Broad-spectrum Antifungal Agent Research Articles

Utilization of Box Behnken Design for the Development and Evaluation of Luliconazole-loaded Nanoemulgel

Pharmaceutical research is always looking for novel ways to distribute drugs and developing these methods to increase the effectiveness of existing medications. The physicochemical characteristics of the drug determine the dose form. Lipophilic API development is a result of recent discoveries in high-throughput screening. A topical broad-spectrum antifungal agent is luliconazole. Due to its lower water solubility, topical administration is limited, and topical absorption is confined. The stratum corneum’s lipid phase’s solubility of the medication also serves as a rate-limiting step for penetration. Because fungal infections impact the skin’s epidermis, dermis, and deeper layers, medicine administration must be tailored to target high drug concentrations at the layers of the epidermis and dermis

Rational modification of melatonin for broad-spectrum antifungal agents discovery.

Natural products, known for their environmental safety, are regarded as a significant basis for the modification and advancement of fungicides. Melatonin, as a low-cost natural indole, exhibits diverse biological functions, including antifungal activity. However, its potential as an antifungal agent has not been fully explored. In this study, a series of melatonin derivatives targeting the mitogen-activated protein kinase (Mps1) protein of fungal pathogens were synthesized based on properties of melatonin, among which the trifluoromethyl-substituted derivative Mt-23 exhibited antifungal activity against seven plant pathogenic fungi, and effectively reduced the severity of crop diseases, including rice blast, Fusarium head blight of wheat and gray mold of tomato. In particular, its EC50 (5.4 µM) against the rice blast fungus Magnaporthe oryzae is only one-fourth that of isoprothiolane (22 µM), a commercial fungicide. Comparative analyzes revealed that Mt-23 simultaneously targets the conserved protein kinase Mps1 and lipid protein Cap20. Surface plasmon resonance assays showed that Mt-23 directly binds to Mps1 and Cap20. In this study, we provide a strategy for developing antifungal agents by modifying melatonin, and the resultant melatonin derivative Mt-23 is a commercially valuable, eco-friendly and broad-spectrum antifungal agent to combat crop disease.

First report of amphotericin B resistant Candida haemulonii isolated from the ICU of a referral hospital in Indonesia

Background and Objectives: Amphotericin B is a broad-spectrum antifungal agent commonly used to treat Candida haemulonii infection. C. haemulonii was isolated from patients reported to be intrinsically resistant to amphotericin B, en- coded by the ERG2 and ERG11 genes. However, there have been limited studies concerning amphotericin B-resistant C. haemulonii in Indonesia. The objective of this study is to explore the phenotypic and genotypic characteristics (ERG2 and ERG11) of C. haemulonii isolated from the ICU of a referral hospital in Indonesia.
 Materials and Methods: Identification and susceptibility tests were conducted using VITEK2. Thereafter, DNA was ex-tracted and amplified using conventional PCR followed by DNA sequencing (Sanger method).
 Results: The results of the phenotypic susceptibility test showed that all C. haemulonii were resistant to amphotericin B. ERG2 and ERG11 sequences showed the same amino acid sequence and corresponded to references that are resistant to amphotericin B.
 Conclusion: The resistant properties of C. haemulonii against amphotericin B found in this study require further exploration, including comparing resistant and sensitive C. haemulonii to amphotericin B. In addition, it is necessary to analyze other genes besides ERG2 and ERG11.

Pharmacokinetics of amphotericin B after accidental overdose in an adult critically ill patient treated with plasmapheresis: a case report and review of literature

