Tioconazole Research Articles

Tioconazole-Loaded Transethosomal Gel Using Box-Behnken Design for Topical Applications: In Vitro, In Vivo, and Molecular Docking Approaches.

Tioconazole (TCZ) is a broad-spectrum fungicidal BCS class II drug with reported activity against Candida albicans, dermatophytes, and certain Staphylococci bacteria. We report the use of TCZ-loaded transethosomes (TEs) to overcome the skin's barrier function. TCZ-loaded TEs were fabricated by using a cold method with slight modification. Box-Behnken composite design was utilized to investigate the effect of independent variables. The fabricated TEs were assessed with various physicochemical characterizations. The optimized formulation of TCZ-loaded TEs was incorporated into gel and evaluated for pH, conductivity, drug content, spreadability, rheology, in vitro permeation, ex vivo permeation, and in vitro and in vivo antifungal activity. The fabricated TCZ-loaded TEs had a % EE of 60.56 to 86.13, with particle sizes ranging from 219.1 to 757.1 nm. The SEM images showed spherically shaped vesicles. The % drug permeation was between 77.01 and 92.03. The kinetic analysis of all release profiles followed Higuchi's diffusion model. The FTIR, DSC, and XRD analysis showed no significant chemical interactions between the drug and excipients. A significantly higher antifungal activity was observed for TCZ-loaded transethosomal gel in comparison to the control. The in vivo antifungal study on albino rats indicated that TCZ-loaded transethosomal gel showed a comparable therapeutic effect in comparison to the market brand Canesten®. Molecular docking demonstrated that the TCZ in the TE composition was surrounded by hydrophobic excipients with increased overall hydrophobicity and better permeation. Therefore, TCZ in the form of transethosomal gel can serve as an effective drug delivery system, having the ability to penetrate the skin and overcome the stratum corneum barrier with improved efficacy.

Spectroscopic (FT-IR, FT-Raman, UV-vis and NMR) Investigation, Molecular Structure, Docking and Chemical Reactivity Elucidation of Antifungal Drug Tioconazole

Tioconazole (TCZ), an antifungal medication of the imidazole class has been investigated by experimental FT-IR, FT-Raman, UV–vis and NMR spectroscopy. The optimized molecular geometry and vibrational wave numbers were calculated by DFT/B3LYP/6–311++G(d, p). The detailed vibrational assignments were made by Potential Energy Distribution. The electrophilic and nucleophilic reactive sites were studied using MEP surface. The NBO analysis has been done to describe various hyper-conjugative interactions responsible for the stabilization of the molecule. The 1H NMR chemical shifts were calculated by GIAO method and these matched well with the experimental results, indicating the accuracy of calculation method. TD-DFT calculations have been performed to determine several electronic properties like excitation energies, oscillator strengths, wavelengths and HOMO-LUMO energies. The lipophilicity and aqueous solubility of the compound have been predicted by ALOGPS method. The log P value for TCZ approaches that this compound is less recommended for oral administration. Docking studies have been made to predict the binding orientation, affinity and antifungal activity of the compound.

Development and optimization of a new tioconazole vaginal mucoadhesive film using an experimental design strategy. Physicochemical and biological characterization

A new tioconazole (TCZ) mucoadhesive film, based on a biodegradable chitosan/ hydroxypropyl methylcellulose (CH/HPMC) blend, was developed for treatment of vaginal candidiasis. The formulation was optimized through an I-optimal design (minimizing the integral of the prediction variance across the factor space), where the impact of the proportion of the ingredients and processing variables on the quality of the final product was evaluated. Both, the thickness of the film and the swelling index, which affect patients’ comfort and compliance, were considered. Mechanical testing, such as load at break, elongation at break, and mucoadhesive strength were also included as dependent variables.The optimal mucoadhesive film formulation, which should be obtained at a drying temperature of 30 °C, was found to include the combination of CH and HPMC (forming polymers) at 0.25:0.75 ratio, a mixture of polyethylene glycol 400 and propylene glycol as plasticizers (0.07:0.93, 5% w/w), and TCZ loaded at 15 % w/w.The optimal preparation was subjected to exhaustive characterization studies, which revealed that the drug was entrapped in the polymeric matrix in an amorphous state and that the film exhibited a smooth and uniform surface, demonstrating excellent component compatibility. In vitro tests showed that the formulation has an excellent time to kill value (3 min) and lacks cytotoxicity, suggesting that it should be highly effective and safe.

