Polyene Macrolide Antibiotic Research Articles

Effect of fatty acids on action of polyene antibiotics

Fatty acids cause a decrease in the absorption spectra of the antifungal polyene macrolide antibiotics nystatin, filipin, candicidin, and amphotericin B. For nystatin, filipin, and candicidin, this decrease in absorption could be correlated with the activity of the fatty acids in protecting the yeast Saccharomyces cerevisiae against the action of these antibiotics. With amphotericin B a correlation was observed between the decrease in absorption caused by certain derivatives of fatty acids and the protective action of these derivatives against the activity of amphotericin B on yeast. It is concluded that, like the sterols, fatty acids also interact with the polyene antibiotics and thereby reduce their effective concentrations.

Regulation by aromatic amino acids of the biosynthesis of candicidin by Streptomyces griseus.

The biosynthesis by Streptomyces griseus of candicidin, an aromatic polyene macrolide antibiotic, was inhibited by L-tryptophan, L-phenylalanine and, to a lesser degree, by L-tyrosine. A mixture of the three aromatic amino acids inhibited candicidin biosynthesis to a greater extent than did each amino acid separately. L-Tryptophan strongly inhibited the incorporation of the labelled precursors propionate or 4-aminobenzoic acid into candicidin. Incorporation of propionate into candicidin was 50% inhibited by 2.5 mM-tryptophan. Inhibition by tryptophan did not require protein synthesis as the same effect was observed in cells in which protein synthesis was prevented by chloramphenicol. The inhibitory effect of L-tryptophan was partially reversed by exogenous 4-aminobenzoic acid suggesting that this effect is exerted at the level of 4-aminobenzoic acid synthase.

Field desorption mass spectrometry in structural studies of polyene macrolide antibiotics: isolation and early identification of a pentaene macrolide antibiotic.

Isolation of a pentaene macrolide antibiotic (NSC-277813) from Streptomyces griseus (FCRC-21) fermentation broth is described. Using field desorption mass spectrometry and high resolution field desorption mass spectrometry on the intact and derivatized antibiotic and degradation products, the antibiotic was identified as fungichromin. The application of field desorption mass spectrometry in the identification of polyene antibiotics is discussed.

The synthesis of amides of polyene macrolide antibiotics.

The synthesis of amides of polyene macrolide antibiotics.

Energy dependence and reversibility of membrane alterations induced by polyene macrolide antibiotics in Chlorella vulgaris.

The requirement of metabolic energy for the interaction of polyene macrolide antibiotics with eukaryotic organisms remains a controversial subject (for review see ref. 1) It has been claimed that the lethal binding of these antibiotics to the sterol target component of the hydrophobic core of the membrane, in accordance with the model of de Kruijff and Demel, is an energy-dependent process. When energy production is reduced by removal of all metabolisable substrates or by adding metabolic inhibitors, polyene binding and antifungal effects are also reduced. Metabolic energy may be required to maintain binding site accessibility or to move antibiotic molecules to the active site. The interaction is also restricted at low temperatures, possibly because of the reduced thermal mobilities of the groups concerned with antibiotic uptake. However, it should be emphasised that the interaction of polyene macrolides with artificial lipid membranes is a purely physicochemical process, although the type of permeability pathways induced are similar to those observed in natural membranes. Using Chlorella vulgaris as a model organism, we demonstrate here that the interaction of polyene macrolides with sensitive cells and the induction of lethal membrane permeability changes are energy-dependent processes or purely physicochemical phenomena, depending on the structure of the antibiotic used.

Polyene macrolide antibiotic cytotoxicity and membrane permeability alterations. II. Phenotypic expression in intraspecific and interspecific somatic cell hybrids

Cytotoxicity and membrane permeability alterations induced by the polyene macrolide antibiotics filipin (FIL) and pimaricin (PIM) have been compared in parental intraspecific and interspecific somatic cell hybrids. B82 (mouse) and B1 (hamster) cells were found to be more resistant than RAG (mouse) parental cells to both polyene macrolides as indicated by 24-hour survival, 72-hour viability, and growth rate. Analysis of both intraspecific and interspecific somatic cell hybrids indicated that polyene macrolide resistance was being expressed even in the presence of the polyene macrolide-sensitive (RAG) genome. Where one of the two parental cell types is relatively polyene macrolide resistant, the use of specific polyene macrolides may prove efficacious as half-selective agents in cell hybridization.

