Concentrations Of Clotrimazole Research Articles

Delayed clearance of the pro-carcinogen benzo[a]pyrene in PLHC-1 cells when co-exposed to the antifungal drug clotrimazole and effects on the CYP1A biomarker

Cytochrome P450 1 A (CYP1A) is a key enzyme in the metabolism of the polycyclic aromatic hydrocarbon (PAH) benzo[a]pyrene (BaP) in animals, and a biomarker for environmental PAH exposure. The common antimycotic imidazole drug clotrimazole (CLO) has been detected in the aquatic environment and likely co-exists with BaP. Like BaP, CLO can bind to CYP1A enzymes and can act as a CYP1A inhibitor. Co-exposure of BaP with CLO significantly delayed BaP elimination in a fish liver cell line (PLHC-1). Intracellular BaP concentration was 2.4 times higher after 6 h in co-exposed cells, compared to cells exposed to BaP alone. Higher BaP concentrations in cells co-exposed to CLO positively correlated with CLO dose, indicating CLO-mediated delays in BaP clearance. After 24 h, BaP was undetectable irrespective of CLO co-exposure. In contrast, intracellular CLO concentrations remained constant over the 72 h experimental period. Co-exposure of BaP with CLO caused synergistic and time-dependent increases on the CYP1A biomarker both on CYP1A mRNA levels and on CYP1A enzyme activity, in accordance with an apparent delayed BaP elimination in the presence of CLO. These results indicate a toxicokinetic interaction between BaP and CLO on the CYP1A enzyme that delays metabolic clearance of BaP.

Topical delivery systems containing clotrimazole for the management of candidiasis: Effect of different excipients and enhanced antifungal activity of nanovesicles

WHO classified Candida albicans as one of the four critical priority fungi for public health worldwide in 2022. Conventional topical formulations commercially available for the treatment of cutaneous candidiasis are associated with low drug bioavailability at the infection site and the lack of a sustained therapeutic effect. The main objectives of this work were to develop new topical administration systems of clotrimazole (CLT) and study the influence of surfactants on the antifungal inhibitory efficacy. Therefore, the minimum concentration of CLT required to inhibit 50 % of growth (MIC50) was determined, obtaining a value of approximately 15 ng/mL. A non-ionic emulsion type 1, Beeler base cream, hydrogel and liposomes containing CLT were designed, prepared, characterized and their antifungal activity against C. albicans was tested. CLT loaded liposomes were small in size (102 nm), homogeneous (polydispersity index = 0.3) and uncharged (+0.07 mV), showing higher antifungal activity against C. albicans than that of the commercially available cream Canesten®. Furthermore, the antifungal activity of CLT was reduced in combination with surfactants such as Tween-80/Span-80 or Brij-S10. Sodium lauryl sulphate showed a fungicidal effect that disappeared when formulated as part of the Beeler base cream.

Evaluation of parental and transgenerational effects of clotrimazole in Daphnia magna — A multi-parametric approach

Azole antifungals inhibit the cytochrome P450 complex, decreasing the production of ergosterol in fungi, and compromising the biosynthesis of ecdysteroids in crustaceans, which are hormones regulating reproduction and ecdysis. The azole antifungal clotrimazole (CLO) raises environmental concerns due its toxicity. This work evaluated the effects on the number of moults, feeding rate, growth, reproduction, transgenerational reproductive effects on two different generations (F0, parental generation; and F1, organisms born from F0), and energetic balance in Daphnia magna. Neonates (<24 h) were exposed to sublethal concentrations (0, 2.7, and 3.4 mg/L) of CLO, to assess its effects on the moulting process. Neonates were also exposed to environmentally realistic concentrations of CLO (0, 30, 150, 750, and 3750 ng/L) for 24 and 96 h, to assess adverse effects on their feeding behaviour. Effects on energy reserves (fatty acids, glycogen, and protein levels) were also measured in animals exposed to CLO. A reproduction test was carried out to evaluate the amount and size of neonates from F0 and F1 generations. CLO exposure decreased the number of moults, and the size of organisms, but did not alter the feeding pattern of 5 days old individuals. However, neonates (<24 h) exposed to CLO had a significant decrease in their feeding pattern. CLO decreased the fatty acids content in exposed animals, but did not change glycogen and protein. CLO also decreased the size of adult daphnids from the third brood, born from animals exposed in F0; in F1 animals, the size of neonates from the third brood was decreased. This study evidenced the toxic effects caused by CLO on growth, feeding and reproduction of D. magna. Nevertheless, it is not possible to conclude whether the effects are due to the inhibition of cytochrome P450 enzymes, or to unspecific effects caused by general toxic stress and decreased nutrition.