Sublimation thermodynamics of antifungal drugs: Tioconazole, miconazole and climbazole

The temperature dependence of the saturated vapour pressure of the antifungal drugs – tioconazole (TCZ), miconazole (MCZ) and climbazole (CLZ) – was determined by the inert gas carrier transfer method. The standard molar enthalpies of sublimation at T = 298.15 K calculated from these data were 117.2 ± 0.5 kJ·mol-1 for TCZ ± 0.9, 106.2 ± 1.0 kJ·mol-1 for MCZ and 113.3 kJ·mol-1for CLZ. The vaporization and sublimation enthalpies of the compounds were found by the Langmuir and Clausius-Clapeyron equations without determining the vapour pressures of the drugs. Sublimation enthalpies derived by both these methods were in good agreement. It was shown that it was possible to estimate the thermodynamic sublimation functions by applying the space clusterization approach to the studied and structurally related compounds. In addition, the temperature and enthalpy of fusion of the compounds were measured by differential scanning calorimetry.

Tioconazole and Chloroquine Act Synergistically to Combat Doxorubicin-Induced Toxicity via Inactivation of PI3K/AKT/mTOR Signaling Mediated ROS-Dependent Apoptosis and Autophagic Flux Inhibition in MCF-7 Breast Cancer Cells.

Cancer is a complex devastating disease with enormous treatment challenges, including chemo- and radiotherapeutic resistance. Combination therapy demonstrated a promising strategy to target hard-to-treat cancers and sensitize cancer cells to conventional anti-cancer drugs such as doxorubicin. This study aimed to establish molecular profiling and therapeutic efficacy assessment of chloroquine and/or tioconazole (TIC) combination with doxorubicin (DOX) as anew combination model in MCF-7 breast cancer. The drugs are tested against apoptotic/autophagic pathways and related redox status. Molecular docking revealed that chloroquine (CQ) and TIC could be potential PI3K and ATG4B pathway inhibitors. Combination therapy significantly inhibited cancer cell viability, PI3K/AkT/mTOR pathway, and tumor-supporting autophagic flux, however, induced apoptotic pathways and altered nuclear genotoxic feature. Our data revealed that the combination cocktail therapy markedly inhibited tumor proliferation marker (KI-67) and cell growth, along with the accumulation of autophagosomes and elevation of LC3-II and p62 levels indicated autophagic flux blockage and increased apoptosis. Additionally, CQ and/or TIC combination therapy with DOX exerts its activity on the redox balance of cancer cells mediated ROS-dependent apoptosis induction achieved by GPX3 suppression. Besides, Autophagy inhibition causes moderately upregulation in ATGs 5,7 redundant proteins strengthened combinations induced apoptosis, whereas inhibition of PI3K/AKT/mTOR pathway with Beclin-1 upregulation leading to cytodestructive autophagy with overcome drug resistance effectively in curing cancer. Notably, the tumor growth inhibition and various antioxidant effects were observed in vivo. These results suggest CQ and/or TIC combination with DOX could act as effective cocktail therapy targeting autophagy and PI3K/AKT/mTOR pathways in MCF-7 breast cancer cells and hence, sensitizes cancer cells to doxorubicin treatment and combat its toxicity.

Novel Application of Pentabromobenzyl Column for Simultaneous Determination of Eight Antifungal Drugs Using High-performance Liquid Chromatography.

A new, accurate and sensitive reversed-phase high-performance liquid chromatography (RP-HPLC) as an analytical method for the quantitative determination of eight antifungal drugs in spiked human plasma has been described optimized and validated. The analyzed compounds were voriconazole (VOR), luliconazole (LUL), clotrimazole (CLO), tioconazole (TIO), posaconazole (POS), ketoconazole (KET), sertaconazole (SER) and terconazole (TER). The separation of the analyzed compounds was conducted using a novel pentabromobenzyl column known as COSMOSIL PBB-R (150 mm × 4.6 mm I.D., particle size 5 μm). The analysis of the studied drugs was determined within 14 min using a diode array detector and the mobile phase consisted of: 10 mM potassium dihydrogen phosphate buffer (pH 2.1): Methanol (2: 98 v/v). A linear response was observed for all compounds in the range of concentration studied. Sample preparation was done through liquid-liquid extraction using diethyl ether. This proposed method was validated in terms of linearity, limit of quantification, limit of detection, accuracy, precision and selectivity. The method was successfully applied for the determination of these drugs in their pharmaceutical formulations and in human plasma samples.

