ABSTRACT
Quenching of anthracycline fluorescence by a series of spin-labeled fatty acids was used to probe the transverse location of the drug in phosphatidylcholine bilayers in the form of small unilamellar vesicles. Stern-Volmer plots of the quenching data indicate that the fluorophore moiety of the anthracycline is intercalated into the hydrocarbon region of the bilayer, with deeper penetration observed in fluid-phase than in solid-phase vesicles. 31P-NMR parameters (T1 and nuclear Overhauser enhancement (NOE] are unaffected by the presence of drug, consistent with a binding site removed from the interfacial region. Comparison of intensity (F0/F) plots with lifetime (tau 0/tau) data shows that the predominant mechanism of anthracycline quenching by membrane-bound nitroxides is static. Since the membrane-bound drug is also accessible to quenching by I-, the binding site in the membrane must create a channel which is accessible to solvent. Two other fluorescent probes, 12-(9-anthroyloxy)stearate (12-AS) and diphenylhexatriene (DPH), were employed to confirm the results obtained with the anthracyclines, giving quenching data representative of their location in the bilayer.
Subject(s)
Antibiotics, Antineoplastic/analysis , Lipid Bilayers/analysis , Carubicin/analysis , Doxorubicin/analysis , Electron Spin Resonance Spectroscopy , Fluorescence , Fluorescent Dyes , Liposomes , Magnetic Resonance Spectroscopy , Phosphorus Isotopes , Spin LabelsABSTRACT
A three component system for separating a mixture of carminomycin, carminomycinone and 13-dihydrocarminomycinone by HPLC was developed. Spherisorb ODS Column, 4.6 X 250 mm, the particle size of 10 micron was used. The impact of the mobile phase composition, temperature during chromatography and buffer solution pH on the capacity factors K' for every of the above compounds was studied. For determining purity of carminomycin dosage forms the procedure with an external standard was applied. The procedure provides routine quantitative assay of carminomycin hydrochloride dosage forms.
Subject(s)
Carubicin/analysis , Daunorubicin/analogs & derivatives , Buffers , Carubicin/analogs & derivatives , Carubicin/isolation & purification , Chromatography, High Pressure Liquid/instrumentation , Chromatography, High Pressure Liquid/methods , Hydrogen-Ion ConcentrationABSTRACT
A flow-through cuvette in which cells attach as a monolayer to a quartz plate was developed for measurement of the light absorbance of anthracyclines in cells. Despite the drawback of a short path-length (of the order of the cell diameter), a dynamic flow-through set-up and baseline storage made it possible to measure intracellular absorbance and obtain spectral data for daunomycin and carminomycin. Stopping the flow and allowing the drug to equilibrate between medium and cells led to a 20% decrease of molar light absorption of cellular anthracycline, which permitted measurement of the total cellular concentration. Furthermore, accumulation and efflux kinetics were determined for H35 rat hepatoma cells. On the basis of the reported formation constant of the iron-complex of carminomycin, which is of the order of 10(34), we expected to find this complex within the cells. However, the spectrum of cellular drug did not show absorbance bands characteristic of the complex. A red shift and hypochromism were found in the daunomycin spectrum after intracellular binding, which corresponds with the spectral change observed after intercalation of daunomycin into DNA.
Subject(s)
Carubicin/analysis , Daunorubicin/analogs & derivatives , Daunorubicin/analysis , Liver Neoplasms, Experimental/analysis , Animals , Antibiotics, Antineoplastic , Carubicin/metabolism , Daunorubicin/metabolism , Liver Neoplasms, Experimental/metabolism , Naphthacenes/analysis , Rats , Spectrophotometry/instrumentation , Spectrophotometry/methodsABSTRACT
A new antitumor antibiotic, 13-deoxycarminomycin, has been isolated from the anthracycline complex produced by Streptomyces peucetius var. carminatus (ATCC 31502), a biochemical mutant of Streptomyces peucetius var. caesius, the doxorubicin-producing microorganism. The new anthracycline, showing antibacterial and cytotoxic activity in vitro, was found active against P-388 murine leukemia.
Subject(s)
Antibiotics, Antineoplastic/analysis , Animals , Antibiotics, Antineoplastic/biosynthesis , Antibiotics, Antineoplastic/pharmacology , Carubicin/analogs & derivatives , Carubicin/analysis , Carubicin/biosynthesis , Carubicin/pharmacology , Chemical Phenomena , Chemistry, Physical , Fermentation , HeLa Cells , Humans , Leukemia P388/drug therapy , Mice , Naphthacenes/analysis , Naphthacenes/biosynthesis , Naphthacenes/pharmacology , Streptomyces/metabolismABSTRACT
Saccharides II and III were obtained on catalytic hydrogenolysis of carminomycins II and III. The saccharides contained daunosamine and a nitrogen-free fragment of unknown structure. Crotonic aldehyde in the form of 2,4-dinitrophenylhydrazone and 1,2-propylenglycol in the form of 3,5-dinitrobenzoate were isolated on acid hydrolysis of the saccharides. The study on the chemical and spectral properties (PMR-spectra) of the saccharides, their N,O-acetates and hydrolysis products suggested the structure of th carbohydrate moiety of carminomycins II and III as a disaccharide of daunosamine and acyclic semiacetal of 2,4-desoxytetrose.
Subject(s)
Carbohydrates/analysis , Carubicin/analysis , Daunorubicin/analogs & derivatives , Chemical Phenomena , Chemistry , Chromatography , Electron Spin Resonance Spectroscopy , Spectrophotometry, InfraredABSTRACT
Carminomycins II and III, the main components of the carminomycin complex were isolated in pure state. Their crystalline exalates and acetate of cardminomycin II were prepared. The PMR spectra of both carminomycins and the 13C-NMR spectra of the oxalates were obtained. The molecular weights of the antibiotics were determined by mass-spectrometry. On the basis of the PMR spectra it was shown that carminomycins II and III had similar structures and differed in the stereoisomerism of the nitrogen-free fragment linked to the amino sugar. This was confirmed by the 13C-NMR spectra. The above fragment (C7H15O3) is analogous to the fragment of baumycins A1 and A2 described earlier.
