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Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology]2024; 56(1); 331-340; doi: 10.1007/s42770-024-01572-y

Anti-Pythium insidiosum activity of three novel triazole compounds: synthesis, pharmacokinetic and toxicological parameters.

Abstract: Pythiosis, caused by Pythium insidiosum, is an infectious and non-transmissible disease affecting horses, dogs, and humans, with no effective drug treatment available. Triazoles are compounds of interest for their potential pharmacological properties against fungi and bacteria. In this study, we synthesized three new triazole compounds (C1, C2, and C3) to assess their in vitro activities against P. insidiosum and their safety on human leukocytes. Susceptibility testing was performed against P. insidiosum isolates (n = 15) to determine the minimum inhibitory concentration (MIC) and minimum oomicidal concentration (MOC). The leukocyte toxicity of triazoles was evaluated by measuring cell viability, morphological aspects, and oxidative stress endpoints. In silico prediction of the compounds absorption, distribution, metabolism, excretion and toxicity (ADMET) was determined using the pkCSM platform. Both triazoles C1 and C2 exhibited anti-Pythium insidiosum activity at concentrations from 2 to 64 µg/mL to MIC and MOC, while C3 MIC was 4-64 µg/mL and MOC 8-64 µg/mL. The three compounds did not induce viability loss and/or morphologic changes to human leukocytes, and showed absence of a pro-oxidant profile. ADMET properties prediction of the compounds was similar to the reference drug fluconazole. This study introduces novel triazole compounds exhibiting anti-P. insidiosum activity at concentrations non-toxic to human leukocytes.
© 2024. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.
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Publication Date: 2024-12-12 PubMed ID: 39666162PubMed Central: PMC11885195DOI: 10.1007/s42770-024-01572-yGoogle Scholar: Lookup
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Summary

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Overview

  • This research article reports the synthesis and evaluation of three new triazole compounds (C1, C2, and C3) for their effectiveness against the pathogen Pythium insidiosum, which causes pythiosis, a difficult-to-treat infectious disease.
  • The study investigates the compounds’ antifungal activity, toxicity toward human leukocytes, and pharmacokinetic profiles using in vitro and in silico methods.

Background

  • Pythiosis is an infectious disease caused by the oomycete Pythium insidiosum, affecting horses, dogs, and humans.
  • Currently, there are no effective drug treatments for pythiosis, making the search for new therapeutic agents urgent.
  • Triazoles are a class of compounds widely researched for antifungal and antibacterial properties due to their ability to inhibit microbial growth.

Objectives

  • Synthesize three novel triazole compounds labeled C1, C2, and C3.
  • Assess their in vitro activity against clinical isolates of Pythium insidiosum.
  • Evaluate their safety on human leukocytes by examining cell viability, morphology, and oxidative stress.
  • Predict pharmacokinetic and toxicological properties (ADMET) using computational tools.

Methods

  • Synthesis: Chemical synthesis of the three triazole derivatives C1, C2, and C3 (specific synthetic routes are implied but not detailed in the abstract).
  • In vitro susceptibility testing: Fifteen different isolates of P. insidiosum were tested to determine:
    • Minimum Inhibitory Concentration (MIC): lowest concentration that inhibits visible growth.
    • Minimum Oomicidal Concentration (MOC): lowest concentration that kills the organism.
  • Leukocyte toxicity testing: Human leukocytes were exposed to the compounds to measure:
    • Cell viability (survival and metabolic activity).
    • Morphological changes (microscopic examination for damage or alterations).
    • Oxidative stress endpoints (measurement of reactive oxygen species or related biomarkers to assess potential pro-oxidant effects).
  • In silico ADMET prediction: Using the pkCSM platform to predict:
    • Absorption, distribution, metabolism, excretion profiles.
    • Potential toxicity risks.
  • Comparison: The properties of the novel compounds were compared to fluconazole, a known antifungal triazole drug.

Results

  • Antifungal Activity:
    • C1 and C2 showed anti-Pythium activity with MIC and MOC values ranging from 2 to 64 µg/mL.
    • C3 had MIC values of 4-64 µg/mL and MOC values of 8-64 µg/mL, indicating slightly less potency compared to C1 and C2.
  • Leukocyte Toxicity:
    • None of the three compounds caused a significant decrease in leukocyte viability.
    • No observable morphological changes were noted in treated leukocytes, indicating low cytotoxicity.
    • The compounds did not induce oxidative stress or a pro-oxidant effect, an important safety parameter.
  • ADMET Profiling:
    • Predicted pharmacokinetic and toxicological parameters were similar to fluconazole, suggesting the compounds may have favorable drug-like properties.
    • This indicates reasonable absorption and distribution potential, metabolism pathways, and low toxicity risks, though experimental validation is needed.

