Antifungal Ketoconazole Research Articles

Structural modeling of cytochrome P450 51 from a deep-sea fish points to a novel structural feature in other CYP51s

Cytochromes P450 (CYP), enzymes involved in the metabolism of endogenous and xenobiotic substrates, provide an excellent model system to study how membrane proteins with unique functions have catalytically adapted through evolution. Molecular adaptation of deep-sea proteins to high hydrostatic pressure remains poorly understood. Herein, we have characterized recombinant cytochrome P450 sterol 14α-demethylase (CYP51), an essential enzyme of cholesterol biosynthesis, from an abyssal fish species, Coryphaenoides armatus. C. armatus CYP51 was heterologously expressed in Escherichia coli following N-terminal truncation and purified to homogeneity. Recombinant C. armatus CYP51 bound its sterol substrate lanosterol giving a Type I binding spectra (KD 15 μM) and catalyzed lanosterol 14α-demethylation turnover at 5.8 nmol/min/nmol P450. C. armatus CYP51 also bound the azole antifungals ketoconazole (KD 0.12 μM) and propiconazole (KD 0.54 μM) as determined by Type II absorbance spectra. Comparison of C. armatus CYP51 primary sequence and modeled structures with other CYP51s identified amino acid substitutions that may confer an ability to function under pressures of the deep sea and revealed heretofore undescribed internal cavities in human and other non-deep sea CYP51s. The functional significance of these cavities is not known. PrologueThis paper is dedicated in memory of Michael Waterman and Tsuneo Omura, who as good friends and colleagues enriched our lives. They continue to inspire us.

A Physiologically Based Pharmacokinetic Model of Ketoconazole and Its Metabolites as Drug–Drug Interaction Perpetrators

The antifungal ketoconazole, which is mainly used for dermal infections and treatment of Cushing's syndrome, is prone to drug-food interactions (DFIs) and is well known for its strong drug-drug interaction (DDI) potential. Some of ketoconazole's potent inhibitory activity can be attributed to its metabolites that predominantly accumulate in the liver. This work aimed to develop a whole-body physiologically based pharmacokinetic (PBPK) model of ketoconazole and its metabolites for fasted and fed states and to investigate the impact of ketoconazole's metabolites on its DDI potential. The parent-metabolites model was developed with PK-Sim® and MoBi® using 53 plasma concentration-time profiles. With 7 out of 7 (7/7) DFI AUClast and DFI Cmax ratios within two-fold of observed ratios, the developed model demonstrated good predictive performance under fasted and fed conditions. DDI scenarios that included either the parent alone or with its metabolites were simulated and evaluated for the victim drugs alfentanil, alprazolam, midazolam, triazolam, and digoxin. DDI scenarios that included all metabolites as reversible inhibitors of CYP3A4 and P-gp performed best: 26/27 of DDI AUClast and 21/21 DDI Cmax ratios were within two-fold of observed ratios, while DDI models that simulated only ketoconazole as the perpetrator underperformed: 12/27 DDI AUClast and 18/21 DDI Cmax ratios were within the success limits.

In silico prediction of Antifungal compounds from Natural sources towards Lanosterol 14-alpha demethylase (CYP51) using Molecular docking and Molecular dynamic simulation

An increase in the occurrence of fungal infections throughout the world, as well as the rise of novel fungal strains and antifungal resistance to commercially available drugs, suggests that new therapeutic choices for fungal infections are needed. The purpose of this research was to find new antifungal candidates or leads of secondary metabolites derived from natural sources that could effectively inhibit the enzymatic activity of Candida albicans lanosterol 14-alpha demethylase (CYP51) while also having good pharmacokinetics. In silico prediction of the drug-likeness, chemo-informatics and enzyme inhibition indicate that the 46 compounds derived from fungi, sponges, plants, bacteria and algae sources have a high novelty to meet all five requirements of Lipinski's rules and impede enzymatic function. Among the 15 candidate molecules with strong binding affinity to CYP51 investigated by molecular docking simulation, didymellamide A-E compounds demonstrated the strongest binding energy against the target protein at −11.14, −11.46, −11.98, −11.98, and −11.50 kcal/mol, respectively. Didymellamide molecules bind to comparable active pocket sites of antifungal ketoconazole and itraconazole medicines by hydrogen bonds forming to Tyr132, Ser378, Met508, His377 and Ser507, and hydrophobic interactions with HEM601 molecule. The stability of the CYP51-ligand complexes was further investigated using molecular dynamics simulations that took into account different geometric features and computed binding free energy. Using the pkCSM ADMET descriptors tool, several pharmacokinetic characteristics and the toxicity of candidate compounds were assessed. The findings of this study revealed that didymellamides could be a promising inhibitor against these CYP51 protein. However, there is still a need for further in vivo and in vitro studies to support these findings.

