Synthesis of Novel Triazine-Based Chalcones and 8,9-dihydro-7H-pyrimido[4,5-b][1,4]diazepines as Potential Leads in the Search of Anticancer, Antibacterial and Antifungal Agents.

This study presents the synthesis of four series of novel hybrid chalcones (20,21)a-g and (23,24)a-g and six series of 1,3,5-triazine-based pyrimido[4,5-b][1,4]diazepines (28-33)a-g and the evaluation of their anticancer, antibacterial, antifungal, and cytotoxic properties. Chalcones 20b,d, 21a,b,d, 23a,d-g, 24a-g and the pyrimido[4,5-b][1,4]diazepines 29e,g, 30g, 31a,b,e-g, 33a,b,e-g exhibited outstanding anticancer activity against a panel of 60 cancer cell lines with GI50 values between 0.01 and 100 µM and LC50 values in the range of 4.09 µM to >100 µM, several of such derivatives showing higher activity than the standard drug 5-fluorouracil (5-FU). On the other hand, among the synthesized compounds, the best antibacterial properties against N. gonorrhoeae, S. aureus (ATCC 43300), and M. tuberculosis were exhibited by the pyrimido[4,5-b][1,4]diazepines (MICs: 0.25-62.5 µg/mL). The antifungal activity studies showed that triazinylamino-chalcone 29e and triazinyloxy-chalcone 31g were the most active compounds against T. rubrum and T. mentagrophytes and A. fumigatus, respectively (MICs = 62.5 µg/mL). Hemolytic activity studies and in silico toxicity analysis demonstrated that most of the compounds are safe.

[Optimization of variables in photodynamic antifungal assays against Candida species]. / Optimización de variables en ensayos de actividad antifúngica fotodinámica frente a especies de Candida.

Oral infections due to yeasts of the genus Candida are very common in patients with predisposing factors, such as physiological conditions or certain underlying diseases. Moreover, oral candidiasis can also evolve into disseminated forms. In this work, strains of the genus Candida were studied to establish the optimal radiation conditions for photosensitizing compounds, in a photodynamic antifungal therapy protocol. A total of 39 isolates were evaluated. The strains were exposed to twelve dyestuffs, eight radiation sources and three different exposure times. Orthotoluidine blue exhibited good photodynamic activity, in combination with exposure to a 20W reflector lamp LED (light-emitting diode) light for 60minutes. When considering the difficulties of using conventional antifungal drugs, the emergence of resistant strains, recurrences, and adverse reactions of certain commonly used drugs, the photodynamic antifungal therapy is a promising strategy for the treatment of localized infections.

Water Extract from Inflorescences of Industrial Hemp Futura 75 Variety as a Source of Anti-Inflammatory, Anti-Proliferative and Antimycotic Agents: Results from In Silico, In Vitro and Ex Vivo Studies.

Industrial hemp (Cannabis sativa) is traditionally cultivated as a valuable source of fibers and nutrients. Multiple studies also demonstrated antimicrobial, anti-proliferative, phytotoxic and insecticide effects of the essential oil from hemp female inflorescences. On the other side, only a few studies explored the potential pharmacological application of polar extracts from inflorescences. In the present study, we investigated the water extract from inflorescences of industrial hemp Futura 75 variety, from phytochemical and pharmacological point of view. The water extract was assayed for phenolic compound content, radical scavenger/reducing, chelating and anti-tyrosinase effects. Through an ex vivo model of toxicity induced by lipopolysaccharide (LPS) on isolated rat colon and liver, we explored the extract effects on serotonin, dopamine and kynurenine pathways and the production of prostaglandin (PG)E2. Anti-proliferative effects were also evaluated against human colon cancer HCT116 cell line. Additionally, antimycotic effects were investigated against Trichophyton rubrum, Trichophyton interdigitale, Microsporum gypseum. Finally, in silico studies, including bioinformatics, network pharmacology and docking approaches were conducted in order to predict the putative targets underlying the observed pharmacological and microbiological effects. Futura 75 water extract was able to blunt LPS-induced reduction of serotonin and increase of dopamine and kynurenine turnover, in rat colon. Additionally, the reduction of PGE2 levels was observed in both colon and liver specimens, as well. The extract inhibited the HCT116 cell viability, the growth of T. rubrum and T. interdigitale and the activity of tyrosinase, in vitro, whereas in silico studies highlighting the inhibitions of cyclooxygenase-1 (induced by carvacrol), carbonic anhydrase IX (induced by chlorogenic acid and gallic acid) and lanosterol 14-α-demethylase (induced by rutin) further support the observed pharmacological and antimycotic effects. The present findings suggest female inflorescences from industrial hemp as high quality by-products, thus representing promising sources of nutraceuticals and cosmeceuticals against inflammatory and infectious diseases.

