Treatment Of Cutaneous Fungal Infections Research Articles

Amphotericin B-loaded natural latex dressing for treating Candida albicans wound infections using Galleria mellonella model

Amphotericin B (AmB) is the gold standard for antifungal drugs. However, AmB systemic administration is restricted because of its side effects. Here, we report AmB loaded in natural rubber latex (NRL), a sustained delivery system with low toxicity, which stimulates angiogenesis, cell adhesion and accelerates wound healing. Physicochemical characterizations showed that AmB did not bind chemically to the polymeric matrix. Electronic and topographical images showed small crystalline aggregates from AmB crystals on the polymer surface. About 56.6% of AmB was released by the NRL in 120 h. However, 33.6% of this antifungal was delivered in the first 24 h due to the presence of AmB on the polymer surface. The biomaterial's excellent hemo- and cytocompatibility with erythrocytes and human dermal fibroblasts (HDF) confirmed its safety for dermal wound application. Antifungal assay against Candida albicans showed that AmB-NRL presented a dose-dependent behavior with an inhibition halo of 30.0 ± 1.0 mm. Galleria mellonella was employed as an in vivo model for C. albicans infection. Survival rates of 60% were observed following the injection of AmB (0.5 mg.mL−1) in G. mellonella larvae infected by C. albicans. Likewise, AmB-NRL (0.5 mg.mL−1) presented survival rates of 40%, inferring antifungal activity against fungus. Thus, NRL adequately acts as an AmB-sustained release matrix, which is an exciting approach, since this antifungal is toxic at high concentrations. Our findings suggest that AmB-NRL is an efficient, safe, and reasonably priced ($0.15) dressing for the treatment of cutaneous fungal infections.

Formulation and Characterization of Griseofulvin Liposome Loaded Topical Film

This research paper explores the formulation and characterization of a liposome-loaded topical film containing griseofulvin, an antifungal drug. Liposome-based drug delivery systems offer enhanced drug concentration, prolonged residence time, and reduced side effects. This study aims to improve the efficacy of griseofulvin by incorporating it into liposomes and formulating it into a topical film for transdermal delivery. Liposomes were used as carriers to encapsulate Griseofulvin and incorporated into a topical film for ease of application. The paper reviews the principles of liposome-based drug delivery, transdermal film technology, and the potential benefits of this approach. The work plan includes preformulation studies, liposome preparation and evaluation, formulation of liposome-loaded topical films, and evaluation of film properties. The results demonstrated that the developed formulation has the potential to improve the delivery and efficacy of Griseofulvin in the treatment of cutaneous fungal infections.

Dissolving microneedle patches loaded with amphotericin B microparticles for localised and sustained intradermal delivery: Potential for enhanced treatment of cutaneous fungal infections

Fungal infections affect millions of people globally and are often unreceptive to conventional topical or oral preparations because of low drug bioavailability at the infection site, lack of sustained therapeutic effect, and the development of drug resistance. Amphotericin B (AmB) is one of the most potent antifungal agents. It is increasingly important since fungal co-infections associated with COVID-19 are frequently reported. AmB is only administered via injections (IV) and restricted to life-threatening infections due to its nephrotoxicity and administration-related side effects. In this work, we introduce, for the first time, dissolving microneedle patches (DMP) loaded with micronised particles of AmB to achieve localised and long-acting intradermal delivery of AmB for treatment of cutaneous fungal infections. AmB was pulverised with poly (vinyl alcohol) and poly (vinyl pyrrolidone) to form micronised particles-loaded gels, which were then cast into DMP moulds to form the tips. The mean particle size of AmB in AmB DMP tips after pulverisation was 1.67 ± 0.01 μm. This is an easy way to fabricate and load microparticles into DMP, as few steps are required, and no organic solvents are needed. AmB had no covalent chemical interaction with the excipients, but the crystallinity of AmB was reduced in the tips. AmB was completely released from the tips within 4 days in vitro. AmB DMP presented inhibition of Candida albicans (CA) and the killing rate of AmB DMP against CA biofilm inside porcine skin reached 100% within 24 h. AmB DMP were able to pierce excised neonatal porcine skin at an insertion depth of 301.34 ± 46.86 μm. Ex vivo dermatokinetic and drug deposition studies showed that AmB was mainly deposited in the dermis. An in vivo dermatokinetic study revealed that the area under curve (AUC0-inf) values of AmB DMP and IV (Fungizone® bolus injection 1 mg/kg) groups were 8823.0 d∙μg/g and 33.4 d∙μg/g, respectively (264-fold higher). AmB remained at high levels (219.07 ± 102.81 μg/g or more) in the skin until 7 days after the application of AmB DMP. Pharmacokinetic and biodistribution studies showed that AmB concentration in plasma, kidney, liver, and spleen in the AmB DMP group was significantly lower than that in the IV group. Accordingly, this system addressed the systemic side effects of intravenous injection of AmB and localised the drug inside the skin for a week. This work establishes a novel, easy and effective method for long-acting and localised intradermal drug delivery.

