nystatin-a1 has been researched along with Hemolysis* in 20 studies
20 other study(ies) available for nystatin-a1 and Hemolysis
Article | Year |
---|---|
Genomic and Targeted Approaches Unveil the Cell Membrane as a Major Target of the Antifungal Cytotoxin Amantelide A.
Amantelide A, a polyhydroxylated macrolide isolated from a marine cyanobacterium, displays broad-spectrum activity against mammalian cells, bacterial pathogens, and marine fungi. We conducted comprehensive mechanistic studies to identify the molecular targets and pathways affected by amantelide A. Our investigations relied on chemical structure similarities with compounds of known mechanisms, yeast knockout mutants, yeast chemogenomic profiling, and direct biochemical and biophysical methods. We established that amantelide A exerts its antifungal action by binding to ergosterol-containing membranes followed by pore formation and cell death, a mechanism partially shared with polyene antifungals. Binding assays demonstrated that amantelide A also binds to membranes containing epicholesterol or mammalian cholesterol, thus suggesting that the cytotoxicity to mammalian cells might be due to its affinity to cholesterol-containing membranes. However, membrane interactions were not completely dependent on sterols. Yeast chemogenomic profiling suggested additional direct or indirect effects on actin. Accordingly, we performed actin polymerization assays, which suggested that amantelide A also promotes actin polymerization in cell-free systems. However, the C-33 acetoxy derivative amantelide B showed a similar effect on actin dynamics in vitro but no significant activity against yeast. Overall, these studies suggest that the membrane effects are the most functionally relevant for amantelide A mechanism of action. Topics: Actin Cytoskeleton; Animals; Antifungal Agents; Cell Membrane; Cell Membrane Permeability; Drug Resistance, Fungal; Ergosterol; Erythrocytes; Hemolysis; Liposomes; Macrolides; Nystatin; Saccharomyces cerevisiae; Sheep | 2021 |
Generation of tetramycin B derivative with improved pharmacological property based on pathway engineering.
Polyene antibiotics, including amphotericin, nystatin, pimaricin, and tetramycin, are important antifungal agents. Increasing the production of polyenes and generation of their improved analogues based on the biosynthetic pathway engineering has aroused wide concern in application researches. Herein, tetramycin and nystatin, both of which share most of acyl-CoA precursors, are produced by Streptomyces hygrospinosus var. beijingensis CGMCC 4.1123. Thus, the intracellular malonyl-CoA is found to be insufficient for PKSs (polyketide synthases) extension of tetramycin by quantitative analysis in this wild-type strain. To circumvent this problem and increase tetramycin titer, the acyl-CoA competing biosynthetic gene cluster (BGC) of nystatin was disrupted, and the biosynthetic genes of malonyl-CoA from S. coelicolor M145 were integrated and overexpressed in nys-disruption mutant strain (SY02). Moreover, in order to specifically accumulate tetramycin B from A, two copies of tetrK and a copy of tetrF were introduced, resulting in elevating tetramycin B fermentration titer by 122% to 865 ± 8 mg/L than the wild type. In this optimized strain, a new tetramycin derivative, 12-decarboxy-12-methyl tetramycin B, was generated with a titer of 371 ± 26 mg/L through inactivation of a P450 monooxygenase gene tetrG. Compared with tetramycin B, the new compound exhibited higher antifungal activity against Saccharomyces cerevisiae and Rhodotorula glutinis, but lower hemolytic toxicity to erythrocyte. This research provided a good example of employing biosynthetic engineering strategies for fermentation titer improvement of polyene and development of the derivatives for medicinal applications. Topics: Animals; Antifungal Agents; Biosynthetic Pathways; Erythrocytes; Fermentation; Hemolysis; Horses; Macrolides; Metabolic Engineering; Multigene Family; Nystatin; Rhodotorula; Saccharomyces cerevisiae; Streptomyces | 2020 |
Molecular Umbrellas Modulate the Selective Toxicity of Polyene Macrolide Antifungals.