An FDA-Approved Antifungal, Ketoconazole, and Its Novel Derivative Suppress tGLI1-Mediated Breast Cancer Brain Metastasis by Inhibiting the DNA-Binding Activity of Brain Metastasis-Promoting Transcription Factor tGLI1.

Simple SummaryBreast cancer is the most commonly diagnosed cancer in American women, and metastasis accounts for the majority of breast cancer-related deaths. The common metastatic sites for breast cancer includes the bones, lungs, brain, and liver. Breast cancer brain metastasis (BCBM) patients have dismal prognoses, primarily due to the lack of understanding of the molecular mechanisms driving breast cancer cell colonization to the brain. In breast cancer, truncated glioma-associated oncogene homolog 1, tGLI1, promotes preferential metastasis to the brain through the upregulation of the cancer stem cell subpopulation and the activation of astrocytes. Whether tGLI1 is an actionable therapeutic target for any cancer type has not yet been investigated. Herein, we identified an FDA-approved antifungal, ketoconazole (KCZ), and its novel derivative, KCZ-7, to antagonize tGLI1 transcriptional activity, suppress cancer stem cells, and inhibit BCBM, rendering tGLI1, for the first time, as an actionable therapeutic target for the prevention and treatment of BCBM.The goal of this study is to identify pharmacological inhibitors that target a recently identified novel mediator of breast cancer brain metastasis (BCBM), truncated glioma-associated oncogene homolog 1 (tGLI1). Inhibitors of tGLI1 are not yet available. To identify compounds that selectively kill tGLI1-expressing breast cancer, we screened 1527 compounds using two sets of isogenic breast cancer and brain-tropic breast cancer cell lines engineered to stably express the control, GLI1, or tGLI1 vector, and identified the FDA-approved antifungal ketoconazole (KCZ) to selectively target tGLI1-positive breast cancer cells and breast cancer stem cells, but not tGLI1-negative breast cancer and normal cells. KCZ’s effects are dependent on tGLI1. Two experimental mouse metastasis studies have demonstrated that systemic KCZ administration prevented the preferential brain metastasis of tGLI1-positive breast cancer and suppressed the progression of established tGLI1-positive BCBM without liver toxicities. We further developed six KCZ derivatives, two of which (KCZ-5 and KCZ-7) retained tGLI1-selectivity in vitro. KCZ-7 exhibited higher blood–brain barrier penetration than KCZ/KCZ-5 and more effectively reduced the BCBM frequency. In contrast, itraconazole, another FDA-approved antifungal, failed to suppress BCBM. The mechanistic studies suggest that KCZ and KCZ-7 inhibit tGLI1’s ability to bind to DNA, activate its target stemness genes Nanog and OCT4, and promote tumor proliferation and angiogenesis. Our study establishes the rationale for using KCZ and KCZ-7 for treating and preventing BCBM and identifies their mechanism of action.

Interaction of Antifungal Drugs with CYP3A- and OATP1B-Mediated Venetoclax Elimination.

Venetoclax, a BCL-2 inhibitor used to treat certain hematological cancers, exhibits low oral bioavailability and high interpatient pharmacokinetic variability. Venetoclax is commonly administered with prophylactic antifungal drugs that may result in drug interactions, of which the underlying mechanisms remain poorly understood. We hypothesized that antifungal drugs may increase venetoclax exposure through inhibition of both CYP3A-mediated metabolism and OATP1B-mediated transport. Pharmacokinetic studies were performed in wild-type mice and mice genetically engineered to lack all CYP3A isoforms, or OATP1B2 that received venetoclax alone or in combination with ketoconazole or micafungin. In mice lacking all CYP3A isoforms, venetoclax AUC was increased by 1.8-fold, and pretreatment with the antifungal ketoconazole further increased venetoclax exposure by 1.6-fold, despite the absence of CYP3A. Ensuing experiments demonstrated that the deficiency of OATP1B-type transporters is also associated with increases in venetoclax exposure, and that many antifungal drugs, including micafungin, posaconazole, and isavuconazole, are inhibitors of this transport mechanism both in vitro and in vivo. These studies have identified OATP1B-mediated transport as a previously unrecognized contributor to the elimination of venetoclax that is sensitive to inhibition by various clinically-relevant antifungal drugs. Additional consideration is warranted when venetoclax is administered together with agents that inhibit both CYP3A-mediated metabolism and OATP1B-mediated transport.

