ABSTRACT
The prevalence of Candida albicans infection has increased during the past few years, which contributes to the need for new, effective treatments due to the increasing concerns regarding antifungal drug toxicity and multidrug resistance. Butyl isothiocyanate (butylITC) is a glucosinolate derivative, and has shown a significant antifungal effect contrary to Candida albicans. Additionally, how butylITC affects the virulence traits of C. albicans and molecular mode of actions are not well known. Present study shows that at 17.36 mM concentration butylITC inhibit planktonic growth. butylITC initially slowed the hyphal transition at 0.542 mM concentration. butylITC hampered biofilm development, and inhibits biofilm formation at 17.36 mM concentration which was analysed using metabolic assay (XTT assay) and Scanning Electron Microscopy (SEM). In addition, it was noted that butylITC inhibits ergosterol biosynthesis. The permeability of cell membranes was enhanced by butylITC treatment. Moreover, butylITC arrests cells at S-phase and induces intracellular Reactive Oxygen Species (ROS) accumulation in C. albicans. The results suggest that butylITC may have a dual mode of action, inhibit virulence factors and modulate cellular processes like inhibit ergosterol biosynthesis, cell cycle arrest, induces ROS production which leads to cell death in C. albicans.
Subject(s)
Antifungal Agents , Biofilms , Candida albicans , Cell Membrane , Isothiocyanates , Oxidative Stress , Reactive Oxygen Species , Candida albicans/drug effects , Candida albicans/physiology , Biofilms/drug effects , Antifungal Agents/pharmacology , Isothiocyanates/pharmacology , Oxidative Stress/drug effects , Cell Membrane/drug effects , Cell Membrane/metabolism , Reactive Oxygen Species/metabolism , Microbial Sensitivity Tests , Cell Cycle/drug effects , Hyphae/drug effects , Hyphae/growth & development , Ergosterol/metabolismABSTRACT
Candida albicans is the primary etiological agent associated with candidiasis in humans. Unrestricted growth of C. albicans can progress to systemic infections in the worst situation. This study investigates the antifungal activity of Hydroxychloroquine (HCQ) and mode of action against C. albicans. HCQ inhibited the planktonic growth and yeast to hyphal form morphogenesis of C. albicans significantly at 0.5 mg/ml concentration. The minimum inhibitory concentrations (MIC50) of HCQ for C. albicans adhesion and biofilm formation on the polystyrene surface was at 2 mg/ml and 4 mg/ml respectively. Various methods, such as scanning electron microscopy, exploration of the ergosterol biosynthesis pathway, cell cycle analysis, and assessment of S oxygen species (ROS) generation, were employed to investigate HCQ exerting its antifungal effects. HCQ was observed to reduce ergosterol levels in the cell membranes of C. albicans in a dose-dependent manner. Furthermore, HCQ treatment caused a substantial arrest of the C. albicans cell cycle at the G0/G1 phase, which impeded normal cell growth. Gene expression analysis revealed upregulation of SOD2, SOD1, and CAT1 genes after HCQ treatment, while genes like HWP1, RAS1, TEC1, and CDC 35 were downregulated. The study also assessed the in vivo efficacy of HCQ in a mice model, revealing a reduction in the pathogenicity of C. albicans after HCQ treatment. These results indicate that HCQ holds for the development of novel antifungal therapies.