Penicillin production by mutants ofPenicillium chrysogenum resistant to polyene macrolide antibiotics

Polyene-resistant mutants ofPenicillium chrysogenum Wis. 54–1255 have been obtained by stepwise selection in increasing concentrations of polyene antibiotics. From the parent strain, sensitive to 10 μg/ml of polyene antibiotics, mutants resistant to fungimycin (1.3 mg/ml), amphotericin B (0. 5 mg/ml), or nystatin (167 μg//ml) were obtained. Their penicillin production is different from that of the parent strain and in particular some of the fungimycin-resistant mutants produce higher levels of penicillin.

Methyl esters of trimethylammonium derivatives of polyene macrolide antibiotics.

Methyl esters of trimethylammonium derivatives of polyene macrolide antibiotics.

Selecting interspecific human-mouse and Chinese hamster-mouse hybrids using a new half-selection technique with a polyene antibiotic.

Interspecific human-mouse and Chinese hamster-mouse hybrids were isolated from polyethylene glycol fused cells by a new half-selection technique employing a structurally modified polyene macrolide antibiotic, amphotericin B methyl ester (AME), and HAT media. Unfused parental cells were killed as a result of innate sensitivity to AME or their genetic deficiency, absence of thymidine kinase (TK-) or hypoxanthine guanine-phosphoribosyl transferase (HGPRT-). In contrast, hybrid colonies were isolated after two to three weeks growth in three or four changes of HAT-AME media and subsequent growth in HAT media alone. The ability of hybrid cells to proliferate using this selective protocol indicates that genetic complementation resulted, and polyene antibiotic resistance was expressed as a dominant phenotypic property in the hybrids. Hybrid selection was dependent on: (1) the number of cells of each parental cell type co-cultivated; (2) the level of polyene antibiotic administered; and (3) the time interval before selection was initiated. The half-selection technique described in this report is simple to use, very effective in eliminating unfused parental cells and increases the potential types of hybrids which can be formed. Only one parental cell type need contain a biochemical defect, whereas the second parental type can be genetically normal.

The structure of lienomycin, a novel type of polyene macrolide antibiotic

The structure of lienomycin, a novel type of polyene macrolide antibiotic

The structure of a novel sugar component of polyene macrolide antibiotics: 2,6-dideoxy-L-ribohexopyranose.

A novel carbohydrate has been isolated after acidic hydrolysis of nystatin A3, candidinin and polyfungin B and its structure established as 2,6-dideoxy-L-ribohexopyranose.

Investigation of polyene macrolide antibiotic-induced permeability changes in vesicles by 31P nuclear magnetic resonance.

The permeability of egg yolk lecithin (EYL) vesicles to Pr3+ has been measured by 31P nuclear magnetic resonance (nmr) spectroscopy. Measurable Pr3+ leakage into the internal aqueous compartment of EYL vesicles at ambient (21 degrees C) temperature required the presence of small (7--10 mol%) amounts of dicetyl phosphate (DCP). The permeability of DCP-containing vesicles is decreased by incorporation of sterol (cholesterol greater than ergosterol approximately 5.6-dihydroergosterol greater than zymosterol) into the lipid bilayer. Addition of the polyene macrolide antibiotic, nystatin, to DCP-containing EYL vesicles with and without sterol resulted in increased Pr3+ permeability at the three temperatures studied (21--37.5 degrees C). Permeability changes observed upon addition of nystatin to sterol-impregnated, DCP-containing vesicles varied with sterol structure: ergosterol approximately 5,6-dihydroergosterol greater than cholesterol approximately zymosterol. These results are compared with other polyene macrolide induced permeability changes on model and natural membrane systems. Permeability changes induced by nystatin in sterol-free EYL vesicles were generally greater than for comparable sterol-containing vesicles. This is attributed to a nonspecific interaction of the antibiotic with the latter vesicles.

