Ergosterol Biosynthesis Inhibitors Research Articles

Biofumigant potential and inhibition mechanism of Trichoderma asperelloides SKRU-01 volatile organic compounds for controlling aflatoxigenic Aspergillus parasiticus and Aspergillus flavus in stored peanuts

Microbial volatile organic compounds (VOCs) have emerged as promising alternatives to synthetic fungicides for managing stored grain. In this study, the inhibitory effects of Trichoderma asperelloides SKRU-01 VOCs on hyphal growth and spore germination of Aspergillus parasiticus TISTR 3276 and A. flavus PSRDC-4 were investigated. The results demonstrated significant suppression of hyphal growth and spore germination in both strains by SKRU-01 VOCs. By using solid-phase microextraction (SPME) coupled with gas chromatography-mass spectrometry (GC-MS), the SKRU-01 VOCs were identified to contain 23 compounds, with acetophenone being predominant (14.41%). Antifungal experiments revealed that acetophenone, at a minimum inhibitory concentration (MIC) of 1.0 μL/mL (solid media assay) or 1.0 μL/L (vapor phase assay), completely inhibited the growth of both strains. Moreover, the application of SKRU-01 VOCs effectively reduced mold density and aflatoxin B1 (AFB1) production in peanuts and also exhibited superior efficacy compared to acetophenone at a 1/2 MIC concentration. Acetophenone exhibited inhibitory effects on both fungal strains through multiple mechanisms of action. This included its ability to inhibit ergosterol biosynthesis, reduce the aflatoxin inducer methylglyoxal, impair antioxidant defense enzymes, and disrupt non-enzymatic defense molecules. Overall, this research highlights the biofumigant potential of SKRU-01 VOCs in stored grain management and provides novel insights into their antifungal mechanism against A. parasiticus and A. flavus. Furthermore, these findings contribute to the development of sustainable strategies for grain preservation and underscore the broader applications of microbial VOCs in agricultural and food safety practices.

Synthesis, Antifungal Ergosterol Inhibition, Antibiofilm Activities, and Molecular Docking on β-Tubulin and Sterol 14-Alpha Demethylase along with DFT-Based Quantum Mechanical Calculation of Pyrazole Containing Fused Pyridine-Pyrimidine Derivatives.

Multidrug-resistant fungal infections have become much more common in recent years, especially in immune-compromised patients. Therefore, researchers and pharmaceutical professionals have focused on the development of novel antifungal agents that can tackle the problem of resistance. In continuation to this, a novel series of pyrazole-bearing pyrido[2,3-d]pyrimidine-2,4(1H,3H)-dione derivatives (4a-4o) have been developed. These compounds have been screened against Candida albicans, Aspergillus niger, and Aspergillus clavatus. The synthesized compounds were characterized by well-known spectroscopic techniques, i.e., IR, 1H NMR, 13C NMR, and mass spectrometry. In vitro antifungal results revealed that compound 4n showed activity against C. albicans having MIC value of 200 μg/mL. To know the plausible mode of action, the active derivatives were screened for anti-biofilm and ergosterol biosynthesis inhibition activities. The compounds 4h, 4j, 4k, and 4n showed greater ergosterol biosynthesis inhibition than the control DMSO. To comprehend how molecules interact with the receptor, studies of molecular docking of 4k and 4n have been performed on the homology-modeled protein of β-tubulin. The molecular docking revealed that the active compounds 4h, 4j, 4k, 4l, and 4n interacting with the active site amino acid of sterol 14-alpha demethylase (PDB ID: 5v5z) indicate one of the possible modes of action of ergosterol inhibition activity. The synthesized compounds 4c, 4e, 4h, 4i, 4j, 4k, 4l, and 4n inhibited biofilm formation and possessed the potential for anti-biofilm activity. DFT-based quantum mechanical calculations were carried out to optimize, predict, and compare the vibration modes of the molecule 4a.

Exploration of Baicalein-Core Derivatives as Potent Antifungal Agents: SAR and Mechanism Insights.

