NL7900217A - Antibiotics 20-deoxy-narasin and 20-deoxy-epi-17-narasin - useful as antibacterials, anticoccidials, antivirals, feed utilisation improvers, cardiotonic agents etc. - Google Patents

Abstract

20-Deoxynarasin of formula (I), and 20-doxy-epi-17-narasin, and their salts are new. The cpds. are useful as antibacterials effective against anaerobic strains and against Mycoplasma spp.; antivirals esp. active against rhinovirus type 3, vaccinia virus, herpes virus and influenza A virus; anticoccidials active against Eimeria tenella; and for improving feed utilisation by animals esp. ruminants. Also potentially effective against swine dysentery, as cardiotonic agents, as inhibitors of AtPase and as ionophores for selective removal of a cation from a medium, esp. Ag ions from photographic solns. and toxic cations from industrial wastes, or as components of a membrane useful for selective transport of cations against a concn. gradient. (I) and (II) are found in the antibiotic complex produced by cultivating Streptomyces aureofaciens Suggar NRRL 11181. Other factors are also formed, esp. narasin, adn these are sepd. by solvent extraction, chromatography etc.

Description

N / 28. 909-Kp / vdM * 1 f *

Eli Lilly and Company in Indianapolis, Indiana, United States of America.

Deoxynarasin antibiotics, method for their preparation and their use.

The invention relates to cc-deoxynarasin antibiotics, to their use as an antibacterial and anticocidal agent, and to improving feed utilization in ruminants.

5 There is always a need for new improved antibiotics in veterinary medicine. For example, coccidiosis is always a major problem in poultry breeding rows. Coccidiosis results from infection by one or more species of Eimeria or Isospora (for a summary 10 see Lund and Farr in "Diseases of Poultry", fifth edition,

Biester and Schwarte, Eds., Iowa State University Press,

Ames, Iowa, 1965, pp. 1056-1096). The economic losses due to coccidiosis are high, leading to a high demand for better anticocidal agents.

15 Promoting the growth of ruminants, such as cattle and sheep, is another economically desirable goal of veterinary science. Of particular interest is growth promotion, which is obtained by improving the utilization of the feed. The mechanism of utilizing the main feed portion (hydrocarbons) of fodder is known. The micro-organisms present in the rumen of the animal break down the hydrocarbons into monosaccharides, which are then converted into pyruvate derivatives. The pyruvates are metabolized by microbiological processes and converted into acetates, butyrates or propionates, commonly referred to as volatile fatty acids (WZ). For a further discussion, see Ling in "Physiology of Digestion and Metabolism in the Ruminant", Phillipson et al., Eds., 30 Oriel Press, Newcastle-upon-Tyne, England, 1970, pages 408-790 02 1 7 410.

2

The utilization of WZ is discussed in McCullough / Feedstuffs, June 19, 1971, page 19? Eskeland et al., In J. An. Sci. 33, 282 (1971)? and Church et al., in "Digestive 5 Physiology and Nutrition of Ruminants," Vol. 2, 1971, p.

622 and 625. Although acetates and butyrates are used, the use of propionates has been found to be better. Moreover, it has been found that when a too small amount of propionate is available, the animals develop ketosis.

As a result, a beneficial compound is a compound which stimulates the animals to form a greater amount of propionates from the hydrocarbons, with the result that the utilization of the hydrocarbons is improved and, at the same time, the formation of ketosis is reduced. The 20-deoxynarasin and the 20-deoxy-epi-17-narasin are new polyether antibiotics. They are most closely related to the known polyether antibiotic narasin (antibiotic A-28086 factor A; U.S. Pat. Nos. 4,035,481 and 4,038,384). The documents described in these patents and 20 by Occolowitz et al. / Biomed. Mass Spectrometry 3, 272-277 (1976) J proposed structure for narasin was recently confirmed by H. Seto et al. / J. Antibiotics 30 (6) 530-532 (1977) _J, which structure appears to correspond to formula 3 of the formula sheet.

25 According to Seto and colleagues. narasin is closely related to salinomycin (salinomycin = 4-dimethylnarasin). More recently, J.W. Westley and colleagues. drawn attention to C-17 epimers of deoxy- (0-8) -salinomycin / J. Antibiotics 30 (7) 610-612 (1977) /. Although the epimers of deoxy-salinomycin are analogous to the epimers of deoxynarasin of the invention, there is no known chemical method by which the deoxynarasin pimers can be prepared from the deoxysalinomycin pimers.

The invention includes the deoxynarasinantibioti-35 cum complex, consisting of at least 2 separate factors 790 02 1 7 3, namely 20-deoxynarasin and 20-deoxy-epi-17-narasin. The complex is prepared by cultivating a strain of the organism Streptomyces aureofaciens Duggar, NRRL 11181.

The term "antibiotic complex," as used in fermentation practice and in this specification, does not refer to a chemical complex, but to a mixture of collectively formed individual antibiotic factors. As is known to those skilled in the art of antibiotic preparation by fermentation, the ratio of the individual factors in an antibiotic complex may vary depending on the fermentation conditions employed.

The pharmaceutically acceptable salts of 20-deoxynarasin and 20-deoxy-epi-17-narasin are also part of the invention. In order to simplify discussions on utilization, the term "deoxynarasin antibiotic" is used, which term refers to a representative from the group of deoxynarasin-antibiotic complex, 20-deoxynarasin, 20-deoxy-epi-17-narasin and the 20 pharmaceutically acceptable salts of 20-deoxynarasin and 20-deoxy-epi-17-narasin.

The present deoxynarasin antibiotics inhibit the growth of the pathogenic organisms in animals and plants. In one aspect of this activity, the present deoxynarasin antibiotics can be used as anticocidal agents. In addition, deoxynarasin antibiotics have been shown to improve the utilization of the feed administered in ruminants.

The deoxynarasin antibiotic complex includes 20-30 deoxynarasin and 20-deoxy-epi-17-narasin, which are obtained from the fermentation in the form of a mixture. The compounds 20-deoxynarasin and 20-deoxy-epi-17-narasin are isolated from the deoxynarasin complex and isolated as separate compounds as described below. The deoxynarasin complex additionally contains minor factors which are removed by the procedures described. The deoxy-narasin antibiotic complex is soluble in most organic solvents, but not in water.

20-deoxynarasin.

20-deoxynarasin, one of the factors of the present complex, has the same absolute configuration as narasin. The structure of 20-deoxynarasin is similar to that of formula 1 of the formula sheet.

20-deoxy-epi-17-narasin.

The structure of 20-deoxy-epi-17-narasin, the other factor of the present complex, has formula 2 of the formula sheet.