Design and Characterization of Chitosan Nanoformulations for the Delivery of Antifungal Agents.

Among different Candida species triggering vaginal candidiasis, Candida albicans is the most predominant yeast. It is commonly treated using azole drugs such as Tioconazole (TIO) and Econazole (ECO). However, their low water solubility may affect their therapeutic efficiency. Therefore, the aim of this research was to produce a novel chitosan nanocapsule based delivery system comprising of TIO or ECO and to study their suitability in vaginal application. These systems were characterized by their physicochemical properties, encapsulation efficiency, in vitro release, storage stability, cytotoxicity, and in vitro biological activity. Both nanocapsules loaded with TIO (average hydrodynamic size of 146.8 ± 0.8 nm, zeta potential of +24.7 ± 1.1 mV) or ECO (average hydrodynamic size of 127.1 ± 1.5 nm, zeta potential of +33.0 ± 1.0 mV) showed excellent association efficiency (99% for TIO and 87% for ECO). The analysis of size, polydispersity index, and zeta potential of the systems at 4, 25, and 37 °C (over a period of two months) showed the stability of the systems. Finally, the developed nanosystems presented fungicidal activity against C. albicans at non-toxic concentrations (studied on model human skin cells). The results obtained from this study are the first step in the development of a pharmaceutical dosage form suitable for the treatment of vaginal candidiasis.

New approaches to identification and characterization of tioconazole in raw material and in pharmaceutical dosage forms

Tioconazole (TCZ), a broad-spectrum antifungal agent, has significant activity against Candida albicans and other Candida species, and therefore, it is indicated for the topical treatment of superficial mycoses. The main goal of this work is to report an exhaustive identification and characterization procedure to improve and facilitate the online quality control and continuous process monitoring of TCZ in bulk material and loaded in two different dosage forms: ovules and nail lacquer. The methodologies were based on thermal (differential scanning calorimetry (DSC), melting point, and thermogravimetry (TG)), spectroscopic (ultraviolet (UV), Raman, near infrared (NIR), infrared spectroscopy coupled to attenuated total reflectance (FTIR-ATR), and nuclear magnetic resonance (NMR)), microscopic and X-ray diffraction (XRD). The TCZ bulk powder showed a high crystallinity, as observed by XRD, with a particles size distribution (3–95 µm) resolved by microscopic measurements. TCZ melting point (82.8 °C) and a degradation peak centered at 297.8 °C were obtained by DSC and DTG, respectively. An unambiguous structure elucidation of TCZ was obtained by mono- and two- dimensional 1H and 13C NMR spectral data analysis. The FTIR-ATR, Raman and NIR spectra of both the raw material and the commercial products were analyzed and their characteristic bands were tabulated. The best methods for TCZ identification in ovules were DSC, TG, XRD, NIR and Raman, while NIR and FTIR-ATR were the most appropriate techniques to analyze it in the nail lacquer. DSC, TG, DRX, Raman, FTIR-ATR and NIR spectroscopy are effective techniques to be used in online process analysis, because they do not require sample preparation, and they are considerably sensitive to analyze complex samples.