Regulatory Requirements and Innovation: A Comparison of the Dermatologic Antifungal Drug Product Markets in Brazil and United States

Development of novel dermatological topical products for the treatment of cutaneous fungal infections is a constant necessity, especially in developing countries. Through public health policies, many developing countries have facilitated in the last decades the entry of generic products, which can be superficially seen as a threat to innovation. To verify whether regulatory requirements, or the waiving of some requirements, could have an impact on innovation, we performed a detailed technical comparison of the dermatologic antifungal markets of Brazil and of the United States, taking Brazil as an example of a developing country with more lenient requirements regarding the registration of generic topical drug products. The official databank of ANVISA (DATAVISA) and of US Food and Drug Administration (Orange Book) were assessed for valid topical dermatological antifungal drug products registered. The Brazilian market has a greater number of registered drug products encompassing a greater variety of drug substances than the US, but the latter comprises more products with novel technologies. In both countries, cream was the predominant dosage form and imidazoles were the major substance group. Ketoconazole was the lead active substance in Brazil and ciclopirox was the lead drug in the US. Generic products dominated both markets. Despite the great number of registered products, the Brazilian market lacks the latest technologies, reflecting that the ease of generics registration is not accompanied by innovation.

Biochemical and Molecular Effects of <i>Phoenix dactylifera</i> and <i>Ziziphus spina-christi</i> Extracts on <i>Candida albicans</i>

Our overall knowledge of the medicinal uses of plants suggests that natural compounds could be useful in the treatment of cutaneous fungal infections in tropical regions. Furthermore, the possibilities of treatment using plant extracts may be even broader than is already known when one considers plants that have not been extensively studied in this context, such as the regional species Phoenix dactylifera and Ziziphus spina-christi. This study compared the antimicrobial activity of Phoenix dactylifera and Ziziphus spina-christi extracts in terms of their biochemical and molecular effects on Candida albicans (ATCC CA 10231). These effects included altered levels of intracellular sterols, changes in the permeability of the cell membrane, and changes in the TEF1: QRTTEF1, CaERG1: ERG1, CdERG12: CdERG1, and ERG25: ERG25 genes. Scanning electron microscopy (SEM) was used to identify morphological characteristics, and energy-dispersive X-ray spectroscopy (EDAX) analyses were conducted. In treated samples, the SEM and EDAX analyses showed cell cavities and shrinkage of the cell wall, and the number of cells was reduced to only a few abnormal cells as compared with that in the untreated samples. Yttrium was detected in the cells treated with Z. spina-christi, and high levels of osmium were detected in the cells treated with P. dactylifera. Compared with control cells, cells exposed to the concentration 150 μl/ml of Z. spina-christi extract had an average sterol concentration that was nearly 3 times higher, while the concentration was 5.5 times higher for cells treated with the 150 μl/ml of P. dactylifera extract. The ethanol extracts affected the permeability of C. albicans cell membrane. Gene sequencing showed gaps and mismatches in the ERG1, ERG12, and ERG25 genes after treatment with P. dactylifera and Z. spina-christi extracts compared with that in the controls. The results were highly significant (p ≤ 0.01). We conclude that the ethanol extracts of P. dactylifera and Z. spina-christi have antimicrobial activity through several mechanisms in the yeast cell.

