A stepwise optimization strategy to formulate in situ gelling formulations comprising fluconazole-hydroxypropyl-beta-cyclodextrin complex loaded niosomal vesicles and Eudragit nanoparticles for enhanced antifungal activity and prolonged ocular delivery

Fungal keratitis and endopthalmitis are serious eye diseases.Fluconazole(FL)is indicated for their treatment,but suffers from poor topical ocular availability.This study was intended to improve and prolong its ocular availability.FL niosomal vesicles were prepared using span 60.Also,polymeric nanoparticles were prepared using cationic Eudragit RS100 and Eudragit RL100.The investigated particles had adequate entrapment efficiency(EE%),nanoscale particle size and high zeta potential.Subsequently,formulations were optimized using full factorial design.FL-HP-β-CD complex was encapsulated in selected Eudragit nanoprticles(FL-CD-ERS1)and niosmal vesicles.The niosomes were further coated with cationic and bioadhesive chitosan(FL-CD-Nios-ch).EE%for FL-CD-ERS1 and FL-CD-Niosch formulations were 76.4%and 61.7%;particle sizes were 151.1 and 392 nm;also,they exhibited satisfactory zeta potential+40.1 and+28.5 m V.In situ gels were prepared by poloxamer P407,HPMC and chitosan and evaluated for gelling capacity,rheological behavior and gelling temperature.To increase the precorneal residence time,free drug and selected nano-formulations were incorporated in the selected in situ gel.Release study revealed sustained release within 24 h.Permeation through excised rabbits corneas demonstrated enhanced drug flux and large AUC0-6 h in comparison to plain drug.Corneal permeation of selected formulations labeled with Rhodamine B was visualized by Confocal laser microscopy.Histopathological study and in vivo tolerance test evidenced safety.In vivo susceptibility test using Candida albicans depicted enhanced growth inhibition and sustained effect.In this study the adopted stepwise optimization strategy combined cylodextrin complexation,drug nano-encapsulation and loading within thermosenstive in situ gel.Finally,the developed innovated formulations displayed boosted corneal permeation,enhanced antifungal activity and prolonged action.

Treatment of pulmonary coccidioidomycosis after successful hepatitis C therapy in a patient with fluconazole induced hepatotoxicity

A patient with hepatitis C infection and cavitary pul- monary coccidioidomycosis is reported. Treatment of hepatitis C was associated with resolution of flucona- zole-induced hepatotoxicity. Successful treatment of he- patitis C enabled the patient to tolerate increaseing doses of fluconazole. This case highlights that hepatic toxicity of fluconazole can improve after successful treatment of hepatitis C.

Fluconazole Prophylaxis in Neonates (Non-Systematic) Literature Review

Background: Nosocomial infection remains an important contributing factor for morbidity and mortality in neonates. Coagulase-negative staphylococci have emerged as the predominant pathogens of late onset sepsis. This is followed by staphylococcus aurous, gram negative bacilli, and fungi. Old studies noted that mortality due to candidemia was higher in infants weigh less than 2000 g after being exposed to risk factors. The prophylactic use of fluconazole for the prevention of IC in extremely low birth weight was first reported in 2001. Methods: Current guidelines from Europe and North America that refer to the treatment of fungal infections are included. Literature search was performed using Medline, Scopus and Cochrane Central Register of Controlled Trials through March, 2016. Conclusion: Mortality was not different in early studies. However, recent studies concluded that mortality was reduced in the fluconazole arms. Risk-based approach towards fluconazole prophylaxis seems to be safe and effective.