Amphotericin B is a broad-spectrum antifungal agent that is used in the treatment of systemic fungal infections. We describe the case of a 62-year-old female patient with recent aneurysmal subarachnoid hemorrhage who was treated for suspected ventriculitis and a fungal coinfection. Instead of liposomal amphotericin B (L-AmB), 465 mg (5 mg/kg) amphotericin B deoxycholate (DOC) was inadvertently administered, leading to refractory shock with multiple organ failure and requiring mechanical ventilation. Since an overdose of amphotericin B can lead to fatal consequences and has a half-life of 15 days, plasmapheresis was started. The serum concentration decreased from 1.32 µg/mL to 0.62 µg/mL before plasmapheresis, demonstrating a mean half-life of 49 hours. After two plasmapheresis sessions, the serum concentration further dropped to 0.26 µg/mL, demonstrating a mean half-life of 17 hours. In contrast, the third plasmapheresis session had no effect on serum concentration. The patient made a full recovery, potentially facilitated by enhanced amphotericin B elimination through plasmapheresis. Positive outcomes were previously reported in two adult patients treated with plasmapheresis. However, other reports without plasmapheresis described fatal outcomes in adult patients, albeit with a twofold overdose compared to the two patients successfully treated with plasmapheresis. Moreover, plasmapheresis itself carries risks such as hypocalcemia, metabolic alkalosis, and coagulation deficits. Consequently, the role of plasmapheresis in amphotericin B overdose is still debated.

Stability and Analytical Characterization of Voriconazole as Measured by Immunoassay.

Voriconazole is a broad-spectrum triazole antifungal agent recommended for invasive fungal diseases, including invasive aspergillosis. Therapeutic drug monitoring via voriconazole target trough concentration is important to ensure efficacy while preventing toxicity. Our aim was to determine the stability of voriconazole as adapted and measured by an immunoassay. Plasma from patient samples (n = 45) evaluated by a liquid chromatography with tandem mass spectrometry (LC-MS/MS) method was compared against an ARK immunoassay method, adapted and optimized on the Abbott Alinity c analyzer. Stability of voriconazole and analytical performance of ARK immunoassay was assessed, including functional sensitivity, limit of blank (LoB), limit of detection (LoD), and limit of quantification (LoQ), linearity, and precision. ARK voriconazole immunoassay was highly correlated (Pearson R = 0.988) to the LC-MS/MS method, with an average bias of 0.09 mg/L (2%). CV at LoQ of 0.5 mg/L was 3.7% while the functional sensitivity was established at 0.05 mg/L. Overall imprecision with liquid quality control material obtained from ARK was 5.0%, 6.3%, and 5.9% at 1 mg/L, 5 mg/L, and 10 mg/L, respectively. Limit of blank and LoD were 0.02 mg/L and 0.05 mg/L, respectively. Voriconazole in lithium heparin plasma separator tube declines over time, with a decrease that is more evident near or above toxic concentrations. Voriconazole collected in gel separation tubes declines over time, possibly due to absorptive properties. Voriconazole measurements by immunoassay and LC-MS/MS demonstrated acceptable comparability with sufficient level of sensitivity and precision.

SAR study of N′-(Salicylidene)heteroarenecarbohydrazides as promising antifungal agents

Clinically available antifungal drugs have therapeutic limitations due to toxicity, narrow spectrum of activity, and intrinsic or acquired drug resistance. Thus, there is an urgent need for new broad-spectrum antifungal agents with low toxicity and a novel mechanism of action. In this context, we have successfully identified several highly promising lead compounds, i.e., aromatic N′-(salicylidene)carbohydrazides, exhibiting excellent antifungal activities against Cryptococcus neoformans, Candida albicans, Aspergillus fumigatus and several other fungi both in vitro and in vivo. Building upon these highly promising results, 71 novel N′-(salicylidene)heteroarenecarbohydrazides 5 were designed, synthesized and their antifungal activities examined against fungi. Based on the SAR study, four highly promising lead compounds, i.e., 5.6a, 5.6b, 5.7b and 5.13a were identified, which exhibited excellent potency against C. neoformans, C. albicans and A. fumigatus, and displayed impressive time-kill profiles against C. neoformans with exceptionally high selectivity indices (SI ≥ 500). These four lead compounds also showed synergy with clinical antifungal drugs, fluconazole, caspofungin (CS) and amphotericin B against C. neoformans. For the SAR study, we also employed quantitative structure–activity relationship (QSAR) analysis by taking advantage of the accumulated data on a large number of aromatic and heteroaromatic N′-(salicylidene)carbohydrazides, which successfully led to rational design and selection of promising compounds for chemical synthesis and biological evaluation.