Photodynamic activity of Tagetes minuta extracts against superficial fungal infections.

Candida and dermatophyte species are the most common causes of superficial mycoses because their treatment can be difficult due to limitations of current antifungal drugs in terms of toxicity, bioavailability, interactions, narrow-spectrum activity, and development of resistance. Photodynamic therapy (PDT) involves the topical administration of a photosensitizer in combination with light of an appropriate wavelength and molecular oxygen that produces reactive oxygen species (ROS), which promote damage to several vital components of the microorganism. Tagetes species are known as a source of thiophenes, biologically active compounds whose antifungal activity is enhanced by irradiation with UVA. The present investigation evaluated Tagetes minuta extracts as a photosensitizer on growth of Candida and dermatophytes and their effect on Candida virulence factors. T. minuta root hexane and dichloromethane extracts demonstrated high photodynamic antifungal activity. Bioautographic assays and chromatographic analysis revealed the presence of five thiophenes with reported photodynamic antifungal activities under UVA. Analysis of ROS production indicated that both type I and II reactions were involved in the activity of the extracts. In addition, the extracts inhibited virulence factors of Candida, such as adherence to epithelial surfaces and germ tube formation and showed efficacy against different Candida morphologies: budding cells, cells with germ tube and biofilms. Results suggested that PDT with T. minuta extracts might become a valuable alternative to the already established antifungal drugs for the treatment of superficial fungal infections.

Approaches to the mechanism of antifungal activity of Zuccagnia punctata-Larrea nitida bi-herbal combination.

BACKGROUND: In our previous study the synergism of four combinations of Zuccagnia punctata (ZpE) and Larrea nitida (LnE) exudates with the reliable statistical-based MixLow method was assessed, and the markers of the most anti-C. albicans synergistic ZpE-LnE bi-herbal combination were quantified according to European Medicines Agency (EMA). PURPOSE: To study the mechanisms of action as well as the cytotoxic properties of the ZpE-LnE most synergistic combination found in the previous work. MATERIALS AND METHODS: Minimum Fungicidal Concentration (MFC) and rate of killing of ZpE-LnE were assessed with the microbroth dilution and the time-kill assays respectively. Morphological alterations were observed with both confocal and fluorescence microscopy on the yeast Schizosaccharomyces pombe. The ergosterol exogenous assay, the quantification of ergosterol, the sorbitol as well as glucan synthase (GS) and chitin synthase (ChS) assays were used to detect the effects on the fungal membrane and cell wall respectively. The capacity of ZpE-LnE of inhibiting Candida virulence factors was assessed with previously reported methods. The effect of ZpE-LnE and of ZpE or LnE alone on cell viability was determined on human hepatoma cells line Huh7. RESULTS: ZpE-Ln E was fungicidal killing C. albicans in a shorter time than amphotericin B and produced malformations in S. pombe cells. ZpE-LnE showed to bind to ergosterol but not to inhibit any step of the ergosterol biosynthesis. ZpE-LnE showed a low or moderate capacity of inhibiting GS and ChS. Regarding inhibition of virulence factors, ZpE-LnE significantly decreased the capacity of adhesion to eukaryotic buccal epithelial cells (BECs), did not inhibit the germ tube formation and inhibited the secretion of phospholipases and proteinases but not of haemolysins. ZpE-LnE demonstrated very low toxicity on Huh7 cells, much lower than that each extract alone. CONCLUSION: The fungicidal properties of ZpE-LnE against C. albicans, its dual mechanism of action targeting the fungal membrane's ergosterol as well as the cell wall, its capacity of inhibiting several important virulence factors added to its low toxicity, make ZpE-LnE a good candidate for the development of a new antifungal bi-Herbal Medicinal Product.