One of the best methods for differentiating narasin (A-28086 factor A), 20-deoxynarasin and 20-13 deoxy-epi-17-narasin involves the use of C nuclear spm-13 resonance (nmr spectrometry. Table A includes the C nmr spectral data of these compounds, each in the form of its free acid, performed in deuterium chloroform (data in ppm).

20 TABLE A

narasin 20-deoxynarasin 20-deoxy-epi-l7-narasin 216.5 216.8 218.0 178.4 177.9 177.6 132.0 125.2 '129.2 25 122.0 121.9 125.8 106.5 105.0 107.0 99.6 99.3 99.4 88.5 88.1 85.6 78.4 78.1 78.2 30 77.1 76.6 77.7 75.1 75.0 77 , 0 73.8 73.7 73.3 72.0 72.3 72.6 70.8 71.1 71.1 35 68.5 68.3 69.1 790 02 1 7 5 TABLE A (continued) narasln 20-deoxynarasin 20-deoxy-epi-17-narasin 67.6 56.2 57.7 56.1 49.9 49.3 5 49.9 49.3 48.7 49.3 41.0 38.9 41.1 40.0 36.6 38.7 38, 8 36.4 36.6 36.3 36.2 10 36.2 35.6 35.7 35.5 32.8 33.9 32.9 31.7 33.7 30.9 31.7 32.8 39.5 30.3 30.5 15 29.4 29.6 29.3 29.0 29.0 29.0 28.0 28.1 28.2 26.1 25.8 24.7 24.0 23.7 23.3 20 21.5 21.8 21.8 19.0 18. 8 21.1 18.0 17.5 18.4 16.4 16.3 18.2 15.7 15.7 15.8 25 14.3 14.1 14.3 13.2 13.3 13.7 13.0 13.2 13.5 12.1 12.5 12.5 12.1 12.1 12.1 30 7.0 7.3 7.7 6.3 6.5 6.4

Other good methods for differentiating 20-deoxynarasin from 20-deoxy-epi-17-narasin and from the known A-28086 factors include paper and thin layer chromatography. The R ^ value of narasin (A-28086 factor A), 790 02 1 7 6 A-28086 factor D, 20-deoxynarasin and 20-deoxy-epi-17-narasin in various paper chromatography systems, using Bacillus subtilis ATCC 6633 as a detection organism are listed in Table B.

5 TABLE B

R ^ values A-deoxy-epideoxy solvent system narasin 28086D narasin narasin water: methanol: acetone 10 (12: 3: 1) adjusted to pH 10.5 with NH4OH then lowered to pH 7.5 with H3PO4 0.20 0 .11 0.08 0.21 water: methanol: acetone 15 (12: 3: 1), adjusted to pH 10.5 with NH4OH and then reduced to pH 7.5 with HCl 0.42 0.25 0.21 0 .44 1% methyl isobutyl-20 ketone (MIBK), 0.5% NH4OH in water 0.56 0.38 0.33 0.66 benzene saturated with water 0.51 0.51 0.74 0.55 water: MIBK ethyl acetate (98: 1: 1) 0.77 0.61 0.51 0.68

The R values of these antibiotics in a typical thin layer chromatography (DLC) system, using silica gel, are listed in Table C.

30 790 02 1 7 7

TABLE C

R ^ values A-deoxy-epideoxy-solvent system narasin 28086D naracin narasin 5 ethyl acetate 0.29 0.35 0.42 0.74

The deoxynarasin antibiotics (20-deoxynarasin and 20-deoxy-epi-17-narasin) are soluble in a wide variety of organic solvents, such as methanol, ethanol, dimethyl formamide, dimethyl sulfoxide, ethyl acetate, chloroform, acetone and benzene. However, they are only slightly soluble in non-polar organic solvents, such as hexane, while they are insoluble in water. It should be noted that 20-deoxynarasin as a free acid is unstable in alcohols and changes to 20-deoxy-epi-17-narasin free acid. For example, 20-deoxynarasic acid (435.1 mg) was dissolved in methanol (40 ml), which solution was stored at room temperature for 4 hours. The solution was then evaporated to dryness under vacuum. The residue was redissolved in 2q dioxane and freeze dried, yielding 417.8 mg of 20-deoxy-epi-17-narasic acid.

Another substance, which co-occurs with A-28086-I by chromatographic analysis, is described in U.S. Pat. No. 4,038,384, and is co-formed with the deoxynarasin complex. Although initially precipitated with the deoxynarasin antibiotics, it can be easily removed by silica gel chromatography. On silica gel thin layer chromatography, this substance appears to be less plolar than 20-deoxynarasin or 20-deoxy-epi-17-3Q narasin when using ethyl acetate as a developing solvent. Preferably, vanillin spray reagent (3% vanillin in methanol + 0.5 ml concentrated H 2 SO 4 per 100 ml solution) is used for the detection. After spraying with vanillin and heating, this substance, like A-22 28086-1, appears to give a blue spot, while the deoxynarasin antibiotics give bright yellow spots, which darken later.

The compounds 20-deoxynarasin and 20-deoxy-epi-17-narasin can be converted into their salts. The pharmaceutically acceptable alkali metal, alkaline earth metal and amine salts of 20-deoxynarasin and 20-deoxy-epi-17-narasin are also part of the invention. "Pharmaceutically acceptable salts" are salts in which the toxicity of the compound as a whole to warm blooded animals has not increased compared to the non-salt form.

Examples of suitable alkali metal and alkaline earth metal salts of 20-deoxynarasin and 20-deoxy-epi-17-narasin are those of sodium, potassium, lithium, cesium, rubidium, barium, calcium and magnesium. Suitable amine salts of 20-deoxy-15 narasin and 20-deoxy-epi-17-narasin are the ammonium salts, as well as the primary, secondary and tertiary C 1 -C 2 alkyl ammonium and hydroxy-C 2 -C 20 alkyl ammonium salts. Examples of amine salts are those salts formed by reacting 20-deoxynarasin and 20-deoxy-epi-17-narasin with 20 ammonium hydroxide, methylamine, sec-butylamine, isopropylamine, diethylamine, diisopropylamine, ethanolamine, triethylamine, 3-amino-1-propanol ed

The alkali and alkaline earth metal cationic salts of 20-deoxynarasin and 20-deoxy-epi-17-narasin 25 are prepared using procedures commonly used for the preparation of cationic salts. For example, the free acid form of the antibiotic is dissolved in a suitable solvent, such as acetone or dioxane water, to which solution a solution of the stoichiometric amount of the desired inorganic base is then added. The salt thus formed can be isolated by routine methods such as filtration or evaporation of the solvent.

The salts formed with organic amines can be prepared in a similar manner. For example, the gaseous or 7900217 9 liquid amine can be added to a solution of the antibiotic factor in a suitable solution of, for example, acetone, after which the solvent and excess amine can be removed by evaporation.

