LD50. In conclusion, the 3,4,5-trihydroxybenzoic acid showed antifungal activity against species of medical importance, mainly dermatophytosis-causing fungi, and the 3,4,5-tris(acetyloxy)benzoic acid showed no increasing antifungal activity and toxicity in relation to the original compound.
KEYWORDS
Antifungal, Toxicity, Candida, Dermatophytes, 3, 4, 5-Trihydroxybenzoic Acid, 3, 4, 5-Tris(Acetyloxy)Benzoic Acid
Antifungal, Toxicity, Candida, Dermatophytes, 3, 4, 5-Trihydroxybenzoic Acid, 3, 4, 5-Tris(Acetyloxy)Benzoic Acid
Cite this paper
Pinto, J. , Silva, D. , Honorato, J. , Menezes, A. , Pinto, E. and Naves, P. (2015) Antifungal Activity and Toxicity of the 3,4,5-Trihydroxybenzoic and 3,4,5-Tris(Acetyloxy)Benzoic Acids. Advances in Microbiology, 5, 517-522. doi: 10.4236/aim.2015.57053.
Pinto, J. , Silva, D. , Honorato, J. , Menezes, A. , Pinto, E. and Naves, P. (2015) Antifungal Activity and Toxicity of the 3,4,5-Trihydroxybenzoic and 3,4,5-Tris(Acetyloxy)Benzoic Acids. Advances in Microbiology, 5, 517-522. doi: 10.4236/aim.2015.57053.
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[1] Brown, G.D., Denning, D.W., Gow, N.A.R., Levitz, S.M., Netea, M.G. and White, T.C. (2012) Hidden Killers: Human Fungal Infections. Science Translational Medicine, 4, 1-9.
http://dx.doi.org/10.1126/scitranslmed.3004404 [2] Delaloye, J. and Calandra, T. (2013) Invasive Candidiasis as a Cause of Sepsis in the Critically Ill Patient. Virulence, 5, 1-9. [3] Ameen, M. (2011) Epidemiology of Superficial Fungal Infections. Clinics in Dermatology, 28, 197-201.
http://dx.doi.org/10.1016/j.clindermatol.2009.12.005 [4] Richardson, M.D. and Warnock, D.W. (2012) Fungal Infection: Diagnosis and Management. 4th Edition, Wiley-Blackwell, Oxford.
http://dx.doi.org/10.1002/9781118321492 [5] Pfaller, M.A. (2012) Antifungal Drug Resistance: Mechanism, Epidemiology and Consequences for Treatment. The American Journal of Medicine, 125, 3-13.
http://dx.doi.org/10.1016/j.amjmed.2011.11.001 [6] De Pauw, B.E. and Picazo, J.J. (2008) Present Situation in the Treatment of Invasive Fungal Infection. International Journal Antimicrobial Agents, 32, 167-171.
http://dx.doi.org/10.1016/S0924-8579(08)70020-7 [7] Pinto, E., Vale-Silva, L., Cavaleiro, C. and Salgueiro, L. (2009) Antifungal Activity of the Clove Essential Oil from Syzygium aromaticum on Candida, Aspergillus and Dermatophyte Species. Journal Medical Microbiology, 58, 1454-1462.
http://dx.doi.org/10.1099/jmm.0.010538-0 [8] Denning, D.W. and Hope, W.W. (2010) Therapy for Fungal Diseases: Opportunities and Priorities. Trends in Microbiology, 18, 195-204.
http://dx.doi.org/10.1016/j.tim.2010.02.004 [9] Montanari, C.A. and Bolzani, V.S. (2001) Planejamento racional de fármacos baseado em produtos naturais. Quimica Nova, 24, 105-111.
http://dx.doi.org/10.1590/S0100-40422001000100018 [10] Rosso, R. (2005) Avaliacao das propriedades antioxidantes de derivados ésteres do ácido gálico. M.Sc. Thesis, Faculdade de Farmácia, Universidade Federal de Santa Catarina, 145 p. [11] Molina-Salinas, G.M. and Said-Fernández, S. (2006) A Modified Microplate Cytotoxicity Assay with Brine Shrimp Larvae (Artemia salina). Pharmacologyonline, 3, 633-638. [12] Kaur, M., Velmurugan, B., Rajamanickam, S., Agarwal, R. and Agarwal, C. (2009) Gallic Acid, an Active Constituent of Grape Seed Extract, Exhibits Anti-Proliferative, Pro-Apoptotic and Anti-Tumorigenic Effects against Prostate Carcinoma Xenograft Growth in Nude Mice. Pharmaceutical Research, 26, 2133-2140.
http://dx.doi.org/10.1007/s11095-009-9926-y [13] Silverstein, R.M, Webster, F.X. and Kiemle, D.J. (2006) Identification Espectrométrica de Compostos Organicos. 7th Edition, LTC, Rio de Janeiro. [14] Costa, E.S.S., Dolabela, M.F., Póvoa, M.M., Oliveira, D.J. and Müller, A.H. (2009) Estudos farmacognósticos, fitoquímicos, atividade antiplasmódica e toxicidade em Artemia salina de extrato etanólico de folhas de Montrichardia linifera (Arruda) Schott, Araceae. Revista Brasileira de Farmacognosia, 19, 834-838.
http://dx.doi.org/10.1590/S0102-695X2009000600006 [15] Rahman, A., Choudhary, M.I. and Thomsen, W.J. (2005) Bioassay Techniques for Drug Development. Taylor & Francis e-Library, Singapure. [16] Nascimento, J.E., Melo, A.F.M., Lima e Silva, T.C., Veras Filho, J., Santos, E.M., Albuquerque, U.P. and Amorim, E.L.C. (2008) Estudo fitoquímico e bioensaio toxicológico frente a larvas de Artemia salina Leach. de três espécies medicinais do gênero Phyllanthus (Phyllanthaceae). Revista de Ciências Farmacêuticas Básica e Aplicada, 29, 145-150. [17] Baker, R.H. (1948) A Side Reaction in the Williamson Synthesis. Journal of the American Chemical Society, 70, 3857-3859. http://dx.doi.org/10.1021/ja01191a094 [18] CLSI—Clinical and Laboratory Standards Institute (2008) Reference Method for Broth Dilution Antifungal Susceptibility Testing of Yeasts. Approved Standard M27-A3, 3rd Edition, CLSI, Wayne, 33 p. [19] CLSI—Clinical and Laboratory Standards Institute (2008) Reference Method for Broth Dilution Antifungal Susceptibility Testing of Filamentous Fungi. Approved Standard M38-A2, 2nd Edition, Wayne, 37 p. [20] Leal, P.C., Mascarello, A., Derita, M., Zuljan, F., Nunes, R.J., Zacchino, S. and Yunes, R.A. (2009) Relation between Lipophilicity of Alkyl Gallates and Antifungal Activity against Yeasts and Filamentous Fungi. Bioorganic & Medicinal Chemistry Letters, 19, 1793-1796. http://dx.doi.org/10.1016/j.bmcl.2009.01.061 [21] Parra, A.L., Yhebra, R.S., Sardinas, I.G. and Buela, L.I. (2001) Comparative Study of the Assay of Artemia salina L. and the Estimate of the Medium Lethal Dose (LD50 Value) in Mice, to Determine Oral Acute Toxicity of Plant Extracts. Phytomedicine, 8, 395-400. http://dx.doi.org/10.1078/0944-7113-00044