Monomethylsulochrin isolated from biomass extract of Aspergillus sp. against Leishmania amazonensis: In vitro biological evaluation and molecular docking

Leishmaniasis represents a serious world health problem, with 1 billion people being exposed to infection and a broad spectrum of clinical manifestations with a potentially fatal outcome. Based on the limitations observed in the treatment of leishmaniasis, such as high cost, significant adverse effects, and the potential for drug resistance, the aim of the present study was to evaluate the leishmanicidal activity of the compounds pseurotin A and monomethylsulochrin isolated from the biomass extract of Aspergillus sp. The chromatographic profiles of the extract were determined by high-performance liquid chromatography coupled with a diode-array UV-Vis detector (HPLC-DAD-UV), and the molecular identification of the pseurotin A and monomethylsulochrin were carried out by electrospray ionization mass spectrometry in tandem (LC-ESI-MS-MS) and nuclear magnetic resonance (NMR). Antileishmanial activity was assayed against promastigote and intracellular amastigote of Leishmania amazonensis. As a control, cytotoxicity assays were performed in non-infected BALB/c peritoneal macrophages. Ultrastructural alterations in parasites were evaluated by transmission electron microscopy. Changes in mitochondrial membrane potential were determined by flow cytometry. Only monomethylsulochrin inhibited the promastigote growth (IC50 18.04 ± 1.11 µM), with cytotoxicity to peritoneal macrophages (CC50 5.09 91.63 ± 1.28 µM). Activity against intracellular amastigote forms (IC50 5.09 ± 1.06 µM) revealed an increase in antileishmanial activity when compared with promastigotes. In addition to a statistically significant reduction in the evaluated infection parameters, monomethylsulochrin altered the ultrastructure of the promastigote forms with atypical vacuoles, electron-dense corpuscles in the cytoplasm, changes at the mitochondria outer membrane and abnormal disposition around the kinetoplast. It was showed that monomethylsulochrin leads to a decrease in the mitochondrial membrane potential (25.9%, p = 0.0286). Molecular modeling studies revealed that monomethylsulochrin can act as inhibitor of sterol 14-alpha-demethylase (CYP51), a therapeutic target for human trypanosomiasis and leishmaniasis. Assessed for its drug likeness, monomethylsulochrin follows the Lipinski Rule of five and Ghose, Veber, Egan, and Muegge criteria. Furthermore, monomethylsulochrin can be used as a reference in the development of novel and therapeutically useful antileishmanial agents.

[1]  L. Goulart,et al.  Mitochondrial dysfunction on Leishmania (Leishmania) amazonensis induced by ketoconazole: insights into drug mode of action , 2022, Memorias do Instituto Oswaldo Cruz.

[2]  Fernando Almeida-Souza,et al.  Carajurin Induces Apoptosis in Leishmania amazonensis Promastigotes through Reactive Oxygen Species Production and Mitochondrial Dysfunction , 2022, Pharmaceuticals.

[3]  R. Menna-Barreto,et al.  Is the mitochondrion a promising drug target in trypanosomatids? , 2022, Memorias do Instituto Oswaldo Cruz.

[4]  Jeremy D. DeBarry,et al.  VEuPathDB: the eukaryotic pathogen, vector and host bioinformatics resource center , 2021, Nucleic Acids Res..

[5]  W. de Souza,et al.  Development of new dinuclear Fe(III) coordination compounds with in vitro nanomolar antitrypanosomal activity. , 2021, Dalton transactions.

[6]  Fernando Almeida-Souza,et al.  Antileishmanial Activity of Flavones-Rich Fraction From Arrabidaea chica Verlot (Bignoniaceae) , 2021, Frontiers in Pharmacology.

[7]  Fernando Almeida-Souza,et al.  Carajurin: a anthocyanidin from Arrabidaea chica as a potential biological marker of antileishmanial activity. , 2021, Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie.

[8]  C. R. Alves,et al.  Axenic amastigotes of Leishmania species as a suitable model for in vitro studies. , 2021, Acta Tropica.

[9]  D. C. Miguel,et al.  Dihydroartemisinin, an active metabolite of artemisinin, interferes with Leishmania braziliensis mitochondrial bioenergetics and survival , 2021, Parasitology Research.

[10]  Renata Mondêgo-Oliveira,et al.  Vernonia brasiliana (L.) Druce induces ultrastructural changes and apoptosis-like death of Leishmania infantum promastigotes. , 2020, Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie.