Plant phenolics and terpenoids as adjuvants of antibacterial and antifungal drugs.

BACKGROUND: The intensive use of antibacterial and antifungal drugs has dramatically increased the microbial resistance and has led to a higher number of difficult-to-eradicate infections. Combination therapy with two or more antimicrobial drugs has emerged some years ago to overcome the issue, but it has proven to be not completely effective. Natural secondary metabolites of MW ≤ 500 represent promising adjuvants for antimicrobials and have been the object of several researches that have increased in the last two decades. PURPOSE: The purpose of this Review is to do a literature search of the natural compounds that showed high enhancing capacity of antibacterials' and antifungals' effects against planktonic bacteria and fungi and to analyze which are the natural products most used in combination with a focus on polyphenols and terpenoids. RESULTS: One hundred of papers were collected for reviewing. Fifty six (56) of them deal with combinations of low MW natural products with antibacterial drugs against planktonic bacteria and forty four (44) on natural products with antifungal drugs against planktonic fungi. Of the antibacterial adjuvants, 41 (73%) were either polyphenols (27; 48%) or terpenes (14; 25%). The remaining 15 papers (27%), deal with different class of natural products. Since most natural potentiators belong to the terpene or phenolic structural types, a more detailed description of the works dealing with these type of compounds is provided here. Bacterial and fungal resistance mechanisms, the modes of action of the main classes of antibacterial and antifungal drugs and the methodologies most used to assess the type of interactions in the combinations were included in the Review too. CONCLUSIONS AND PERSPECTIVES: Several promising results on the potentiation effects of antifungals' and antibacterials' activities by low MW natural products mainly on polyphenols and terpenes were reported in the literature and, in spite of that most works included only in vitro assays, this knowledge opens a wide range of possibilities for the combination antimicrobial therapy. Further research including in vivo assays and clinical trials are required to determine the relevance of these antimicrobial enhancers in the clinical area and should be the focus of future studies in order to develop new antimicrobial combination agents that overpass the drawbacks of the existing antibiotics and antifungals in clinical use.

Hybrid Molecules Containing a 7-Chloro-4-aminoquinoline Nucleus and a Substituted 2-Pyrazoline with Antiproliferative and Antifungal Activity.

Twenty-four new hybrid analogues (15-38) containing 7-chloro-4-aminoquinoline and 2-pyrazoline N-heterocyclic fragments were synthesized. Twelve of the new compounds were evaluated against 58 human cancer cell lines by the U.S. National Cancer Institute (NCI). Compounds 25, 30, 31, 36, and 37 showed significant cytostatic activity, with the most outstanding GI50 values ranging from 0.05 to 0.95 µM. The hybrid compounds (15-38) were also evaluated for antifungal activity against Candida albicans and Cryptococcus neoformans. From the obtained results some structure-activity relationships were outlined.

Photodynamic inactivation of oropharyngeal Candida strains.