Synthesis, antimicrobial activity and molecular docking studies of novel hydantoin derivatives as potential phospholipase A2 inhibitors

This work presents the synthesis, characterization and evaluation of a series of new multifunctional N-substituted hydantoin derivatives for their antibacterial and antifungal activity. Elemental analysis, 1H NMR, 13C NMR, and mass spectroscopy were used to confirm the newly synthesized compounds' structure. The antibacterial and antifungal properties of each synthesized molecule were examined. The examined compounds showed significant to moderate antimicrobial activity against the tested Gram-positive, Gram-negative, and fungal strains with MIC values ranging from 10.3 to 84.2 µM. Compounds 22 (MIC: 11.7–13.5 µM) and 25 (MIC: 10.2–11.9 µM) demonstrated an impressive MIC value against the tested bacterial and fungal strains when compared to the reference medications fluconazole (MIC: 11.7–14.5 µM) and streptomycin (MIC: 14.5 µM), which are broad-spectrum antibiotics and antifungal agents, respectively. Additionally, all of the compounds were tested for their ability to inhibit the phospholipase A2 (PLA2) enzyme, with the IC50 values ranging from 8.53 to 65.14 µM. When compared to the reference drug ursolic acid (IC50: 12.58 µM), compounds 22 (IC50: 10.27 µM) and 25 (IC50: 8.53 µM) were shown to be the most potent PLA2 inhibiting compounds in this series. The findings of the Molecule description with Drug-Likeness Prediction demonstrated that all the compounds are in a linear correlation with Lipinski's rule of five, demonstrating good drug-likeness qualities. The inhibitory effects of the most potent compounds (18, 19, 22 and 25) against the target PLA2 protein (PDB ID: 2H4C) were explained by molecular docking studies. The molecular docking results were in good accord with the experimental findings, and as these compounds had superior binding affinities within the active pocket, they may be classified as potent inhibitors of specific targets.

SP-141 targets Trs85 to inhibit rice blast fungus infection and functions as a potential broad-spectrum antifungal agent

Rice blast is a devastating disease worldwide, threatening rice production and food security. The blast fungus Magnaporthe oryzae invades the host via the appressorium, a specialized pressure-generating structure that generates enormous turgor pressure to penetrate the host cuticle. However, owing to ongoing evolution of fungicide resistance, it is vitally important to identify new targets and fungicides. Here, we show that Trs85, a subunit of the transport protein particle III complex, is essential for appressorium-mediated infection in M. oryzae. We explain how Trs85 regulates autophagy through Ypt1 (a small guanosine triphosphatase protein) in M. oryzae. We then identify a key conserved amphipathic α helix within Trs85 that is associated with pathogenicity of M. oryzae. Through computer-aided screening, we identify a lead compound, SP-141, that affects autophagy and the Trs85–Ypt1 interaction. SP-141 demonstrates a substantial capacity to effectively inhibit infection caused by the rice blast fungus while also exhibiting wide-ranging potential as an antifungal agent with broad-spectrum activity. Taken together, our data show that Trs85 is a potential new target and that SP-141 has potential for the control of rice blast. Our findings thus provide a novel strategy that may help in the fight against rice blast.

Validation of RP-HPLC UV method for determination ketoconazole in rabbit plasma: An application to the pharmacokinetic study

Ketoconazole is an imidazole broad-spectrum antifungal agent used for systemic and local infections. The pharmacokinetic information for ketoconazole is relatively limited. The validated method for determining ketoconazole in plasma rabbit is not yet reported. The HPLC-UV method is simple, rapid, cost-effective, sensitive, and only requires a small blood sampling. The chromatographic system consisted of a reversed-phase C18 column (250 x 4.6 mm, 5 µm) at a flow rate of 1 ml/min and detection wavelength of 240 nm, and the retention time of about 5 minutes for ketoconazole and 11 min for itraconazole as internal standard. The calibration curve presented linearity in the 0.05-8 µg/ml with R2=0.9969, which indicated good linearity. The method's precision was obtained by intra-day and inter-day repeatability studies, and accuracy was examined by %diff. The result is

Phytochemical Screening, Bioactive Compound Quantification by HPLC, and Antifungal Properties of Andrographis paniculata Extracts Against Plant Pathogenic Fungi