Biocompatible microemulsions for improved dermal delivery of sertaconazole nitrate: Phase behavior study and microstructure influence on drug biopharamaceutical properties

The aim of this study was development of biocompatible topical microemulsions (MEs) for incorporation and improved dermal delivery of sertaconazole nitrate (SN). For this purpose, phase behavior and microstructure of pseudo-ternary glycereth-7-caprylate/caprate (Emanon EV-E, EV)/cosurfactant/Capryol™ 90/water systems were investigated. Furhermore, the influence of these properties on the drug skin delivery was also assessed. Expansion of ME single-phase regions with the use of short chain alcohols was a consequence of the more fluid interface when compared to other investigated systems, which was confirmed by electron paramagnetic resonance spectroscopy – EPR. The chosen bicontinuous to inverted bicontinuous formulations were assessed against the ME based on polysorbate 80 as referent sample. Despite incorporation of SN within the selected formulations induced similar alternations in electrical conductivity, viscosity and pH values, obtained EPR spectra suggested different SN localization: within the oil phase (for most of the EV based formulations), or interacting with the interface (polysorbate 80 based formulation). Due to higher in vitro drug release (12.24%–18.53%), ex vivo SN penetration into porcine ear skin (dermal retention Enhancement Ratio (ERD) ranged from 2.66 to 4.25) and pronounced antifungal activity, the chosen MEs represent promising vehicles for dermal delivery of SN in treatment of cutaneous fungal infections. The biopharmaceutical and skin performance differences obtained with different formulations were possible to be explained on the basis of their physicochemical characteristics.

In-vitro antifungal susceptibility testing of lanoconazole and luliconazole against Aspergillus flavus as an important agent of invasive aspergillosis

The incidence of Aspergillus infections has recently increased remarkably in certain tropical and sub-tropical countries, with Aspergillus flavus being identified as the leading cause of infections after A.fumigatus. Lanoconazole (LAN) and luliconazole (LUL) are currently approved for topical treatment of cutaneous fungal infections. We aimed the in-vitro antifungal susceptibility testing of two imidazole, LANand LUL against A. flavus. One hundred and eighty-seven clinical and environmental A.flavus were tested originating from different climate zones of Iran between 2008 and 2015. The identification of all isolates was confirmed by using PCR-sequencing of β-tubuline ribosomal DNA gene. In-vitro antifungal susceptibility test was performed using CLSI guidelines against LAN, LUL, itraconazole (ITC), voriconazole (VRC), posaconazole (POS), Isavuconazole (ISA), amphotericin B (AMB), 5-flucytosine (5FC), caspofungin (CAS) and anidulafungin (AFG). The minimum inhibitory concentration (MIC) and minimum effect concentration (MEC) values were evaluated according to CLSI M38-A2 guidelines. The geometric mean MICs for tested antifungals, in increasing order, were: 0.009μg/mL for LUL (ranging from 0.004 to 0.062), 0.02μg/mL for LAN (ranging from 0.004 to 0.125), POS (0.10), ISA (0.16), ITC (0.24), VRC (0.27), AMB (1.8) and 5FC (63.06) μg/mL. The mean value of MECs for AFG and CAS were 0.06 and 0.07, respectively. Overall, LUL and LAN showed the lowest MIC against all isolates of A.flavus. Further studies are required to evaluate the in-vivo efficacy of these agents, and the possibility of using these agents in systemic infections.

Formulation and Evaluation of Nystatin Microparticles as a Sustained Release System