Antifungal polyene macrolide antibiotics Amphotericin B (AmB) and Nystatin (NYS) were conjugated through the ω-amino acid linkers with diwalled "molecular umbrellas" composed of spermidine-linked deoxycholic or cholic acids. The presence of "umbrella" substituents modulated biological properties of the antibiotics, especially their selective toxicity. Some of the AmB-umbrella conjugates demonstrated antifungal in vitro activity comparable to that of the mother antibiotic but diminished mammalian toxicity, especially the hemolytic activity. In contrast, antifungal in vitro activity of NYS-umbrella conjugates was strongly reduced and all these conjugates demonstrated poorer than NYS selective toxicity. No correlation between the aggregation state and hemolytic activity of the novel conjugates was found. Topics: Amphotericin B; Antifungal Agents; Fungi; HEK293 Cells; Hemolysis; Hep G2 Cells; Humans; Mycoses; Nystatin; Polyenes | 2018 |
Improved recovery and biological activities of an engineered polyene NPP analogue in Pseudonocardia autotrophica.
NPP A1 produced by Pseudonocardia autotrophica is a unique disaccharide-containing polyene macrolide. NPP A1 was reported to have higher water solubility and lower hemolytic toxicity than nystatin A1 while retaining its antifungal activity. An engineered NPP A1 analogue, NPP A2, was generated by inactivation of the nppL gene, encoding a P450 monooxygenase in P. autotrophica. The resulting compound exhibited the corresponding chemical structure of NPP A1 but lacked a C10 hydroxyl group. In this study, newly developed crystallization recovery methods for NPP A2 purification, followed by an evaluation of in vitro antifungal activity and hemolytic activity, were performed. The crystallization methods were designed to eliminate the undesired viscous impurities encountered during the NPP A2 purification process, resulting in improved purity from 5.3 to 83.5% w/w. NPP A2 isolated from the improved purification process also exhibited two times higher antifungal activity and 1.8 times higher hemolytic toxicity than those of NPP A1. These results suggest that the minor structural modification of disaccharide-containing polyene macrolides, such as removing a C10 hydroxyl group, might require an alternative recovery process, such as crystallization, to confirm its improved biological activity. Topics: Actinomycetales; Antifungal Agents; Disaccharides; Hemolysis; Macrolides; Nystatin; Polyenes | 2017 |
Structural analysis and biosynthetic engineering of a solubility-improved and less-hemolytic nystatin-like polyene in Pseudonocardia autotrophica.
Polyene antibiotics such as nystatin are a large family of very valuable antifungal polyketide compounds typically produced by soil actinomycetes. Previously, using a polyene cytochrome P450 hydroxylase-specific genome screening strategy, Pseudonocardia autotrophica KCTC9441 was determined to contain an approximately 125.7-kb region of contiguous DNA with a total of 23 open reading frames, which are involved in the biosynthesis and regulation of a structurally unique polyene natural product named NPP. Here, we report the complete structure of NPP, which contains an aglycone identical to nystatin and harbors a unique di-sugar moiety, mycosaminyl-(α1-4)-N-acetyl-glucosamine. A mutant generated by inactivation of a sole glycosyltransferase gene (nppDI) within the npp gene cluster can be complemented in trans either by nppDI-encoded protein or by its nystatin counterpart, NysDI, suggesting that the two sugars might be attached by two different glycosyltransferases. Compared with nystatin (which bears a single sugar moiety), the di-sugar containing NPP exhibits approximately 300-fold higher water solubility and 10-fold reduced hemolytic activity, while retaining about 50% antifungal activity against Candida albicans. These characteristics reveal NPP as a promising candidate for further development into a pharmacokinetically improved, less-cytotoxic polyene antifungal antibiotic. Topics: Actinomycetales; Antifungal Agents; Biotechnology; Candida albicans; Fungal Proteins; Genetic Engineering; Hemolysis; Microbial Sensitivity Tests; Multigene Family; Mutation; Nystatin; Polyenes; Solubility; Structure-Activity Relationship | 2012 |
New nystatin-related antifungal polyene macrolides with altered polyol region generated via biosynthetic engineering of Streptomyces noursei.
