Itraconazole loaded nanoemulsions: Hansen solubility & charge-driven permeation and GastroPlus based predicted In vivo performance.

Targeting tumor-associated macrophages (TAMs) represents a promising strategy for next-generation immunotherapy. This study investigates the underlying mechanisms of TAM repolarization from the pro-tumorigenic M2 phenotype to the anti-tumorigenic M1 phenotype induced by itraconazole (ITZ). M2 macrophages derived from THP-1 cells were used in all experiments, and ITZ was administered at a concentration of 10-5 M. Morphological changes were monitored via time-lapse imaging. Single-cell RNA sequencing (scRNA-seq) and triple-color immunostaining for organelles, cholesterol, and the M1 marker interleukin (IL)-1β were performed with and without ITZ treatment. For inhibition studies, β-cyclodextrin polyrotaxane (βCD-PRX), a compound that removes cholesterol from lysosomes, was used. The effects were assessed by time-lapse imaging and western blot analysis of the M2 marker CD163. Following ITZ treatment, a subpopulation of M2 macrophages exhibited morphological changes, shedding dendrites and migrating, indicative of an M1-like phenotype. Additionally, intracellular lipid droplets enlarged and swelled. scRNA-seq analysis revealed that M2 macrophages with reduced lysosomal vesicle biogenesis transitioned to an M1-like phenotype and identified 1,142 significantly enriched pathways, including M1-related signaling activation and cholesterol metabolism and transport pathways. Immunofluorescence analysis confirmed that macrophages shifting toward an M1-like phenotype expressed IL-1β, with enlarged intracellular lipid droplets identified as cholesterol-containing lysosomes. Notably, M2 macrophages that had adopted an M1-like morphology in response to ITZ reverted to their original M2-like shape and exhibited increased CD163 expression following βCD-PRX treatment. Inhibition of lysosomal cholesterol release by ITZ reprogrammed M2 macrophages into an M1-like phenotype, revealing a novel mechanism that may serve as a foundation for developing innovative TAM-targeted immunotherapies.

High dose dry powder inhalation of itraconazole nanocrystals: Impact of drug load and inhalation device.

Triazole antifungals are clinically established agents for the prophylaxis and treatment of invasive fungal diseases in patients with hematologic malignancies. To address the requirements of clinical therapeutic drug monitoring, we developed and validated a novel ultra-high-performance liquid chromatography tandem mass spectrometry method for the simultaneous quantification of three triazole antifungals and their metabolites in human plasma: itraconazole (ICZ), hydroxy-itraconazole (ICZ-OH), voriconazole (VCZ), voriconazole N-oxide (VCZ N-oxide), and posaconazole. The five target analytes were successfully separated using a BEH C18 column (2.1 × 50mm, 1.7 µm) maintained at 40°C, with gradient elution employing mobile phase A (5.0mM ammonium acetate in water containing 0.1% formic acid) and mobile phase B (100% acetonitrile [ACN]) at a constant flow rate of 0.4mL/min. A positive ion pattern was chosen for quantification under multiple reaction monitoring. Following the addition of 10 µL of internal standard, 50 µL of plasma underwent protein precipitation with ACN, and the resulting supernatant was diluted for subsequent analysis. Method validation followed Food and Drug Administration guidelines and Chinese Pharmacopoeia regulations, demonstrating acceptable accuracy, precision, matrix effects, recovery, and stability. The calibration curves exhibited excellent linearity over the range of 0.1-10µg/mL for all five analytes, with correlation coefficients (r2) ≥ 0.9962, while the lower limit of quantification and limit of detection were established at 0.1and 0.03µg/mL, respectively. The intra- and inter-day coefficients of variation were below 8.7% at all concentration levels, and the accuracy was 90.0%-113.0%. This methodology was successfully applied for TDM of three triazole antifungals and their metabolites in 150 patients with hematologic malignancies. Therefore, the method demonstrates good analytical performance, establishing its reliability for clinical TDM of triazole antifungals.