QbD steered fabrication of Pullulan-Terminalia catappa-Carbopol®971P film forming gel for improved rheological, textural and biopharmaceutical aspects

In present work, a film forming gel (FFG) was developed through ingenious amalgamation of polymers: Pullulan, Terminalia catappa and Carbopol®971P ® for cutaneous delivery of clotrimazole (CTZ) employing D-optimal mixture design. The developed FFG possess pseudoplastic, viscoelastic, thixotropic characteristics leading to good spreadability (35.71 ± 1.72 g·s, work of shear; 452.73 ± 8.23 g, firmness). Upon solvent evaporation, FFG converted in situ into bioadhesive film (81.90 ± 3.24 g) leading to longer residence on skin surface, prolonged delivery and ~1.3 fold enhanced CTZ skin retention as compare to commercial cream as evident from biopharmaceutical analysis, which is ideal for skin infections treatment. The simulation analysis suggested ≥10 μg/mL (MIC against C. albicans) CTZ concentration maintained for 2 times the days in rat skin as well as human skin as compared to commercial cream. Overall, the developed FFG system ascertained to be promising delivery system for treatment of chronic skin conditions.

Susceptibility to Clotrimazole of Candida spp. Isolated from the Genitourinary System-A Single Center Study.

The aim of this study was to determine the susceptibility to clotrimazole of 125 isolates of Candida spp. originated from the genitourinary system of hospitalized patients as well as outpatients, tested in the mycological laboratory of Wroclaw Medical University in the years 1999–2018. The minimal inhibitory concentrations of clotrimazole and fluconazole were determined with the use of the microdilution method according to EUCAST, and the MFC was determined by subsequent subculture on Sabouraud agar. For the tested population of Candida yeasts, the MIC values of clotrimazole ranged from 0.008 to 8 mg/L, and MIC90 was 1 mg/L, whereas MIC50 was 0.008 mg/L. The minimal fungicidal concentration ranged between 1 and >8 mg/L. The great majority of the isolates (88%; 110/125) displayed MIC < 1 mg/L and were classified as WT (wild-type), whereas MIC ≥ 1 mg/L was determined for 2/61 (3.2%) isolates of C. albicans, 9/38 (23.6%) of C. glabrata, 1/2 of C. tropicalis, and 3/3 of C. guilliermondii. Six isolates (four of C. glabrata and two of C. albicans), defined as non-WT for clotrimazole, were classified as resistant to fluconazole, according to CBP from EUCAST. The isolates with elevated MIC to clotrimazole originated mostly from patients of the pediatric hematology unit, and their proportion in this population amounted to 17.8% (13 out of 73 isolates). In contrast, among strains from ambulatory patients, the highest observed MIC value was 1 mg/L (1 out of 37 isolates; 2.7%). The data obtained correlate well with those of most published studies on the in vitro susceptibility of Candida spp. to clotrimazole, which is usually very high. However, the existence of reports regarding the growing prevalence of resistant isolates has also to be noted. These results support the need for routinely checking the susceptibility of Candida clinical isolates to this imidazole derivative.

Clotrimazole Fluidizes Phospholipid Membranes and Localizes at the Hydrophobic Part near the Polar Part of the Membrane.