Conclusions

  • The study successfully identified three novel triazole compounds with inhibitory and lethal activity against Pythium insidiosum at concentrations that are not toxic to human white blood cells.
  • These compounds have promising profiles that could make them candidates for further drug development against pythiosis, addressing the current lack of effective treatments.
  • In silico ADMET predictions reinforce the potential for these compounds to be developed into therapeutics, but further in vivo studies and clinical evaluations are necessary.

Significance and Future Directions

  • This research opens a new avenue for treatment options targeting pythiosis, a disease with limited therapeutic choices.
  • The novel triazole compounds warrant further investigation through animal models and clinical trials to validate efficacy and safety.
  • Additional studies could include exploring their mechanism of action, pharmacodynamics, and potential resistance development.

Cite This Article

APA
Fernandes CM, Prestes AS, Ianiski LB, Maciel AF, Noro BG, da Silva FD, Vizzotto BS, Botton SA, Schumacher RF, Pereira DIB, Barbosa NV. (2024). Anti-Pythium insidiosum activity of three novel triazole compounds: synthesis, pharmacokinetic and toxicological parameters. Braz J Microbiol, 56(1), 331-340. https://doi.org/10.1007/s42770-024-01572-y

Publication

Brazilian journal of microbiology : [publication of the Brazilian Society for Microbiology]
ISSN: 1678-4405
NlmUniqueID: 101095924
Country: Brazil
Language: English
Volume: 56
Issue: 1
Pages: 331-340

Researcher Affiliations

Fernandes, Carolina Martins
  • Department of Biochemistry and Molecular Biology, Federal University of Santa Maria, Santa Maria, RS, Brazil.
Prestes, Alessandro de Souza
  • Department of Biochemistry and Molecular Biology, Federal University of Santa Maria, Santa Maria, RS, Brazil.
Ianiski, Lara Baccarin
  • Post Graduate Program in Pharmaceutical Sciences, Federal University of Santa Maria, Santa Maria, RS, Brazil.
Maciel, Aline Fontanella
  • Post Graduate Program in Pharmaceutical Sciences, Federal University of Santa Maria, Santa Maria, RS, Brazil.
Noro, Bruna Godoy
  • Molecular Biology Laboratory, Franciscan University, Santa Maria, RS, Brazil.
da Silva, Fernanda D'Avila
  • Department of Biochemistry and Molecular Biology, Federal University of Santa Maria, Santa Maria, RS, Brazil.
Vizzotto, Bruno Stefanello
  • Molecular Biology Laboratory, Franciscan University, Santa Maria, RS, Brazil.
Botton, Sônia de Avila
  • Post Graduate Program in Pharmaceutical Sciences, Federal University of Santa Maria, Santa Maria, RS, Brazil.
Schumacher, Ricardo Frederico
  • Department of Chemistry, Federal University of Santa Maria, Santa Maria, RS, Brazil.
Pereira, Daniela Isabel Brayer
  • Department of Microbiology and Pathology, Federal University of Pelotas, Pelotas, RS, Brazil.
Barbosa, Nilda Vargas
  • Department of Biochemistry and Molecular Biology, Federal University of Santa Maria, Santa Maria, RS, Brazil. nvbarbosa@yahoo.com.br.

MeSH Terms

  • Triazoles / pharmacology
  • Triazoles / chemical synthesis
  • Triazoles / pharmacokinetics
  • Triazoles / toxicity
  • Triazoles / chemistry
  • Humans
  • Microbial Sensitivity Tests
  • Pythium / drug effects
  • Leukocytes / drug effects
  • Cell Survival / drug effects
  • Pythiosis / drug therapy
  • Animals
  • Antifungal Agents / pharmacology
  • Antifungal Agents / chemical synthesis
  • Antifungal Agents / pharmacokinetics

Conflict of Interest Statement

Declarations. Ethical approval: Protocol was approved by the Ethics Committee for Research with Human from Federal University of Santa Maria (protocol number 67825122.1.0000.5346). Competing interests: The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

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