Corneal fungal infections are a leading cause of blindness worldwide; however, poor ocular drug absorption limits current topical antifungal treatments. Itraconazole (ICZ), a potent antifungal agent, exhibits low aqueous solubility and limited permeability. This study aimed to develop self-emulsifying drug delivery systems (SEDDS) to enhance the solubility, permeability, and ocular cell uptake of ICZ, providing a more effective topical therapy. ICZ solubility was evaluated in various vehicles (Tween 80, Tween 60, Span 20, coconut oil, and olive oil). Ten optimized SEDDS formulations were prepared, with particle sizes ranging from 514 to 1,384nm (tenfold dilution). In vitro drug permeation was assessed using Franz diffusion cells with a parallel artificial membrane permeability assay. Cellular uptake was evaluated in ocular cell lines, and drug diffusion kinetics were analyzed using the Higuchi model. Formulation stability was assessed over a 6-month period. Formulation F1 achieved the highest permeation (96.71% ± 1.99%), followed by F3 (96.33% ± 3.24%) and F2 (80.98% ± 2.85%), whereas ICZ-PEG showed minimal permeation (11.55% ± 2.80%). The Higuchi model indicated diffusion-controlled transport. Cellular uptake was highest for F3, followed by F1 and F2, with approximately 50.3-, 38.6-, and 12.3-fold higher uptake than ICZ-PEG, respectively. All ICZ-SEDDS formulations remained stable for > 6months. ICZ-SEDDS markedly improved the solubility, permeability, and ocular cell uptake of ICZ compared with ICZ-PEG. The superior performance of formulation F3 highlights the potential of SEDDS as an effective strategy for overcoming limitations in topical antifungal therapy for corneal fungal infections.

Candida glabrata often resides within macrophages, creating challenges for antifungal treatment. This study investigates the impact of antifungal agents on the immune response of C. glabrata within macrophages to understand its immune evasion mechanisms. Standard C. glabrata strains ATCC2001 and a clinical strain (17K1152) were pretreated with micafungin (MCF), itraconazole (ICZ), and amphotericin B (AmB) for 24 hours and then co-cultured with macrophages for 6, 12, 24, and 48 hours. Transmission electron microscopy (TEM) analysis was performed, and cytokine levels (interleukin [IL]-6, granulocyte-macrophage colony-stimulating factor [GM-CSF]) were measured using enzyme-linked immunosorbent assay kits. C. glabrata showed the greatest susceptibility to MCF, followed by AmB, with ICZ having the least effect. TEM analysis revealed MCF-induced damage to both the cell wall and membrane, whereas AmB disrupted only the membrane. ICZ showed less noticeable damage. MCF-pretreated yeast cells were more readily damaged in macrophages than those pretreated with AmB or ICZ. IL-6 secretion patterns were similar in untreated and ICZ-treated groups, while AmB initially decreased IL-6 secretion before recovery, and MCF consistently increased IL-6 levels. GM-CSF secretion showed comparable trends for untreated and ICZ groups, with MCF and AmB exhibiting fluctuations. Echinocandins enhanced macrophage immune activity by modifying C. glabrata's cell wall and increasing pro-inflammatory cytokine secretion, impairing its survival within host cells. Azole pretreatment had minimal impact on yeast replication and survival, reflecting resistance. Polyenes-treated yeast evade innate immunity by reducing pro-inflammatory cytokine secretion and modulating macrophage recruitment.IMPORTANCECandida glabrata is a major cause of bloodstream infections and is often resistant to antifungal treatment. Understanding how different antifungal drugs affect its survival within macrophages is critical for improving therapy. This study demonstrates that echinocandins enhance macrophage-mediated killing by damaging the fungal cell wall and increasing pro-inflammatory cytokine secretion, whereas azoles have minimal effect, and polyenes may aid immune evasion. These findings provide mechanistic insights into antifungal resistance and host-pathogen interactions, informing strategies for more effective treatment of C. glabrata infections.