Baicalein (BE), the major component of Scutellaria Baicalensis, exhibited potently antifungal activity against drug-resistant Candida albicans, and strong inhibition on biofilm formation. Therefore, a series of baicalein-core derivatives were designed and synthesized to find more potent compounds and investigate structure-activity relationship (SAR) and mode of action (MoA). Results demonstrate that A4 and B5 exert a more potent antifungal effect (MIC80 = 0.125 μg/mL) than BE (MIC80 = 4 μg/mL) when used in combination with fluconazole (FLC), while the MIC80 of FLC dropped from 128 μg/mL to 1 μg/mL. SAR analysis indicates that the presence of 5-OH is crucial for synergistic antifungal activities, while o-dihydroxyls and vic-trihydroxyls are an essential pharmacophore, whether they are located on the A ring or the B ring of flavonoids. The MoA demonstrated that these compounds exhibited potent antifungal effects by inhibiting hypha formation of C. albicans. However, sterol composition assay and enzymatic assay conducted in vitro indicated minimal impact of these compounds on sterol biosynthesis and Eno1. These findings were further confirmed by the results of the in-silico assay, which assessed the stability of the complexes. Moreover, the inhibition of hypha of this kind of compound could be attributed to their effect on the catalytic subunit of 1,3-β-d-glucan synthase, 1,3-β-d-glucan-UDP glucosyltransferase and glycosyl-phosphatidylinositol protein, rather than inhibiting ergosterol biosynthesis and Eno1 activity by Induced-Fit Docking and Molecular Dynamics Simulations. This study presents potential antifungal agents with synergistic effects that can effectively inhibit hypha formation. It also provides new insights into the MoA.

Angelica archangelica essential oil loaded chitosan nanoemulsion as edible coating for preservation of table grape fruit against Botrytis cinerea contamination and storage quality deterioration

The present study deals with the encapsulation of Angelica archangelica essential oil into chitosan nanoemulsion (AAEO-NE) and evaluation of its effectiveness as novel coating agent with innovative controlled release delivery system for protection of table grape (Vitis vinifera L.) fruit against Botrytis cinerea contamination, and quality deterioration over 30 d of storage (25 ± 2 °C, Relative humidity 85%). The AAEO-NE was characterized through dynamic light scattering (DLS), scanning electron microscopy (SEM), and atomic force microscopy (AFM). The mean particle size was tended to increase from 53.69 to 98.63 nm, while ζ-potential was decreased from + 32.3 to + 23.5 mV. Fourier transform infrared spectroscopy (FTIR), and X-ray diffractometry (XRD) analyses suggested intermolecular interaction followed by effective encompassment of AAEO into chitosan nanomatrix. In vitro release study confirmed the biphasic and controlled delivery of AAEO, essentially required for its long term effectiveness in view of maintaining postharvest quality of the treated fruit. The antifungal efficacy of unencapsulated AAEO against B. cinerea was found as 5.5 g L−1, and noticed to be enhanced (2.5 g L−1) after encapsulation into chitosan nanoemulsion. The inhibition of ergosterol biosynthesis followed by subsequent enhancement of ions leakage after treatment of B. cinerea cells with AAEO-NE suggested plasma membrane as major site of antifungal activity. During in vivo investigation, AAEO-NE coating inhibited the contamination of B. cinerea and maintained the quality attributes like weight, titrable acidity, total soluble solids, phenolic content, and pH of table grape fruit. Additionally, the preservation of enzymatic antioxidants, reduced respiration rate, and better sensory qualities of coated table grape fruit recommend the functional potentiality of chitosan as biocompatible wall matrix to encapsulate AAEO and application as novel nano-green smart coating based fungitoxicant and shelf-life enhancer in fruit and agricultural industries.