Oropharyngeal candidiasis (OPC) is an infection frequent in immunocompromised patients. Photodynamic therapy is an alternative to conventional treatments, based on the utilization of compounds that inhibit or kill microorganisms only under the effect of light, process known as Photodynamic Inactivation (PDI). In the present study, PDI of Candida spp. by the natural product α-terthienyl (α-T) was investigated following the guidelines of CLSI M27-A3, under UV-A light irradiation. The optimal values of two variables, exposure irradiation time (ET) and distance to the irradiation source (DIS) were established by employing Design Expert Software (DES). For this purpose, a panel of Candida strains isolated from OPC (C. albicans, C. tropicalis, C. parapsilosis and C. krusei) was employed and optimal values were 5 min (ET) and between 6.06 and 6.43 cm (DIS) with a desirability factor of 0.989. α-T plus UV-A light in the optimal conditions caused a complete reduction in viable cells in 5 min which was demonstrated by viable cells reduction assays and confocal microscopy after vital staining (propidium iodide/fluorescein diacetate). The germ tube formation of C. albicans was inhibited by α-T at sub-inhibitory concentrations. Results showed that α-T plus UV-A light could constitute an alternative for OPC treatments at the optimal conditions determined here.

Cytotoxic and Antifungal Activities of Diverse α-Naphthylamine Derivatives.

Diverse α-naphthylamine derivatives were easily prepared from corresponding aldimines derived from commercially available α-naphthaldehyde and anilines or isomeric pyridinecarboxyaldehydes and α-naphthylamine. The secondary amines obtained were tested as possible antifungal and cytotoxic agents. The diverse N-aryl-N-[1-(1-naphthyl)but-3-enyl]amines obtained were active (IC(50) < 10 µg/mL) against breast (MCF-7), non-small cell lung (H-460), and central nervous system (SF-268) human cancer cell lines, while N-(pyridinylmethyl)-naphthalen-1-amines resulted in activity against (MIC 25-32 µg/mL) some human opportunistic pathogenic fungi including yeasts, hialohyphomycetes, and dermatophytes.

Antifungal and cytotoxic activities of some N-substituted aniline derivatives bearing a hetaryl fragment.

Diverse N-substituted anilines bearing hetaryl fragments were easily prepared from corresponding aldimines derived from commercially available aromatic aldehydes and anilines. 2-Furyl substituted anilines showed very good antifungal activities against dermatophytes, particularly against Trichophyton rubrum (MIC=3.12-6.25microg/mL). In addition, all active compounds, 45-47, 73, and 74, were tested for cytotoxic activities against breast (MCF-7), lung (H-460), and central nervous system (SF-268) human cancer cell lines with the NCI-anticancer-drug screen. The activity of amines described in this paper, along with the low toxicity of most of them, shows promise for the future development of non-toxic new antimycotic agents.

Identificación y epidemiología de las distintas especies de Malassezia en la ciudad de Rosario / Identification and epidemiology of the different species of Malassezia in the city of Rosario, Argentina

Las especies del género Malassezia han sido exhaustivamente estudiadas en estos últimos años, tanto desde el punto de vista fenotípico como desde las pruebas fisiológicas y características genotípicas. En tal sentido, se han incorporado las especies M. Globosa, M. slooffiae, M. restricta y M. obtusa a las ya conocidas M. furfur, M. pachydermatis y M. sympodialis. El objetivo de este trabajo fue, teniendo en cuenta estos estudios taxonómicos, identificar las distintas especies de Malassezia en muestras provenientes de pacientes con pitiriasis versicolor, dermatitis seborreica y foliculitis que consultaron al Servicio de Dermatología, en el Hospital Provincial "Centenario" de Rosario. Las especies más frecuentemente halladas en la población estudiada fueron M. sympodialis, seguida por M obtusa, M. slooffiae y, con menor frecuencia, M furfur.

Botanical (morphological, micrographic), chemical and pharmacological characteristics of Pfaffia species (Amaranthaceae) native to South Brazil

Some parameters for the quality control of P. glomerata and P. paniculata roots using their botanical and chemical characteristics are presented. it was also carried out an in vitro pharmacological screening to evaluate some biological properties of P. glomerata that could be related to its popular use as "tonic". Relating to biological assays, ethanolic extract from P. glomerata roots did not presented antiviral, antiproliferative, antifungal or MAO inhibitory activities. The cytotoxicity evaluation of P. glomerata determined that `IC IND. 50ï is >2,000 µg/mL. The main morphological and micrographic characteristics of P. glomerata and P. paniculata roots are described in this paper in order to aid in their unequivocal identification