Multivariate analysis of tioconazole — TCNQ charge transfer interaction: Kinetics, thermodynamics and twofold response optimization

Charge-transfer complex (CTC) formation between tioconazole (TCZ) as an n-electron donor and 7, 7, 8, 8-tetracyanoquinodimethane (TCNQ) as a π-acceptor was studied spectrophotometrically with an accompanying kinetic and thermodynamic investigation. Multivariate data analysis via a set of experimental designs was executed for this purpose. A 23 - two-level full factorial design (FFD) was used for inspecting the proposed variables while a face-centered central composite design (FCCCD) was used to adjust the levels of variables proved to be significant. Two responses were quantified as a result of this interaction; complex I (Y1, measured at 743 nm) and complex II (Y2, measured at 842 nm). Derringer's function and overlaid contour plots were used to concurrently optimize both responses. Benesi–Hildebrand equation was applied to determine of formation constant (K), and the molar absorptivity (Ɛ) of the formed complex. Different thermodynamic parameters; the standard Gibbs free energy change (∆G°), the standard enthalpy of formation (∆H°) and the standard entropy change (∆S°) were determined for the reaction product. The proposed method was validated regarding the linearity, intra-, and inter-day precision and accuracy, limit of detection, limit of quantification and following the ICH standards. The proposed method was also applied for the determination of TCZ in its pharmaceutical preparations. Having a higher molar absorptivity and higher formation constant, complex II was of choice for all subsequent measurements. Application of Benesi–Hildebrand equation supported the formation of 1: 1 CTC. Thermodynamic study revealed the endothermic characters and the spontaneity of formation of the CTC at high temperature.

HPLC-UV vs. UPLC-DAD for Estimation of Tinidazole, Benzyl Alcohol and Hydrocortisone Acetate Simultaneously with Tioconazole and Its Related Impurities in Bulk and Pharmaceutical Formulations

Tioconazole (TCN) (anti fungal) is commonly co-formulated with tinidazole (TND) (antiprotozoal), hydrocortisone acetate (HYD) (anti infalmmatory) and benzyl alcohol (BZL) (preservative) in different pharmaceutical preparations. Tioconazole has three related impurities namely; tioconazole related impurities A, B and C (TCN-RA), (TCN-RB) and (TCN-RC), respectively. As the main purpose of a test for related substances is to control degradation impurities and to limit impurities arising during the synthesis of active ingredients and on the other hand, high efficiency and less elution are the basic requirements of high-speed chromatographic separation. So, two simple, precise, and accurate reversed-phase HPLC-UV and UPLC-DAD methods have been developed and compared for simultaneous estimation of TND, BZL, HYD and TCN in bulk and pharmaceutical formulations in the presence of TCN related impurities. A Eurospher C18 (250 mm × 4.6 mm, 5 μm) for HPLC method and an Eclipse plus C18 (100 × 2.1 mm, 1.7 μm) for UPLC method were used. An isocratic mobile phase consisted of 20 mM anhydrous KH2PO4 (containing 0.2% TEA v/v), pH adjusted to 6 with orthophosphoric acid: acetonitrile, (25:75, v/v) for HPLC method and a gradient mobile phase of 20 mM anhydrous KH2PO4 (containing 0.2% TEA v/v), pH adjusted to 6 with orthophosphoric acid and acetonitrile mixed in different proportions for UPLC method, were prepared. Flow rate was set at 1.0 mL min-1 and 0.3 mL min-1 for HPLC and UPLC methods, respectively and the detection was performed for both methods at 295 nm, 210 nm, 245 nm and 225 nm for TND, BZL, HYD and TCN and its related impurities, respectively. The propsed UPLC-DAD method had advantages, of being economic on the large scale with relatively much satisfactory results for linearity, sensitivity and specificity. The two proposed methods are applicable for the purity assessment of TCN in bulk, vaginal tablets and creams.

Pharmaceuticals from the whole root of Nauclea latifolia (Rubiaceae)

The fruit, leaf, stem and root of Nauclea latifolia have been found to posses antimicrobial properties. This work shows the characterization and antimicrobial studies of pharmaceuticals from the whole root of the plant. Soxhlet extraction of the whole root was done using HPLC grade n-hexane, ethylacetate and methanol in that order. Silica gel and sephadex column chromatography were used to isolate pure fractions. Three compounds were isolated. NMR spectroscopic analyses were used to elucidate the structures of the compounds as: strictosamide, quinovic acid and 3,4 dihydroxybenzoic acid. The compounds were found to have antibacterial and antifungal activities against eleven (11) human pathogenic bacteria namely: Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Streptococcus mutans, Streptococcuss pyogenes, Klebsiella pneumonia, Salmonella typhii, Proteus vulgaris, Enterobacter aerogenes, Staphylococcus albus and Bacillus subtilis and three human pathogenic fungi namely: Candida albicans, Aspergillus flavus and Aspergillus niger . The average diameters of zones of inhibition of the isolated compounds were comparable with those of the standard antibiotics (gentamicin for bacteria and tioconazole for fungi). The minimum inhibitory concentration (MIC), minimum bactericidal concentration (MBC) and minimum fungicidal concentration (MFC) of the three compounds confirmed they were active even at low concentrations.