Polyene macrolide antibiotics, including nystatin and amphotericin B, possess fungicidal activity and are being used as antifungal agents to treat both superficial and invasive fungal infections. Due to their toxicity, however, their clinical applications are relatively limited, and new-generation polyene macrolides with an improved therapeutic index are highly desirable. We subjected the polyol region of the heptaene nystatin analogue S44HP to biosynthetic engineering designed to remove and introduce hydroxyl groups in the C-9-C-10 region. This modification strategy involved inactivation of the P450 monooxygenase NysL and the dehydratase domain in module 15 (DH15) of the nystatin polyketide synthase. Subsequently, these modifications were combined with replacement of the exocyclic C-16 carboxyl with the methyl group through inactivation of the P450 monooxygenase NysN. Four new polyene macrolides with up to three chemical modifications were generated, produced at relatively high yields (up to 0.51 g/liter), purified, structurally characterized, and subjected to in vitro assays for antifungal and hemolytic activities. Introduction of a C-9 hydroxyl by DH15 inactivation also blocked NysL-catalyzed C-10 hydroxylation, and these modifications caused a drastic decrease in both antifungal and hemolytic activities of the resulting analogues. In contrast, single removal of the C-10 hydroxyl group by NysL inactivation had only a marginal effect on these activities. Results from the extended antifungal assays strongly suggested that the 9-hydroxy-10-deoxy S44HP analogues became fungistatic rather than fungicidal antibiotics. Topics: Animals; Antifungal Agents; Biosynthetic Pathways; Candida albicans; Erythrocytes; Hemolysis; Horses; Macrolides; Magnetic Resonance Spectroscopy; Microbial Sensitivity Tests; Models, Molecular; Molecular Structure; Nystatin; Polyenes; Polymers; Streptomyces | 2011 |
Improved antifungal polyene macrolides via engineering of the nystatin biosynthetic genes in Streptomyces noursei.
Seven polyene macrolides with alterations in the polyol region and exocyclic carboxy group were obtained via genetic engineering of the nystatin biosynthesis genes in Streptomyces noursei. In vitro analyses of the compounds for antifungal and hemolytic activities indicated that combinations of several mutations caused additive improvements in their activity-toxicity properties. The two best analogs selected on the basis of in vitro data were tested for acute toxicity and antifungal activity in a mouse model. Both analogs were shown to be effective against disseminated candidosis, while being considerably less toxic than amphotericin B. To our knowledge, this is the first report on polyene macrolides with improved in vivo pharmacological properties obtained by genetic engineering. These results indicate that the engineered nystatin analogs can be further developed into antifungal drugs for human use. Topics: Animals; Antifungal Agents; Base Sequence; Candida albicans; Genes, Bacterial; Genetic Engineering; Hemolysis; Humans; Male; Mice; Nystatin; Polyenes; Polymers; Streptomyces; Structure-Activity Relationship | 2008 |
Precise size determination of amphotericin B and nystatin channels formed in erythrocyte and liposomal membranes based on osmotic protection experiments.
The colloid osmotic nature of the cell lysis can be prevented by adding osmotic protectants of appropriate sizes to the outer medium. We introduced inorganic and organic electrolytes as protectants to determine the precise channel sizes of the polyene antibiotics, amphotericin B and nystatin, in addition to the sugars so far widely used for this purpose. Because colloid osmotic cell lysis is evidenced by the loss of membrane permeability barriers for small sizes of ions, such as K(+), preceding hemolysis, we firstly simultaneously monitored the time response of the K(+) efflux and hemolysis induced by amphotericin B by combining a fiber-optic spectrometer with a K(+)-selective electrode. Based on this experiment, we evaluated the sizes of channels of the polyene antibiotics formed in the erythrocyte membrane using the radii of hydrated ions calculated from a modified Stokes' law, as well as the radii of sugars. The radii of channels formed by amphotericin B and nystatin were found to be in a very narrow range of 0.36 - 0.37 nm. Similar experiments were performed using calcein-loaded liposomes containing cholesterol or ergosterol, and the radii of channels formed in these liposomal membranes were also found to be the same as when formed in an erythrocyte membrane. The present results demonstrated that introducing the sizes of hydrated ions can afford a more precise channel size than the use of sugars alone. Topics: Amphotericin B; Animals; Carbohydrates; Electrolytes; Erythrocytes; Hemolysis; Liposomes; Nystatin; Osmosis; Potassium; Sheep; Time Factors; Water | 2008 |
Analysis of the mycosamine biosynthesis and attachment genes in the nystatin biosynthetic gene cluster of Streptomyces noursei ATCC 11455.