Comparison of the Inhibitory Effects of Clotrimazole and Ketoconazole against Human Carboxylesterase 2.

Both clotrimazole and ketoconazole have been verified to have an inhibitory effect on CYP3A4. hCE2 is an enzyme closely related to the side effects of several anti-cancer drugs. However, the interactions between hCE2, clotrimazole, and ketoconazole remain unclear. The objective of this study was to investigate and compare the inhibition behaviors of the two antifungal agents, ketoconazole and clotrimazole, on the human liver microsome hCE2 and to explore their underlying mechanism. The inhibitory effects were investigated in human liver microsomes (HLMs) using fluorescein diacetate (FD), N-(2-butyl-1,3-dioxo-2,3-dihydro-1H-phenalen-6-yl)-2-chloroacetamide (NCEN) and irinotecan (CPT- 11) as substrates of hCE2. Clotrimazole significantly inhibited the hCE2 activity, which was manifested by attenuated fluorescence when the substrates were FD and NCEN. The inhibitory effect of clotrimazole towards hCE2 was much stronger than that of ketoconazole, and the inhibitory behaviors displayed substrate-dependent inhibition. The IC50 value of clotrimazole, with CPT-11 as the substate, increased by 5 and 37 times more than that with FD and NCEN, respectively. Furthermore, the inhibitions of clotrimazole towards hCE2-mediated hydrolysis of FD, NCEN, and CPT-11 were all in competitive mode with the Ki values of 0.483 μM, 8.63 μM, and 29.0 μM, respectively. Molecular docking result of clotrimazole binding to hCE2 illustrated that clotrimazole could efficiently orient itself in the Z site cavity of hCE2. Clotrimazole displayed a strong inhibitory effect against hCE2, which might be used as a potential combined agent co-administrated with CPT-11 to alleviate the hCE2-mediated severe side effects.

Evaluation of susceptibility and response in the surface of agents of surface mycoses (Trichophyton mentagrophytes; T. tonsurans) to antifungal drugs of interest in a medical clinic

Introduction: The resistance of fungal species to drugs usually used in clinics is of great interest in the medical field. Objective: To evaluate susceptibility and in vitro response of species of Trichophyton spp. to antifungal drugs of interest in clinical medicine. Methods: 12 samples of clinical isolates from humans were used, nine of T. mentagrophytes and three of T. tonsurans. Susceptibility tests were performed according to the agar diffusion (AD) and broth microdilution (BM) methods. Results: In the AD method, the species T. tonsurans presented a percentage of sensitivity of 33% in relation to amphotericin B and 66% to itraconazole, with 100% resistance to ketoconazole and fluconazole. T. mentagrophytes also showed 100% resistance to ketoconazole in this technique, with 11% sensitivity to ketoconazole, 22% to itraconazole and 22% of samples classified as sensitive dose dependent. In the MC method, the species T. tonsurans presented a sensitivity percentage of 66%, 55% and 33% in relation to ketoconazole, fluconazole and itraconazole, respectively. The T. mentagrophytes species presented sensitivity percentages of 11%, 11%, 33% and 55% for amphotericin B, itraconazole, ketoconazole and fluconazole, respectively. Conclusion: There was resistance in vitro of the species of T. mentagrophytes and T. tonsurans against the antifungal fluconazole and relative resistance against ketoconazole in the AD method. In BM, however, important percentages of sensitivity were observed for the two species analyzed in relation to the antifungals fluconazole and ketoconazole when compared to itraconazole and amphotericin B.

Objective: Antifungal activity of total ethanolic extract and fractions of Usnea varieties were proved against Aspergillus niger, Penicillium digitatum, and Rhizopus nigricans fungi. Methods: To perform the study of relative antifungal activity (AFR), flu

Objective: Antifungal activity of total ethanolic extract and fractions of Usnea varieties were proved against Aspergillus niger, Penicillium digitatum, and Rhizopus nigricans fungi.
 Methods: To perform the study of relative antifungal activity (AFR), fluconazole and ketoconazole were used as standards. The inhibition coefficient 50 (IC50) for each extract and the fractions was determined.
 Results: It was established that the highest activity was presented by the acetone fraction with a value of 58.53. In the study of antifungal activity, the fraction that presented the best activity against P. digitatum was ethanolic with a value of 44.33, while for R. nigricans was petrol extract, with a value of 75.35 and finally for A. niger was the total extract with a value of 35.48.
 Conclusions: Comparing the values obtained from the extract and the different fractions resulted in the dichloromethane fraction showing the best values.