Clotrimazole (1-[(2-chlorophenyl)-diphenylmethyl]-imidazole) is an azole antifungal drug belonging to the imidazole subclass that is widely used in pharmacology and that can be incorporated in membranes. We studied its interaction with 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) phospholipid vesicles by using differential scanning calorimetry and found that the transition temperature decreases progressively as the concentration of clotrimazole increases. However, the temperature of completion of the transition remained constant despite the increase of clotrimazole concentration, suggesting the formation of fluid immiscibility. 1H-NMR and 1H NOESY MAS-NMR were employed to investigate the location of clotrimazole in 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC) phospholipid membranes. In the presence of clotrimazole, all the resonances originating from POPC were shifted upfield, but mainly those corresponding to C2 and C3 of the fatty acyl, chains suggesting that clotrimazole aromatic rings preferentially locate near these carbons. In the same way, 2D-NOESY measurements showed that the highest cross-relaxation rates between protons of clotrimazole and POPC were with those bound to the C2 and C3 carbons of the fatty acyl chains. Molecular dynamics simulations indicated that clotrimazole is located near the top of the hydrocarbon-chain phase, with the nitrogen atoms of the imidazole ring of clotrimazole being closest to the polar group of the carbonyl moiety. These results are in close agreement with the NMR and the conclusion is that clotrimazole is located near the water–lipid interface and in the upper part of the hydrophobic bilayer.

Imidazole antifungal drug clotrimazole alters the behavior, brain acetylcholinesterase and oxidative stress biomarkers in African catfish Clarias gariepinus (Burchell 1822)

This study was carried out to determine the effect of clotrimazole (CTZ), an imidazole fungicide on behavior, brain acetylcholinesterase, lipid peroxidation and oxidative stress parameters in Clarias gariepinus juveniles. Fish were acutely exposed to five nominal concentrations of CTZ and control to assess the behavioral effects on fish. To determine the effects on brain acetylcholinesterase, lipid peroxidation and oxidative stress parameters, fish were exposed to three sub-lethal concentrations vis 7.76, 3.89 and 1.94 mg/L which corresponds to 20 10 and 5% of 96 h LC50 value of CTZ respectively and a control for 21 days and allowed to recover for 7 days. The brain, liver and gills were sampled weekly. Fish exposed to different concentrations of the drug displayed behavioral responses such as reduced swimming rate, mucus secretion, decrease in both the feeding frequency and rate. A duration dependent increase in the levels of brain acetylcholinesterase was observed among the exposed groups. The result of the sub-lethal exposure revealed concentration and duration significant increase in lipid peroxidation (LPO), catalase activity (CAT), superoxide dismutase (SOD), reduced glutathione (GSH), glutathione reductase (GR) and glutathione peroxide (GPx) in both the liver and gill tissues of the exposed groups compared to the control. The effects of the drug on many of the observed parameters did not wane after the 7-day withdrawal period. This study revealed that CTZ has a negative impact on the observed parameters, thus providing additional evidence of its toxic effect on non-target aquatic species, especially fish.

Biomimetic clotrimazole-loaded PLGA films with enhanced adhesiveness for controlled drug release

Biomimetic adhesive surfaces have a number of potential applications in the pharmaceutical and biomedical fields. Fabrication techniques must be adapted to biocompatible and biodegradable materials required for controlled drug release applications. In this study biomimetic adhesive poly(lactic-co-glycolic acid) (PLGA) films loaded with different concentrations of clotrimazole (CTZ) were prepared without combining other adhesive excipients as a controlled release system for potential local oral drug delivery. The films were fully characterized from morphological point of view, and CTZ-loaded biomimetic films exhibited adequate surface pH values, high drug encapsulation efficiency, and loading content. The adhesion strength of the obtained films was significantly higher compared to a flat film reference under different contact conditions. Thermal analysis indicated a decrease of drug crystallinity upon incorporation into PLGA films. The in vitro release of CTZ from PLGA biomimetic films was tested in simulated saliva, and it exhibited an initial burst release, accompanied by a sustained release phase over 10 days. Finally, the mucoadhesive properties of the obtained films was studied using agar/mucin plate as a representative mucosal substrate, and the results demonstrated superior mucoadhesion potential of CTZ-loaded biomimetic film in comparison to its flat counterpart. Having demonstrated the ability to load CTZ into PLGA biomimetic films with enhanced adhesion capacity, the potential use in local oral drug delivery applications warrants further in vitro and in vivo investigations.