Dissolvable microarray patches (MAP) represent a promising drug delivery platform; however, the absence of standardised protocols for their preclinical evaluation poses a significant barrier to regulatory approval and clinical translation. Physiologically Based Pharmacokinetic (PBPK) modelling is a powerful tool for predicting drug kinetics following MAP application, addressing key challenges associated with in vitro and in vivo studies such as experimental variability, complex study design, and data extrapolating across different populations. However, adapting PBPK models for dissolvable MAP is inherently complex due to the interplay between microneedle geometry, drug release kinetics, and skin physiology. In this study, an existing dermal PBPK model in MoBi® was optimised for dissolvable MAP by incorporating microneedle geometry and in vitro release profiles of MAP formulations containing the antihistamines loratadine (LOR) and chlorpheniramine maleate (CPM), as well as the antifungal drug itraconazole (ITZ). Model refinement involved systematically optimising input parameters related to skin thickness and drug-skin diffusion, partitioning and binding, to enhance predictive accuracy. Validation was performed using in vitro permeation testing with porcine skin for CPM and LOR MAP, alongside in vivo preclinical studies in pigs for ITZ MAP. The optimised model demonstrated robust predictive performance across the diverse drug molecules and experimental conditions investigated, highlighting its value as a powerful tool to accelerate preclinical MAP development.

Objective: To develop and optimize itraconazole (ICZ) nanoembedded microparticles (NMPs) for pulmonary delivery to enhance the treatment of aggressive pulmonary fungal infections, such as aspergillosis, in immunocompromised patients with chronic respiratory conditions by improving ICZ solubility, dissolution, and lung-specific drug delivery. Methods: Itraconazole nanocrystals (INCs) were formulated using ultrasonic processing with Poloxamer-188 or Brij 58 as stabilizers to enhance solubility. Quality by Design (QbD) principles were applied to evaluate the effects of formulation and process variables on ICZ solubility and dissolution. Optimized INCs were lyophilized into NMPs using α-Lactose Monohydrate USP as a matrix, sifted to a particle size of <5 µm, and subjected to micromeritic and dissolution studies. The pharmacokinetic performance of NMPs was compared to commercially available oral ICZ formulations by assessing plasma drug concentrations. Results: Optimized INCs, sonicated with Poloxamer-188 at 50% amplitude for 15 minutes, achieved a particle size of 174.4 nm, solubility of 0.31 mg/mL, and a dissolution time of 22.97 minutes to reach 90% of the dose. NMPs exhibited suitable properties for inhalation, disintegrating in the secondary bronchi to release INCs that rapidly penetrated alveolar fluids. Compared to oral formulations, NMPs showed a faster tmax, higher Cmax, and increased plasma ICZ bioavailability at equivalent doses, with significantly higher drug concentrations in the lungs and sustained effective levels. Conclusion: ICZ-loaded NMPs for pulmonary delivery offer a promising approach for treating pulmonary fungal infections, providing enhanced solubility, rapid dissolution, and superior lung-targeted drug delivery compared to oral formulations, potentially improving therapeutic outcomes in vulnerable populations.

Polymeric films are effective for topical drug delivery due to their flexibility and adaptability, which are advantageous for patients who have difficulty with traditional dosage forms. Films can offer extended residence times and controlled drug release based on their composition. Gellan gum (GG) and pectin (P) are promising materials due to their mucoadhesive properties and pH-responsive behavior. In this study, GG/P films, crosslinked with Ca2+ and/or Al3+ containing or not itraconazole (ITZ), were developed for the potential treatment of oral candidiasis. Films were produced by ionotropic gelation method followed by solvent casting and assessed for macroscopic appearance, thickness, surface pH, mechanical properties, saliva uptake, erosion, and mucoadhesion. The films’ surface pH ranged from 4.7 to 5.5, with a thickness of <1 mm. Average saliva uptake was 9.32 mL/g, and the erosion varied from 13% to 44%, with films based on GG:P (4:1) and crosslinked with Ca2+ showing the lowest value. Films exhibited elongation before perforation from 0.78% to 2.43% and breaking point from 17.44 to 66.44 mPa. Films showed high mucoadhesive capacity, and the incorporation of ITZ did not affect this property, resulting in greater mechanical strength and lower erosion. The set of data proved that the developed platform is promising for the proposed application.

Multiple-component covalent organic framework coated stainless steel fiber for efficient microextraction and determination of triazole antifungal drugs in human plasma.