The common issues that occurred in plant disease had been studied for many years and fungi were determined as the major microbial agent that typically reduces the crop yield quality and economic losses. The increasing of antimicrobial resistance fungi and chemical fungicides has urged the development of new antifungal agents from medicinal plants such as Andrographis paniculata which is riches in phytochemical compounds. Hence, this research was carried out to investigate the phytochemical compounds and antifungal properties of a botanical extract derived from A. paniculata to inhibit fungal growth. A maceration technique was used where the extracts were soaked in methanol solvent at room temperature (±28°C) for 72 hours. The prepared samples were studied for phytochemical screening, and it shows the presence of alkaloids, flavonoids, saponins, tannins, and terpenoids. Whereas the quantitative estimation of andrographolide compound was conducted by reversed-phase high performance liquid chromatography (HPLC) method. The antifungal test was tested on Fusarium spp., Colletotrichum spp., Neoscytalidium dimidiatum, and Thielaviopsis paradoxa through agar disc diffusion. The result revealed that it exhibits significant fungal growth inhibition. Therefore, this study presented the versatile use of A. paniculata extract in developing its role as a novel broad-spectrum antifungal agent against plant disease.

Rapidly Resolving Orbital Pseudotumor in Systemic Lupus Erythematosus

Rapidly Resolving Orbital Pseudotumor in Systemic Lupus Erythematosus

Comparison between Ethylcellulose and Polymethacrylate Topical Nanosponges Loaded with Butenafi ne Hydrochloride

In this study, nanosponges (NS) loaded with butenafi ne hydrochloride (a broad-spectrum benzylamine antifungal agent) were successfully prepared by emulsion solvent diff usion method using ethylcellulose or polymethacrylate as retarder polymers. PVA was used as a stabilizer. NS 18 formulations were formulated with various ratios of 1:1, 1:2, and 1:3 (%, w/w), using diff erent PVA concentrations of 0.25, 0.5, and 0.75 (%, w/w). The prepared formulations were tested for particle size in terms of diameter, which ranged from (73.6 ± 15.4–991 ± 11.2 nm) for ethylcellulose NS, and from (116 ± 0.96–1439.6 ± 237.4 nm) for polymethacrylate NS. The entrapment effi ciency of the selected formulas (F1, F10) were (74.12 ± 2.60%) and (72.89 ± 1.41%). The selected formulas F1 and F10 were also evaluated for their polydispersity, yield, and in-vitro drug release. The release of BFNS powder in F10 (90.42 ± 1.81%) was higher than that of ECNS F1 powder (78.53 ± 3.24%) within 24 hours. Evaluation tests of the scanning electron microscope (SEM), Fourier transform infrared ray (FTIR), and powder X-ray diff raction (PXRD) confi rmed the NS’ spherical and porous features, the absence of any drug polymer interaction, and the stability of the drug in the delivery system. The two formulas of F1 and F10 were found to exhibit prolonged drug release, which minimizes side eff ects and dosing frequency of BF drug application.

Development of miconazole nitrate nanoparticles loaded in nanoemulgel to improve its antifungal activity

Miconazole is a synthetic derivative of imidazole, a medication with a broad-spectrum antifungal agent that is used to treat localized vaginal, skin, and nail infections. The aim of the study was to develop an innovative technique to improve the permeability and efficacy of topical miconazole nitrate. A nanoemulgel of miconazole nitrate was formulated by the incorporation of a nanoemulsion and a hydrogel. The nanoemulsion was first optimized using a self-emulsifying technique, and the drug was then loaded into the optimum formulation and evaluated prior to mixing with the hydrogel. Miconazole nitrate nanoemulgel formulations were evaluated for their physical characteristics and antifungal activity. Based on the results, the formulation with 0.4 % Carbopol showed the highest release profile (41.8 mg/ml after 2 h); thus, it was chosen as the optimum formulation. A cell diffusion test was performed to examine the ability of the Miconazole nitrate nanoemulgel to penetrate the skin and reach the bloodstream. Percentage cumulative drug releases of 29.67 % and 23.79 % after 6 h were achieved for the MNZ nanoemulgel and the commercial cream, Daktazol, respectively. The antifungal activity of the novel MNZ nanoemulgel formulation was tested against Candida albicans and compared to Daktazol cream and almond oil; the results were: 40.9 ± 2.3 mm, 25.4 ± 2.7 mm and 18 ± 1.9 mm, respectively. In conclusion, a novel MNZ nanoemulgel showing superior antifungal activity compared to that of the commercial product has been developed. This nanotechnology technique is a step toward making pharmaceutical dosage forms that has a lot of promise.