A preferred method for the preparation of a desired salt of one of the deoxynarasin antibiotics is an appropriate choice of isolation method, such as eg adjusting the pH of the mixture with a suitable base or adding an appropriate cationic salt to the solvent, which the extraction is used.

It is known in veterinary pharmacy that the shape of an antibiotic is insignificant in the treatment of an animal with the antibiotic. In most cases, it has been found that due to the conditions present in the animal, the drug passes into a different form than the form in which it was administered. The salt form in which the drug is administered is therefore not significant for the treatment method. However, the salt form can be chosen for reasons of economy, convenience and toxicity.

The present new antibiotics are prepared by culturing a deoxynarasin-producing strain of Streptomyces aureofaciens under submerged aerobic conditions in an appropriate culture medium until significant antibiotic activity is obtained. The antibiotics are recovered using various well known isolation and purification procedures.

The organism to be used for the preparation of deoxynarasin antibiotics was obtained by N-methyl-N'-nitro-N-nitrosoguanidine-induced mutation of Strepto-30 myces aureofaciens NRRL 8092. The taxonomic basis on which S. aureofaciens NRRL 8092 was classified as the strain of Streptomyces aureofaciens Duggar is described in U.S. Patent 4,038,384.

The Streptomyces aureofaciens culture, suitable for the preparation of the deoxynarasin antibiotics, has been deposited 790 02 1 7 * 10 and is part of the culture collection of the Northern Regional Research Center, US. Dept. of Agriculture, Agricultural Research Service, Peoria, Illinois, 61604 and is available to the public under No. NRRL 11181. Deposit 5 was made on October 6, 1977. A -17 deposit of the same culture was made at the American Type Culture Collection October 26, 1978, under No. ATCC 31445.

The culture identified as NRRL 11181 was isolated and mutated from a culture of 10 Streptomyces aureofaciens originally isolated from a soil sample from Mount Ararat in Turkey. The original soil isolate was grown on agar plates, after which subcultures were obtained by removing the individual colonies of the microorganism from the surface of the agar plate. One of the isolates from a culture of the original soil isolate was the culture identified as NRRL 5758.

The above-mentioned naturally selected culture was plated, from which a particular colony was selected. This natural selection method was repeated 8 times, after which a ninth-generation colony isolate was chosen for further selection.

The ninth generation isolate was grown in a chemically defined agar medium with 1% galactose as the sole carbon source. A spore suspension was prepared by ultrasonic stirring of the agar medium, loosening the spores, while minimizing the hyphal fragments by filtration of the suspension through a Whatman No. 1 filter paper.

The spore suspension was grown in a complex liquid medium, the pH of which was adjusted to 8.8, which medium contained 2 mg / ml N-methyl-N1-nitro-N-nitrosoguanidine, at 30 ° C for 72 hours. The cultures grown on the medium were then collected and ground to homogeneous.

The homogenized culture was diluted and spread on agar plates and incubated at 30 ° C until individual colonies could be distinguished. One of the individually selected colonies was used as the starting material for a new culture, identified here as NRRL 8092.

The culture identified as NRRL 8092 was grown in a chemically defined agar medium with glucose as the sole carbon source. A suspension of both spores and hyphal fragments was prepared by ultrasonic stirring. of the agar medium. The mixed spore and hyphal suspension was then treated with -2 mg / ml N-methyl-N'-nitro-N-nitrosoguanidine in a complex liquid medium at a pH of 8.8 for 96 hours on a 30 ° C shaker, after which the culture was collected, homogenized and plated as described above. One of the individual colonies from these agar plates was grown to yield a culture identified as NRRL 11181.

It will be apparent to those skilled in the art that it is now possible to develop additional strains under the present invention which have substantially the same biosynthetic properties as S. aureofaciens NRRL 11181 (ie the ability to produce 20-deoxynarasin and 20-deoxy -epi-17-narasin) by subjecting S. aureofaciens NRRL 5758, NRRL 8092 or NRRL 11181 to mutagenic treatment. Although N-methyl-N'-nitro-N-nitrosoguanidine was used to obtain S. aureofaciens NRRL 11181, other mutagens may also be used, such as UV rays, X-rays, high-frequency waves, radioactive rays and other chemical agents. substances, triggering such mutagenesis. Therefore, part of the invention is directed to a process for the production of deoxynarasin antibiotic complex, for which a Strep tomyces aureofaciens having practically the same biosynthetic properties of NRRL 11181 is grown.

. 790 02 1 7 12

The culture medium used for growing Streptomyces aureofaclens NRRL 11181 can be one of a wide variety of media. Certain culture media, however, are preferred for economical production, optimum yield, and ease of product isolation. For example, preferred carbohydrate sources in large scale fermentation are tapioca, dextrin and sucrose, although glucose, corn starch, fructose, mannose, maltose, lactose and the like can also be used. Corn oil, peanut oil, soybean oil and fish oil 10 are other suitable carbon sources. A preferred stitch. substance source is enzyme hydrolysed casein, although peptone, soybean meal, cottonseed meal, amino acids such as glutamic acid and the like are also suitable. Among the food inorganic salts which can be incorporated into the culture medium are to be mentioned the usually soluble salts which are capable of sodium ions, magnesium ions, calcium ions, ammonium ions, chloride ions, carbonate ions, sulfate ions, nitrate ions, ed ions.

In addition, essential trace elements necessary for the growth and development of the organism must be included in the culture medium. These trace elements usually exist as impurities in other components of the medium in such an amount that the growth requirements of the organism are met.

It may be necessary to add small amounts (i.e. 0.2 ml / l) of an anti-foaming agent, such as polypropylene glycol, to fermentation media at large scale fermentation, in order to prevent foaming.

Although not essential, antibiotic production is promoted by the addition of a small amount of an oil, such as soybean oil.

Submerged aerobic fermentation in tanks is preferred for the production of significant amounts of the deoxynarasin antibiotics. Smaller amounts can be obtained by using shake flask cultures. Due to delays in antibiotic production, usually accompanied by inoculation of large tanks with the spores of the organism, it is preferable to use a vegetative inoculum. The vegetative inoculum is prepared by inoculating a small volume of culture medium containing spores or mycelial fragments of the organism to yield a fresh, actively growing culture of the organism. The vegetative inoculum is then transferred to a larger tank. The medium used for the growth of the vegetative inoculum can be the same as the medium used for larger fermentations, but other media can also be used.

The deoxynarasin-producing organism can be grown at temperatures of 20-40 ° C. Optimal deoxynarasin production takes place at temperatures of 27-30 ° C.