Abstract 5025: Antifungal ketoconazole inhibits tumor-specific transcription factor tGLI1 leading to suppression of breast cancer stem cells and brain metastasis

Abstract Breast cancer is the second leading cause of brain metastases in women; patients with breast cancer brain metastasis (BCBM) survive only 6-18 months following diagnosis. Current standard of care options for BCBM include stereotactic radiosurgery, whole-brain radiotherapy, and surgical resection. Cancer stem cells are thought to be one of the driving forces behind not only distant metastasis, but also late-stage recurrence. The hedgehog pathway has been identified as an important mediator of stem cells, however, FDA-approved therapies targeting this pathway have demonstrated limited efficacy in breast cancer clinical trials. Despite our current knowledge of breast cancer stem cells (BCSC), there remains a significant challenge in managing patients with metastatic breast cancer, underscoring the need to identify novel, actionable targets. Our laboratory discovered an alternative splice variant of glioma-associated oncogene homolog 1 (GLI1), termed truncated GLI1 (tGLI1), that is a tumor-specific gain-of-function transcription factor and terminal effector of the hedgehog pathway that is preferentially expressed in most BCBM samples and recurrent gliomas. Recent results established that tGLI1 promotes BCSC and is associated with preferential metastasis to the brain and radioresistance, justifying tGLI1 as an ideal therapeutic target for BCBM patients. To identify tGLI1-targeting agents, we screened 1,520 compounds across three commercial drug libraries and found ketoconazole (KCZ), an FDA-approved imidazole antifungal and component of previously studied anti-neoplastic regimens, selectively killed tGLI1-expressing breast cancer cells with increased efficacy against BCSC in vitro. tGLI1 knockdown abolished the ability of KCZ to target BCSC, indicating that KCZ-mediated suppression of BCSC is dependent on tGLI1. Intracardiac mouse studies showed KCZ selectively inhibited circulating tGLI1-positive breast cancer cells from developing into brain metastases and suppressed the progression of existing brain metastases. Mass spectrometry demonstrated KCZ effectively penetrated the blood-brain barrier (BBB) and blood-tumor barrier (BTB). Mechanistic studies suggest that KCZ-dependent cell kill is, in part, mediated through disruption of the functional tGLI1-STAT3 interaction. Furthermore, modification of KCZ side chains produced derivative compounds that retained tGLI1-selectivity in both in vitro models of BCSC and in vivo models of BCBM with increased BBB penetrance. Collectively, our preclinical results demonstrate that KCZ is an effective inhibitor of BCSC and brain metastasis of tGLI1-positive breast cancer. Based on these promising preclinical data, we opened a window-of-opportunity study in patients with BCBM and recurrent gliomas to determine if KCZ treatment alters tGLI1 signaling in humans (NCT03796273). Citation Format: Daniel L. Doheny, Sherona R. Sirkisoon, Tadas Rimkus, Dongqin Zhu, Noah R. Aguayo, Alexandria Harrison, Marlyn Anguelov, Sara Manore, Fei Xing, Linda J. Metheny-Barlow, Kounosuke Watabe, Alexandra Thomas, Adrianna Henson Masters, Roy Strowd, Hui-Wen Lo. Antifungal ketoconazole inhibits tumor-specific transcription factor tGLI1 leading to suppression of breast cancer stem cells and brain metastasis [abstract]. In: Proceedings of the Annual Meeting of the American Association for Cancer Research 2020; 2020 Apr 27-28 and Jun 22-24. Philadelphia (PA): AACR; Cancer Res 2020;80(16 Suppl):Abstract nr 5025.

