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[Erythromycin and oleandomycin concentration in the body of angina patients taking the preparations orally].

The antibiotic levels in patients suffering from angina were studied with the agar-diffusion method. The drugs were used orally in single and multiple doses (every 6 hours): erythromycin in doses of 4000 and 6000 gamma/kg and oleandomycin in doses of 4000 and 8000 gamma/kg. When erythromycin was used in a single dose of 4000 gamma/kg, its levels in the objects tested, i.e. the blood serum, saliva, mucus from the tonsil surface and tissue did not reach the concentrations corresponding to the sensitivity levels of the microbes to the antibiotic (0.75 gamma/ml). When the drug was used every 6 hours, the concentration increased. Still, it reached the above level only in the blood and tonsil surface mucus. Only when the antibiotic was used repeatedly in doses of 6000 gamma/kg, its levels in the examined objects reached 0.75 gamma/ml. When oleandomycin was used in single and repeated doses of 4000 gamma/kg, its levels were always lower than those providing sensitivity of beta-hemolytic streptococci to it (1.0 gamma/ml). The antibiotic use in doses of 8000 gamma/kg provided the required level in all the object examined.

Administration, Oral↗

[Sensitivity to the differential dose of erythromycin and oleandomycin of various holarctic strains of tularemia microbe isolated in the Kazakh SSR].

The presence of strains of holarctic race of tuleremia microbe, both sensitive and resistant to the differential doses of erythromycine and oleandomycine was revealed at the territoryn of Kazakh SSR. The existence of such strains does not permit to differentiate them into geographical races by the sensitivity to erythromycine and oleandomycine.

Climate↗

[Quantitative determination of anhydroleandomycin and glycol in oleandomycin phosphate].

A spectrophotometric method for determination of inactive admixtures in oleandomycin phosphate-anhydrooleandomycin was developed. The method was based on measuring optical density of methanol solutions of oleandomycin and its admixtures at a wave length of 235 nm of UV-spectrum. The minimun detectable amount was 0.5 per cent. A method of thin-layer chromatography in a buffered fixed layer was developed for determination of the 2nd inactive admixture, i.e. glycole. The chromatogrammes were developed with xanthydrol. The minimum detectable amount was 3 mug.

Anhydrides↗

[Study of the process of inactivating oleandomycin phosphate preparations].

Ageing of oleandomycin preparations was studied. Certain biologically inactive components accumulating on storage of the preparations were isolated and investigated. A scheme for ageing of oleandomycin phosphate preparations is described. The scheme includes cleaving up of the water molecule at C11 and cleaving by the epoxide ring at the early stages and hydrolysis by the glycoside bonds at the later stages.

Chemical Phenomena↗

[Cyclic adenosine-3',5-monophosphate in Streptomyces antibioticus and its possible role in regulating oleandomycin biosynthesis and the growth of the culture].

When glucose is substituted for sucrose in the fermentation medium for Streptomyces antibioticus, the pH of the cultural broth becomes more acidic, the rate of protein synthesis in the mycelium rises, and the rate of oleandomycin synthesis decreases abruptly. The dynamics of cAMP (cyclic monophosphate) accumulation was studied in the process of biosynthesis by the culture in different media. Most of the synthesized cAMP (80-90%) was shown to be excreted into the medium. Glucose stimulates cAMP synthesis and excretion from the mycelium by a factor of 1.5-3. No distinct correlation was found between cAMP content in S. antibioticus cells and the level of oleandomycin biosynthesis. A correlation between changes in the concentration of exocellular cAMP and protein synthesis in the mycelium suggests that the excreted cAMP may be involved in regulating the growth of the culture producing the antibiotic.

Culture Media↗

[Prenatal effect of oleandomycin on the development of the immunogenesis organs].

The ante- and postnatal development of the thymus gland and mesenteric lymph nodes of the rats receiving oleandomycin in the period of placentation and organogenesis (the 8th-14th days of embryogenesis) was studied by using anatomical, histological and electron microscopical methods. Oleandomycin produced inhibition of lymphopoiesis against the background of an increase of eosinophilic and neutrophilic granulocytes, reactive changes of the microcirculatory bed and ultrastructural lesions of the blood capillary wall.

Animals↗

[Multiplication of a DNA fragment in Streptomyces antibioticus--producer of oleandomycin].