As usual in aerobic immersed culture processes, sterile air 20 is blown through the culture medium. For efficient growth of the organism, the volume of air used in tank production should preferably be above 0.1 volume of air per volume of culture medium per minute. For efficient antibiotic production, the volume of air used in tank production is preferably above 0.25 volume of air per volume of culture medium per minute. Large amounts of dissolved oxygen do not adversely affect the production of the antibiotics.

The production of the antibiotics can be monitored during the fermentation by taking samples of the culture mixtures or extracts of the mycelial solid and testing them for antibiotic activity as compared to known organisms sensitive to the antibiotics. A suitable organism for testing the present antibiotics is Bacillus subtilis ATCC 35 6633. The bioassay is conveniently performed using a paper disc test on agar plates.

Another suitable method is the turbidometric test on a semi-automatic system (Autoturb microbiological assay system, Elanco), described by N.R.

5 Kuzel and F.W. Kavanaugh in J. Pharmaceut. Sci. 60 (5), 764 and 767 (1971). The following test parameters are used in the investigation of deoxynarasinanti biotics: Staphylococcus aureus (H-Heatley) NRRL B-314 in a nutrient culture medium (pH 7) incubated for 4 hours at 37 ° C. The samples to be tested and the standard are dissolved in methanol-water (1: 1). The standard, A-28086 factor A, is presented to the Autoturb carousel at concentrations of 1, 2, 3, 4 and 5 mcg / ml.

The original pH of the inoculated culture medium varies with the medium used. In general, the pH should be between 6.0 and 7.5. The pH at which the organism is recovered, at the end of the fermentation, is slightly higher, between 6.5 and 8.0.

Generally, an antibiotic activity is detectable after the second day of fermentation. Maximum production of antibiotic activity generally occurs between about the fourth and the tenth day.

After their production under submerged aerobic fermentation conditions, the previously described deoxy-narasin antibiotics can be recovered from the fermentation broth using methods commonly used in the fermentation technique. The antibiotics produced during the fermentation occur in both the mycelial mass and in the filtered medium. The maximum recovery of the deoxynarasin antibiotics is therefore obtained by a combination of methods such as filtration, extraction and adsorption chromatography. A preferred solvent for separating the deoxynarasin antibiotics from the whole or filtered fermentation medium is ethyl acetate, although other commonly used solvents are also satisfactory.

A particularly advantageous method for the separation of the deoxynarasin antibiotics is to lower the pH of the entire fermentation broth to about 3/0. At this pH 5, the deoxynarasin antibiotics can be easily separated with the mycelial mass by filtration. This method has been described for the recovery of the related antibiotics A-28086 factors A, B and D and salinomycin, by Boeck and Berg in U.S. Patent 4,009,262. Another advantageous aspect of this method includes the addition of a bicarbonate, such as eg sodium bicarbonate / to the whole medium in an amount of 1 g / l. Using this method, the deoxynarasin antibiotics can be easily isolated with the mycelial mass in salt form. Preferably, methanol is used as the solvent for the separation of the antibiotics from the mycelial mass, but other lower alcohols and ketones can also be used.

Azeotropic distillation can also be beneficial in the recovery of the deoxynarasin antibiotics. In this method, an organic solvent, which forms a suitable azeotrope with water, is added to the aqueous fermentation medium. This solvent culture mixture is subjected to azeotropic distillation to remove at least half of the water from the mixture, leaving a water-solvent mixture in which the deoxynarasin antibiotics are dissolved in the organic solvent. Insoluble by-products can be isolated by suitable techniques, such as filtration or centrifugation. Then, the deoxynarasin antibiotics can be recovered from the organic solution by known procedures such as evaporation of the solvent, precipitation by addition of a non-solvent or extraction.

Organic solvents which form suitable azeotropes with water are butyl alcohol, amyl alcohol, hexyl 780 02 1 7 16 alcohol, benzyl alcohol, butyl acetate, amyl acetate, 1,2-dichloroethane, 3-pentanone, 2-hexanone, benzene , cyclohexanone, tuluene, the xylenes, etc.

There is a special advantage in azeotropic distillation recovery in large scale fermentation processes. Both water and solvent, which are removed during azeotropic distillation, can be separated using known techniques and then reused. The water thus removed is free from impurities.

A further purification of the deoxynarasinanti biotics includes additional extraction and adsorption procedures. Advantageously, adsorption materials such as silica gel, Ό carbon, Florisil (magnesium silicate, Floridin Co., P.O.

Box 989, Tallahassee, Fla.) Etc. are used.

Alternatively, the culture solids, including media components and mycelium, can be used without extraction or separation, but preferably after removal of water, as a source of the deoxynarasin-antibiotic-cum complex. For example, after the production of deoxynarasin antibiotic activity, the culture medium can be dried by freeze-drying or tumble-drying and then directly mixed with the feed premix.

In another aspect, after the production of deoxynarasin activity in the culture medium, the mycelium can be isolated and dried to yield a product which can be used directly in a feed premix. When the mycelium is isolated for such purposes, the addition of calcium carbonate (ca. 10 30 g / l) has been found to aid filtration and yield an improved dry product.

Under the conditions used hitherto, 20-deoxynarasin and 20-deoxy-epi-17-narasin are recovered as the major factors from the Streptomyces aureofaciens 35 strain, as described above and designated NRRL 11181.

780 02 17 17

However, some minor factors can be recovered from S. aureofaciens NRKL 11181 in amounts too small to be characterized. Although the ratio of the factors varies depending on the fermentation and isolation conditions employed, generally more 20-deoxy-epi-17-narasin than 20 deoxynarasin is recovered.

The compounds 20-deoxynarasin and 20-deoxy-epi-17-narasin are separated from each other and isolated as separate compounds using known methods such as column chromatography, thin layer chromatography, counter-current distribution, etc. For example, column chromatography over silica gel used to separate 20-deoxynarasin and 20-deoxy-epi-17-narasin, eluting the column with various solvent mixtures. For example, by using benzene-ethyl acetate in a silica gel column, first 20-deoxy-epi-17-narasin is eluted and then 20-deoxynarasin. Silica gel thin layer chromatography, using 100% ethyl acetate solvent, is a good method for monitoring the elution rate.

The present deoxynarasin antibiotics are antibacterial agents. The relative microbiological activity of 20-deoxy-epi-17-narasin (free acid) is described in Table D. Using the conventional disk diffusion method.