Newfangled Topical Film-Forming Solution for Facilitated Antifungal Therapy: Design, Development, Characterization, and In Vitro Evaluation.

Luliconazole is a broad-spectrum topical antifungal agent that acts by altering the synthesis of fungi cell membranes. Literature suggests that the recurrence of fungal infection can be avoided by altering the pH of the site of infection. Studies have also suggested that fungi thrive by altering skin pH to be slightly acidic, i.e., pH 3-5. The current study is aimed to design, develop, characterize, and evaluate an alkaline pH-based antifungal spray solution for antifungal effects. Luliconazole was used as an antifungal agent and an alkaline spray was formulated for topical application by using Eudragit RS 100, propylene glycol (PG), water, sodium bicarbonate, and ethanol via solubilization method. Herein, sodium bicarbonate was used as an alkalizing agent. Based on DSC, FTIR, PXRD, scanning electron microscopy (SEM), and rheological analysis outcomes, the drug (luliconazole) and polymer were found to be compatible. F-14 formulation containing 22% Eudragit RS 100 (ERS), 1.5% PG, and 0.25% sodium bicarbonate was optimized by adopting the quality by design approach by using design of experiment software. The viscosity, pH, drying time, volume of solution post spraying, and spray angle were, 14.99 ± 0.21 cp, 8 pH, 60 s, 0.25 mL ± 0.05 mL, and 80 ± 2, respectively. In vitro drug diffusion studies and in vitro antifungal trials against Candida albicans revealed 98.0 ± 0.2% drug diffusion with a zone of inhibition of 9 ± 0.12 mm. The findings of the optimized luliconazole topical film-forming solution were satisfactory, it was compatible with human skin, and depicted sustained drug release that suggests promising applicability in facilitated topical antifungal treatments.

Voriconazole Eye Drops: Enhanced Solubility and Stability through Ternary Voriconazole/Sulfobutyl Ether β-Cyclodextrin/Polyvinyl Alcohol Complexes.

Voriconazole (VCZ) is a broad-spectrum antifungal agent used to treat ocular fungal keratitis. However, VCZ has low aqueous solubility and chemical instability in aqueous solutions. This study aimed to develop VCZ eye drop formulations using cyclodextrin (CD) and water-soluble polymers, forming CD complex aggregates to improve the aqueous solubility and chemical stability of VCZ. The VCZ solubility was greatly enhanced using sulfobutyl ether β-cyclodextrin (SBEβCD). The addition of polyvinyl alcohol (PVA) showed a synergistic effect on VCZ/SBEβCD solubilization and a stabilization effect on the VCZ/SBEβCD complex. The formation of binary VCZ/SBEβCD and ternary VCZ/SBEβCD/PVA complexes was confirmed by spectroscopic techniques and in silico studies. The 0.5% w/v VCZ eye drop formulations were developed consisting of 6% w/v SBEβCD and different types and concentrations of PVA. The VCZ/SBEβCD systems containing high-molecular-weight PVA prepared under freeze-thaw conditions (PVA-H hydrogel) provided high mucoadhesion, sustained release, good ex vivo permeability through the porcine cornea and no sign of irritation. Additionally, PVA-H hydrogel was effective against the filamentous fungi tested. The stability study revealed that our VCZ eye drops provide a shelf-life of more than 2.5 years at room temperature, while a shelf-life of only 3.5 months was observed for the extemporaneous Vfend® eye drops.

Myxobacterial Outer Membrane β-1,6-Glucanase Induced the Cell Death of Fusarium oxysporum by Destroying the Cell Wall Integrity.