Design, Synthesis and Biopharmacological Profile Evaluation of New 2-((4- Chlorophenoxy)Methyl)-N-(Arylcarbamothioyl)Benzamides with Broad Spectrum Antifungal Activity

The emerging antifungal resistance represents a major challenge for the treatment of severe fungal infections, highlighting the need to develop novel and efficient antifungal compounds. This study aimed to synthesize new title compounds and screen them for their antifungal activity in order to generate highly accurate structure - activity relationships of 2-((4-chlorophenoxy)methyl)-N-(arylcarbamothioyl)benzamides and their de novo derivatives and to unveil some of their mechanisms of action by flow cytometry and fluorescence microscopy. The presence of functional groups was confirmed for nine new 2-((4- chlorophenoxy) methyl)-N-(arylcarbamothioyl)benzamides, using experimental and in silico methods. The antifungal activity was assessed against a broad spectrum of 26 yeast and filamentous fungal strains, using qualitative and quantitative assays. The results showed that Candida kefyr has been the most susceptible to all tested compounds, while 1b and 1f induced a strong inhibitory effect on the filamentous fungi Alternaria rubi, Aspergillus ochraceus and A. niger strains growth. The derivative 1c in subinhibitory concentrations alsoincreased the susceptibility of Candida albicans clinical strains to azoles. Predicted drug likeness and pharmacokinetics profiles of most active compounds were compared with the standard antifungal ketoconazole. Furthermore, the potentially more potent 1c and 1f derivatives were designed and studied regarding the chemical structure-biological activity relationship and pharmacokinetics profiles versus ketoconazole. The study confirms that the new benzamide derivatives exhibited an improved pharmacokinetics profile and a good antifungal activity, acting at least by increasing membrane permeability of fungal cells. Our results are recommending them as promising candidates for the development of novel therapeutic alternatives.

<p>A novel mechanism of action of ketoconazole: inhibition of the <em>NorA</em> efflux pump system and biofilm formation in multidrug-resistant <em>Staphylococcus aureus</em></p>

Background: The rapid emergence of antimicrobial resistance among Gram-positive organisms, especially staphylococci, has become a serious clinical challenge. Efflux machinery and biofilm formation are considered two of the main causes of antimicrobial resistance and therapy failure.Aim: Our study aims to evaluate the antibiofilm and efflux pump inhibitory activity of the antifungal ketoconazole against multidrug-resistant (MDR) Staphylococcus aureus.Methods: Ketoconazole was tested for its effect on the following: minimum inhibitory concentrations (MICs) of ciprofloxacin, norfloxacin, levofloxacin, and ethidium bromide (EtBr) by the broth microdilution method, the efflux of EtBr by NorA-positive MDR S. aureus, and the relative expression of NorA, NorB, and NorC efflux pump genes. Docking studies of ketoconazole were performed using 1PW4 (glycerol-3-phosphate transporter from Escherichia coli which was the representative structure from the major facilitator superfamily).Results: Ketoconazole significantly decreased the MICs of levofloxacin, ciprofloxacin, norfloxacin, and EtBr (a substrate for efflux pump) by 8 to 1024-fold (P<0.01) and decreased the efflux of EtBr. Furthermore, a time-kill assay revealed that combinations of levofloxacin with ketoconazole or carbonyl cyanide m-chlorophenylhydrazone showed no growth for the tested strains after 24 h in comparison to the effect of levofloxacin alone. Docking studies and the ability of ketoconazole to diminish the relative expression of NorA gene in comparison to control (untreated strains) confirmed its action as an efflux pump inhibitor. Conclusion: The findings showed that the antifungal ketoconazole has no antibacterial activity but can potentiate the activity of the fluroquinolones against MDR S. aureus via inhibiting efflux pump and biofilm formation in vitro.

Chinese herbal remedy found to contain steroids and antifungals

A 14-year-old boy with early-onset guttate psoriasis vulgaris presented at our clinic with several reddish-purple, horizontal streaks on the distal part of his shins. Previously, he had been treated with narrow-band ultraviolet B, topical corticosteroids ranging from mild (hydrocortisone 17-butyrate cream 1 mg/g) to moderate (mometasone furoate cream 1 mg/g and betamethasone valerate ointment 1 mg/g), and tacrolimus ointment 1 mg/g—but the response to treatment was not long lasting. The family consulted a licensed practitioner of traditional Chinese medicine who prescribed a so-called herbal cream (figure). The patient applied the cream topically on the psoriatic lesions twice a day. After 9 months of doing this, he developed a new rash on his shins that was later confirmed to be striae distensae (figure). We suspected that the cause was exposure to a corticosteroid—but the patient had not been using a corticosteroid either topically or systemically during this time. The so-called herbal cream was suspected as a possible source; however, the content label only listed herbal ingredients including Chinese sumac, Sophora flavescens, Stemona tuberosa Lour. Nevertheless, the location of the striae was very unusual, and as a simple test, the herbal cream was applied on the upper arm of the senior physician; after 12 h, a strong vasoconstriction effect was seen (figure), which further raised our suspicion that the cream contained a corticosteroid. We therefore sent it for analysis at the Department of Official Medicines Control Laboratory, Swedish Medical Products Agency, Uppsala, Sweden. The laboratory used liquid chromatography-tandem mass spectrometry and nuclear magnetic resonance, and reported that the cream contained both a potent corticosteroid (0·065% clobetasol propionate) and the antifungals ketoconazole and miconazole. We then reported this herbal product to the Danish Medicines Agency to ensure that it was dealt with appropriately.