The poor solubility of active compounds and the stability of solid dispersions containing processed drugs are among the main challenges in pharmaceutical research. This study investigates the potential for improving the solubility of itraconazole (ITR) through solid dispersion formulations using two different carriers (Polyvinylpyrrolidone K30 and Eudragit E 100) at two API : polymer weight ratios (1:2 and 1:5). Two different solvent-based methods for preparing solid dispersions – supercritical carbon dioxide (scCO2) and electrospinning (Esp) – were introduced. The results confirmed improvement in ITR solubility and dissolution, as well as changes in particle shape and size for both weight ratio formulations. After processing, a decrease in ITR crystallinity was observed. The results indicated that an increased amount of carriers led to better results. Consequently, these formulations were selected for stability studies. The samples were packed into aluminum sachets and stored in climate chambers under long-term (25°C, 60%RH) and accelerated (40°C, 75%RH) conditions for one month. We demonstrated the scCO2 samples were resistant to storage; neither the temperature nor humidity affected the properties of solid dispersions. In contrast, samples obtained via electrospinning were more sensitive to storage conditions.

Fungal keratitis (FK), a severe ocular infection caused primarily by Fusarium spp. and Aspergillus spp., remains a major cause of blindness worldwide. Current treatment relies on antifungal agents, though emerging azole resistance in Aspergillus fumigatus complicates therapeutic outcomes. We isolated a clinical FK strain and assessed its antifungal susceptibility using broth microdilution and E-test methods, followed by sequencing of the cyp51A gene's promoter region and coding sequence (CDS) to identify resistance-associated mutations. The isolate, confirmed as A. fumigatus (Aftr46-001) via ITS and β-tubulin sequencing, exhibited elevated minimum inhibitory concentrations (MICs) 16 µg/mL for itraconazole (ITC), 32 µg/mL for isavuconazole (ISA), 32 µg/mL for voriconazole (VRC), and 1 µg/mL for posaconazole (POS) by broth microdilution. E-test results revealed an ITR MIC of 12 µg/mL, ISA MIC 32 µg/mL, VRC MIC 32 µg/mL, and POS MIC of 2 µg/mL. The isolate exhibited MICs values at or above the CLSI and EUCAST epidemiological cutoff values (ECVs): 1 µg/mL for ITC, ISA, and VRC according to CLSI (with insufficient data to establish a POS ECV), and 1 µg/mL for ITC and VRC, 2 µg/mL for ISA, and 0.25 µg/mL POS per EUCAST criteria. Genotypic analysis identified a 46-nucleotide tandem repeat in the cyp51A promoter and two nonsynonymous mutations (Y46F, F70L). This represents the first report of a cyp51A TR46/Y46F/F70L-harboring A. fumigatus strain isolated from FK, with potential agricultural environmental origins, suggesting that this mutation may have become a potential driver of azole-resistant FK in areas with high agricultural exposure, urgently needing to be included in local treatment guidelines and active surveillance systems. The isolate's resistance to all tested azoles underscores the clinical challenge posed by this genotype. Following this combined intervention, the patient's left eye vision improved from counting fingers at 1 m upon admission to a final visual acuity of 0.3, indicating that combined surgery and local high-dose drug delivery can serve as an effective strategy for drug-resistant FK. These findings highlight the importance of monitoring azole-resistant A. fumigatus strains to guide clinical treatment. The global increase in azole drug resistance further highlights the urgency and importance of rapid and accurate pathogen identification and drug resistance testing in the current environment.

Sporothrix brasiliensis is the leading cause of feline sporotrichosis in Brazil, an emerging zoonosis. Itraconazole (ITZ) is the first-line therapy; however, therapeutic failures and reports of ITZ-resistant isolates emphasize the need for new therapeutic options. This study evaluated the in vitro susceptibility profile of 25 Brazilian S. brasiliensis isolates to amorolfine hydrochloride (AMR) and ITZ. AMR evidenced fungicidal activity in 76% and fungistatic activity in 24% of isolates. All isolates were susceptible to ITZ. These findings support AMR as a promising antifungal candidate against S. brasiliensis and highlight the importance of continued surveillance of ITZ susceptibility, especially in endemic regions of Brazil.