The β-1,6-glucan is the key linker between mannoproteins in the outermost part of the cell wall and β-1,3-glucan/chitin polysaccharide to maintain the rigid structure of the cell wall. The β-1,6-glucanase GluM, which was purified from the fermentation supernatant of Corallococcus sp. EGB, was able to inhibit the germination of Fusarium oxysporum f. sp. cucumerinum conidia at a minimum concentration of 2.0 U/mL (0.08 μg/mL). The survival rates of GluM-treated conidia and monohyphae were 10.4% and 30.7%, respectively, which were significantly lower than that of β-1,3-glucanase treatment (Zymolyase, 20.0 U/mL; equate to 1.0 mg/mL) (72.9% and 73.9%). In contrast to β-1,3-glucanase treatment, the high-osmolarity glycerol (HOG) pathway of F. oxysporum f. sp. cucumerinum cells was activated after GluM treatment, and the intracellular glycerol content was increased by 2.6-fold. Moreover, the accumulation of reactive oxygen species (ROS) in F. oxysporum f. sp. cucumerinum cells after GluM treatment induced apoptosis, but it was not associated with the increased intracellular glycerol content. Together, the results indicate that β-1,6-glucan is a promising target for the development of novel broad-spectrum antifungal agents. IMPORTANCE Phytopathogenic fungi are the most devastating plant pathogens in agriculture, causing enormous economic losses to global crop production. Biocontrol agents have been promoted as replacements to synthetic chemical pesticides for sustainable agriculture development. Cell wall-degrading enzymes (CWDEs), including chitinases and β-1,3-glucanases, have been considered as important armaments to damage the cell wall. Here, we found that F. oxysporum f. sp. cucumerinum is more sensitive to β-1,6-glucanase GluM treatment (0.08 μg/mL) than β-1,3-glucanase Zymolyase (1.0 mg/mL). The HOG pathway was activated in F. oxysporum f. sp. cucumerinum cells after GluM treatment, and the intracellular glycerol content was significantly increased. Moreover, the decomposition of F. oxysporum f. sp. cucumerinum cell wall by GluM induced the burst of intracellular ROS and apoptosis, which eventually leads to cell death. Therefore, we suggest that the β-1,6-glucan of the fungal cell wall may be a better antifungal target compared to the β-1,3-glucan.

Fabrication of Clotrimazole Microparticles Using Polyethylene Glycol 6000 and Beeswax

Background Microparticles are one of drug delivery systems designed for sustained and controlled release of drugs for a long period of time. Clotrimazole is a broad-spectrum antifungal agent, which is generally used for treatment of Candida albicans and others fungal infections. Objective The present study aims to fabricate Clotrimazole microparticles using beeswax (lipophilic coating) and polyethylene glycol (hydrophilic coating). Methods In this study, Clotrimazole microparticles by polyethylene glycol (PEG 6000) and beeswax were prepared using spray drying technique. Morphology and size of the microparticles were assessed using optical microscopy and scanning electron microscopy. Particle size analyses were performed using Zetasizer technique. The drug release rate was measured using ultraviolet-visible spectroscopy. To study the interactions between microparticles and carriers, Fourier transform infrared spectroscopy (FTIR) was used. Results Size of the microparticles was below 1000 µm. The microparticles prepared from beeswax and PEG 6000 had quasicrystalline and crystalline morphologies, respectively. The maximum drug release rate in 6 hours was 80.53% in formulation F3 (microparticles prepared by beeswax) and 79.29 in formulation F6 (microparticles prepared by PEG 6000). The FTIR results showed probable interactions between clotrimazole microparticles and beeswax. Conclusion Both beeswax and PEG 6000 polymers result in similar rate of drug release; however, since beeswax has interactions with clotrimazole, PEG 600 formulation seems to be more appropriate for fabrication of clotrimazole microparticles.

The environmentally friendly approaches based on the heterojunction interface of the LaFeO3/Fe2O3@g-C3N4 composite for the disposable and laboratory sensing of triclosan.

Triclosan (TCS) is a polychlorinated phenoxy phenol (PCPPs) used as a disinfectant and a broad-spectrum antibacterial and antifungal agent in personal hygiene products. TCS easily forms diphenyl ethers and dioxins, which are persistent organic pollutants. This work used a double approach for the TSC sensing: a) screen-printed (SPE) electrochemical platform for on-site application, modified with lanthanum iron oxide and graphitic carbon nitride composite (LaFeO3/Fe2O3@g-C3N4/SPE); and b) carbon paste electrode (CPE), modified with the same material and used in laboratory conditions. Linear range from 0.1 μM to 10 μM, the limit of detection (LOD) of 29 nM and the limit of quantification (LOQ) of 91 nM were obtained for CP electrode in BRBS pH 8. SPE showed the best analytical parameters in BRBS at pH 3, with a linear range from 0.3 μM to 7 μM, LOD of 0.09 μM and LOQ of 0.28 μM. Furthermore, the influence of potential interferents was investigated and proven to be negligible. Determination of TSC was performed to estimate the environmental impact of this compound as well as the practical usefulness of the proposed sensor in the real sample analysis, confirmed with a HPLC analysis.