IN VITRO ANTIBACTERIAL ACTIVITY OF AZOLE ANTIFUNGALS AGAINST CARBAPENEM-RESISTANT ENTEROBACTERIACEAE

Introduction: The frequency of serious bacterial infections has increased due to the high prevalence of HIV, contributing to the increasing rates of multi-drug organisms which include carbapenem-resistant Enterobacteriaceae (CRE). This has resulted in higher use of immunosuppressive and cytotoxic drugs to treat serious bacterial infections and optimal treatment for infections caused by CRE remains unknown. The benefits of azole antifungals and its derivatives have become very topical due to their diverse spectrum of pharmacological properties, but its efficacy has not been tested in the South African context. The aim of this study was to determine the antibacterial effects of azole antifungals against CRE. Method: Different concentrations of azole antifungals (ketoconazole, metronidazole and fluconazole) were used to prepare sensitivity discs and four pathogenic strains of CRE ( K. pneumoniae, E. coli, S. marcesens and C. freundii ). These were obtained from Lancet Laboratory in Durban and were used to determine the antibacterial activity of the selected azole antifungals, using: Disc Diffusion, Modified Agar Diffusion and Minimum Inhibition Concentration (MIC) method, as described by Bauer et al. 1966. Results: Antimicrobial Susceptibility Testing revealed that azoles have no inhibition activity against CRE test organisms and biochemical tests also demonstrated that azoles have no adverse effects on the CRE organisms. Conclusion: Although the results obtained in this study indicated no activity of azoles against CRE, clinical studies are still necessary for the modification of the azole substituents and synthesis of azole derived drugs to confirm and optimise the antibacterial efficacy of these compounds.

Synthesis and antimicrobial activity of new 2-((1-furan-2-yl)ethylidene)hydrazono)-4-phenylthiazol-3(2H)-amine derivatives and their schiff bases with 4-nitrobenzaldehyde

A new series of 2-((1-furan-2-yl)ethylidene)hydrazono)-4-substitutedphenylthiazol-3(2H)-amines (2a–2o) and their Schiff bases (3a–3o) from 4-nitrobenzaldehyde were synthesized. The chemical structures of all the synthesized compounds were confirmed by their IR, 1H-NMR, 13C-NMR spectroscopy and mass spectrometry. They were screened for their antimicrobial and antifungal activities. Additionally, in vitro cytotoxic acivity of the most active antifungal compound (3o) and ketoconazole was determined in NIH/3T3 cells by MTT assay. Compound 2i (4-{3-Amino-2-[(1-(furan-2-yl)ethylidene)hydrazono]-2,3-dihydrothiazol-4-yl}phenol) showed the greatest antifungal activity among the newly synthesized derivatives. Schiff bases (3c-3n) displayed an undeniable fungicidal action against Candida parapsilosis ATCC 22019 as intense as the reference ketoconazole. In addition, the most active Schiff base 3o (2-[(1-(Furan-2-yl)ethylidene)hydrazono]-N-(4-nitrobenzylidene)-4-(2,3,4-trichloro phenyl)thiazol-3(2H)-amine) showed the highest antifungal activity against both Candida krusei ATCC 6258 and Candida parapsilosis ATCC 22019, and was as potent as ketoconazole. Moreover, compound 3o was found to be non-cytotoxic against NIH/3T3 cells.