THE INFLUENCE OF MEDICINAL AND AROMATIC PLANTS EXTRACTS ON THE ANTIFUNGAL EFFECT OF FLUCONAZOLE REGARDING C. ALBICANS AND C. TROPICALIS ISOLATED FROM PATIENTS WITH DENTURE STOMATITS

Candida albicans is one of the most common and virulent representatives of Candida fungi which causes fungal lesions of the mucous membranes predominantly in people with compromised immune system. Candida resistance to classical antifungals such as polyenes, azoles, allylamines is a serious practical problem. Imidazole and triazole are the most commonly used antifungal agents for candidiasis today. Their mechanism of action lies in the inhibition of ergosterol biosynthesis which is indispensable for maintaining the structural integrity of fungal cell membranes. Activation of elimination systems is often associated with changes in the structure of fungal membranes, leading to a decrease in the supply of azoles into the fungal cell Screening of antifungal activity and synergism of antifungal activity of 166 aqueous-ethanol plant extracts with Fluconazole in relation to 2 clinical strains of Candida fungi (C.) (C. albicans and C. tropicalis) with different mechanisms of azole resistance isolated from patients with prosthetic stomatitis was conducted by means of agar diffusion method. The expressed direct antifungal activity of extracts of Calendula officinalis L. inflorescences, flowers of Limonium platyphyllum Lincz., flowers of Grataegus monogyna Jacq., roots of Potentilla repens L., the grass of Melilotus albus Medik, leaves of Peucedanum ruthenicum Bieb, leaves of Rhus coriaria L. and aerial part of Polythridum commune Hedw was determined in reference to the tested strains. Synergism of antifungal action with 1/4, 1/8 and 1/32 of Fluconazole MIC (minimal inhibiting concentration) in relation to both test strains of yeast-like Candida fungi was demonstrated by the flowers of Calendula officinalis L, aerial part of Melilotus albus Medik. The main purpose of the screening was to discriminate extracts in order to select those that would be promising for further testing by a more accurate сheckerboard titration method. The efflux mechanism of resistance to triazoles (in C. tropicalis test strain) is more susceptible to modification by biologically active compounds of medicinal plants. Regarding C. albicans test strain which combines the efflux mechanism of resistance with mutation(s) of ergosterol biosynthesis enzymes, the modifying effect of plant extracts is much less frequent and less pronounced. Therefore, it may be assumed that mutations of ergosterol biosynthesis enzymes provide high stability of the phenotypic manifestation of Candida azole resistance. Should that be the case, determining the azole resistance mechanism of Candida clinical isolates may be important for clinical practice. Combination therapy is more beneficial than monotherapy because it may provide more effective destruction or suppression of pathogens. The combined action of synergistic agents is quite easy to achieve in case of localized superficial lesions (which occur on the mucous membranes of the prosthetic bed in candidal prosthetic stomatitis): for example, using a systemic antifungal agent and a topical agent that contains a modifier of resistance of plant origin fungi. Synergistic interactions will increase the therapeutic efficacy of drugs, reduce the likelihood of resistance occurrence or further development, and will reduce dose-related toxicity.

The Zanthoxylum armatum fruit's oil exterminates Candida cells by inhibiting ergosterol biosynthesis without generating reactive oxygen species.

Candida spp. is a significant cause of topical and fungal infections in humans. In addition to Candida albicans, many non-albicans species such as C. krusei, C. glabrata, C. parapsilosis, C. tropicalis, C. guilliermondii cause severe infections. The main antifungal agents belong to three different classes, including azoles, polyenes, and echinocandins. However, resistance to all three categories of drugs has been reported. Therefore, there is an urgent need to search for other alternatives with antifungal activity. Many herbal extracts and compounds from natural sources show excellent antifungal activity. In this study, we used an oil extract from the fruits of Zanthoxylum armatum, which showed significant antifungal activity against various Candida spp. by two different methods-minimum inhibitory concentration (MIC) and agar diffusion. In addition, we attempted to explore the possible mechanism of action in C. albicans. It was found that the antifungal activity of Z. armatum oil is fungicidal and involves a decrease in the level of ergosterol in the cell membrane. The decrease in ergosterol level resulted in increased passive diffusion of a fluorescent molecule, rhodamine6G, across the plasma membrane, indicating increased membrane fluidity. The oil-treated cells showed decreased germ tube formation, an important indicator of C. albicans' virulence. The fungal cells also exhibited decreased attachment to the buccal epithelium, the first step toward invasion, biofilm formation, and damage to oral epithelial cells. Interestingly, unlike most antifungal agents, in which the generation of reactive oxygen species is responsible for killing, no significant effect was observed in the present study.