Evaluation of Dosages Regimen of Fluconazole in Patients of Candidemia with Gender by PK/PD and Mote Carlo Simulation

Candidemia is one of the four most common nosocomial blood infections and is the most common hospital-acquired fungemia in a recent multi-institutional study from the US. The mortality of Candidemia can be up to 50%. Fluconazole is a triazole derivative widely used for the treatment of invasive candidiasis. It was recommended as first-line drugs for the treatment and prevention of mycoses. In our study, we aimed to optimise the dosage of fluconazole with gender against Candida spp. based on pharmacokinetic/pharmacodynamics (PK/PD) analysis. We collected the published data about pharmacokinetic parameters of fluconazole and the MIC distribution of Candida spp. on fluconazole. We decided to evaluate the gender between males and females with the pharmacokinetics of fluconazole. Using probability of target attainment (PTA) and cumulative fraction of response (CFR) as indexes, crystal ball software 11.1.2.4 was used for Monte Carlo simulation of different dosage regimens of different males and females. For C. albicans, C. tropicalis and C. lusitaniae, when doses of regimen are 100 mg IV, 200 mg IV and MIC was lower than 1 g/ml, PTA was higher than 90%. For C. tropicalis, each dosing regimen and MIC was less than 2 g/ml. PTA was higher than 90%. As C. glabrata, C. parapsilosis, C. krusei, C. guilliermondii for PTA with more than 90%, MIC of fluconazole 200 mg were less than 32 g/ml, 64 g/ml and 64 g/ml, respectively. For the different dosage regimens 100 mg IV and 200 mg IV of fluconazole for Candida spp., it is desirable that fluconazole dosage regimens take into account both the gender of the patient.

Synthesis,Crystal Structure and Magnetic Property of a Novel Cu(Ⅱ)-Fluconazole-azide Complex

The title compound formulated as Cu2（C13H11N6OF2）（N3）3CH3OH was synthesized and structurally characterized by elemental analysis,IR and single-crystal X-ray diffraction.In the structure of the complex,the deprotonated fluconazole and azide anion link two copper centers to construct a binuclear SBU and the azide anion exhibits a μ1,1-coordination mode.Each triazole group of fluconazole links two SBUs and the compound exhibits a chain-like architecture with strong antiferromagnetism.

Simple,Inexpensive and Ecologically Friendly Derivative Spectrophotometric Fluconazole Assay from Nail Lacquer Formulations

Nail lacquers represent new drug form specifically designed to treat infected nail plate. They are complex organic solutions with specific assaying problems due to the high content of the polymer and plasticizer. Furthermore, there is a lack of assaying methods of active substances from this type of formulations in scientific literature. We developed derivative UV-spectrophotometric method for determination of fluconazole content in antifungal nail lacquer formulations. The method was validated for specificity, linearity, precision (repeatability), intermediate precision and accuracy (recovery). The method is specific, linear in the range of 99.53 - 497.65 μg/ml, precise and showed good recovery (98.79% - 101.77% from all six developed formulations). Besides, it is inexpensive, simple and nontoxic, i.e. ecologically acceptable. This method can be used for assaying fluconazole from this type of formulations.

Relationship between antifungal resistance of fluconazole resistant Candida albicans and mutations in ERG11 gene

Background The cytochrome P450 lanosterol 14a-demethylase （Ergllp） encoded by ERG11 gene is the primary target for azole antifungals. Changes in azole affinity of this enzyme caused by amino acid substitutions have been reported as a mechanism of azole antifungal resistance. This study aimed to investigate the relationship between amino acid substitutions in Erg11 p from fluconazole resistant Candida a/bicans （C. albicans） isolates and their cross-resistance to azoles. Methods Mutations in ERG11 gene were screened in 10 clinical isolates of fluconazole resistant C. albicans strains. DNA sequence of ERG11 was determined by PCR based DNA sequencing. Results In the 10 isolates, 19 types of amino acid substitutions were found, of which 10 substitutions （F72S, F103L, F1451, F198L, G206D, G227D, N349S, F416S, F422L and T482A） have not been reported previously. Mutations in ERG11 gene were detected in 9 isolates of fluconazole resistant C. albicans, but were not detected in 1 isolate. Conclusions Although no definite correlation was found between the type of amino acid substitutions in Ergllp and the phenotype of cross-resistance to azoles, the substitutions F72S, F1451 and G227D in our study may be highly associated with resistance to azoles because of their special location in Erg11p.