Subject(s)
Carubicin/analysis , Daunorubicin/analogs & derivatives , Novobiocin/analogs & derivatives , Acetates/analysis , Chemical Phenomena , Chemistry , Chromatography, Gel , Chromatography, Thin Layer , Electron Spin Resonance Spectroscopy , Magnetic Resonance Spectroscopy , Mass Spectrometry , Molecular Weight , Novobiocin/analysis , Novobiocin/isolation & purification , Oxalates/analysisABSTRACT
Three components differing by their properties from the carminomycins described earlier were isolated from the carminomycin complex. Comparison of the IR and UV spectra, as well as chromatographic and physicochemical properties of 2 of them showed that they were dihydrocarminomycin and its aglycone or dihydrocarminomycinone, which was prepared earlier by synthesis. The third component was a chromophore belonging to 1,4,6-trihydroxyanthraquinone. Investigation of its IR spectrum, physicochemica properties and PMR spectrum showed it to be carboxymethylethylcarminomycinone identical to epsilon-rodomycinone. The data were confirmed by 13C-NMR spectrometry.
Subject(s)
Carubicin/analysis , Daunorubicin/analogs & derivatives , Chemical Phenomena , Chemistry, Physical , Chromatography, Gel , Crystallization , Electron Spin Resonance Spectroscopy , Glycosides/analysis , Magnetic Resonance Spectroscopy , Spectrophotometry, Infrared , Spectrophotometry, UltravioletABSTRACT
The intracellular distribution of important chemotherapeutic antibiotics belonging to the anthracycline group (e.g. adriamycin) can be detected by laser flow cytometry. The indirect method is based on the interference of these compounds with the binding of propidium iodide to the nuclear DNA. While in the direct method, the intracellular fluorescence of these antibiotics is excited and detected with a laser beam in a flow system. The present report demonstrates the use of these two methods for intracellular detection and quantitation of a number of important anthracyclines.
Subject(s)
Antibiotics, Antineoplastic/analysis , Cytological Techniques , Animals , Carubicin/analysis , Cell Line , DNA/metabolism , Daunorubicin/analogs & derivatives , Daunorubicin/analysis , Doxorubicin/analogs & derivatives , Doxorubicin/analysis , Humans , Lasers , Leukemia , Mice , Nogalamycin/analysis , Propidium/metabolism , Spectrometry, FluorescenceABSTRACT
Interaction of 14-bromine derivatives of daunorubicin and carminomycin, as well as their aglycones with secondary amines, such as piperidine, N-methylpiperazine and morpholine was studied with a purpose of preparing new potentially antitumor antibiotics. It was found that reaction in acetone or dioxan at a temperature of 50--60 degrees C resulted in formation of 14-amino derivatives of the respective bromides. 14-Piperidinyl- and 14-(N-methylpiperazinyl)-daunorubicin, 14-piperidinyl-daunorubicinon and 14-piperidinyl-carminomycinon were prepared. The structure of the new substances was confirmed by the IR, UV and NMR spectra. The antimicrobial activity of 14-amino derivatives amounted to 10--35 per cent of the activity of the initial antibiotics, i.e. daunorubicin and carminomycin.
Subject(s)
Antibiotics, Antineoplastic/chemical synthesis , Carubicin/chemical synthesis , Daunorubicin/analogs & derivatives , Bacillus/drug effects , Carubicin/analogs & derivatives , Carubicin/analysis , Daunorubicin/analysis , Daunorubicin/chemical synthesisABSTRACT
A mutant strain of Streptomyces peucetius produced an anthracycline antibiotic whose structure has been established to be 4-O-demethyl-13-dihydrodaunorubicin (4), by application of spectroscopic methods and chemical degradation. A new synthesis of 4-O-demethyl-daunorubicin (carminomycin I, 2) starting from daunomycinone, together with the comparison of the antitumor activity of the anthracycline glycosides 2 and 4 are also reported.
Subject(s)
Antibiotics, Antineoplastic/pharmacology , Carubicin/pharmacology , Animals , Antibiotics, Antineoplastic/therapeutic use , Carubicin/analogs & derivatives , Carubicin/analysis , Carubicin/chemical synthesis , Cells, Cultured , Chemical Phenomena , Chemistry , HeLa Cells , Humans , Leukemia L1210/drug therapy , MiceABSTRACT
The effect of the aeration rate on biosynthesis of carminomycin by Actinomadura carminata and biochemical changes in the fermentation broth on the use of 3 complex media of different composition was studied. The carminomycin-producing organism can grow and produce the antibiotic within the ranges of the aeration changes from 0.98 to 18.56 mgO2/1-min. A decrease in the maximum rate of oxygen dissolution up to 0.98 mgO2/1-min resulted in some decrease in the activity level. Intensive aeration, i.e. 18.56 mgO2/1-min induced suppression of the antibiotic production by 25 per cent.
Subject(s)
Antibiotics, Antineoplastic/biosynthesis , Carubicin/biosynthesis , Culture Media/metabolism , Nocardia/metabolism , Aerobiosis , Carubicin/analysis , Time FactorsABSTRACT
An actinomycete strain No. 85 was isolated from a soil sample on media with bleomycin. It was described as a representative of a new species. Actinomyces cremeospinus sp. nov. An antibiotic substance identical with carminomycin, an antitumor antibiotic was isolated from the culture fluid of the strain.