25 TABLE D

inhibition zone _ test organism_ 300 mcg / slice 30 mcg / slice

Staphylococcus aureus 3055 16.9 13.8 ”” 30741 19.5 15.4 30 ”“ 31302 19.0 17.2

Streptococcus pyogenes (Group A) 12.0 10.0 s £; (D) 17.0 14.6

Diplococcus pneumoniae 16.0 13.0 Penicillin G resistant 2 35 Methicillin resistant 790 02 1 7 »18

Deoxynarasin antibiotics have been shown to inhibit the growth of anaerobic bacteria. The minimum inhibitory concentrations (MRC), at which 20-deoxy-epi-17-narasin (free acid) inhibits the growth of various anaerobic bacteria, determined by standard agar dilution methods, are listed in Table E. The end points were read after 24 hour incubation period.

TABLE E

_aerobic bacteria _ MRC (mcg / ml) 10 Actinomyces israelii W855 8

Clostridium perfringens 81 16

Clostridium septicum 1128 16

Eubacterium aerofaciens 1235 16

Peptococcus asaccharolyticus 1302 8 15 Peptococcus prevoti 1281 4

Peptostreptococcus anaerobius 1428 4

Peptostreptococcus intermedius 1264 4

Propionibacterium acnes 79 2

Bacteroides fragilis 111 128 20 The activity against mycoplasma is another suitable aspect of the antimicrobial activity of the deoxynarasin antibiotics. Mycoplasma species, also known as pleuropneumonia-like (PPLO) organisms, are pathogenic in humans and various animals. There is a great need for agents against PPLO organisms, especially in poultry farms. The minimum inhibitory concentrations (MRC) of 20-deoxy-epi-17-narasin (free acid) against mycoplasma species, as determined by in vitro dilution culture, are listed in Table F.

30 TABLE F

organism MRC (mcg / ml) M. gallisepticum 25.0 M. hyorhinis 50.0 M. synoviae 50.0 35 The deoxynarasin antibiotics are also anti-viral agents. For example, 20-deoxy-epi-17-narasin is active against rhinovirus type 3, vaccinia virus, herpes virus and influenza A virus, as demonstrated by in vitro plaque suppression experiments, as described by Siminoff, Applied 5 Microbiology 9 £ 1 / .66-72 (1961).

According to the invention, therefore, a deoxyna-rasin antibiotic can be administered orally, topically or parenterally to humans and animals for the control of viruses. Suitable dosage amounts for the prevention or treatment of viral diseases range from 1-5 mg / kg body weight of human or animal, depending on the type of virus and whether the drug is used prophylactically or therapeutically.

In addition, solutions of a deoxynara-15 sin antibiotic, preferably in conjunction with a surfactant, can be used to clean the in vitro host on which viruses such as polio or herpes are present. Solutions containing 1-1500 mcg / ml of a deoxynarasin antibiotic are suitable for fighting viruses.

The acute toxicities of 20-deoxy-epi-17-narasin (free acid) and 20-deoxynarasin (Na salt) when administered intraperitoneally in the mouse and expressed as LD ^ q are respectively. 201 mg / kg and 5 mg / kg.

The anticoccid activity is an important property of the present deoxynarasin antibiotics. E.g.

20-deoxynarasin (Na salt) and 20-epi-17-narasin (free acid) have been shown to be active in vitro against Eimeria tenella, the protozoan organism, which usually accompanies coccidiosis, at concentrations as low as 0.2 ppm.

In a feeding experiment in young chicks, the deoxynarasin antibiotics have been shown to have anti-cococidal activity in vivo. In this experiment, 20-deoxynarasin (Na salt) was administered in amounts of 100 ppm to the feed of chicks infected with Eimeria tenella, with the result that the chicks did not die, a greater weight gain of 790 02 1 7 20 and the number of damage in the chicks decreased. The results of this experiment are reported in Table G.

TABLE · G

5% morta-weight damage 1) 2) treatment_lity increase (g) went infected controls 37.5 114 4.00 20-deoxynarasin (100ppm) 0 185 0.13 1) two pens with four chicks each per treatment .

10 2) maximum possible score = 4.00.

For the prevention or treatment of coccidiosis in poultry, an effective anticocidal amount of deoxynarasin antibiotics should be administered to birds, preferably orally, daily. The deoxynarasinanti-15 biotic can be administered in various forms, with administration with a physiologically acceptable carrier, preferably the bird-consumed form, being preferred. Although a number of factors have to be taken into account in the determination of an appropriate concentration of deoxynarasin antibiotic, this concentration is generally 0.003 - 0.03% by weight of the drug-free feed and preferably 0.004-0.02 wt%.

The invention also includes the anticoccidial feed mixtures for poultry, consisting of poultry feed and 35-180 g of a deoxynarasin antibiotic per ton.

The ability to improve feed utilization in animals is another important property of deoxynarasin antibiotics. For example, deoxynarasin antibiotics improve feed utilization in ruminants with a developed rumen function. As explained above, the efficiency of carbohydrate utilization in ruminants by the present treatment, which stimulates the rumen flora of the animals, is improved to produce propionate derivatives instead of acetates or butyrates. The efficiency of the feed utilization can be followed by monitoring the production and concentration of propionate compounds in the rumen fluid, using the methods described in U.S. Patent 4,038,384.

The results of in vitro experiments with 20-deoxy-narasin (Na salt) and 20-deoxy-epi-17-narasin (free acid), which measured the ratio of volatile fatty acid (WZ) concentrations in treated flasks to concentrations in control flasks, 10 tones, are shown in Table H.

TABLE H

ratio of treated to control dose molar% molar% molar% total

compound_ mcg / ml acetate propionate butyrate WZ

15 20-deoxy-epi-17-narasin 10 0.9093 1.2665 * 0.6560 1.0325 "5 0.9237 1.1836 * 0.8201 1.0487" 2 0.9581 1.0858 0.9414 1.0717 2 0-deoxynarasin 1 0.8727 1.4597 * 0.3942 1.2096 "0.3 0.9084 1.3799 * 0.45882 1.1829 20" 0.1 0.9504 1.2148 * 0, 6870 1.1892

Statistically significant (P ^ 0.01) by the two-tailed LSD test (R.G.D. Steel and J.H. Torrie, "Principles and Procedures of Statistics", McGraw-Hill, New York, N.Y., 1960, p. 106).

The present deoxynarasin antibiotics are particularly suitable for increasing the amount of propionates and additionally for improving feed utilization when administered orally to ruminants in an amount of 0.05-5.0 mg / kg / day. The most favorable results are obtained at an amount of 0.1-2.5 mg / kg / day. A preferred method of administration of an antibiotic according to the invention comprises mixing it with the feed. However, the antibiotic can also be administered by other means, e.g. in the form of tablets, drinks, large pills or capsules. The composition of these dosage forms can be accomplished using methods known in veterinary pharmacy. Each individual dosage unit should contain an amount of a compound of the invention, which amount is directly related to the correct daily dose for the animal to be treated.