Evaluation of the disc diffusion method with a comparison study for fluconazole susceptibility of Candida strains

To performance susceptibility testing of antifungal agents Due to the increasing number of resistant strains, susceptibility testing of antifungal agents is gaining importance Methods We compared the results of standard macrotube dilution reference method with two different microdilution methods, as well as the disc diffusion method in order to test the susceptibility of 150 Candida strains to fluconazole Results Overall correlation between microdilution and macrodilution methods was 86% It was 91% between the Minimal Inhibitory Concentrations obtained from macrodilution and disc diffusion zone diameters Conclusion The disc diffusion test was evaluated as a low-cost, reproducible, and efficient way of assessing the in vitro susceptibility of Candida strains to fluconazole

Preparation of fluconazole buccal tablet and influence of formulation expedients on its properties

The aim of present study was to prepare buccal tablets of fluconazole for oral candidiasis.The dosage forms were designed to release the drug above the minimum inhibitory concentration for prolonged period of time so as to reduce the frequency of administration and to overcome the side effects of systemic treatment.The buccal tablets were prepared by using Carbopol 71G and Noveon AA-1 by direct compression method.Microcry stalline cellulose was used as the filler and its effect was also studied.The prepared dosage forms were evaluated for physicochemical properties,in vitro release studies and mucoadhesive properties using sheep buccal mucosa as a model tissue.Tablets containing 50% of polymers(Carbopol & Noveon) were found to be the best with moderate swelling along with favorable bioadhesion force,residence time and in vitro drug release.The in vitro drug release studies revealed that drug released for 8 h,which in turn may reduce dosing frequency and improved patient compliance in oral candidiasis patients.

Resistant mechanisms of Candida albicans to fluconazole

Objective To detect the resistant mechanisms o f Candida albicans to fluconazole (FCZ) at molecular biology lev el, since the resistance me chanisms of azole antifungal agents have been the focus of attention these years . Methods Thirty two FCZ resistant C.albicans were selected as our test strains (MICs≥64?μg/ml). With 14 α demethylase gene (ERG16 ge ne, target enzyme encoding gene of azoles) as our object, we chose six sets of prim ers from the ERG16 gene to amplify the interested fragments, and conducted South ern blot hybridization, restriction fragment length polymorphism (RFLP), and sin gle strand conformation polymorphism (SSCP) analysis for the fragments which wer e amplified by the six sets of primers, and pre resistant sensitive strains wer e used as controls. Three representative fragments, A66, D66 and E78, were selec ted to be cloned and sequenced. Results The polymerase chain reaction (PCR) amplification showed that s everal tested strains were negative for some primers. However, our Southern blot analysis reminded that their resistance did not result from the lack of targ et enzyme coding gene. SSCP analysis s howed that differences were noted between the resistant and sensitive strains an d inter resistant strains. Statistical analysis showed that the most variable s equence lied in the amplifier of the sixth pair of primer, and all the tested 32 strains showed positive results. In the 11 mutation points we found, five resu lted in amino acid alterations. It is likely that one or more mutational alterations (alone or in combination) might lead to the expression of an enzyme highly resi stant to the inhibitory action of FCZ which in turn is responsible for the FCZ r esistant trait in these strains. Conclusion One or more mutational alterations might lead to the azole r esistant trait in this strains.

Extractive Spectrophotometric Determination of Fluconazole by Ion-pair Complex Formation with Bromocresol Green

An extraction-spectrophotometric method for the determination of trace amounts of fluconazole was described. Fluconazole was effectively extracted as a 1 ： 1 ion-pair complex with bromocresole green （BCG） at pH 3.0 into chloroform, followed by spectrophotometric determination at 420 nm. Beer＇s law was obeyed over the range of 4-50 μg.mL^-1 of fluconazole with a detection limit of 3.7 μg.mL^-1 . The method is simple, rapid and sensitive. The procedure was applied to the determination of fluconazole in pharmaceutical preparations as well as its recovery from a blood serum sample.