The polyene macrolide antibiotic nystatin produced by Streptomyces noursei contains a deoxyaminosugar mycosamine moiety attached to the C-19 carbon of the macrolactone ring through the beta-glycosidic bond. The nystatin biosynthetic gene cluster contains three genes, nysDI, nysDII, and nysDIII, encoding enzymes with presumed roles in mycosamine biosynthesis and attachment as glycosyltransferase, aminotransferase, and GDP-mannose dehydratase, respectively. In the present study, the functions of these three genes were analyzed. The recombinant NysDIII protein was expressed in Escherichia coli and purified, and its in vitro GDP-mannose dehydratase activity was demonstrated. The nysDI and nysDII genes were inactivated individually in S. noursei, and analyses of the resulting mutants showed that both genes produced nystatinolide and 10-deoxynystatinolide as major products. Expression of the nysDI and nysDII genes in trans in the respective mutants partially restored nystatin biosynthesis in both cases, supporting the predicted roles of these two genes in mycosamine biosynthesis and attachment. Both antifungal and hemolytic activities of the purified nystatinolides were shown to be strongly reduced compared to those of nystatin, confirming the importance of the mycosamine moiety for the biological activity of nystatin. Topics: Animals; Blotting, Western; Carbohydrate Dehydrogenases; Chromatography, High Pressure Liquid; Chromatography, Liquid; Genetic Vectors; Glycosyltransferases; Hemolysis; Hexosamines; Horses; Mass Spectrometry; Molecular Structure; Multigene Family; Nystatin; Polymerase Chain Reaction; Recombinant Proteins; Streptomyces; Transaminases | 2007 |
Selective lysis of erythrocytes infected with the trophozoite stage of Plasmodium falciparum by polyene macrolide antibiotics.
The continuous increase in strains of the human malaria parasite Plasmodium falciparum resistant to most front-line antimalarial compounds is reason for grave clinical concern. The search for new drugs led us to investigate a number of membrane active polyene macrolide antibiotics, such as amphotericin B, nystatin, filipin and natamycin. The interaction of these compounds with sterols in bilayer cell membranes can lead to cell damage and ultimately cell lysis. The malaria parasite modifies the host erythrocyte membrane by changing the protein and lipid composition and thus the infected cell could be a selective target for membrane active compounds. We found that erythrocytes infected with the trophozoite stage of P. falciparum were particularly susceptible to lysis by amphotericin B (Fungizone) and, to a lesser extent, nystatin, as determined by ELISA and various microscopy assays. Liposomal amphotericin B (AmBisome) displayed a similar specificity for parasitised erythrocytes, but complete lysis required a longer incubation period. In contrast, filipin and natamycin did not distinguish between normal and parasite-infected erythrocytes, but lysed both at similar concentrations. In addition, when added to ring-stage cultures, the amphotericin B preparations and nystatin produced a marked disruption in parasite morphology in less than 2 h without an accompanying permeabilisation of the infected host cell, suggesting a second plasmodicidal mode of action. The results imply that selected polyene macrolide antibiotics or their derivatives could find application in the treatment of severe malaria caused by of P. falciparum. Topics: Amphotericin B; Animals; Anti-Bacterial Agents; Antiprotozoal Agents; Dose-Response Relationship, Drug; Enzyme-Linked Immunosorbent Assay; Erythrocyte Membrane; Erythrocytes; Filipin; Hemolysis; Humans; Life Cycle Stages; Natamycin; Nystatin; Plasmodium falciparum | 2006 |
A kinetics study of pig erythrocyte hemolysis induced by polyene antibiotics.