Brazil is an endemic region for both human and feline sporotrichosis, with Sporothrix brasiliensis being the main etiological agent. Currently, few effective antifungal agents are available for treating this mycosis in cats, and therapeutic studies remain limited. Itraconazole (ITZ) is the first-line drug; however, its effectiveness is variable. To evaluate the use of ITZ combined with β-glucans (Euglena gracilis) in feline sporotrichosis, a prospective, uncontrolled interventional study was conducted in 29 cats. Clinical cure was achieved in 21 animals (72.4%) with a median treatment duration of 10 weeks. Most of these cats presented with nasal region lesions, nasal mucosa involvement, and respiratory signs, which are commonly associated with poor therapeutic outcomes. Treatment failure occurred in 5 animals (17.2%), and 3 (10.3%) were lost to follow-up. No deaths were recorded during the study. Adverse drug reactions (ADRs) were observed in 2 cats (6.9%). These findings suggest that β-glucans may be a complementary strategy in the treatment of feline sporotrichosis, particularly in cases involving nasal lesions and respiratory signs, and may also contribute to the prevention of ADRs associated with conventional therapy.

We report the first case of phaeohyphomycosis caused by the dematiaceous dimorphic fungus Exophiala spinifera in a free-roaming domestic cat from the Americas, specifically diagnosed in the Central-West Region of Brazil, which progressed to euthanasia due to severe clinical deterioration. A comprehensive diagnostic approach was employed, integrating epidemiological, clinical, laboratory, mycological, and molecular analyses to confirm the fungal infection. In addition to the fungal infection, the cat tested seropositive for feline immunodeficiency virus (FIV) and was suspected of having seromucous adenocarcinoma. The response of E. spinifera to itraconazole (ITZ) therapy and the post-mortem findings are also presented. This case highlights the importance of considering Exophiala species in the differential diagnosis of fungal infections in animals. The findings contribute to a better understanding of the clinical presentation, diagnosis, and potential ecological implications of E. spinifera, offering valuable insights for veterinarians, researchers, and clinicians involved in the management of zoonotic mycoses.

Background: The limited aqueous solubility of BCS Class II drugs, exemplified by itraconazole (ITR), continues to hinder their bioavailability and therapeutic performance following oral administration. The present study investigated the development of amorphous solid dispersions (ASDs) of ITR via continuous manufacturing technologies, such as hot melt extrusion (HME) and spray drying (SD), to improve drug release. Methods: Polymer selection was guided by Hansen solubility parameter (HSP) analysis, film casting, and molecular modeling, leading to the identification of aminoalkyl methacrylate copolymer type A (Eudragit® EPO), polyvinyl caprolactam–polyvinyl acetate–polyethylene glycol graft copolymer (Soluplus®), and hypromellose acetate succinate HG (AQOAT® AS-HG) as suitable carriers. ASDs were prepared at drug-to-polymer ratios of 1:1, 1:2, and 2:1. Comprehensive characterization was performed using ATR-FTIR, NMR, DSC, PXRD, SEM, PLM, and contact angle analysis. Results: HME demonstrated higher process efficiency, solvent-free operation, and superior dissolution enhancement compared to SD. Optimized HME-based ASDs were formulated into tablets. The ITR–Eudragit® EPO formulation achieved 95.88% drug release within 2 h (Weibull model, R2 > 0.99), while Soluplus® and AQOAT® AS-HG systems achieved complete release, best described by the Peppas–Sahlin model. Molecular modeling confirmed favorable drug–polymer interactions, correlating with the formation of stable complex and enhanced release performance. Conclusions: HME-based continuous manufacturing provides a scalable and robust strategy for improving the oral delivery of poorly water-soluble drugs. Integrating predictive modeling with experimental screening enables the rational design of ASD formulations with optimized dissolution behavior, offering potential for improved therapeutic outcomes in BCS Class II drug delivery.