DVL, lectin from Dioclea violacea seeds, has multiples mechanisms of action against Candida spp via carbohydrate recognition domain

Lectins are proteins of non-immunological origin with the ability to bind to carbohydrates reversibly. They emerge as an alternative to conventional antifungals, given the ability to interact with carbohydrates in the fungal cell wall inhibiting fungal growth. The lectin from D. violacea (DVL) already has its activity described as anti-candida in some species. Here, we observed the anti-candida effect of DVL on C. albicans, C. krusei and C. parapsilosis and its multiple mechanisms of action toward the yeasts. Additionally, it was observed that DVL induces membrane and cell wall damage and ROS overproduction. DVL was also able to cause an imbalance in the redox system of the cells, interact with ergosterol, inhibit ergosterol biosynthesis, and induce cytochrome c release from the mitochondrial membrane. These results endorse the potential application of DVL in developing a new antifungal drug to fight back against fungal resistance.

A semi purified hydroalcoholic fraction from Caesalpinia bonduc seeds causes ergosterol biosynthesis inhibition in Candida albicans resulting in cell membrane damage.

Candida species are currently developing resistance to prevailing commercially available drugs, which raises an instantaneous need to discover novel antifungals. To cope with this shocking situation, phytochemicals are the richest, safest, and most potent source of excellent antimicrobials with broad-spectrum activity. The aim of the current study is to explore the anticandidal potential of the various fractions purified from the hydroalcoholic extract of C. bonduc seed. Out of five fractions purified from the hydroalcoholic extract, fraction 3 (Fr. 3) recorded the best activity against C. albicans (8μg/mL) and thus this species was chosen for further mechanism of action studies. The phytochemical examination reveals that Fr. 3 was found to contain steroids and triterpenoids. This was further supported by LC-QTOF-MS and GCMS analyses. Our findings show that Fr. 3 targets the ergosterol biosynthesis pathway in C. albicans by inhibiting the lanosterol 14-α demethylase enzyme and downregulating expression of its related gene ERG11. Molecular docking outcomes disclosed favorable structural dynamics of the compounds, implying that the compounds present in Fr. 3 would be able to successfully bind to the lanosterol 14-α demethylase, as evidenced by the docked compounds' strong interaction with the target enzyme's amino acid residues. Considering virulence factors, the Fr. 3 recorded significant antibiofilm activity as well as germ-tube reduction potential. Furthermore, Fr. 3 enhances the production of intracellular reactive oxygen species (ROS). This suggests that the antifungal activity of Fr. 3 was associated with membrane damage and the induction of ROS production, resulting in cell death. Fluorescence microscopic analysis of PI stained Candida further showed changes in the plasma membrane permeability, which causes severe loss of intracellular material and osmotic balance. This was demonstrated by the potassium ion leakage and release of genetic materials. Finally, the erythrocyte lysis assay confirmed the low cytotoxicity of Fr. 3. Both in silico and in vitro results suggest that Fr. 3 has the potential to propel forward novel antifungal drug discovery programmes.

Antifungal activity of Allyl isothiocyanate by targeting signal transduction pathway, ergosterol biosynthesis, and cell cycle in Candida albicans.

In recent years, the inclusion of Candida albicans on the list of infections that pose a threat due to drug resistance has urged researchers to look into cutting-edge and effective antifungal medications. In this regard, the current study investigated the probable mode of action of allyl isothiocyanate (AITC) against Candida albicans. In this study, planktonic assay, germ tube inhibition assay, adhesion, and biofilm formation assay were performed to check the growth and virulence factors. Furthermore, ergosterol assay, reactive oxygen production analysis, cell cycle analysis, and quantitative real-time polymerase chain reaction analysis were performed with the aim of finding the mode of action. A biomedical model organism, like a silkworm, was used in an in vivo study to demonstrate AITC anti-infective ability against C. albicans infection. Allyl isothiocyanate completely inhibited ergosterol biosynthesis in C. albicans at 0.125 mg/ml. Allyl isothiocyanate produces reactive oxygen species in both planktonic and biofilm cells of C. albicans. At 0.125 mg/ml concentration, AITC arrested cells at the G2/M phase of the cell cycle, which may induce apoptosis in C. albicans. In quantitative real-time polymerase chain reaction analysis, it was found that AITC inhibited virulence factors, like germ tube formation, at 0.125 mg/ml concentration by downregulation of PDE2, CEK1, TEC1 by 2.54-, 1.91-, and 1.04-fold change, respectively, and upregulation of MIG1, NRG1, and TUP1 by 9.22-, 3.35-, and 7.80-fold change, respectively. The in vivo study showed that AITC treatment successfully protected silkworms against C. albicans infections and increased their survival rate by preventing internal colonization by C. albicans. In vitro and in vivo studies revealed that AITC can be an alternative therapeutic option for the treatment of C. albicans infection.