Nystatin is the drug of choice for treatment of cutaneous fungal infections with main disadvantage that is the need for multiple applications to achieve complete eradication which may reduce patient compliance. Microparticles offer a solution for such issue as they are one of sustained release preparations that achieve slow release of drug over an extended period of time. The objectives of this study were to fabricate nystatin-loaded chitosan microparticles with the ultimate goal of prolonging drug release and to analyze the influence of polymer concentration on various properties of microparticles. Microparticles were prepared by chemical cross-linking method using glutaraldehyde as cross-linking agent. Five formulas, namely N1C1, N1C2, N1C3, N1C4 and N1C5, were prepared and the effect of drug to polymer ratio was studied with respect to drug loading, encapsulation efficiency, particle size and morphology. Furthermore the prepared microparticles were subjected to various physico-chemical studies, such as drug- polymer compatibility by Fourier Transform Infrared Spectroscopy (FTIR) and in-vitro drug release characteristics. Microparticles obtained from N1C1, N1C2 and N1C3 formulas were regular in shape with mean particle size ranging between 1µm and 10µm. N1C5 formula was resulted in particles with irregular shape while N1C4 showed a blend of microparticles and deformed particles. The effect of chitosan concentration on drug loading and entrapment efficiency was studied. The results showed increment in these parameters that was directly proportional to the increment in polymer concentration. Percentage yield showed a significant increment which was related to the increment in the ratio of chitosan used during the study. FTIR results showed no interactions between nystatin and chitosan. DSC studies proved the crystalline nature of nystatin and chitosan. On other hand, the thermogram of loaded microparticles showed the absence of endothermic peak corresponding to nystatin which may indicate the loss of the crystalline nature of the drug presented inside the microparticles. In- vitro release studies resulted in 95.6% release of nystatin for N1C1 after 15 hours. N1C1 appeared to be promising in formulating microparticles that provide nearly complete release of the drug within15 hours. This formula can be selected in future work to be formulated as topical gel that prolongs the release of nystatin.
 Keywords: Nystatin, Chitosan, Glutaraldehyde, Chemical cross-linking.

A study of the treatment of cutaneous fungal infection in animal model using photoactivated composite of methylene blue and gold nanoparticle

BackgroundOnychomycosis is a widespread public health problem, in which T. rubrum and T. mentagrophytes is the commenest causative organisms. Current medical therapy has many drawbacks and side effects. Methylene blue (m.b) photodynamic therapy (pdt) proved efficacy but with lengthy sessions. ObjectivesOptimizing methylene blue photodynamic therapy by combination of methylene blue photosensitizer and gold nanoparticles (aunps) in a composite as gold nanoparticles are efficient delivery systems and efficient enhancers of photosensitizers for antifungal photodynamic therapy. Materials and methodsEighty newzealand rabbit (Oryctolagus cuniculus) were used and categorized in eight equal groups as follows; healthy and infection control, composite photodynamic therapy and five comparative groups. Photodynamic therapy was initiated at day three to five post inoculation, for four sessions forty eight hours apart. Each group divided and light exposure at two fluencies; 80J and 100J. All groups were investigated macroscopically and microscopically (histopathology and scanning electron microscope) also flowcytometry assessment for cell death and X-ray analysis for gold nanoparticles accumulation in brain and liver tissues were determined. ResultsRecovery from infection approaching 96% in gold nanoparticles+light group, around 40% in methylene blue photodynamic therapy and 34% in composite photodynamic therapy. The observed findings confirmed by apparent decrease of apoptosis, however small amounts of gold nanoparticles detected in brain and liver. ConclusionLight stimulated gold nanoparticles is a promising tool in treatment of onychomycosis.

The anti-dermatophyte activity of Commiphora molmol

Context: Commiphora molmol Engl (Burseraceae) or myrrh has been traditionally used for the treatment of skin fungal infections.Objective: This study evaluates the antifungal activity of myrrh ethanol extract and essential oil against skin dermatophytes.Materials and methods: The antifungal evaluations were performed by the food poisoning technique (250 ppm) and micro-broth dilution assay (800–6.25 µg/mL) against Trichophyton rubrum, T. mentagrophytes, Microsporum canis, M. gypseum, and T. verrucosum. The chemical composition of myrrh oil and ethanol extract was analyzed by GC and GC-MS.Results: Furanoeudesma 1,3-diene and menthofuran were the main components of myrrh oil, while 2-tert-butyl-1,4-naphthoquinone, benzenemethanol,3-methoxy-α-phenyl, and curzerene were the main components of myrrh ethanol extract. The inhibitory effect of myrrh oil and ethanol extract against dermatophytes were 43.1–61.6% and 12.5–27.5%, respectively. The MIC and MFC values of myrrh oil were 25–100 and 25–200 µg/mL while these amounts for ethanol extract were 25–400 and 25–400 µg/mL, respectively. Therefore, myrrh oil had higher antifungal activity than that of the ethanol extract. Both extracts showed good anti-elastase activity.Conclusion: The results of our investigation confirmed the traditional uses of C. molmol as a poultice for the treatment of cutaneous fungal infections.