Development of an UPLC-MS/MS Method for Quantitative Analysis of Clotrimazole in Human Plasma Samples

An ultra-performance liquid chromatography-tandem mass spectrometry method was developed for the quantification of clotrimazole (CTZ) plasma levels after intravaginal administration of the drug given at approved dosages. Plasma samples were extracted by liquid–liquid extraction and a single chromatographic run could be completed within about 2 min. The method was linear over the investigated range (0.488–250 ng/mL) with all the correlation coefficients, R2, greater than 0.9903. All data were in the range of ±15.0% with respect to the nominal concentration for high QC and medium QC, and in the range ±20% with respect to the nominal concentration for low QC. This rapid and sensitive method was validated and could be applied to human plasma samples from a healthy volunteer, showing that the assay is able to detect plasma concentrations of CTZ in the range of those found after the administration of the drug at approved dosages in the clinical setting.

Ecotoxicological effects of the azole antifungal agent clotrimazole on the macrophyte species Lemna minor and Lemna gibba

Pharmaceuticals are a large and diverse group of compounds used to treat, prevent and diagnose disease. Among these, a group that has been recently detected in the aquatic environment is that of the azole compounds, commonly used as antifungals. Clotrimazole (CLO) is a nonbiodegradable persistent azole compound, with broad-spectrum antifungal activity for which virtually no toxicological data are available, especially towards aquatic plants. The few existent data point to a documented interference with cytochrome P450 system of exposed organisms. Therefore, the aim of this paper was to evaluate the ecotoxicological effects of the fungicide CLO on two aquatic macrophyte species, namely, Lemna minor and Lemna gibba. To attain this purpose, an acute assay (96 h) was performed with both species being exposed to CLO, in a concentration range of 0 to 5 μg L−1. The analyzed endpoints were levels of chlorophyll a and b, total, carotenoids, catalase (CAT) and glutathione -s-transferases activities (GSTs). In general, CLO exposure caused some minor alterations in L. minor and L. gibba pigment contents. Antioxidant enzymes exhibited a different pattern in both species, since the highest concentrations of CLO caused an increase on CAT activity, and a decrease on GSTs activity in L. minor, and the opposite in L. gibba, reflected by a decrease on CAT activity and an increase on GSTs activity in all tested concentrations. These results demonstrate that CLO exposure resulted in potential deleterious effects on macrophytes, namely with the involvement of the antioxidant defense mechanisms that were likely deployed to cope with pro-oxidative conditions established by CLO.

Design of Experiments-Assisted Development of Clotrimazole-Loaded Ionic Polymeric Micelles Based on Hyaluronic Acid.

Polymeric micelles based on amphiphilic polysaccharides have some advantages as a carrier of poorly soluble lipophilic drugs thanks to their characteristic "core–shell" structure. Previously, ionic polymeric micelles based on chitosan and fatty acids have been developed. The aim of the present study was the preparation and characterization of hyaluronic acid (HA) derivatives by direct ionic interaction between the HA carboxylic groups and the amine groups of dodecyl amine (DDA) and hexadecyl amine (HDA). The HA–HDA polymeric micelles were loaded with a poorly soluble hydrophobic antifungal drug, clotrimazole (CLO). A 23 full factorial experimental design was used to evaluate the effect of the following factors: HA/HDA ratio from 1:0.25 to 1:0.75, cholesterol (CHOL%) as percentage of HA from 10% to 30%, and preparation temperature from 20 to 40 °C. As dependent variables (responses), nanoparticle dimensions and clotrimazole concentration in the final colloidal dispersion were considered. To optimize the drug final concentration, the design was therefore expanded into a rotatable central composite design (CCD). The effects of the formulation variables and the composition of the optimized formulation were confirmed by a mixture design. Physicochemical characterization of the optimized formulation was performed, confirming the ionic interaction between the polysaccharide and the HDA.