Newly Discovered Antimicrobial Peptide Scyampcin44-63 from Scylla paramamosain Exhibits a Multitargeted Candidacidal Mechanism In Vitro and Is Effective in a Murine Model of Vaginal Candidiasis.

The emergence of azole-resistant and biofilm-forming Candida spp. contributes to the constantly increasing incidence of vulvovaginal candidiasis. It is imperative to explore new antifungal drugs or potential substituents, such as antimicrobial peptides, to alleviate the serious crisis caused by resistant fungi. In this study, a novel antimicrobial peptide named Scyampcin44-63 was identified in the mud crab Scylla paramamosain. Scyampcin44-63 exhibited broad-spectrum antimicrobial activity against bacteria and fungi, was particularly effective against planktonic and biofilm cells of Candida albicans, and exhibited no cytotoxicity to mammalian cells (HaCaT and RAW264.7) or mouse erythrocytes. Transcriptomic analysis revealed four potential candidacidal modes of Scyampcin44-63, including promotion of apoptosis and autophagy and inhibition of ergosterol biosynthesis and the cell cycle. Further study showed that Scyampcin44-63 caused damage to the plasma membrane and induced apoptosis and cell cycle arrest at G2/M in C. albicans. Scanning and transmission electron microscopy demonstrated that Scyampcin44-63-treated C. albicans cells were deformed with vacuolar expansion and destruction of organelles. In addition, C. albicans cells pretreated with the autophagy inhibitor 3-methyladenine significantly delayed the candidacidal effect of Scyampcin44-63, suggesting that Scyampcin44-63 might contribute to autophagic cell death. In a murine model of vulvovaginal candidiasis, the fungal burden of vaginal lavage was significantly decreased after treatment with Scyampcin44-63.

The proteasome regulator Rpn4 controls antifungal drug tolerance by coupling protein homeostasis with metabolic responses to drug stress.

Fungal pathogens overcome antifungal drug therapy by classic resistance mechanisms, such as increased efflux or changes to the drug target. However, even when a fungal strain is susceptible, trailing or persistent microbial growth in the presence of an antifungal drug can contribute to therapeutic failure. This trailing growth is caused by adaptive physiological changes that enable the growth of a subpopulation of fungal cells in high drug concentrations, in what is described as drug tolerance. Mechanistically, antifungal drug tolerance is incompletely understood. Here we report that the transcriptional activator Rpn4 is important for drug tolerance in the human fungal pathogen Candida albicans. Deletion of RPN4 eliminates tolerance to the commonly used antifungal drug fluconazole. We defined the mechanism and show that Rpn4 controls fluconazole tolerance via two target pathways. First, Rpn4 activates proteasome gene expression, which enables sufficient proteasome capacity to overcome fluconazole-induced proteotoxicity and the accumulation of ubiquitinated proteins targeted for degradation. Consistently, inhibition of the proteasome with MG132 eliminates fluconazole tolerance and resistance, and phenocopies the rpn4Δ/Δ mutant for loss of tolerance. Second, Rpn4 is required for wild type expression of the genes required for the synthesis of the membrane lipid ergosterol. Our data indicates that this function of Rpn4 is required for mitigating the inhibition of ergosterol biosynthesis by fluconazole. Based on our findings, we propose that Rpn4 is a central hub for fluconazole tolerance in C. albicans by coupling the regulation of protein homeostasis (proteostasis) and lipid metabolism to overcome drug-induced proteotoxicity and membrane stress.