Enhancing the Antifungal Activity and Ophthalmic Transport of Fluconazole from PEGylated Polycaprolactone Loaded Nanoparticles

Fungal eye infections are caused mainly by an eye injury and can result in serious eye damage. Fluconazole (FLZ), a broad-spectrum antifungal agent, is a poorly soluble drug with a risk of hepatotoxicity. This work aimed to investigate the antifungal activity, ocular irritation, and transport of FLZ-loaded poly (ε-caprolactone) nanoparticles using a rabbit eye model. Three formulation factors affecting the nanoparticle's size, zeta potential, and entrapment efficiency were optimized utilizing the Box-Behnken design. Morphological characteristics and antifungal activity of the optimized nanoparticles were studied. The optimized nanoparticles were loaded into thermosensitive in situ hydrogel and hydroxypropylmethylcellulose (HPMC) hydrogel ophthalmic formulations. The rheological behavior, in vitro release and in vivo corneal transport were investigated. Results revealed that the percentage of poly (ε-caprolactone) in the nanoparticle matrix, polymer addition rate, and mixing speed significantly affected the particle size, zeta potential, and entrapment efficiency. The optimized nanoparticles were spherical in shape and show an average size of 145 nm, a zeta potential of -28.23 mV, and a FLZ entrapment efficiency of 98.2%. The antifungal activity of FLZ-loaded nanoparticles was significantly higher than the pure drug. The developed ophthalmic formulations exhibited a pseudoplastic flow, prolonged the drug release and were found to be non-irritating to the cornea. The prepared FLZ pegylated nanoparticles were able to reach the posterior eye segment without eye irritation. As a result, the developed thermosensitive in situ hydrogel formulation loaded with FLZ polymeric nanoparticles is a promising drug delivery strategy for treating deep fungal eye infections.

208. Clinical outcomes of empirical versus pre-emptive broad spectrum antifungal therapy in patients with acute myelogenous leukemia receiving antimold prophylaxis

Abstract Background Since antimold prophylaxis has been widely used in induction chemotherapy for acute myelogenous leukemia (AML), it should be re-evaluated whether broad spectrum antifungal therapy should be empirically used in prolonged febrile neutropenia. Therefore, we compared clinical outcomes of empirical versus pre-emptive antifungal therapy in patients with AML receiving antimold prophylaxis. Methods From September 2016 to December 2020, all adult AML patients (≥ 18 years) receiving antimold prophylaxis who had febrile neutropenia for ≥ 4 days during induction or re-induction chemotherapy at Seoul National University Hospital were retrospectively reviewed. They were classified into the empirical group (therapeutic broad spectrum antifungal agents had been used without evidence of invasive fungal infection [IFI]) or the pre-emptive group (antimold prophylaxis had been maintained until the emergence of IFI’s evidence). We compared clinical outcomes between the two groups after propensity score matching. Results A total of 229 chemotherapy episodes, 36 in the empirical group and 193 in the pre-emptive group, were analyzed. In the pre-emptive group, broad spectrum antifungal therapy was administered in 45 (23.3%) episodes. After 1:3 matching with age, gender, induction or re-induction chemotherapy, and worst qSOFA score at febrile neutropenia, incidence of proven or probable IFI (0/36 [0%] in the empirical group vs. 5/97 [5.2%] in the pre-emptive group, P=0.323) and all-cause mortality (3/36 [8.3%] in the empirical group vs. 4/97 [4.1%] in the pre-emptive group, P=0.388) were not different between the two groups. Conclusion Clinical outcomes of empirical versus pre-emptive broad spectrum antifungal therapy were comparable in patients with AML receiving antimold prophylaxis. Broad spectrum antifungal therapy could be delayed until the emergence of evidence of IFI, in the current era of antimold prophylaxis. Disclosures All Authors: No reported disclosures.