Nanoethosomal formulation for skin targeting of amphotericin B: an in vitro and in vivo assessment

The present study is envisaged to develop nanoethosomal formulation for enhanced topical delivery of amphotericin B (AmB) for the treatment of cutaneous fungal infections. AmB encapsulated nanoethosomes were prepared using mechanical dispersion method in a strength of 0.1% w/w similar to the strength of marketed topical formulation. Vesicle size of nanoethosomal formulations was found to be in the range of 186 ± 2 to 298 ± 4 nm. The optimized nanoethosomal formulation was further incorporated in gel base to form AmB nanoethogel formulation. Rheological characterization study of nanoethogel demonstrated its viscoelastic nature with high elasticity and resistance to deformation at 37 °C. The yield stress value was found to be 108.05 ± 2.4 and 52.15 ± 0.9 Pa for nanoethogel and marketed gel formulation, respectively. The nanoethogel formulation exhibited 2.7- and 3.5-fold higher steady state transdermal flux and skin deposition of AmB, respectively, in comparison to marketed formulation. Confocal laser scanning microscopy (CLSM) study also revealed enhanced skin permeation and deposition with nanoethogel formulation. In vivo study showed that topical application of nanoethogel does not exhibit any skin irritation as tested by Draize test. The developed formulation, in comparison to the marketed gel, demonstrated a remarkable increase in the antifungal activity against Candida albicans. It is thus corroborated from the above results that nanoethosomal formulation represents an efficacious carrier for effective topical delivery of AmB.

Inkjet printing for pharmaceutical applications

Miconazole is an imidazole used for treatment of fungal infections that exhibits poor solubility in polar solvents (e.g., aqueous solutions). Microneedles, small-scale lancet-shaped devices that are commonly used for delivery of pharmacologic agents and vaccines, were made out of an acid anhydride copolymer using visible light dynamic mask micro-stereolithography/micromolding and loaded with miconazole using a piezoelectric inkjet printer. The miconazole-coated microneedles showed biodegradation and antifungal activity against the organism Candida albicans (ATCC 90028) on Sabouraud dextrose agar using an in vitro agar plating method. The results of this study demonstrate that piezoelectric inkjet printing may be used load microneedles and other drug delivery devices with pharmacologic agents. Miconazole-loaded microneedles prepared by the visible light dynamic mask micro-stereolithography–micromolding–piezoelectric inkjet printing approach have potential use in transdermal treatment of cutaneous fungal infections.

Oleic acid vesicles: a new approach for topical delivery of antifungal agent

The occurrence of topical fungal infections is increasing nowadays. Cutaneous fungal infections like cutaneous candidiasis are more prominent in patients associated with AIDS. Current available strategies for the treatment of cutaneous fungal infections are creams or gels which show various adverse effects on skin along with systemic absorption. These drawbacks can be overcome by using various novel drug delivery systems. So, the present investigation aims at exploring the potential of fatty acid vesicles (ufasomes) for the topical delivery of clotrimazole. Oleic acid was employed as a fatty material for the preparation of vesicles. Clotrimazole-loaded oleic acid vesicles were prepared using a thin film hydration method. Prepared vesicles were characterized for size, size distribution, shape, thermal behaviour (differential scanning calorimetry), in vitro release, in vitro antifungal activity, in vitro skin permeation and retention studies and for in vivo antifungal activity. Transmission electron microscopic (TEM) images confirmed the formation of vesicular dispersion (ufasomes) of clotrimazole. Oleic acid vesicles possessed high drug entrapment (49.5 ± 1.0%) and optimum size (455 ± 22 nm) along with good colloidal characteristics (polydispersity index = 0.210 ± 0.035 & zeta potential = − 22.45 ± 0.25 mV) at 4:6 drug-to-oleic acid ratio. In vitro drug release study showed sustained release of drug from the vesicular dispersion. Skin permeation and skin retention studies suggested accumulation of drug in the epidermal part of the skin. In vivo study confirmed prolonged release of drug from oleic acid vesicle up to five days indicating its usefulness for long-term therapy. So, it can be concluded from the present study that fatty acid vesicle may be a good approach to treat topical fungal infections.