Cryptococcosis in kidney transplant recipients:Current understanding and practices

Cryptococcosis is the third most commonly occurring invasive fungal disease in solid organ transplant recipients(SOT).It is caused by encapsulated yeast,Cryptococcus species,predominantly Cryptococcus neoformans and Cryptococcus gattii.Though kidney transplant recipients are at the lowest risk of cryptococcosis when compared to other solid organ transplant recipients such as lung,liver or heart,still this opportunistic infection causes significant morbidity and mortality in this subset of patients.Mortality rates with cryptococcosis range from 10%-25%,while it can be as high as 50%in SOT recipients with central nervous system involvement.The main aim of diagnosis is to find out if there is any involvement of the central nervous system in disseminated disease or whether there is only localized pulmonary involvement as it has implications for both prognostication and treatment.Detection of cryptococcal antigen(CrAg)in cerebrospinal fluid or plasma is a highly recommended test as it is more sensitive and specific than India ink and fungal cultures.The CrAg lateral flow assay is the single point of care test that can rapidly detect cryptococcal polysaccharide capsule.Treatment of cryptococcosis is challenging in kidney transplant recipients.Apart from the reduction or optimization of immunosuppression,lipid formulations of amphotericin B are preferred as induction antifungal agents.Consolidation and maintenance are done with fluconazole;carefully monitoring its interactions with calcineurin inhibitors.This review further discusses in depth the evolving developments in the epidemiology,pathogenesis,diagnostic assays,and management approach of cryptococcosis in kidney transplant recipients.

Solid-state characterization and impurities determination of fluconazol generic products marketed in Morocco

In this paper, we report the results of quality control based in pbysicochemical characteriza- tion and impurities determination of three samples of fluconazole drug substances marketed in Morocco. These samples were supplied by different pharmaceuticals companies. The sample A, as the discovered product, was supplied by Pfizer, while samples B and C （generics）, were manufactured by two different Indian industries. Solid-state characterization of the three samples was realized with different physicochemical methods as： X-ray powder diffraction, Fourier-transformation infrared spectroscopy, differential scanning calorimetry. High performance liquid chromatography was used to quantify the impurities in the different samples. The results from the physicochemical methods cited above, showed difference in polymorph structure of the three drug substances. Sample A consisted in pure polymorph II1, sample B consisted in pure polymorph I1, sample C consisted in a mixture of fluconazole Form Ili, form II and the monohydrate. This result was confirmed by differential scanning calorimetry. Also it was demonstrated that solvents used during the re-crystallization step were among the origins of these differences in the structure form. On the other hand, the result of the stability study under humidity and temperature showed that fluconazole polymorphic transformation could be owed to the no compliance with the conditions of storage. The HPLC analysis of these compounds showed the presence of specific

Prevalence of Non-<i>Albicans Candida</i>Infections in Women with Recurrent Vulvovaginal Symptomatology

Background: Candida vulvovaginitis is one of the most frequently diagnosed conditions in women’s care practices. Historically, 90% of cultured yeast species were C. albicans. However, due to a variety of interventions, the proportion of non-albicans Candida (NAC) infections appears to be increasing. We sought to estimate the current prevalence of Candida vulvovaginitis and the species-specific distribution of such infections in recurrent cases. Methods: Women with recurrent vulvovaginal symptomatology referred to an Obstetrics and Gynecology practice were tested by genital fungus culture, Candida-specific polymerase chain reaction (PCR), or both between July 2010 and February 2013. Results: A total of 103 women were tested. Mean age was 45.6 years. Including only their most recent positive test result, 29.1% (30/103) of women tested positive for Candida by any of the above testing measures. Of those, 50% (15/30) tested positive for C. albicans and 50% (15/30) tested positive for a NAC species. Across all visits, 60% (18/30) tested positive for C. albicans, 56.7% (17/30) tested positive for NAC, and 16.7% (5/30) tested positive for both a C. albicans and a NAC species. Among all isolated NAC species, 28.6% (6/21) were determined to be C. glabrata, 23.8% (5/21) C. krusei, 23.8% (5/21) C. parapsilosis, and 23.8% (5/21) other Candida species. Conclusion: Approximately 30% of women with recurrent vulvovaginal symptomatology have detectable Candida strains and it appears that NAC species may cause half of all these infections. This is imperative because NAC infections are usually more difficult to diagnose and are resistant to most treatments.