The kinetics of the hemolysis induced by filipin is of the damage type, indicating the formation of large nonselective perforations of erythrocyte membranes. The process is relatively independent of the ionic composition of the incubation medium, and the differences between the hemolysis induced by filipin in pig and human erythrocytes are not significant. In a sucrose medium, filipin-induced hemolysis is inhibited in humans, whereas it is stimulated in pig erythrocytes. It is suggested that low ionic strength is the reason for the different modifications of complexation of filipin in pig and human erythrocyte membranes in a sucrose medium. The kinetics of the hemolysis induced in pig erythrocytes by amphotericin B and nystatin is of the permeability type, indicating the formation of selective channels in erythrocyte membranes and colloid osmotic hemolysis. The rate of the hemolysis, which is high in a KCl medium, is decreased in all the other media tested (CaCl2, MgCl2, potassium phosphate buffer, K2SO4, sucrose), although there are no changes in the kinetics of hemolysis. The results are interpreted as the formation of highly selective channels at a low concentration of the antibiotics. At increasing concentrations, channels of decreasing selectivity occur. The resistances of pig erythrocytes to amphotericin B and nystatin are lower than those of human erythrocytes. Topics: Amphotericin B; Animals; Anti-Bacterial Agents; Erythrocytes; Filipin; Hemolysis; Humans; Kinetics; Nystatin; Polyenes; Swine; Time Factors | 2003 |
The mechanism of the hemolytic activity of polyene antibiotics.
The kinetics of the filipin-, amphotericin B- and nystatin-induced hemolysis of human erythrocytes were investigated. Filipin-induced hemolysis is of the damage type. It is an all-or-none process, partly inhibited by Ca2+ or Ba2+ but not by Mg2+, Na+ or SO42-. The hemolytic activity of filipin is explained by the formation of large aggregates within the erythrocyte membrane in the form of large perforations, permeable to substances of low molecular weight as well as to macromolecules, including hemoglobin. In isotonic KCl solution, both amphotericin B and nystatin, at low concentrations, form smaller aggregates within the membranes. As a result, the permeability of the membranes to KCl increases and hemolysis occurs. However, the kinetics of the hemolysis induced by the two polyenes is complex. The process shows some features of the permeability type and some of the damage type. It is suggested that amphotericin B and nystatin may simultaneously form a number of transport systems, differing in their molecular organisation and hemolytic activity. Their participation in erythrocyte membrane permeability can be modified by small changes in membrane organisation and the chemical composition of the incubation medium. In isotonic solutions of divalent cation chlorides, and at higher antibiotic concentration, additional aggregates, allowing divalent cations to permeate, appear. These structures do not permit SO4(2-) to permeate. Topics: Amphotericin B; Anti-Bacterial Agents; Cations; Erythrocyte Membrane; Filipin; Hemolysis; Humans; Isotonic Solutions; Nystatin; Osmolar Concentration; Polyenes; Time Factors | 2002 |
The kinetics of colloid osmotic hemolysis. I. Nystatin-induced lysis.
A kinetic model of colloid osmotic hemolysis for cation-permeable cells has been developed. The model consists of three essential components. The first is a set of flux equations, under the assumption that the membrane potential is equal to the chloride equilibrium potential and that cation fluxes are described by the Goldman flux equation. The second is the osmotic equilibrium model of Freedman and Hoffman that takes into account the non-ideal osmotic behavior of erythrocytes. The third is an empirical relation between hemolysis and cell volume, developed from the lysis behavior in hypoosmotic media. Model simulations are compared with lysis experiments using the antibiotic nystatin to raise cation permeability. The form of the kinetics and inhibition of lysis by sucrose are described well by the model. In additional lysis experiments at different external pH the small pH dependence is accounted for by the model. Topics: Colloids; Hemolysis; Humans; Hydrogen-Ion Concentration; Kinetics; Mathematics; Nystatin; Osmotic Fragility; Osmotic Pressure | 1985 |
Cell volume and osmotic properties of erythrocytes after complement lysis measured by flow cytometry.