Inkjet Printing of Amphotericin B onto Biodegradable Microneedles Using Piezoelectric Inkjet Printing

The delivery of amphotericin B, a pharmacologic agent with activity against a broad spectrum of fungi as well as against parasitic protozoa, has been complicated by the fact that amphotericin B exhibits poor solubility in aqueous solutions at physiologic pH levels. In this study, piezoelectric inkjet printing was used to modify the surfaces of Gantrez 169 BF microneedles (Ashland, Covington, KY). These amphotericin B-loaded microneedles demonstrated activity against Candida parapsilosis in a radial diffusion assay. The results of this study suggest that a combination of visible light dynamic mask microstereolithography, micromolding, and piezoelectric inkjet printing may be used to prepare amphotericin B-loaded microneedles with antifungal properties. It is envisioned that microneedles containing amphotericin B may be used for transdermal delivery of pharmacologic agents for the treatment of cutaneous fungal infections as well as cutaneous leishmaniasis.

Effect of Oral Administration of Zinc Sulphate with Simultaneous Use of Nonspecific Immunostimulation on the Course of Trichophytosis in Beef Cattle

Abstract The paper assesses the treatment of cutaneous fungal infection in young beef cattle caused by Trichophyton verrucosum using orally administered zinc sulphate for 28 d at 2 g per animal per day and subcutaneous application of 2.5 mg/kg levamisole administered three times every 7 d. The study was conducted on animals at the age of 110 to 122 d, which were divided into two experimental groups: group I (animals suffering from ringworm with low levels of zinc in serum) and group II (animals suffering from ringworm with normal levels of zinc in serum). The remaining animals (control group) were divided into three subgroups: K (clinically healthy animals, which had normal serum levels of zinc), KI (animals infected with ringworm and showing a reduced level of zinc), and KII (animals suffering from ringworm with normal serum zinc levels). During the study the following parameters were investigated: clinical signs of fungal infection, zinc content in serum, percentage of phagocytic cells, and phagocytic index. The symptoms of the disease were observed only in group II on day 56. In group I, an increase in the percentage of phagocytic cells was observed, as compared to the original values, on days 7, 21, and 56 and a fall in this parameter was noted on days 14 and 28. In group II, there was a decrease on day 14 and an increase in comparison to the original values, followed by a gradual fall till day 56 of the experiment. In group I, phagocytic index remained on a stable level till day 56 and in group II until day 28, and subsequently began to increase gradually. It was found that oral supplementation of zinc sulphate combined with nonspecific immunostimulation may be applied in fighting skin fungal infection in cattle, and that zinc and levamisole may exhibit synergism affecting the evolution of nonspecific immunity.

The off-Patent Anti-Fungal Ciclopirox Olamine Displays Preclinical Activity in Leukemia and Myeloma.