Isolation and characterization of amphotericin B-resistant cell lines in Chinese hamster cells

Amphotericin B is a polyene macrolide antibiotic which interacts specifically with steroids in mammalian cell membranes. Amphotericin B-resistant (AMB r) lines of stable phenotype have been isolated from cultured Chinese hamster (V79) cells. Three AMB r clones (AMB r-1, -2 and -3) isolated independently after treatment with nitrosoguanidine were resistant to ≥150 μg/ml of the antibiotic, while DNA synthesis as well as the colony-forming ability of the parental V79 cells was blocked by >80% of control in the presence of 20–50 μg/ml amphotericin B. The AMB r cell line also exhibited increased resistance to other polyene macrolide antibiotics such as nystatin and pentamycin. Other agents, however, such as cytosine arabinoside or ricin, blocked DNA synthesis in AMB r cells to the same extent as in V79 cells. The amphotericin B resistance phenotype was stably retained even after AMB r cells were cultured in the absence of the drug for over 200 generations. The content of free cholesterol or its esters was significantly decreased in all three resistant clones. Furthermore, cholesterol synthesis from acetate as well as mevalonate was partly defective in AMB r cells, compared with that in V79 cells.

ChemInform Abstract: PIMARICIN. VIII. STRUCTURAL AND CONFIGURATIONAL STUDIES BY ELECTRON IMPACT AND FIELD DESORPTION MASS SPECTROMETRY, CARBON‐13 (25.2 MHZ) AND PROTON (270 MHZ)‐NMR SPECTROSCOPY

Attempts to obtain a molecular ion from the polyene macrolide antibiotic pimaricin by EI and FD mass spectrometry were unsuccessful The loss of carbon dioxide and a varying number of water molecules from the molecular ion made a molecular-weight determination impossible 13C NMR spectroscopy of N-acetylpimaricin, its dodecahydroderivative, and of HH-2, a hydrogenation-hydrogenolysis product of N-acetylpimaricin, confirmed that the antibiotic has structure 3 containing a hemi-ketalic ring and lacking an OH group at C-8. The value of the anomeric coupling constant, Jc-1-h-1, indicates that the mycosamine moiety is β-glycosidically bound to the aglycone. The structure and configuration of the antibiotic have been studied by analysis of the 270 MHz 1H-NMR spectra of N acetylpimaricin and HH-2. By comparison of the two spectra and by extensive decoupling experiments, all signals in the spectrum of N-acetylpimaricin have been assigned to the protons in structure 3. From chemical-shift, coupling constant, and integral values, it was deduced that the mycosamine ring is pyranoid with a chair conformation, the hemi-ketal is 6 membered and occupies a chair conformation with the substituents in equatorial positions, the epoxy protons as well as the olefinic protons are trans to each other, and the antibiotic is diastereomerically pure.

Sequential Expression of Macromolecule Biosynthesis and Candicidin Formation in Streptomyces griseus

Streptomyces griseus did not produce the polyene macrolide antibiotic candicidin during the initial growth phase characterized by rapid RNA synthesis. the absence of candicidin production when RNA or protein synthesis was inhibited by rifampicin or chloramphenicol suggests a transcriptionally controlled late formation of the candicidin synthases. Phosphate levels in the medium control the rate of DNA, RNA and protein synthesis. Depletion of phosphate appears to trigger the onset of candicidin synthesis after a drastic reduction of the rate of RNA synthesis. Changes in the ATP pool during the fermentation suggest that ATP may be the intracellular effector controlling the onset of antibiotic synthesis.

Effect of polyene macrolide antibiotics on invertebrate tissue culture cells.

The effect of amphotericin B (AB) and amphotericin B methyl ester (AME) on viability and DNA synthesis in three insect cell lines, Trichoplusia ni (Tn) Carpocapsa pomonella 169 (Cp) and Aedes aegypti (Aa), has been evaluated. In all cases AME was less toxic than AB and inhibited DNA synthesis to a lesser degree than AB. However, the three cell lines differed in their response to the two polyene macrolide antibiotic preparations. Tn and Aa cell responded in a similar manner when exposed to either AME or AB, whereas Cp cells were more sensitive to both antibiotics than Tn or Aa cells.