Two strains of Str. antibioticus producing oleandomycin were studied. Strain 326 was obtained from the initial laboratory strain and strain 1607 from strain 326 as a result of multistage selection aimed at increasing the antibiotic production capacity. Extrachromosomal ring DNA could be identified in strain 1607. The identified DNA was designated as eSA1 or extrachromosomal element of Streptomyces antibioticus 1. The molecular weight of this DNA is 21.3 Md. It is represented by 1 copy per chromosome. No eSA1 was isolated from strain 326 at CsCl-EtBr gradient. Hybridization studies according to Southern with the use as a probe of 32-eSA1 DNA showed that strain 326 contained 1 copy of eSA1 per chromosome in the integrated state. The hybridization studies, electron microscopy and analysis of the total DNA in CsC1-EtBr gradient showed that eSA1 in strain 1607 was tandemly multireplicated in the chromosome content. In the autonomous state its number was approximately equal to 1 copy per chromosome. The presence of eSA1 in strain 1607 in the autonomous state probably results from its segregation during homologous recombination due to tandem multireplication. The data are indicative of multiplication in strain 1607 of the chromosome fragment 23.3 Md in size. It is suggested that an increase in the oleandomycin production capacity of strain 1607 is associated with multiplication of the DNA fragment (eSA1) containing the genes determining production of the antibiotic.

Chromosomes, Bacterial↗

[Effect of conditions of agitation and aeration on the biosynthesis of oleandomycin].

The data on the effect of the aeration and agitation conditions on biosynthesis of oleandomycin in 0.75, 3 and 50 m3 apparatus are presented. The relationship between the biosynthetic parameters, oxygen dissolution rate and specific power consumption for agitation was studied. It was shown that the values of the specific power consumption for agitation were not acceptable for scaling up the process of oleandomycin biosynthesis.

Bacteriological Techniques↗

[Conditions for grading the process of oleandomycin biosynthesis].

The data on the study of the relation between the activity of the oleandomycin biosynthesis and the operating and physical parameters of the fermenter are presented. A dependence for computation and selection of the aeration and agitation conditions of the oleandomycin biosynthesis in apparatus of different capacity is described.

Air↗

[Components of a native solution that displace the equilibrium distribution of oleandomycin during extraction].

A marked decrease in the coefficient of oleandomycin distribution in the system of the fermentation broth filtrate-butyl acetate was observed during the transfer from the 1st to the following extraction stages. It was supposed that the oleandomycin fermentation broth filtrate contained admixtures affecting significantly the antibiotic distribution between the aqueous phase and organic solvent.

Acetates↗

[Use of aluminum salts for purifying oleandomycin culture broth].

It was shown that "nepheline coagulant"--Al2 (SO4)3-18H2O was effective in coagulating the admixtures in oleandomycin fermentation broth. The use of the "nepheline coagulant" in an amount of 1 to 3 per cent (by the weight) of the volume of the oleandomycin fermentation broth provided butyl acetate extraction of the antibiotic without formation of stable emulsion. Addition of the "nepheline coagulant" in an amount of 1 to 2 per cent (by the weight) to the fermentation broth markedly increased the rate of the fermentation broth filtration.

Aluminum↗

In vitro interaction of rat liver cytochromes P-450 with erythromycin, oleandomycin and erythralosamine derivatives. Importance of structural factors.

Several derivatives of the erythromycin, erythralosamine and oleandomycin series have been prepared. Their abilities to bind to rat liver microsomal cytochrome P-450 and to lead to the formation of stable 456 nm absorbing cytochrome P-450-metabolite complexes after their oxidative microsomal metabolism in vitro have been compared. The obtained data confirmed that cytochrome P-450 induced in rats either by macrolides or by 16 alpha-pregnenolone carbonitrile were the major isozymes involved in the binding of macrolides to liver microsomes and in metabolite-complex formation. They showed that (i) hydrophobicity was in general a beneficial factor for these two properties, (ii) the presence of a bulky substituent in position 3 of erythromycin dramatically decreased their affinity for these isozymes, and (iii) the simultaneous presence of bulky substituents in position 2' and 3 prevented iron-metabolite complex formation. These results led to the selection of two compounds, erythralosamine-2'-benzoate and erythralosamine-2',3-diacetate, which exhibited a particularly high affinity for macrolide inducible cytochrome P-450 and were very good precursors of cytochrome P-450-iron-metabolite complex formation.

Animals↗

In vivo effects of erythromycin, oleandomycin and erythralosamine derivatives on hepatic cytochrome P-450.

Rats have been treated with several derivatives of the erythromycin, erythralosamine or oleandomycin series, in order to compare their ability to induce cytochrome P-450 and to form stable 456 nm-absorbing cytochrome P-450 metabolite complexes. The data obtained confirm that the cytochromes P-450 induced in rats by various macrolides are similar to that induced by pregnenolone 16 alpha-carbonitrile: the cytochrome P-450 IIIA1 isozyme. It showed that: (i) formation of a stable inhibitory 456 nm-absorbing cytochrome P-450 complex is not a prerequisite for cytochrome P-450 induction but enhances induction by stabilization of the IIIA isozyme. Therefore, the best inducers lead also to the maximal in vivo amounts of cytochrome P-450 metabolite complex (except for 2'MBEM); (ii) affinity for cytochrome P-450 IIIA1 is not directly involved for induction; and (iii) hydrophobicity favors induction and formation of complexes. Structural factors are also involved.