The invention further relates to feed mixtures suitable for fattening ruminants, such as cattle and sheep. These feed mixtures contain 10 feeds and 1-30 g deoxynarasin antibiotic per ton.

Pig dysentery is a common disease in the United States and other countries, which means annual large losses to the world's pig herd. From US patent 3,947,586 it follows that polyether antibiotics are suitable for the prevention and treatment of pig dysentery. Since the deoxynarasin antibiotics are new representatives of the polyether antibiotics group, they should also be effective in preventing and treating swine dysentery.

For the prevention or treatment of dysentery in pigs, an effective amount of a deoxynarasin antibiotic is preferably included in the feed. The present deoxynarasin antibiotics are effective for the prevention or treatment of swine dysentery in oral administration to pigs, in an amount of 35-150 g per ton of the active compound. An amount of about 100 g of active compound per ton is particularly preferred. Although the preferred method of administration comprises mixing the compound with the feed, the compound may also be administered differently, eg in the form of tablets, oral solution, large pills or capsules. Each individual unit dose should contain an amount of the present antibiotic directly related to the correct daily dose for the animal to be treated.

The invention further comprises a feed mixture for pigs, 790 02 1 7 23, consisting of pig food and an effective amount of a deoxynarasin antibiotic. As discussed above, an effective amount corresponds to 35-150 g of deoxynarasin antibiotic per ton of feed.

5 Deoxynarasin antibiotics are antiviral agents and are also active against anaerobic bacteria, such as Clostridium perfringens. The deoxynarasin antibiotics can therefore be used advantageously for the treatment or prevention of enteritis in chickens, pigs, cattle and sheep and in the treatment of enterotoxemia in ruminants.

Some deoxynarasin compounds (20-deoxynara-sin and its salts) have ion-binding and ion-transporting properties and are therefore ionophores (ion-15 carriers) (see BC Pressman, Alkali Metal Chelators - The Ionophores, in "Inorganic Biochemistry" , Vol. 1, GL Eichhorn, Elsevier, 1973). These compounds can be used when the selective removal of a particular cation is desired. Examples of such applications include the removal and recovery of silver ions from solutions in photography, the removal of toxic cations from industrial waste liquids before such liquids are released into the environment and for desalination of seawater. A deoxynarasin compound can be used as a component of an ion-specific electrode (see O. Kedem et al., U.S. Patent 3,753,887). These compounds modify the cation permeability of both natural and artificial membranes. Therefore, a deoxynarasin compound can be used as a component in a membrane, which is used for the selective transport of cations against a concentration gradient. A potential application of this property lies in the recovery of heavy metals and precious metals on a commercial basis. (See E. L. Cussler, D. F. Evans and 35 Sister M. A. Matesick, Science 172, 377 (1971)).

790 02 1 7 24

In addition, 20-deoxynarasin and its salts are active as inhibitors of the enzyme ATPase. ATPase, an alkali metal sensitive enzyme, present in cell membranes, is involved in the energy required for active transport.

5 "Active transport" refers to the energy-required series of operations, where intracellular and extracellular fluids retain their compositions. ATPase inhibitors reduce the energy required for active transport. In vitro experiments have shown that 20-10 deoxynarasin (Na salt) inhibits the cation transport ATPase in liver mitochondria at a semi-effective concentration of 0.065 mcg / ml.

The 20-deoxynarasin and its salts are potential cardiotonic agents. For example, in experiments conducted using isolated guinea pig atria, 20-deoxynarasin increased cardiac contractility. Response to this test is expressed as a percentage of the maximum contractile tension, which is revealed -4 at a dose of norepinephrine (10 M). The compound -5 20-deoxynarasin (Na salt) was found to produce an average increase (+ standard error) in contractual tension of 43.8 + 1.4 (n = 4)% at a 10 molar concentration. For a detailed description of this test, reference is made to U.S. Patent 3,985,893.

The invention therefore encompasses the method of increasing the contractile force of the heart muscles of warm-blooded mammals, for which an effective non-toxic dose of 20-deoxynarasin or a pharmacologically acceptable salt thereof is administered. An effective non-toxic dose is a dose of 30-500 mcg / kg body weight. A preferred dose is 30-100 mcg / kg body weight. In this method, the antibiotic is administered parenterally, eg by intravenous infusion. A suitable mode of administration is the drip method, wherein the anti-35 biotic is incorporated into a standard IV solution, such as a dextrose solution.

The compound 20-deoxynarasin is preferably administered at a dose of less than 100 mcg / kg, until the desired increase in contractile force is observed. Thereafter, the amount of 20-deoxynarasin may be administered, controlled by the infusion rate necessary to maintain the desired response. As with the clinical administration of other inotropic agents, the administered dose of 20-deoxynarasin may vary in a given clinical case, according to factors such as individual 20-deoxynarasin tolerance, the nature of the heart defect, eg degree of damage to the heart muscle and the age and general physical condition of the patient.

The method of the invention, using the positive inotropic agent 20-deoxynarasin, can be used in a wide variety of clinical situations, which can be classified as cardiogenic shock. Such conditions include, for example, myocardial infarction, congestive heart defect and postoperative cardiogenic shock.

In order to further illustrate the operation of the invention, a number of examples will now follow.

EXAMPLE I

a. Shake - cob fermentation.

A culture of Streptomyces aureo-faciens NRRL 11181 was prepared and stored on an agar plate having the following composition: ingredient amount K2HPO4 2 g 30 MgSO4-7H20 0.25 g NH4 NO3 2 g

CaCOg 2.5 g

FeSO4-7H20 0.001 g

MnCl2-7H20 0.001 g 35 ZnS04-7H20 0.001 g 790 02 1 7 26 ingredient amount of glucose 10 g agar 20 g deionized water to 1 liter 5 pH (not set) 7.7

The plate was inoculated with Streptomyces aureofaciens NRRL 11181 and the inoculated plate incubated at 30 ° C for 7 days. The mature culture was covered with sterile bovine serum and scraped with a sterile spatula to loosen the spores. The resulting bovine serum suspension of spores and mycelial fragments was lyophilized to a maximum of 6 tablets.

One of the thus lyophilized tablets was used to inoculate 50 ml of a vegetative medium of the following composition: ingredient amount of glucose 20 g of soybean meal 15 g of corn liquid 10 g of 20 CaCOg 2 g of tap water 1 liter of pH, adjusted to 6.5 by addition of 5N NaOH. The inoculated vegetative medium was incubated in a 250 ml Erlenmeyer flask at 30 ° C for 24-48 hours on a rotary shaker rotating at 5 cm diameter arc at 250 rpm.