[11]  Hongxing Zhang,et al.  Molecular Dynamics Investigations of Binding Mechanism for Triazoles Inhibitors to CYP51 , 2020, Frontiers in Molecular Biosciences.

[12]  A. Heck,et al.  A new perspective on fungal metabolites: identification of bioactive compounds from fungi using zebrafish embryogenesis as read-out , 2019, Scientific Reports.

[13]  Jian Wang,et al.  Construction of antifungal dual-target (SE, CYP51) pharmacophore models and the discovery of novel antifungal inhibitors , 2019, RSC advances.

[14]  Fernando Almeida-Souza,et al.  Endlicheria bracteolata (Meisn.) Essential Oil as a Weapon Against Leishmania amazonensis: In Vitro Assay , 2019, Molecules.

[15]  A. L. Abreu-Silva,et al.  Carapa guianensis Aublet (Andiroba) Seed Oil: Chemical Composition and Antileishmanial Activity of Limonoid-Rich Fractions , 2018, BioMed research international.

[16]  A. L. Abreu-Silva,et al.  In vitro activity of Morinda citrifolia Linn. fruit juice against the axenic amastigote form of Leishmania amazonensis and its hydrogen peroxide induction capacity in BALB/c peritoneal macrophages , 2018, BMC Research Notes.

[17]  Torsten Schwede,et al.  SWISS-MODEL: homology modelling of protein structures and complexes , 2018, Nucleic Acids Res..

[18]  M. Monteiro,et al.  Phloroglucinol derivatives from Hypericum species trigger mitochondrial dysfunction in Leishmania amazonensis , 2018, Parasitology.

[19]  H. Schwalbe,et al.  Georatusin, a Specific Antiparasitic Polyketide-Peptide Hybrid from the Fungus Geomyces auratus. , 2018, Organic letters.

[20]  A. Converti,et al.  In vitro leishmanicidal, antibacterial and antitumour potential of anhydrocochlioquinone A obtained from the fungus Cochliobolus sp. , 2017, Journal of Biosciences.

[21]  Olivier Michielin,et al.  SwissADME: a free web tool to evaluate pharmacokinetics, drug-likeness and medicinal chemistry friendliness of small molecules , 2017, Scientific Reports.

[22]  N. Waterhouse,et al.  Measuring Mitochondrial Transmembrane Potential by TMRE Staining. , 2016, Cold Spring Harbor protocols.

[23]  A. L. Abreu-Silva,et al.  Ultrastructural Changes and Death of Leishmania infantum Promastigotes Induced by Morinda citrifolia Linn. Fruit (Noni) Juice Treatment , 2016, Evidence-based complementary and alternative medicine : eCAM.

[24]  M. Dos Santos,et al.  Benzophenone derivatives as cysteine protease inhibitors and biological activity against Leishmania(L.) amazonensis amastigotes. , 2015, Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie.

[25]  Douglas E. V. Pires,et al.  pkCSM: Predicting Small-Molecule Pharmacokinetic and Toxicity Properties Using Graph-Based Signatures , 2015, Journal of medicinal chemistry.

[26]  J. McKerrow,et al.  Targeting Ergosterol Biosynthesis in Leishmania donovani: Essentiality of Sterol 14alpha-demethylase , 2015, PLoS neglected tropical diseases.

[27]  D. Scariot,et al.  Cell death and ultrastructural alterations in Leishmania amazonensis caused by new compound 4-Nitrobenzaldehyde thiosemicarbazone derived from S-limonene , 2014, BMC Microbiology.

[28]  Xiaocui Liu,et al.  Antimicrobial Metabolites from the Endophytic Fungus Aspergillus sp. of Eucommia ulmoides , 2014, Chemistry of Natural Compounds.

[29]  R. S. Menna-Barreto,et al.  The Double-Edged Sword in Pathogenic Trypanosomatids: The Pivotal Role of Mitochondria in Oxidative Stress and Bioenergetics , 2014, BioMed research international.

[30]  W. de Souza,et al.  A novel alkyl phosphocholine-dinitroaniline hybrid molecule exhibits biological activity in vitro against Leishmania amazonensis. , 2013, Experimental parasitology.

[31]  M. Dos Santos,et al.  Synthesis and biological evaluation against Leishmania amazonensis of a series of alkyl-substituted benzophenones. , 2013, Bioorganic & medicinal chemistry.