Subject(s)
Antifungal Agents , Biofilms , Candida albicans , Candidiasis , Hydroxychloroquine , Microbial Sensitivity Tests , Candida albicans/drug effects , Antifungal Agents/pharmacology , Animals , Biofilms/drug effects , Mice , Candidiasis/drug therapy , Candidiasis/microbiology , Hydroxychloroquine/pharmacology , Ergosterol/metabolism , Reactive Oxygen Species/metabolism , Antimalarials/pharmacology , Hyphae/drug effects , Hyphae/growth & development , Gene Expression Regulation, Fungal/drug effects , Cell Cycle/drug effects , Fungal Proteins/genetics , Fungal Proteins/metabolismABSTRACT
In the recent years, occurrence of candidiasis has increased drastically which leads to significant mortality and morbidity mainly in immune compromised patients. Glucosinolate (GLS) derivatives are reported to have antifungal activities. Ethyl isothiocyanate (EITC) and its antifungal activity and mechanism of action is still unclear against Candida albicans. The present work was designed to get a mechanistic insight in to the anti-Candida efficacy of EITC through in vitro and in vivo studies. EITC inhibited C. albicans planktonic growth at 0.5 mg/ml and virulence factors like yeast to hyphal form morphogenesis (0.0312 mg/ml), adhesion to polystyrene surface (0.0312 mg/ml) and biofilm formation (developing biofilm at 2 mg/ml and mature biofilm at 0.5 mg/ml) effectively. EITC blocked ergosterol biosynthesis and arrested C. albicans cells at S-phase. EITC caused ROS-dependent cellular death and nuclear or DNA fragmentation. EITC at 0.0312 mg/ml concentration regulated the expression of genes involved in the signal transduction pathway and inhibited yeast to hyphal form morphogenesis by upregulating TUP1, MIG1, and NRG1 by 3.10, 5.84 and 2.64-fold, respectively and downregulating PDE2 and CEK1 genes by 15.38 and 2.10-fold, respectively. EITC has showed haemolytic activity at 0.5 mg/ml concentration. In vivo study in silk worm model showed that EITC has toxicity to C. albicans at 0.5 mg/ml concentration. Thus, from present study we conclude that EITC has antifungal activity and to reduce its MIC and toxicity, combination study with other antifungal drugs need to be done. EITC and its combinations might be used as alternative therapeutics for the prevention and treatment of C. albicans infections.
Subject(s)
Candida albicans , Candidiasis , Humans , Antifungal Agents/pharmacology , Candidiasis/drug therapy , Candidiasis/microbiology , Isothiocyanates/pharmacology , Isothiocyanates/therapeutic use , BiofilmsABSTRACT
Andrographis paniculata is home to a rich variety of molecules especially andrographolide and its derivatives. Clinical properties of the andrographolide are multifarious and include: analgesic, antipyretic, antiretroviral, antiproliferative, antimalarial, antithrombotic, antihyperglycemic, antiurolethial, antilesihmaniasis, hepatoprotective, immune-modulatory, protective against alcohol induced toxicity and cardioproetcive activity and anticancer activity. Andrographolide, neoandrographolide, dehydroandrographolide and several natural and synthetic derivatives of it: 14-deoxy-11,12-didehydroandrographolide and 14-deoxyandrographolide, dehydroandrographolide succinic acid monoester (DAMS), 14-ά-lipoyl andrographolide (AL-1), 14-acetyl-3,9-isopropyl-ideneandrographolide, 14-acetylandrographolide, 3,14,19-triacetylandrographolide, and 3,9-isopropyl-idene andrographolide, are shown to possess significant antiviral activity against HIV, influenza A, HBV, HCV, HPP and HSV. Studies on SARS CoV 2 is restricted to in silico molecular docking studies on viral targets and selected host target proteins. The main targets of andrographolide and its derivatives are fusion and adsorption of virus to the host cell, binding to viral receptor and co-receptor, enzymes involved in DNA/RNA/Genome replication by the virus, translation, post-translation and reverse transcription. Andrographolide as a drug is yet to reach its full therapeutic potential since this molecule shows low bioavailability. Andrographolide therapy is in need of an appropriate delivery system that may increase its bioavailability. Further high-quality studies are needed to firmly establish the clinical efficacy of the plant.