Biosensor-Enabled Discovery of CaERG6 Inhibitors and Their Antifungal Mode of Action against Candida albicans.

Fungal infections caused by opportunistic pathogens, such as Candida albicans, are generally underappreciated by the public in spite of their high mortality rates. Antifungal arsenals are extremely limited. Herein, based on biosynthetic pathway comparison and functional characterization, CaERG6, a crucial sterol 24-C-methyltransferase involved in the biosynthesis of ubiquitous ergosterol in C. albicans, was set up as an antifungal target. CaERG6 inhibitors were identified from the in-house small-molecule library by a biosensor-based high-throughput screening. The CaERG6 inhibitor NP256 (palustrisoic acid E) is a potential antifungal natural product that acts by inhibiting ergosterol biosynthesis, downregulating the gene expression level in hyphal formation, blocking biofilm formation, and disrupting morphological transition in C. albicans. NP256 enhances C. albicans susceptibility to some known antifungals significantly. The present study demonstrated the CaERG6 inhibitor NP256 as a potential class of antifungal compound for monotherapy or combinatory therapy.

In silico and in vitro investigation of the antifungal activity of trimetallic Cu–Zn-magnetic nanoparticles against Fusarium oxysporum with stimulation of the tomato plant's drought stress tolerance response

Fusarium oxysporum is the fungus responsible for Fusarium wilt. Tomatoes and other plants acquire Fusarium wilt through their root systems. Occasionally, fungicides applied to the soil are used to combat the disease; however, some strains have developed resistance. Carboxymethyl cellulose (CMC) trimetallic magnetic zinc and copper nanoparticles CMC-Cu-Zn-FeMNPs are one of the most promising antifungal agents against a wide range of fungi. One of the most important aspects of using magnetic nanoparticles is their ability to target cells, which confirms the drug's potent fungicidal activity. Using a UV-spectrophotometer, the characterization of synthesized CMC-Cu-Zn-FeMNPs revealed four peaks at226,271, 321 and 335 nm, as well as spherical nanoparticles with a mean size of 5.905 nm and a surface potential of −61.7 mv. In this study, CMC-Cu-Zn-FeMNPs were used to inhibit the growth of F. oxysporum by interfering with the ergosterol production metabolic pathway. Molecular docking experiments demonstrated that the nanoparticles were able to bind to sterol 14-alpha demethylase responsible for inhibiting ergosterol biosynthesis. Real-time PCR analysis showed that the nanoparticles upregulated tomato plants and other assessed parameters under drought stress and downregulated the velvet complex and virulence factors of F. oxysporum on plants. The study results indicate that CMC-Cu-Zn-FeMNPs may be a promising and eco-friendly solution with low potential of accumulation and easy to collected alternative to conventional chemical pesticides that can have negative impacts on the environment and human health. Furthermore, it could provide a sustainable solution for managing Fusarium wilt disease, which can significantly reduce tomato yield and quality.

In vivo and in vitro evaluation of pulmonary administration of itraconazole nanostructured lipid carriers for pulmonary aspergillosis

Objective: Pulmonary aspergillosis which is a secondary complication of fungal pneumonia is widely considered to have an increasing incidence and high mortality. Itraconazole (Itz) can inhibit ergosterol biosynthesis to treat pulmonary aspergillosis. Nevereless Itz's clinical application is limited because of its poor water solubility low oral bioavailability and systemic hepatotoxicity. In this study Itz-loaded nanostructured lipid carriers (Itz-NLCs) were developed to improve the in vitro permeability and bioavailability of Itz via pulmonary administration. Methods: Itz-NLCs were prepared by the emulsification-evaporation method using oleic acid and glycerol monostearate as liquid and solid lipids respectively. Results: The Itz-NLCs were optimized with tiny particle size uniform distribution and excellent entrapment efficiency (EE 97.57% ± 0.45%). A Xenopus alveolar membrane was used in the permeation study and the cumulative permeation percentage of Itz was 10% for Itz-NLCs at 8 h which was 2.50-fold higher than that for Itz suspensions (4% P < 0.001). A rabbit pharmacokinetic investigation revealed that Itz-NLCs have an 83.05% absolute bioavailability after intratracheal instillation. Conclusions: The purpose of Itz-NLCs is to enhance the bioavailability and permeability of Itz in vitro for administration via the lungs