Synthesis, biological evaluation and structure-activity relationships of self-assembled and solubilization properties of amphiphilic quaternary ammonium derivatives of quinuclidine

This work deals with development of polyfunctional biocompatible cationic surfactant systems based on bioactive saturated bicyclic alkaloid quinuclidine. It is focused on the effect of the chemical structure of surfactants on their aggregation behavior, their physicochemical estimation of solubility of model water insoluble dye Orange OT and drugs, quercetin and rutin, microbiology and cytotoxicity. Quaternary ammonium derivatives of quinuclidine (Q-Nuc-n) with different hydrophobicity (R = CnH2n+1, where n = 14, 16, 18) were synthesized. Self-assembly of Q-Nuc-n was investigated by tensiometry, conductometry, spectrophotometry, fluorimetry and dynamic light scattering. The critical micelle concentration, thermodynamic and adsorption parameters at water-air interface, size and aggregation numbers of Q-Nuc-n micelles were determined. The looser packing of surfactant molecules in Q-Nuc-n micelles compared to its analogues, quaternized derivatives of 1,4‑diazabicyclo[2.2.2]octane (DABCO-n), was established. The hydrophobic dye Orange OT and drugs quercetin and rutin were solubilized in micellar Q-Nuc-n solutions better than in solutions of classical surfactant CTAB and its analogue DABCO-n. Solubilization capacity of Q-Nuc-18 is 5 times higher than that of classical surfactant CTAB. Q-Nuc-18 1.95 μg·mL−1 has also bactericidal and fungicidal activity 2 times (against S. aureus 209P) and 8 times (against B. cereus 8035) higher than antibiotics Norfloxacin and antifungal Ketoconazole. Q-Nuc-16 has the highest bactericidal activity. It is 6 times (against S. aureus 209P) and 15 times (against B. cereus 8035) higher than the bactericidal activity value of Norfloxacin. Synthesized cationic surfactants based on quinuclidine are new multifunctional biocompatible compounds with high potential in nanomedicine and biotechnology.

Uses of 2-(Thiophene-2-carbonylcarbamothioylthio)acetic Acid as a Good Synthon for Construction of Some New Thiazole and Annulated Thiazole Derivatives.

The reaction of thiophene-2-carbonyl isothiocyanate 2 with thioglycolic acid gave 2-(thiophene-2-carbonylcarbamothioylthio)acetic acid 3. Compound 3 was subjected to some selected reactions with sulphuric acid as well as benzaldehyde, piperonal and isatin under different reaction conditions. The products obtained were new derivatives of thiazole and annulated thiazole derivatives bearing thiophene moiety in some cases. The structures of the new synthesized compounds were confirmed on the basis of their microanalytical and spectral properties. Some compounds were tested for their antimicrobial activity against six selected microorganisms using the standard antibacterial Gentamycin and antifungal Ketoconazole as references.

No Relationship of Anti-Androgens to Alzheimer's Disease or Cognitive Disorder in the MedWatch Database.

Background:Two large studies suggest that risk is not increased. But other studies have found increased risk of Alzheimer’s disease and impaired cognition.Objective:To determine whether androgen deprivation therapy increases the risk of impaired cognition or Alzheimer’s disease in men with prostate cancer.Methods:We used data from MedWatch, the Food and Drug Administration (FDA) Safety Information and Adverse Event Reporting Program. Machine-readable data from MedWatch, including adverse drug reaction reports from manufacturers, are part of a public database. We used the online tool OpenVigil 2.1 to query the database. OpenVigil calculates proportional reporting ratios (PRRs) from adverse drug reaction reports to determine whether the combination of drug and adverse event are related. For example, PRR = 2 indicates that the adverse reaction is two times more frequent in users of the drug than in the general population.Results:We analyzed adverse event reporting data for these androgen-deprivation drugs: The luteinizing hormone releasing hormone (LHRH) agonists leuprolide, goserelin triptorelin, histrelin; the anti-androgens flutamide, nilutamide, enzalutamide, and bicalutamide; the LHRH antagonist degarelix; the CYP17 inhibitor abiraterone; the anti-fungal ketoconazole, which is also an anti-androgen administered to men with advanced prostate cancer.Conclusion:Our analysis of FDA MedWatch adverse event data reports does not support the idea that androgen deprivation therapy per se is associated with Alzheimer’s disease or cognitive dysfunction. Perhaps the prostate cancer itself, or the stress it imposes on the man who has it, may be detrimental to mood and intellect, increasing susceptibility to Alzheimer’s disease and cognitive disorder.