[32]  B. Strukelj,et al.  Endophytic fungi: the treasure chest of antibacterial substances. , 2012, Phytomedicine : international journal of phytotherapy and phytopharmacology.

[33]  R. Menna-Barreto,et al.  Mitochondrial damage contribute to epigallocatechin-3-gallate induced death in Leishmania amazonensis. , 2012, Experimental parasitology.

[34]  Marcus D. Hanwell,et al.  Avogadro: an advanced semantic chemical editor, visualization, and analysis platform , 2012, Journal of Cheminformatics.

[35]  David M. Shackleford,et al.  Pharmacological Characterization, Structural Studies, and In Vivo Activities of Anti-Chagas Disease Lead Compounds Derived from Tipifarnib , 2012, Antimicrobial Agents and Chemotherapy.

[36]  W. de Souza,et al.  Tomatidine promotes the inhibition of 24-alkylated sterol biosynthesis and mitochondrial dysfunction in Leishmania amazonensis promastigotes , 2012, Parasitology.

[37]  W. Gerwick,et al.  Antiparasitic and Anticancer Constituents of the Endophytic Fungus Aspergillus sp. strain F1544 , 2012, Natural product communications.

[38]  P. Kaye,et al.  Leishmaniasis: complexity at the host–pathogen interface , 2011, Nature Reviews Microbiology.

[39]  M. Waterman,et al.  Sterol 14alpha-demethylase (CYP51) as a therapeutic target for human trypanosomiasis and leishmaniasis. , 2011, Current topics in medicinal chemistry.

[40]  J. C. Bakowska,et al.  Determination of Mitochondrial Membrane Potential and Reactive Oxygen Species in Live Rat Cortical Neurons , 2011, Journal of visualized experiments : JoVE.

[41]  E. E. Almeida-Amaral,et al.  Reactive Oxygen Species Production and Mitochondrial Dysfunction Contribute to Quercetin Induced Death in Leishmania amazonensis , 2011, PloS one.

[42]  R. Moo-Puc,et al.  Design, synthesis, and in vitro antiprotozoal, antimycobacterial activities of N-{2-[(7-chloroquinolin-4-yl)amino]ethyl}ureas. , 2010, Bioorganic & medicinal chemistry.

[43]  S. Sundar,et al.  Drug Resistance in Leishmaniasis , 2010, Journal of global infectious diseases.

[44]  C. Dardonville,et al.  New benzophenone-derived bisphosphonium salts as leishmanicidal leads targeting mitochondria through inhibition of respiratory complex II. , 2010, Journal of medicinal chemistry.

[45]  S. Sundar,et al.  Miltefosine in the treatment of leishmaniasis: Clinical evidence for informed clinical risk management , 2007, Therapeutics and clinical risk management.

[46]  G. Turner,et al.  Identification of a Hybrid PKS/NRPS Required for Pseurotin A Biosynthesis in the Human Pathogen Aspergillus fumigatus , 2007, Chembiochem : a European journal of chemical biology.

[47]  H. Nakhasi,et al.  Transcriptome analysis during the process of in vitro differentiation of Leishmania donovani using genomic microarrays , 2007, Parasitology.

[48]  P. Myler,et al.  Analysis of the Leishmania donovani transcriptome reveals an ordered progression of transient and permanent changes in gene expression during differentiation. , 2007, Molecular and biochemical parasitology.

[49]  R. Tan,et al.  Anti-Helicobacter pylori metabolites from Rhizoctonia sp. Cy064, an endophytic fungus in Cynodon dactylon. , 2004, Fitoterapia.

[50]  P Willett,et al.  Development and validation of a genetic algorithm for flexible docking. , 1997, Journal of molecular biology.

[51]  A. O. Feitosa,et al.  Study on Experimental Leishmanicidal Activity and in silico of Cytochalasin B , 2018 .

[52]  Fernando Almeida-Souza,et al.  In vitro evaluation of (-)α-bisabolol as a promising agent against Leishmania amazonensis. , 2015, Experimental parasitology.

[53]  A. O. Feitosa,et al.  Chemical constituents of Aspergillus sp EJC08 isolated as endophyte from Bauhinia guianensis and their antimicrobial activity. , 2013, Anais da Academia Brasileira de Ciencias.

[54]  P. Le Pape,et al.  Antileishmanial polyphenols from Garcinia vieillardii. , 2008, Fitoterapia.