Synthesis of 2-(6-substituted quinolin-4-yl)-1-(4-aryl-1H-1,2,3-triazol-1-yl) propan-2-ol as potential antifungal and antitubercular agents

The intense pressure of resistance to antibiotics leads the whole world's health security to drastic conditions due to Mycobacterial and fungal infections. To search for potent antifungal and antitubercular lead compounds, the synthesis of a new series of 2-(6-substituted quinolin-4-yl)-1-(4-aryl-1H-1,2,3-triazol-1-yl)propan-2-ol, (9a-l) derivatives by click reaction of 1-azido-2-(quinolin-4-yl)propan-2-ol, (7a-d) and substituted ethynylbenzene (8a-c) is reported. The structures of the novel synthesized 2-(6-substituted quinolin-4-yl)-1-(4-aryl-1H-1,2,3-triazol-1-yl)propan-2-ol derivatives were characterized by 1H NMR, 13C NMR spectroscopic and Mass spectrometric analysis. The synthesized compounds were examined for antifungal activity against C. albicans (NCIM 3100), A. niger (ATCC 504) and antitubercular activity against Mycobacterium tuberculosis H37Rv (MTB) (ATCC 25177). Eleven derivatives showed good to excellent antifungal activity against A. niger with MIC 7.81–62.5 ​μg/mL. To study the plausible antifungal mode of action, compounds 9e, 9f, 9g, 9i, and 9k were further evaluated for the ergosterol biosynthesis inhibition activity. All these compounds showed ergosterol biosynthesis inhibition activity. All twelve derivatives of 2-(6-substituted quinolin-4-yl)-1-(4-aryl-1H-1,2,3-triazol-1-yl)propan-2-ol, (9a-l) showcased excellent antitubercular activity against M. tuberculosis H37Rv with MIC 1.6–3.2 ​μg/mL. No significant cytotoxic activity was displayed by these novel derivatives against l929 mouse fibroblast cells. The potential antifungal and antitubercular activities compelled the conclusion that the 2-(6-substituted quinolin-4-yl)-1-(4-aryl-1H-1,2,3-triazol-1-yl)propan-2-ol derivatives could assist in the development of lead compounds for the treatment of tuberculosis and fungal infections.

Synthesis of novel thiazol-2(3H)-imine derivatives as ergosterol biosynthesis inhibitors, and elucidation of their structures using a 2D NMR technique

In this study, new imidazole-2,3-dihydrothiazole derivatives were synthesized.

Synthesis, antimicrobial and ergosterol biosynthesis inhibition activity of clubbed 1,1′-biphenyl-pyrazole derivatives

Synthesis, antimicrobial and ergosterol biosynthesis inhibition activity of clubbed 1,1′-biphenyl-pyrazole derivatives.

Antifungal and antiaflatoxigenic mechanism activity of freeze-dried culture filtrate ofStreptomyces philanthiRL-1-178 on the two aflatoxigenic fungi and identification of its active component