Subject(s)
Andrographis , Antiviral Agents , Diterpenes , Plant Extracts/pharmacology , Andrographis/chemistry , Antiviral Agents/pharmacology , Diterpenes/pharmacology , Molecular Docking Simulation , SARS-CoV-2/drug effectsABSTRACT
Fluoroquinolines, the widely used antibacterial antibiotics, have been shown to interact with human DNA topoisomerases supporting their use as repurposed cancer drugs in humans. In this communication molecular docking of eleven Fluoroquinolines against predicted structure of Candida albicans DNA Topoisomerase II is reported for the first time. C. albicans topoisomerase II structure prediction was done by using homology modeling tool. Ligand preparation and molecular docking with C. albicans topoisomerase II were done by using Autodock tool. These antibiotics formed hydrogen bond with good binding affinity at ARG 841, GLN803, ALA840 amino acid residues in the active site of C. albicans Topoisomerase II. We hypothesize that DNA toposiomerases may be the targets of Fluroquinoline group of antibiotics in C. albicans causing inhibition of growth.
ABSTRACT
Candidiasis involving the biofilms of Candida albicans is a threat to immunocompromised patients. Candida biofilms are intrinsically resistant to the antifungal drugs and hence novel treatment strategies are desired. The study intended to evaluate the anti-Candida activity of allyl isothiocyanate (AITC) alone and with fluconazole (FLC), particularly against the biofilms. Results revealed the concentration-dependent activity of AITC against the planktonic growth and virulence factors of C. albicans. Significant (p <0.05) inhibition of the biofilms was evident at < or =1 mg/ml concentrations of AITC. Notably, a combination of 0.004 mg/ml of FLC and 0.125 mg/ml of AITC prevented the biofilm formation. Similarly, the preformed biofilms were significantly (p <0.05) inhibited by the AITC-FLC combination. The fractional inhibitory concentration indices ranging from 0.132 to 0.312 indicated the synergistic activity of AITC and FLC against the biofilm formation and the preformed biofilms. No hemolytic activity at the biofilm inhibitory concentrations of AITC and the AITC-FLC combination suggested the absence of cytotoxic effects. The recognizable synergy between AITC and FLC offers a potential therapeutic strategy against biofilm-associated Candida infections.
Subject(s)
Antifungal Agents/pharmacology , Biofilms/drug effects , Candida albicans/drug effects , Fluconazole/pharmacology , Isothiocyanates/pharmacology , Biofilms/growth & development , Candida albicans/cytology , Candidiasis/drug therapy , Candidiasis/microbiology , Drug Combinations , Drug Resistance, Fungal , Drug Resistance, Multiple, Fungal/drug effects , Drug Synergism , Erythrocytes/drug effects , Humans , Hyphae/cytology , Hyphae/drug effects , Microbial Sensitivity Tests , Microbial Viability/drug effects , Microscopy, Electron, Scanning , Plankton/drug effects , Virulence FactorsABSTRACT
DNA relaxation is an important step in DNA replication. DNA topoisomerases play a major role in DNA relaxation. Hence these enzymes are important targets for cancer drugs. DNA topoisomerase inhibitors bind to the transient enzyme-DNA complex and inhibit DNA replication. Various inhibitors of topoisomerase I and II are prescribed as drugs. Topoisomerase II is considered as an important target for the development of anticancer drugs. In this study we have demonstrated molecular docking of thirteen fluoroquinolines with human DNA topoisomerase II alpha (a) and beta (b). Fluoroquinolines are broad spectrum antibacterial antibiotics and it is highly effective against various bacterial infections. Some of the fluoroquinolines like moxifloxacin exert antifungal as well as anti-cancer activity. It forms complexes with topoisomerase II a and are responsible for stoppage DNA replication. Molecular docking studies showed that fluoroquinolines has shown formation of hydrogen bond and good binding affinity with human Topo2a and Topo2b. Hence FQs may inhibit the activity of enzyme topoisomerase by binding at its active site. Ofloxacin, sparafloxacin, ciprofloxacin and moxifloxacin are predicted to be the most potent inhibitors among the thirteen FQs docked. GLN773, ASN770, LYS723 and TRP931 amino acid residues of Topo2a are involved in binding with FQs while ASP479, SER480, ARG820, ARG503, LYS456 and GLN778 amino acid residues of Topo2b are involved in binding with FQs. Our in silico study suggests that fluoroquinolines could be repositioned as DNA topoisomerase II inhibitors hence can be used as anticancer drugs. In vitro and in vivo experiments need to be done to confirm their efficacy.