Off-patent drugs with previously unrecognized anti-cancer activity could be rapidly repurposed for this new indication given their prior testing for safety and toxicity. To identify such compounds, we developed, automated and conducted a cell-based chemical screen of 4800 off-patent drugs and chemicals. From this chemical screen, we identified the off-patent antimicrobial, ciclopirox olamine (CPX) that is currently used for the topical treatment of cutaneous fungal infections, but has not been previously evaluated as a systemic agent for the treatment of malignancy. As an anti-fungal agent, the mechanism of action of CPX is not well understood, but appears related to binding intracellular iron and inhibiting iron containing enzymes. To explore its efficacy and mechanism of action as an anticancer agent, leukemia, myeloma, and solid tumor cell lines were treated with increasing concentrations of CPX. 72 hours after incubation, cell viability was measured by the MTS assay. CPX decreased cell viability in 5/9 leukemia, 3/6 myeloma, and 3/5 solid tumor cells with an LD50 < 5 uM, a concentration that is pharmacologically achievable based on prior animal studies investigating CPX as an anti-fungal. Cell death was confirmed by the presence of a subG1 peak by flow cytometry after staining cells with propidium iodide. In contrast, CPX was less toxic to MRC 5, LF1, and GMO 5757 non malignant fibroblasts with an LD50 > 20 uM. Next, we evaluated CPX in combination with cytarabine and daunorubicin, standard chemotherapeutic agents used in the treatment of AML. In AML cell lines, CPX synergistically enhanced the cytotoxicity of cytarabine as determined by the median effect isobologram analysis. Specifically, the combination indices (CI) at the EC50, 75 and 90 were 0.18, 0.19, and 0.24, respectively, where a CI < 1 denotes synergy. In contrast, the addition of CPX to daunorubicin produced only additive effects. Given the effects in leukemia cells lines, we evaluated the effects of oral CPX in 3 mouse models of leukemia. Sublethally irradiated NOD-SCID mice were injected subcutaneously with OCI-AML2 or K562 human leukemia cells or intraperitoneally with MDAY-D2 murine leukemia cells. After tumor implantation, mice were treated with CPX (25mg/kg) in water or water alone by oral gavage. Oral CPX decreased tumor weight and volume in all 3 mouse models by up to 65% compared to control without evidence of weight loss or gross organ toxicity. Mechanistically, CPX arrested cells in the G1/S phase of the cell cycle and downregulated the expression of survivin, Cyclin D1, and the transcription factors YY1 and FTII-D prior to the onset of cell death. Consistent with effects as an anti-fungal, CPX bound intracellular iron in the malignant cells and its ability to bind intracellular iron was functionally important for its cytotoxicity. In contrast to CPX, deferoxamine, a more avid extracellular iron chelator, was not significantly cytotoxic with an IC25 > 10uM. The highest demand for intracellular iron occurs during the late G1 and S phases due, in part, to the activity of the iron-requiring enzyme ribonucleotide reductase. Therefore, we examined the effects of CPX on the activity of ribonucleotide reductase. By electron paramagnetic resonance (EPR), CPX inhibited ribonucleotide reductase at concentrations associated with cell death. Cell lines resistant to CPX-mediated inhibition of ribonucleotide reductase were also resistant to CPX-induced cell death, supporting a mechanism of action linked to ribonucleotide reductase. Thus, in summary, the off-patent anti-fungal agent CPX induces cell death through its ability to bind intracellular iron. Its ability to inhibit the iron-containing enzyme ribonucleotide reductase appears functionally important for its mechanism of action. CPX displays previously unrecognized anti-cancer activity at concentrations that are pharmacologically achievable. Thus, CPX could be rapidly repurposed for the treatment of malignancies including leukemia and myeloma.

Clotrimazole/betamethasone diproprionate: a review of costs and complications in the treatment of common cutaneous fungal infections.

The use of antifungal/corticosteroid combinations as topical therapy for dermatophytoses has been criticized as being less effective, more expensive, and the cause of more adverse cutaneous reactions than antifungal monotherapy. The combination of clotrimazole and betamethasone diproprionate (Lotrisone) is a mix of an azole antifungal and a high-potency corticosteroid, and is one of the most widely prescribed of these combinations. Our objective was to describe the beneficial and deleterious effects of Lotrisone in the treatment of common cutaneous fungal infections and its relative cost-effectiveness. We did a literature review documenting clinical trial data and adverse reactions to Lotrisone and collected a cost analysis of topical antifungal prescribing data over a 2-month period from a large midwestern staff-model health maintenance organization (HMO). Lotrisone is approved by the U.S. Food and Drug Administration (FDA) for the treatment of tinea pedis, tinea cruris, and tinea corporis in adults and children more than 12 years of age. Treatment is limited to 2 weeks in the groin area and 4 weeks on the feet. The most concerning adverse effects of Lotrisone were reported in children and included treatment failure, striae distensae, hirsuitism, and growth retardation. This combination was also reported to have decreased efficacy in clearing candidal and Trichophyton infections as compared to single-agent antifungals. Lotrisone was considerably more expensive than clotrimazole alone and was found to account for more than 50% of topical antifungal expenditures as prescribed by primary care physicians, but only 7% of topical antifungals prescribed by dermatologists. We found that Lotrisone was shown to have the potential to induce many steroid-related side effects and to be less cost effective than antifungal monotherapy. This combination should be used judiciously in the treatment of cutaneous fungal infections and may not be appropriate for use in children.