The changes of volume distribution curves of erythrocytes during and after lysis by complement or nystatin or in hypotonic buffers were measured by flow cytometry. Biconcave and spheroidal ghosts were observed after complement lysis and spheroidal ghosts were seen only after nystatin and hypotonic lysis. The spheroidal ghosts derived from red cells lysed by complement or nystatin were permeable to sucrose; those from hypotonic lysis were sucrose-impermeable. Spheroidal ghosts after complement lysis remained permeable for sucrose whereas spheroidal ghosts after nystatin lysis resealed after removal of the drug by washing. Biconcave ghosts produced by complement lysis were almost impermeable to sucrose initially and therefore responded to osmotic changes, but they became sucrose-permeable upon prolonged incubation at 37 degrees C. The rate of sucrose equilibration increased as the stability of the biconcave shape diminished with increasing numbers of C5b-9 complexes. At 850 C5b-9 complexes/ghost, the biconcave shape and impermeability for sucrose were completely lost. The results support the hypothesis that complement C5b-9 complexes, in addition to the interaction with the lipid bilayer, may interact with the cytoskeleton of the erythrocyte membrane. Topics: Animals; Complement System Proteins; Erythrocyte Indices; Erythrocyte Membrane; Erythrocytes; Flow Cytometry; Hemolysis; Humans; Hypotonic Solutions; Nystatin; Osmotic Fragility; Sheep; Time Factors | 1983 |
The preparation and comparative in vitro biological properties of derivatives formed in the reaction of nystatin with N-glucuronamides.
Topics: Amides; Antifungal Agents; Erythrocytes; Esters; Glucuronates; Hemolysis; Humans; In Vitro Techniques; Nystatin; Saccharomyces cerevisiae | 1980 |
Haloprogin: mode of action studies in Candida albicans.
Topics: Amphotericin B; Animals; Antifungal Agents; Candida albicans; Carbon Radioisotopes; Cell Membrane; Cell Survival; Erythrocytes; Fungal Proteins; Hemolysis; Leucine; Nystatin; Oxygen Consumption; Phenyl Ethers; Potassium; Rats; RNA; Tritium; Uridine | 1974 |
Selective membrane toxicity of the polyene antibiotics: studies on natural membranes.
The effect of polyene antibiotics on Candida albicans, human erythrocytes, and Acholeplasma laidlawii was studied. The results sustain the observations made with lecithin-sterol liposomes. The distribution of double bonds in the membrane sterol nucleus appears to be of major importance in conferring polyene susceptibility; those sterols with the ergosterol nucleus are far more effective than those with a nucleus similar to cholesterol. Different polyenes vary in their membrane selectivity. The clinical implications of these observations are discussed. Topics: Acholeplasma laidlawii; Amphotericin B; Anti-Bacterial Agents; Candida albicans; Cell Membrane Permeability; Cholesterol; Drug Resistance, Microbial; Ergosterol; Erythrocytes; Fatty Acids, Unsaturated; Hemolysis; Lactones; Microbial Sensitivity Tests; Nystatin; Polyenes; Structure-Activity Relationship | 1973 |
Urinary candidiasis following the haemolytic uraemic syndrome.
Topics: Anti-Bacterial Agents; Candida; Candidiasis; Hemolysis; Humans; Infant; Male; Nystatin; Uremia; Urinary Tract Infections | 1971 |
COMPARATIVE RESPONSES OF MAMMALIAN ERYTHROCYTES AND MICROBIAL PROTOPLASTS TO POLYENE ANTIBIOTICS AND VITAMIN A.
Topics: Amphotericin B; Animals; Anti-Bacterial Agents; Bacillus megaterium; Erythrocytes; Hemolysis; Mannitol; Neurospora; Nystatin; Pharmacology; Polyenes; Protoplasts; Rats; Research; Sodium Chloride; Spectrophotometry; Sterols; Sucrose; Vitamin A | 1963 |
The lytic effect of polyene antifungal antibiotics on mammalian erythrocytes.
Topics: Amphotericin B; Animals; Antifungal Agents; Erythrocytes; Hemolysis; Mammals; Nystatin; Polyenes | 1962 |