The incubated vegetative medium (50 ml) described above was used to inoculate 250 ml of one of the following fermentation media: Medium 1: ingredient amount of tapioca dextrin 60 g enzyme-hydrolyzed casein 6 g enzymatic hydrolyzate of 35 casein 2 g 790 02 17 27 ingredient quantity

CaCo ^ 2 g

MgSO 4 · 7H20 0.5 g molasses 15 g 5 purified soybean oil 5.0 ml / 1 tap water 1 liter pH (not set) 6.6 * Stadex 11, A.E. Staley, Decatur, Illinois Amber EHC, Amber Laboratories, Juneau, Vise.

^ Amine A, Sheffield Chemical Co., Norwich,

New York.

Medium 2: ingredient quantity it tapiocadextrin 30 g 15 glucose 15 g enzyme hydrolysed casein 3 g enzymatic casein hydrolyzate 1 g yeast extract 2.5 g 20 CaCO3 2 g

MgS04 * 7H20 1 g molasses 15 g purified soybean oil 5.0 ml / 1 tap water 1 liter 25 pH (not set) 6.4%

Stadex 11, A.E. Staley, Decatur, Illinois

Amber EHC, Amber Laboratories, Juneau, Vise. Amine A, Sheffield Chemical Co., Norwich,

New York.

30 Medium 3? ingredient amount of soybean meal 25 g glucose 20 g

CaCO3 2.0 g 790 02 1 7 28 ingredient quantity

Na2SO4 "10H2O 1.0 g purified soybean oil 20 ml methyl oleate 20 ml 5 FeS04 * 7H20 0.6 g

MnCl2-4H20 0.3 g ascorbic acid 0.018 g deionized water 1 liter pH (not adjusted) 6.5 The fermentation was incubated for 10 days at 30 ° C on a rotary shaker with a 5 cm arc at a rotation speed of 250 rpm.

b. Tank fermentation.

The tank fermentation was carried out using vegetative and fermenting media, as described under a. For the shake flask fermentation. For tank fermentation, 10 ml of the vegetative medium was used to inject 400 ml of a second stage vegetative medium in a 2-liter Erlenmeyer flask. After incubation for 24 hours at 30 ° C, 800 ml of the second stage vegetative medium was used to inoculate 100 liters of fermentation medium into a 165 liters fermentation tank. The pH of the medium after sterilization at 12 ° C for 45 min.

6.8 + 0.1. Fermentation was allowed to continue for 10 days at 30 + 1 ° C. The tank was aerated with sterile air at a rate of 0.5 volume air per volume culture medium per minute while stirring at a speed of 300 rpm using conventional stirrers.

EXAMPLE II

Separation of the deoxynarasin complex.

The pH of the entire fermentation broth (4 liters), obtained by the method described in Example I, using medium 2, was lowered to a pH of 3.0 by adding concentrated HCl while stirring for 790 02 1 7 29 ran for an hour. The resulting solution was filtered through a filter (125 g of Hyflo Super-Cell, a diatomaceous earth, Johns-Manville Corp.). The separated mycelium cake was extracted batchwise, using a mixer with a total of 2 liters of methanol containing 50 g of NaHCO2 per liter. The methanol filtrate was evaporated in vacuo to a volume of about 450 ml. The pH of this solution was adjusted to 7.5 by adding concentrated HCl. The resulting solution was extracted twice with 10 CHClg (500 ml). The CHCl 2 extracts were combined, dried over Na 2 SO 4 and filtered. The filtrate was evaporated in vacuo to yield 2.0 g of crude deoxy-narasin complex.

EXAMPLE III

Isolation of deoxynarasin and epi-deoxynarasin.

Crude deoxynarasin complex (2 g), obtained as described in Example II, was dissolved in a minimal amount of benzene and then applied to a 1.5 x 22 cm column of silica (Merck 7729). The isolated fractions, solvents used, and amounts obtained herein are listed in the following table.

yield fraction volume _ solvent attitude (mg) _ 1 3.0 1 benzene 100% 24 25 2 2.0 1 benzene: ethyl acetate 9: 1 20 3 600 ml "" 4: 1 85 4 300 ml "" 4: 1 5 5 300 ml "4: 1 7 6 900 ml" "4: 1 18 30 7 2.0 1" "4: 1 22 8 300 ml" 4: 1 7 9 450 ml "" 4: 1 22 10 450 ml M 4: 1 9 11 1.2 1 ”" 4: 1 7 35 12 1.2 1 "" 4: 1 5 790 02 1 7 30 yy * (continued table) yield fraction volume solvent_ attitude (mg) _ 13 1.0 1 Benzene: ethyl acetate 4: 1 12 14 1.0 1 "" 3: 1 14 5 15 1.5 1 "" 3: 1 28 16 1.5 1 "" 3: 1 100 17 450 ml " "3: 1 26 18 900 ml" "3: 1 70 19 1.0 1" "3: 1 54 10 20 300 ml ethyl acetate 100% 12 21 150 ml" 100% 180 22 150 ml "100% 125 23 450 ml "100% 170 24 300 ml methanol 100% 40 15 25 450 ml" 100% 1100 obtained after drying over Na 2 SO 4, filtering and evaporating the filtrate to dryness under vacuum.

The fractions were followed by thin layer chromo-2Q matography using an ethyl acetate solvent system. Fractions 16-17 (126 mg) contain 20-deoxy-epi-17-narasin. Fractions 18-23 contain a mixture of 20-deoxynarasin and 20-deoxy-epi-17-narasin.

A mixture of 20-deoxynarasin and 20-deoxy-epi-2 ^ 17-narasin, obtained as described above for fractions 18-23, was chromatographed via preparative thin layer chromatography, using an ethyl acetate solvent system. The mixture (105 mg on one plate and 130 mg on another plate) was dissolved in a small amount and then transferred to a silica gel (Merck) preparative plate. After plate development, 2 separate materials were observed using UV light. Each of the areas representing the 2 materials was removed from the plate and extracted with C 1 C 4 H 2 OH (4: 1). In this system, deoxynarasin 35 moves more slowly than the other component. From the plate containing 105 mg of 79 0 0 2 1 7 31 of the material thereon, 7.5 mg of 20-deoxy-epi-17-narasin and 27 mg of deoxynarasin were obtained. From the 130 mg plate of the mixture thereon, 4.0 mg of 20-deoxy-epi-17-narasin and 56 mg of 20-deoxynarasin were recovered.

EXAMPLE IV

Alternate isolation of 20-deoxynarasin.

The pH of the entire fermentation broth (95 L) was adjusted to 3 by the addition of HCl and the whole was stirred for 1 hour. Then a filter aid (Hyflo Supercel, 3%) was added and the medium was filtered. The separated mycelium cake was extracted twice with about 45 L of acetone with 50 g of NaHCO2 per liter therein. The acetone extracts were combined and concentrated in vacuo to yield about 10 liters of an aqueous solution. The pH of this solution was adjusted to 8.0 by addition of 5N HCl and the resulting solution extracted 3 times with half volumes of CH 2 Cl 2.