The study reports the antifungal and antiaflatoxigenic mechanism activity of freeze-dried culture filtrate of Streptomyces philanthi RL-1-178 (DCF RL-1-178) against two aflatoxigenic strains (Aspergillus parasiticus and A. flavus) and identification of its active component. Significant inhibition in ergosterol biosynthesis by the DCF RL-1-178 appeared on the plasma membrane. Moreover, the DCF RL-1-178 showed dose-dependent inhibition of methylglyoxal (MG) (an aflatoxin inducer) biosynthesis and exhibited a novel antiaflatoxigenic action mechanism. Significant impairments in enzymatic [superoxide dismutase (SOD) and catalase (CAT)] and nonenzymatic [oxidized and reduced glutathione (GSH) and ratio of oxidized and reduced glutathione (GSSG)] anti-oxidative defense molecules were observed in the two aflatoxigenic cells. The active component of the DCF RL-1-178 was identified as natamycin. The natamycin exhibited against A. parasiticus and A. flavus with the minimum inhibitory concentration (MIC) values of 0.5 and 1.0 µg ml-1, respectively, while the minimum fungicidal concentration values were the same (4.0 µg ml-1). The DCF RL-1-178 containing natamycin exhibited the following effects: (1) inhibition of cellular ergosterol biosynthesis on plasma membrane, (2) reduction in MG (aflatoxin inducer) confirmed novel antiaflatoxigenic mechanism of action, and (3) caused remarkable debasement in antioxidant defense enzymes (SOD and CAT) and nonenzymatic defense molecules (GSH and GSSG) revealing biochemical mechanism of action.

Ipconazole Disrupts Mitochondrial Homeostasis and Alters GABAergic Neuronal Development in Zebrafish.

Ipconazole, a demethylation inhibitor of fungal ergosterol biosynthesis, is widely used in modern agriculture for foliar and seed treatment, and is authorized for use in livestock feed. Waste from ipconazole treatment enters rivers and groundwater through disposal and rain, posing potential toxicity to humans and other organisms. Its metabolites remain stable under standard hydrolysis conditions; however, their neurodevelopmental toxicity is unknown. We investigated the potential neurodevelopmental toxicity of ipconazole pesticides in zebrafish (Danio rerio). Our behavioral monitoring demonstrated that the locomotive activity of ipconazole-exposed zebrafish larvae was reduced during early development, even when morphological abnormalities were undetected. Molecular profiling demonstrated that the mitochondrial-specific antioxidants, superoxide dismutases 1 and 2, and the genes essential for mitochondrial genome maintenance and functions were specifically reduced in ipconazole-treated (0.02 μg/mL) embryos, suggesting underlying ipconazole-driven oxidative stress. Consistently, ipconazole treatment substantially reduced hsp70 expression and increased ERK1/2 phosphorylation in a dose-dependent manner. Interrupted gad1b expression confirmed that GABAergic inhibitory neurons were dysregulated at 0.02 μg/mL ipconazole, whereas glutamatergic excitatory and dopaminergic neurons remained unaffected, resulting in an uncoordinated neural network. Additionally, ipconazole-treated (2 μg/mL) embryos exhibited caspase-independent cell death. This suggests that ipconazole has the potential to alter neurodevelopment by dysregulating mitochondrial homeostasis.

A Comprehensive in vitro and in silico Assessment on Inhibition of CYP51B and Ergosterol Biosynthesis by Eugenol in Rhizopus oryzae

Mucormycosis, also known as Zygomycosis, is a disease caused by invasive fungi, predominantly Rhizopus species belonging to the Order of Mucorales. Seeing from the chemistry perspective, heterocyclic compounds with an "azole" moiety are widely employed as antifungal agent for minimising the effect of mucormycosis as a prescribed treatment. These azoles serve as non-competitive inhibitors of fungal CYP51B by predominantly binding to its heme moiety, rendering its inhibition. However, long-term usage and abuse of azoles as antifungal medicines has resulted in drug resistance among certain fungal pathogens. Hence, there is an unmet need to find alternative therapeutic compounds. In present study, we used various in vitro tests to investigate the antifungal activity of eugenol against R. oryzae/R. arrhizus, including ergosterol quantification to test inhibition of ergosterol production mediated antifungal action. The minimum inhibitory concentration (MIC) value obtained for eugenol was 512μg/ml with reduced ergosterol concentration of 77.11 ± 3.25% at MIC/2 concentration. Further, the molecular interactions of eugenol with fungal CYP51B were meticulously studied making use of proteomics in silico study including molecular docking and molecular dynamics simulations that showed eugenol to be strongly interacting with heme in an identical fashion to that shown by azole drugs (in this case, clotrimazole was evaluated). This is the first of a kind study showing the simulation study of eugenol with CYP51B of fungi. This inhibition results in ergosterol synthesis and is also studied and compared with keeping clotrimazole as a reference.