Corrosion inhibition of zinc in 0.1 M hydrochloric acid medium with clotrimazole: Experimental, theoretical and quantum studies

Clotrimazole an antifungal drug was investigated for its corrosion inhibition action on zinc metal in 0.1 M HCl solution. Corrosion rate was determined by weight loss, potentiodynamic polarization and impedance spectroscopic techniques. Electrochemical methods (EIS and polarization) have reported inhibition efficiency of up to 90% for 500 ppm concentration of clotrimazole. Data obtained from both chemical and electrochemical studies showed that the corrosion rate of Zn decreased with increase in inhibitor concentration and also with temperature. Tafel polarization measurements showed that clotrimazole acts as mixed type of corrosion inhibitor and its adsorption on the zinc surface obeys Langmuir isotherm. Thermodynamic activation and quantum chemical parameters were calculated and discussed to describe the mechanism of adsorption. These results were supported by FTIR spectral study of corrosion product formed on the zinc surface in the presence of clotrimazole. Contact angle measurements show increased hydrophobicity of zinc surface, while SEM images of zinc surface confirmed considerable decrease of surface inhomogenities in the presence of clotrimazole.

The selection of the concentration of clotrimazole and povidone-iodineas the main active substances of the “Micromar” antifungal agent

Microsporia is one of the most common skin diseases of cats, most often provoked by the pathogen Microsporum canis. The pathogen of microsporium has highly contagious properties and for a long time remains capable of infection in the environment. It is important for the veterinary doctor not only to carry out complex therapy, but also to prevent the spread of the pathogen in the environment and to prevent the occurrence of secondary pyoderma during the course of microspores. Due to the tendency to the growth of fungal diseases, the development of the pathogen’s resistance existing medicines, there is a need for effective antifungal medicines that have fungicide and fungicidal effect. The development of the antifungal agent “Micromar” will allow to carry out complex treatments for the treatment of microspores in cats. Also, thanks to the properties of basic active ingredient? Th use of “Micromar” will provide acceleration of recovery period. The combination of clotrimazole and povidone iodine will provide an effective antifungal action with an antiseptic effect. Clotrimazole is a broad-spectrum antifungal agent that does not cause pathogen resistance. In turn, povidone iodine will provide antiseptic protection to the affected area of the cat's body with a prolonged effect. The studies were conducted to determine the concentration of clotrimazole and povidone iodine as the main active substances of the antifungal agent “Micromar” in the laboratory in nutrient media during the cultivation of the fungus Microsporum sanis. The results of the studies showed that the pure culture of the pathogen is sensitive to clotrimazole at a concentration of 0.25% and iodine povidone 5%. In appropriate concentrations, it is recommended to use clotrimazole and povidone iodine for the manufacture of the antifungal agent Micromar.

Development of a Method for the Quantification of Clotrimazole and Itraconazole and Study of Their Stability in a New Microemulsion for the Treatment of Sporotrichosis.

Sporotrichosis occurs worldwide and is caused by the fungus Sporothrix brasiliensis. This agent has a high zoonotic potential and is transmitted mainly by bites and scratches from infected felines. A new association between the drugs clotrimazole and itraconazole is shown to be effective against S. brasiliensis yeasts. This association was formulated as a microemulsion containing benzyl alcohol as oil, Tween® 60 and propylene glycol as surfactant and cosurfactant, respectively, and water. Initially, the compatibility between clotrimazole and itraconazole was studied using differential scanning calorimetry (DSC), thermogravimetric analysis (TG), Fourier transform infrared spectroscopy (FTIR), and X-ray powder diffraction (PXRD). Additionally, a simple and efficient analytical HPLC method was developed to simultaneously determine the concentration of clotrimazole and itraconazole in the novel microemulsion. The developed method proved to be efficient, robust, and reproducible for both components of the microemulsion. We also performed an accelerated stability study of this formulation, and the developed analytical method was applied to monitor the content of active ingredients. Interestingly, these investigations led to the detection of a known clotrimazole degradation product whose structure was confirmed using NMR and HRMS, as well as a possible interaction between itraconazole and benzyl alcohol.