Pimaricin—VIII : Structural and configurational studies by electron impact and field desorption mass spectrometry, 13C (25.2MHz) and 1H (270 MHz)-NMR spectroscopy

Attempts to obtain a molecular ion from the polyene macrolide antibiotic pimaricin by EI and FD mass spectrometry were unsuccessful The loss of carbon dioxide and a varying number of water molecules from the molecular ion made a molecular-weight determination impossible 13C NMR spectroscopy of N-acetylpimaricin, its dodecahydroderivative, and of HH-2, a hydrogenation-hydrogenolysis product of N-acetylpimaricin, confirmed that the antibiotic has structure 3 containing a hemi-ketalic ring and lacking an OH group at C-8. The value of the anomeric coupling constant, J c-1- h-1 , indicates that the mycosamine moiety is β-glycosidically bound to the aglycone. The structure and configuration of the antibiotic have been studied by analysis of the 270 MHz 1H-NMR spectra of N acetylpimaricin and HH-2. By comparison of the two spectra and by extensive decoupling experiments, all signals in the spectrum of N-acetylpimaricin have been assigned to the protons in structure 3. From chemical-shift, coupling constant, and integral values, it was deduced that the mycosamine ring is pyranoid with a chair conformation, the hemi-ketal is 6 membered and occupies a chair conformation with the substituents in equatorial positions, the epoxy protons as well as the olefinic protons are trans to each other, and the antibiotic is diastereomerically pure.

Comparison of candicidin, levorin and trichomycin by means of high-performance liquid chromatography

A high-performance liquid chromatographic method for the separation of individual components of polyene macrolide antibiotics has been developed. The three heptaene macrolide antibiotics, candicidin, levorin and trichomycin, have been investigated and compared. In all instances these compounds proved to be complex mixtures. It is demonstrated that candicidin and levorin are identical, whereas trichomycin differs in composition from these two substances.

The effect of fetal bovine serum on polyene macrolide antibiotic cytotoxicity and antifungal activity.

The relationships between fetal bovine serum (FBS) concentration and polyene macrolide antibiotic cytotoxicity to animal cells and to fungi were evaluated. The toxicity of amphotericin B (AB) and its derivative, amphotericin B methyl ester (AME), toward KB cells was found to be directly related to fetal bovine serum concentration. At higher FBS levels, increased concentrations of AB and AME were required to reduce 72-hr KB viable cell numbers of 50% of control values. Similarly, polyene macrolide antibiotic levels required to inhibit the growth of Saccharomyces cerevisiae to 50% of controls, and for obtaining minimum fungicidal concentrations (MFC), were greater when higher levels of FBS were used. In addition, AME was less toxic than AB toward KB cells grown in media containing 2, 5, 10, 15 or 20% FBS, whereas the antifungal activities of AB and AME were similar. AME was also capable of eliminating Candida albicans, Saccharomyces cerevisiae, Aspergillus niger or Fusarium moniliforme from KB cultures at antibiotic levels which exhibited less cell toxicity than did the concentrations of AB required for a similar response. These findings indicate that AME may be a potentially useful antifungal antibiotic for tissue culture systems.

Sterol biosynthesis in antibiotic sensitive and resistant Candida

Sterol composition and biosynthesis in Candida utilis and C. albicans have been investigated. Significantly these Candida species were found to contain 24-methylene and 24-methyl lanosterol. The occurrence of these sterols constitutes the first evidence that C-24 alkylation of lanosterol occurs in yeast. The results of isolation and tracer studies in C. utilis are interpreted in terms of the formation of ergosterol, the major sterol of Candida, via fecosterol ( 8), episterol ( 13), Δ 7-ergosten-3β-ol ( 14) and Δ 5,7-ergosten-3β-ol ( 16). The order in which ergosterol producing transformations occur in these yeasts differs slightly from that observed in Saccharomyces cerevisiae. Reculturing C. albicans on media containing increasing amounts of the polyene macrolide antibiotics nystatin and lucensomycin led to the segregation of cultures resistant to these antibiotics. Analysis of the sterol composition of these cultures revealed that the major sterols of these successively more resistant cultures were successively more primitive biogenetic precursors of ergosterol in the parent strain. This result parallels observations in studies of S. cerevisiae.