Acute middle ear Aspergillus niger infection in a patient with Stevens-Johnson syndrome: a case report

Rationale: Stevens-Johnson syndrome is a disorder mostly attributed to drug use, which can cause different skin problems. Patient concerns: A 31-year-old woman presented with severe pain and pounding in the left ear. Diagnosis: Fluconazole-induced Stevens-Johnson syndrome with fungal infection in ear. Intervention: Surgical and systemic antibiotic therapy. Outcomes: The patient recovered and resumed her normal life. Lessons: The prescription of fluconazole for patients with underlying conditions should be done with caution.

Dinuclear Nickel(Ⅱ) Complex with a Rare Ni(μ_2-O)_3Ni Core: Crystal Structure and Magnetic Studies

The first triple μ2-O-bridged dinuclear Ni（II） complex, [Ni2（Flu）3]（ClO4）·（H2O） （1, HFlu = 2-（2′,4′-difluorophenyl）-1,3-bis（1,2,4-triazol-1-yl）-propan-2-ol）, has been synthesized and structurally and magnetically characterized. Complex 1 crystallizes in monoclinic, space group P21/n with a = 12.7143（3）, b = 21.3392（6）, c = 17.6497（5）, β = 106.142（2）o, V = 4599.8（2）3, Z = 4, μ（MoKα） = 0.975 mm-1, Dc = 1.662 g/cm3, GOF = 0.83, the final R （I 〉 2σ（I）） = 0.0503 and wR （all data） = 0.1288. In 1, three Flu anions encapsulate two nickel（II） ions bridged by three deprotonated μ2-O groups. The central alkoxo and terminal triazolyl group chelate one nickel（II） in a six-membered boat configuration. Variable temperature magnetic susceptibilities show an intramolecular anti-ferromagnetic interaction between the Ni（II） ions with J = -26.7（3） cm-1.

怀孕期间口服抗真菌药会增加流产风险

据美国WebMD医学新闻网（2016-01—27）报道，丹麦的研究者认为，孕妇口服抗真菌药物fluconazole（商品名Diflucan，辉瑞／Pfizer公司）治疗阴道念珠菌病，流产风险显著增加。在妊娠7～22周期间口服fluconazole的3315名妇女中，147人发生自然流产；相对地，13246名对照组中（没有服用flu—conazole的孕妇）中，563人发生自然流产，差异值换算成风险比（HR）为1．48（95％可信区间“CI”为1．23-1．77）。

Prevalence and Sensitivity Patterns of Candidal Infections in Various Tertiary Care Health Subunits of Karachi