Elevation of the resistance of heat-cured acrylic denture base resin against biofilm-forming Candida albicans by incorporating Amphotericin B or Clotrimazole

Aim: The ability of antifungal agents to reduce or eliminate biofilm formation by Candida albicans after incorporating heat-cured acrylic denture base materials was investigated. Materials and Methods: Different concentrations of Amphotericin B (AmB) and Clotrimazole (CT) were incorporated into polymethylmethacrylate (PMMA) specimens (10 mm × 10 mm × 2 mm). C. albicans was stimulated to produce biofilms on the surface of specimens. Biofilm count was determined by crystal violet and transmittance percentage assays. Porosity percentage of PMMA specimens was measured. Results: AmB and CT showed more effective action against C. albicans biofilms at low concentrations (1, 5, and 10 μg/ml). Meanwhile, high concentrations (25 and 50 μg/ml) showed less activity. Porosity percentage was decreased in PMMA containing low concentrations of both antifungal agents. Conclusion: Incorporation of AmB and CT into denture materials has a significant inhibitory effect on the biofilm produced by C. albicans, especially at low concentrations. Decrease in porosity level is another advantage evidenced by incorporating low concentrations of AmB and CT within denture.

Development of a common carp (Cyprinus carpio) pregnane X receptor (cPXR) transactivation reporter assay and its activation by azole fungicides and pharmaceutical chemicals

In mammals, the pregnane X receptor (PXR) is a transcription factor with a key role in regulating expression of several genes involved in drug biotransformation. PXR is present in fish and some genes known to be under its control can be up-regulated by mammalian PXR ligands. Despite this, direct involvement of PXR in drug biotransformation in fish has yet to be established. Here, the full length PXR sequence was cloned from carp (Cyprinus carpio) and used in a luciferase reporter assay to elucidate its role in xenobiotic metabolism in fish. A reporter assay for human PXR (hPXR) was also established to compare transactivation between human and carp (cPXR) isoforms. Rifampicin activated hPXR as expected, but not cPXR. Conversely, clotrimazole (CTZ) activated both isoforms and was more potent on cPXR, with an EC50 within the range of concentrations of CTZ measured in the aquatic environment. Responses to other azoles tested were similar between both isoforms. A range of pharmaceuticals tested either failed to activate, or were very weakly active, on the cPXR or hPXR. Overall, these results indicate that the cPXR may differ from the hPXR in its responses and/or sensitivity to induction by different environmental chemicals, with implications for risk assessment because of species differences.

Spectrophotometric and spectroscopic studies on charge transfer complexes of the antifungal drug clotrimazole

Clotrimazole (CLZ) is an imidazole derivative that is therapeutically used as an antifungal drug. The routine control analysis of CLZ requires rapid, reliable, accurate and precise methods for the quantification of the drug in its pharmaceutical formulations. Spectrophotometric methods are presented for the determination of the antifungal agent that are based on the charge transfer complexation reaction of CLZ with the π-acceptors tetracyanoethylene (TCE) and 7,7′,8,8′-tetracyanoquinodimethane (TCNQ). The formation of coloured complexes was quantitated at 396nm and 842nm for CLZ-TCE and CLZ-TCNQ, respectively, which enabled the development of simple and accurate spectrophotometric methods for the analysis of CLZ in pure drug substances and its pharmaceutical products. Under the optimum reaction conditions, linear relationships with appreciable correlation coefficients (0.9985–0.9994) were found between the absorbance at the relevant maxima and the concentrations of CLZ in the range of 5.00–35.00μgmL−1 for CLZ-TCE and 1.00–25.00μgmL−1 for CLZ-TCNQ. The molar absorptivities and Sandell's sensitivity values were also statistically evaluated. The proposed methods were successfully applied to analyze CLZ in topical creams and sterile solutions with mean recovery percentages in the range of 99.60–100.20%, which indicated no interference from the inactive ingredients. Furthermore, the charge transfer complexes of CLZ were also characterized by FT-IR spectroscopy.