Antimicrobial resistance is a major global problem for public health, indicating the need for the development of new anti-infective drugs, among other actions (i.e., better stewardship, diagnostics, etc.). A common strategy in medicinal chemistry is to modify existing drugs with an organometallic moiety to enhance their efficacy or overcome resistance. One notable example is ferroquine, an organometallic derivative of chloroquine. Here, we describe the design, in-depth characterization, and in vitro evaluation of seven new derivatives of the antifungal drug itraconazole (ITZ) against parasitic and fungal pathogens. ITZ was selected as a privileged scaffold because it targets ergosterol biosynthesis, which is an essential component of cell membranes in fungi and trypanosomatid parasites. While none of the compounds were active against Trypanosoma cruzi and Leishmania infantum, the ferrocenyl derivatives proved to be 1.5- to 1.9-fold more potent than ITZ toward Trypanosoma brucei. Of particular interest, all of the compounds exhibited high antifungal activity against the azole-susceptible clinical isolates. Furthermore, the ferrocenyl-containing compound was the most active against Aspergillus. Despite showing 10-fold lower activity than ITZ, these organometallic derivatives constitute an interesting starting point for further pharmacomodulation since we confirmed that they blocked the ERG11 enzyme, the main target of azoles.

Endometrial cancer (EC) is a common gynecologic malignancy with limited treatment options. This study aimed to evaluate the potential of itraconazole (ITZ), a widely used antifungal drug, as an anti-tumor agent and an adjuvant to immunotherapy for EC. The effects of ITZ on Ishikawa cells were assessed using proliferation assays, apoptosis assays, and invasion assays. The combination of ITZ and immune checkpoint inhibitors (ICIs) was evaluated to determine their synergistic effects on tumor invasion. Tumor-associated macrophages (TAMs) polarization and cytokine levels were analyzed by flow cytometry and enzyme linked immunosorbent assay (ELISA). Western blotting and Real-time reverse transcription polymerase chain reaction (RT-PCR) were used to investigate the impact of ITZ on the Wnt/β-catenin signaling pathway. Finally, in vivo experiments were conducted using a mice tumor model to validate the anti-tumor effects of ITZ and its combination with ICIs. ITZ inhibits Ishikawa cells proliferation and invasion through apoptosis induction. When combined with ICIs, ITZ significantly enhanced the inhibition of tumor invasion, an effect associated with TAMs polarization. ITZ increased IFN-γ secretion, reduced IL-10 levels, and promoted TAMs polarization from the M2 to the M1 phenotype. Mechanistically, ITZ downregulated Wnt-3a and β-catenin expression while upregulating Axin-1, thereby suppressing Wnt/β-catenin signaling in TAMs. In vivo, ITZ and ICIs synergistically reduced tumor volume and weight, shifted TAMs polarization toward the M1 phenotype, and suppressed Wnt/β-catenin signaling. ITZ demonstrated robust anti-tumor activity against EC by inhibiting Ishikawa cells proliferation, invasion, and enhancing the efficacy of ICIs. Through its dual role in directly targeting tumor cells and modulating the tumor microenvironment, ITZ shows promise as a multitargeted therapeutic agent and a valuable adjuvant to immunotherapy for EC.

The active metabolite hydroxyitraconazole has substantially higher in vivo free fraction and free concentrations compared to itraconazole.

Dermatophytosis is a chronic public health issue in India, often requiring systemic antifungals. Itraconazole (ITZ) is commonly used, but conventional forms show variable absorption. Super-bioavailable itraconazole (SUBA-ITZ) offers improved pharmacokinetics, though clinical comparisons are limited. To compare the efficacy, safety, relapse, and recurrence rates of three oral ITZ formulations-Conventional ITZ 100 mg (C-ITZ), SUBA-ITZ 65 mg, and SUBA-ITZ 50 mg-in superficial dermatophytosis. In this open-label, randomised study at a tertiary hospital, 150 patients with confirmed tinea corporis, cruris, or faciei were equally assigned to: Group A: C-ITZ 100 mg BID Group B: SUBA-ITZ 65 mg BID Group C: SUBA-ITZ 50 mg BID Treatment lasted 6 weeks without topical antifungals. Assessments were done at 3 and 6 weeks; relapse and recurrence were monitored telephonically at 3 and 6 months. All groups showed similar clinical response at 6 weeks (BSA, PGA, KOH negativity), with no significant differences. At 3 months, Group C had significantly lower relapse and recurrence (10.2%) than Group B (31.1%, 28.9%) and Group A (23.9%, 26.1%) (p < 0.05). By 6 months, differences were not significant. All three itraconazole formulations demonstrated comparable short-term efficacy in treating superficial dermatophytosis. These findings underscore the need for larger, long-term randomised controlled trials to optimise itraconazole dosing strategies.