In Vitro Antifungal Activity of Clove Oil against Candida Albicans Isolated from Clinical Samples

The main goal of this study was to investigate the antifungal activity of clove oil against candida albicans of vaginal candidiasis in females from Syria. An in vitro study was carried out using the following Candida albicans strains involved in vaginal candidiasis using the well diffusion (WD) testing.Candida albicans (ATCC 90028) and 15 strains were compiled from Aleppo University Hospital. These strains were collected from women having vaginal candidiasis. The antifungal activity of clove oil was determined in the form of inhibition zone using antifungal assay by agar WD testing.In all experiments, the obtained results indicated that clove oil has inhibitory effects on Candida albicans (ATCC 90028) and against15 fungal strains. This study showed that clove oil was active against the tested Candida albicans strains. Clove oil was more effective against Candida albicans compared to the antifungal antibiotics nystatin, ketoconazole and itraconazol. Clove oil may have potential for use in the development of clinically useful antifungal preparations. Therefore, clove oil might be clinically effective in the natural prevention treatment of vaginal candidiasis. &#x0D;

Impact of forbidden foodstuffs on the efficacy of homoeopathic medicines: An in vitro evaluation

Background: In previous screening experiment, out of some homoeopathic medicines in 30 and 200 potencies Mezereum 200 showed maximum inhibition to growth of Candida albicans on comparing the 'Zone of Inhibition' in culture plates treated with homoeopathic medicines keeping standard antifungal Ketoconazole, rectified spirit and distilled water as control by using Disc method. Objective: To observe the impact of various forbidden edible items such as garlic, onion, cardamom, clove, caraway, ginger, fenugreek, black pepper, asafoetida, red chilli, green chilli, turmeric, lemon and camphor on the efficacy of Mezereum 200 in the inhibition of growth of human pathogenic fungus C. albicans. Materials: Isolation of human pathogenic fungus: Sample collected from patients suffering from oral ulcers was incubated for 15 days at 37+1°C for growth of fungus, if any. The growth of Candida albicans was obtained. Preparation of prohibited items: 500 mg of fine powdered prohibited items were dissolved/mixed in 5 ml distilled water. 5 μl of autoclaved solution of each item was mixed with 5 μl of Mezereum 200. Method: Disc method was used to assess the impact of prohibited items on the efficacy of homoeopathic medicine Mezereum 200 in in-vitro conditions against Candida albicans by “Inhibition Zone Technique”. Result: The result of this in-vitro experiment has shown that there is no inhibitory effect of prohibited items in question on Mezereum 200 in causing inhibition to growth of Candida albicans. Conclusion: It is evident from this experiment that there is no interference of prohibited items on the action of homoeopathic medicine.

Enantiospecific Effects of Ketoconazole on Aryl Hydrocarbon Receptor

Azole antifungal ketoconazole (KET) was demonstrated to activate aryl hydrocarbon receptor (AhR). Since clinically used KET is a racemic mixture of two cis-enantiomers (2R,4S)-(+)-KET and (2S,4R)-(−)-KET, we examined the effects of KET enantiomers on AhR signaling pathway. (+)-KET dose-dependently activated AhR in human gene reporter cell line AZ-AHR, and displayed 5–20× higher agonist activity (efficacy), as compared to (−)-KET; both enantiomers were AhR antagonists with equal potency (IC50). Consistently, (+)-KET strongly induced CYP1A1 mRNA and protein in human HepG2 cells, while (−)-KET exerted less than 10% of (+)-KET activity. In primary human hepatocytes, both enantiomers preferentially induced CYP1A2 over CYP1A1 mRNA and protein, and the potency of (+)-KET was slightly higher as compared to (−)-KET. Ligand binding assay with guinea pig liver cytosols revealed that both (+)-KET and (−)-KET are weak ligands of AhR that displaced [3H]-TCDD with comparable potency. Similarly, both enantiomers weakly transformed AhR to DNA-binding form with similar potency, as showed by EMSA, in guinea pig liver cytosolic extracts and nuclear extracts from mouse Hepa-1 cells. We also examined effects of KET on glucocorticoid receptor (GR), a regulator of AhR activity. Both KET enantiomers antagonized GR with similar potency, as revealed by gene reporter assay in AZ-GR cell line and down-regulation of tyrosine aminotransferase mRNA in human hepatocytes. Finally, we demonstrate enantiospecific antifungal activities of KET enantiomers in six Candida spp. strains. In conclusion, the significance of current study is providing the first evidence of enatiospecific effects of cis-enantiomers of ketoconazole on AhR-CYP1A pathway.