Mostly candida resides as an opportunistic organism on epithelial surfaces of human being. However, under auspicious conditions can cause infections including serious life threatening invasive candidiasis with subsequent mortality particularly in immune deficit and hospitalized patients having co-morbids. Limited data are published on the prevalence of candidiasis, based on the researches conducted at few tertiary care settings which are not representing the overall disease burden in our country, Pakistan. Therefore, this study was conducted to evaluate the frequency and sensitivity patterns of candidiasis in our community. Methods: Out of total 1020 specimens, 130 clinical samples were identified as candidal positive, obtained from March to May 2018. These samples were isolated from vagina, oropharynx, urine, tracheal aspirates, pus, blood, tips of the intubations, wounds and fluids of the body cavities. Identification of candida, its species and antifungal sensitivity screening was done by Kirby Bauer’s disk diffusion method according to CLSI guide lines’ (M - 44 A2 series, 2009). Results: A significant majority, 80 (61.5%) of candidal strains were isolated from females with female to male ratio 8:5 and most of these isolates were obtained from high vaginal swabs (43.75%). Four candidal species (Candida albicans 80%, Candida tropicalis 10%, Candida glabrata 9.2% and Candida ciferrii 0.8%) were isolated from all positive specimens. Maximum number of the positive samples 52 (40%) were obtained from ICU patients. Sensitivity test of candidal positive samples revealed that commonly used azole antifungal drugs, fluconazole and voriconazole were highly resistant, with respective 57.7% and 70.8% resistance. Conclusion: Candidiasis is highly prevalent in our clinical set up and more frequently infecting females in comparison to males as most of the positive isolates were retrieved from HVS (high vaginal swabs). Still, C. albicans was found to be the most prevalent specie isolated among all candida samples. Our study also demonstrated that the resistance of most commonly prescribed antifungals, azoles have shown a rapid rise. Therefore, it is recommended that before prescription of antifungal drugs the clinicians should routinely recommend culture and sensitivity testing of samples taken from candida infected individuals.

Comparison of the Effects of Three Different Anti-fungus Drugs on Candida Albicans of Murine Vaginal Mucosa

To compare the therapeutic effects of three different anti-fungal drugs （i.e., terbinafine, fluconazole and intraconazole） in the treatment of experimental vaginitis caused by Candida albicans （C. albicans） in mice, the fungal vaginitis model was established in female ICR mice by intravaginal inoculation of suspension of C. albicans after the animal had been pretreated with estradiol. Mice were divided at random into different groups and then respectively treated with terbinafine, fluconazole and intraconazole given by gastrogavage. The burden of the fungus in the vaginal lavage fluids in the mice of the different groups was measured dynamically at different time points after the beginning of the drug treatment. The fungal burdens in the vaginal lavage fluids taken at different time points from the mice treated with terbinafine were significantly higher than those taken at corresponding time points from mice treated with fluconazole or itraconazole （P〈0.01）. The fungal burdens in the vaginal lavage fluids taken from mice 1 week after the beginning of the treatment with terbinafine remained at a relatively high level. A dramatic drop in the fungal burden was noted in the vaginal lavage fluids taken on the 2nd day of the treatment from mice treated with itraconazole or fluconazole group and the fungal burden on the 3rd day of the treatment in these mice were at a very low level, suggesting that fluconazole or itraconazole were highly effective for the treatment. However, the difference in the therapeutic effect between the two drugs was not significant （P〉0.05）. Itraconazole or fluconazole, but not terbinafine, is very effective for the treatment of fungal vaginitis caused by C. albicans in mice.

白念珠菌唑类药物作用靶酶在菌株唑类耐药机制中的作用

近年来，随着获得性免疫缺陷综合征（ AIDS，艾滋病）患者的不断增多，高效广谱抗生素、糖皮质激素的广泛应用，抗肿瘤治疗的深入开展，以及器官移植、介入治疗的不断实施，白念珠菌感染的发生率呈上升趋势［1-2］。更重要的是，该菌为导致黏膜和血源播散性感染的重要条件致病菌，感染率远高于其他念珠菌［3-4］。治疗白念珠菌引起的感染常用三唑类抗真菌药物，包括氟康唑（ Fluconazole ）、伊曲康唑（Itraconazole）和伏立康唑（ Voriconazole）等［5］。然而长期和重复用药往往导致耐药菌株的出现，特别是对于艾滋病患者口腔黏膜念珠菌病的治疗，唑类耐药现象已经成为严重的问题［3，6］。其中唑类抗真菌药物的靶酶编码基因ERG11的突变和过度表达是白念珠菌产生耐药性的重要原因，且已成为近年来国内外学者研究的热点。现就此项研究进展进行综述。