Animals↗

The effects of triacetyloleandomycin and oleandomycin phosphate on the glucocorticoid receptor in cultured skin fibroblasts.

Triacetyloleandomycin (TAO) has been described as a "steroid-sparing" drug in that poorly controlled asthmatic patients can be stabilized or improved with the addition of TAO despite decreasing dosages of steroids, specifically methylprednisolone (MP). It is not clear whether the beneficial effects of TAO are due to decreased MP clearance or are due to enhanced glucocorticoid effect peripherally. We tested the latter possibility by studying the interaction of TAO and oleandomycin phosphate (OLEO), the active metabolite of TAO in vivo, with glucocorticoid receptors in dispersed, intact cultured human skin fibroblasts. With the use of cells incubated with [3H]dexamethasone at 22 degrees C, we examined the competitive binding properties of TAO and OLEO (at varying concentrations) with and without MP as compared with several other steroids and MP alone. We also studied the effects on cellular glucocorticoid receptor number and affinity when TAO at a concentration of 4 X 10(-5) alone, at 10(-5) M in combination with a receptor saturating concentration of MP (5 X 10(-8) M), with MP alone, or with OLEO (10(-5) M) combined with MP was added to fibroblasts in the growth phase 1 wk before assay. TAO and OLEO did not compete for the binding of [3H]dexamethasone to the fibroblast glucocorticoid receptor, nor did they alter the binding properties of MP. With prolonged cellular exposure, TAO alone did not alter the number of glucocorticoid receptors (per cell) or their affinity for [3H]dexamethasone. Interestingly, prolonged exposure to saturating concentrations of MP alone decreased glucocorticoid receptor density; this effect was not altered by the presence of TAO or OLEO.(ABSTRACT TRUNCATED AT 250 WORDS)

Cell Count↗

Inversion of the anomeric configuration of the transferred sugar during inactivation of the macrolide antibiotic oleandomycin catalyzed by a macrolide glycosyltransferase.

Macrolides are a group of antibiotics structurally characterized by a macrocyclic lactone to which one or several deoxy-sugar moieties are attached. The sugar moieties are transferred to the different aglycones by glycosyltransferases (GTF). The OleI GTF of an oleandomycin producer, Streptomyces antibioticus, catalyzes the inactivation of this macrolide by glycosylation. The product of this reaction was isolated and its structure elucidated. The donor substrate of the reaction was UDP-alpha-D-glucose, but the reaction product showed a beta-glycosidic linkage. The inversion of the anomeric configuration of the transferred sugar and other data about the kinetics of the reaction and primary structure analysis of several GTFs are compatible with a reaction mechanism involving a single nucleophilic substitution at the sugar anomeric carbon in the catalytic center of the enzyme.

Amino Acid Sequence↗

High-performance liquid chromatographic analysis of oleandomycin in serum and urine.

The determination of oleandomycin in serum and urine by high-performance liquid chromatography using erythromycin as internal standard is described. The separation was achieved on a reversed-phase C18 column employing acetonitrile-0.05 M phosphate buffer (30:70), adjusted to pH 7.0, as the mobile phase with UV detection at 200 nm. A solid-phase extraction procedure, combined with a simple phase-separation step was used prior to chromatographic analysis. Linear calibration curves were obtained in the concentration ranges 0.25-5.0 micrograms/ml (serum) and 1.0-25.0 micrograms/ml (urine). Precise quantitative analysis has been achieved at these levels with relative standard deviations of less than 5%.

Chromatography, High Pressure Liquid↗

Development of a capillary electrophoretic method for the separation of the macrolide antibiotics, erythromycin, josamycin and oleandomycin.

Capillary electrophoresis (CE) provides high separation efficiency and thus is suitable for the analysis of complex mixtures of structurally similar compounds. The versatile nature of CE can be realised by controlling the chemistry of the inner capillary wall, by modifying the electrolyte composition and by altering the physicochemical properties of the analyte. A CE method has been developed for the separation of three macrolide antibiotics, erythromycin, oleandomycin and josamycin. A systematic approach was used to maximise analyte differential electrophoretic mobility by manipulating electrolyte pH, molarity and composition. In addition, some instrumental parameters such as capillary length and diameter and applied voltage were varied. The effect of the sample solvent and on-capillary concentrating techniques such as field amplified sample injection were investigated. Also, the influence of the injection of a water plug on the quantity of sample injected was demonstrated. The macrolides were completely resolved in less than 30 min in a 100 cm x 75 microm I.D. fused-silica uncoated capillary with a Z-shaped flow cell of path-length 3 mm. The analysis was performed in a 75 mM phosphate buffer (pH 7.5) with 50% (v/v) methanol and an applied voltage of 25 kV was selected to effect the separation.

Anti-Bacterial Agents↗