Sub-lethal effects and bioconcentration of the human pharmaceutical clotrimazole in rainbow trout (Oncorhynchus mykiss)

The aim of this study was to characterize biomarker responses, haematological profiles, structural changes and uptake in juvenile rainbow trout exposed to clotrimazole (CLO) at three concentrations (0.01 – [lowest environmentally relevant concentration], 1.0 [highest environmentally relevant concentration] and 10 μg L−1) in a semi-static system over a period of 42 days. Antioxidant defence enzymes, which responded to CLO exposure, changed the oxidative stress status of cells, but no differences were observed in lipid peroxidation. Clotrimazole triggered a biphasic response of CYP3A-like activity in liver microsomes, which may indicate a detoxification process in the liver. Histopathological alterations were most pronounced in kidneys and testes in the group exposed to 10 μg L−1. Structural changes in the kidney included tubulonephrosis and hyaline droplet degeneration in the tubular epithelial cells. The relative proportions of germ cells in testes were changed: The number of spermatozoa was reduced, and the spermatogonia and spermatocytes were increased. The highest CLO concentration was detected in fish liver (3710 ng per gram wet tissue) and kidney (4280 ng per gram wet tissue). Depuration half-life was estimated to be 72, 159, and 682 h in liver, muscle, and kidney, respectively.Taken together, these results provide valuable toxicological data on the effects of CLO on aquatic non-target organisms, which could be useful for further understanding of the potential risks in the real aquatic environment.

SU‐F‐T‐677: Synergistic Effect(s) of Clotrimazole On Radiation Cell Survival of A549 Lung Cancer Cells in Glucose Vs. Galactose Media

Purpose:In order to determine the synergistic effect of clotrimazole on radiosensitivity of A549 lung cancer cells, and the effect of oxidative pathways on modulating radiosensitivity, we studied how these cells survived under varying amounts of radiation and clotrimazole as well ass when glucose was switched for galactose media.Methods:The glucose media was used to determine the presence of any synergistic effect of clotrimazole on radiation using values of radiation and clotrimazole concentrations, varying from 0 – 8 Gy and 0 – 20 µM, respectively. As a galactose diet is known to activate oxidative pathways, which do not rely on hexokinase II (HK2), all trials were repeated using galactose media to determine the extent that HK2 unbinding from the mitochondrial membrane plays a role in modulating the observed radiosensitivity. An apoptosis vs. necrosis assay was implemented to find out the modality by which cell death occurred. An intracellular lactate assay was performed to exhibit the extent of anaerobic glycolysis.Results:After running the primary experiments, it was found that in glucose media, the cancer cells showed higher cell kill when clotrimazole was added to the media, followed by the cells being irradiated.Conclusion:Given the preliminary results it is validated that under higher concentrations of clotrimazole, in glucose media, A549 lung cancer cells exhibit a lower amount of survival. While all results have not yet been gathered. We anticipate that in galactose media the A549 cells will exhibit this effect to a much smaller degree, if at all.

Antifungal activity of clotrimazole against Candida albicans depends on carbon sources, growth phase and morphology.

Vulvovaginal candidiasis, a superficial infection caused predominantly by the pathogenic fungus Candida albicans, is frequently treated with clotrimazole. Some drug formulations contain lactate for improved solubility. Lactate may modify C. albicans physiology and drug sensitivity by serving as a carbon source for the fungus and/or affecting local pH. Here, we explored the effects of lactate, in combination with pH changes, on C. albicans proliferation, morphology and clotrimazole sensitivity. Moreover, we determined the influence of growth phase and morphology per se on drug sensitivity. We showed that utilization of lactate as a carbon source did not promote fast fungal proliferation or filamentation. Lactate had no influence on clotrimazole-mediated killing of C. albicans in standard fungal cultivation medium but had an additive effect on the fungicidal clotrimazole action under in vitro vagina-simulative conditions. Moreover, clotrimazole-mediated killing was growth-phase and morphology dependent. Post-exponential cells were resistant to the fungicidal action of clotrimazole, whilst logarithmic cells were sensitive, and hyphae showed the highest susceptibility. Finally, we showed that treatment of pre-formed C. albicans hyphae with sublethal concentrations of clotrimazole induced a reversion to yeast-phase growth. As C. albicans hyphae are considered the pathogenic morphology during mucosal infections, these data suggest that elevated fungicidal activity of clotrimazole against hyphae plus clotrimazole-induced hyphae-to-yeast reversion may help to dampen acute vaginal infections by reducing the relative proportion of hyphae and thus shifting to a non-invasive commensal-like population. In addition, lactate as an ingredient of clotrimazole formulations may potentiate clotrimazole killing of C. albicans in the vaginal microenvironment.