Physicochemical characterization of the structure and desorption relationship of tioconazole-assembled gold nanoparticles investigated by density functional theory and Raman spectroscopy

Abstract The physicochemical characterization of the structure and desorption relationship in tioconazole (TN)-assembled gold nanoparticles (AuNPs) was studied using density functional theory (DFT) calculations and Raman spectroscopy. UV–vis absorption spectroscopy and surface-enhanced Raman scattering (SERS) were performed to examine the loading of TN on the metal nanoparticle surfaces. From the SERS spectra, TN appeared to adsorb on AuNPs mainly by way of its imidazole and 2-chlorothiophene unit. On the basis of the DFT calculations of the 6-metal cluster atom model, the binding energies of TN via the 2-chlorothiophene unit were predicted to be smaller than those for the imidazole coordination. The hydrodynamic diameters of TN-conjugated AuNPs were reduced to be in the range of ∼165 nm after the PEGylation and to be suitable for the cellular uptake. The TN adsorbate was supposed to release from the AuNPs in the intracellular fungal component of glutathione after treatment with the fungi, consistent with the DFT calculations. Cell viability tests of Penicillium digitatum, Penicillium expansum, and Aspergillus nidulans before and after treatment with the TN-assembled AuNPs were examined to test the feasibility of developing an efficient drug delivery system. After 24 h treatment, TN-adsorbed AuNPs exhibited enhanced fungal cell killing according to a Student t -test ( p

Increase of activity of tioconazole against resistant microorganisms by the addition of butylated hydroxyanisole

Tioconazole (TCZ) killed resistant Candida albicans in less than 3 min, after the addition of butylated hydroxyanisole (BHA), a phenolic antioxidant, at subinhibitory concentrations. The bactericidal activity was also rapid against resistant Escherichia coli. BHA increased the TCZ activity in RPMI 1640 medium 18 times against ten strains of resistant C. albicans as judged by MIC and increased the activity 43 times against ten resistant E. coli strains. BHA at subinhibitory concentrations promoted the reduction of C. albicans virulence by reducing 180 times the hyphal cells of TCZ and decreasing hydrophobicity. The synergy could be due to changes in cellular permeability because of increased leakage of cellular enzymes.

Sertaconazole: In-Vitro Antifungal Activity Against Vaginal and Other Superficial Yeast Isolates

The in vitro susceptibilities of 183 clinical yeast isolates to sertaconazole (STZ) were compared to their susceptibilities to clotrimazole (CTZ), econazole (ECZ), ketoconazole (KTZ), miconazole (MNZ), fluconazole (FLZ), itraconazole (ITZ), tioconazole (TCZ), amphotericin B (AMB) and flucytosine (5FC) by using a commercial agar diffusion method. Strains were isolated from vaginal and other superficial clinical samples (18 species of Candida and five strains belonging to other yeast genera). Only one strain (0.5%) was resistant to STZ out of 87.4% of susceptible strains (n=160). The percentage of susceptible strains was higher than those obtained with the other agents evaluated and the percentage of resistant strains was lower than for most of the other antifun-gals. The pattern of susceptibility of C. albicans to STZ, TCZ, ITZ and CLZ was similar and superior to the pattern of susceptibility of this species to MNZ, ECZ, FLZ, 5FC and KTZ. C. dubliniensis was more susceptible to STZ, MNZ, MNZ, FLZ, ITZ, CLZ than to TCZ, ECZ, 5FC, AMB or KTZ. Ten susceptible strains to STZ were resistant to FLZ and one strain was resistant to ITZ. The overall antifungal activity of STZ in vitro against a wide range of clinically important yeasts from vaginal and cutaneous samples indicates the therapeutic potential of this agent for the treatment of infections caused by these fungi. However, the activity of STZ and the clinical value of in vitro data need to be verified in human clinical trials.