The CH2Cl2 extracts were combined and then evaporated in vacuo to leave an oily residue. This residue was taken up in 500 ml top phase of the solvent system hexane: methanol: water (10: 7: 1).

The upper phase was extracted 6 times with 300 ml of the lower phase. These extracts were combined and then concentrated in vacuo to a residue. This residue was dissolved in dioxane. The dioxane solution was lyophilized to yield 19.4 g of deoxynarasin complex.

Various samples thus obtained were combined (40 g), then dissolved in toluene and finally transferred to a silica gel column in a liquid chromatograph (Waters' Associates Prep. LC / System 500).

The column was eluted with a toluene: ethyl acetate (9: 1) solvent system at a flow rate of 250 ml / min, collecting fractions of 250 ml volume. The fraction content was followed by thin layer chromatography. Fractions 37-52 were combined and concentrated in vacuo under 790 02 1 7 32, yielding a residue, which was redissolved in dioxane and lyophilized, yielding 7.8 g pure material, rich in 20-deoxynarases. This material was re-chromatographed on the 5 Waters' Prep. LC / system 500, as described above. After eluting 50 fractions with the toluene: ethyl acetate (9: 1) solvent system, the elution solvent was changed to 100% ethyl acetate. Fractions 51-53 were combined and evaporated in vacuo to give a residue, which was then dissolved in dioxane and lyophilized to yield 1.12g of 20-deoxynarasin as its sodium salt.

EXAMPLE V

Preparation of 20-deoxynarasin, free acid.

15-Deoxynarasin sodium salt (200mg) dissolved in ethyl acetate (10ml). This solution was washed with 0.1 N HCl (10 ml) and then twice with water (5 ml). The resulting organic layer was evaporated to dryness, yielding a residue, which was then dissolved in dioxane and freeze-dried, yielding 139.6 mg of 20-deoxynarasin, free acid, as a white solid.

EXAMPLE VI

Chicken food for coccidiosis treatment.

A balanced high energy feed for chickens was prepared, intended for rapid weight gain, which feed had the following composition: component% kg ground yellow corn 50 450 30 soybean meal, solvent-extracted, finely ground, 50 % protein 30.9 278 animal fat (beef fat) 6.5 58.5 dried fish meal with soluble components (60% protein) 5.0 45 35 corn soluble components 4.0 36 790 02 1 7 33 constituent% kg dicalcium phosphate, feed quality 1.8 16.2 calcium carbonate 0.8 7.2 vitamin premix (consisting of vitamins 5 A, D, E, K and B ^ / cil ° l: i-ne / niacin, panothenic acid, riboflavin, biotin, with glucose bulking agent) 0.5 4.5 salt (NaCl) 0.3 2.7 trace mineral premix, (consisting of MnSO 4, 10 ZnO, KI, FeSO 4, CaCC> 3) 0.1 0.9 2-amino- 4-hydroxybutter hour (hydroxy analog of methionine) 0.1 0.9

Deoxynarasin antibiotic complex, 20-deoxynarasin or 20-deoxy-epi-17-narasin (about 0.01 wt%) was mixed with this feed according to standard feed mixing techniques.

Chicken feed with water ad libitum, is protected against the action of coccidiosis.

EXAMPLE VII Improved beef feed.

20 A high quality balanced feed was prepared as follows: ingredient_% kg finely ground corn 67.8 610.2 ground corncob 10 90 25 dehydrated alfalfa flour, 17% protein 5 45 soybean meal, solvent extracted, 50% protein 10 90 molasses 5 45 urea 0.6 5.4 30 dicalcium phosphate, feed quality 0.5 4.5 calcium carbonate 0.5 4.5 sodium chloride 0.3 2.7 spore mineral premix 0.03 0.27 vitamin A and D_ premix 0, 07 0.63 φφ 35 vitamin E premix 0.05 0.45 790 02 1 7 34 ingredient__% kg calcium propionate 0.15 1.35 * 0.45 kg contains: 2,000,000 IU vitamin A; 227,200 I.U. vitamin D9 and 385.7 g of soybean food with 1% oil in it.

5 corn ingredients, dry grains, with soluble ingredients with 20,000 I.U. d-α-tocopheryl acetate per 0.45 kg.

Deoxynarasin antibiotic complex, 20-deoxynarasin or 20-deoxy-epi-17-narasin (about 0.004 wt%) was mixed with this feed according to standard techniques. The mixed feed was compressed into tablets. With an average daily feed of about 7 kg per animal, this corresponded to about 300 mg of antibiotics per animal per day.

EXAMPLE VIII Improved pig food.

15 A premix was prepared according to standard methods, using the following ingredients: ingredient g / kg active compound 150.0 20 calcium silicate 20.0 calcium carbonate (oyster shell meal) 830.0 total weight: 1000 g

This premix was added to a commercial pig feed, using standard feed mixing techniques, to yield a final active compound concentration of 100 g / ton.

30 790 02 1 7

Claims (14)

A deoxynarasin antibiotic complex, consisting of 20-deoxynarasin and 20-deoxy-epi-17-narasin or a pharmacologically acceptable salt thereof. 2. 20-Deoxynarasin of formula 1 of the formula sheet and its pharmaceutically acceptable salts. 3. 20-Deoxy-epi-17-narasin of Formula 2 of the formula sheet and its pharmaceutically acceptable salts. Process for the preparation of the deoxy-10 narasin antibiotic complex, as defined in claim 1, characterized in that a Streptomyces aureofaciens is cultivated with practically the same biosynthetic properties as NRRL 11181 in a culture medium containing assimilable sources. of carbohydrate, nitrogen and inorganic salts, under submerged aerobic fermentation conditions, until a significant amount of antibiotic activity is obtained, followed by recovery of the antibiotic complex in the form of the acids or in the form of their pharmaceutically acceptable salts. 5. A method according to claim 4, characterized in that Streptomyces aureofaciens is the organism indicated as NRRL 11181. 6. A method according to claim 4 or 5, characterized in that the deoxynarasin antibiotic complex, or the pharmaceutically acceptable salts thereof, are isolated from the culture medium. 7. A method according to claim 6, characterized in that 20-deoxynarasin, 20-deoxy-epi-17-nara-sinin or a pharmaceutically acceptable salt thereof is isolated from the deoxynarasin-antibiotic complex. 8. A method of improving feed utilization in ruminants, characterized in that an effective propionate-increasing amount of deoxynarasin antibiotic complex, 20-deoxy-790 02 1 7 narasin, 20-deoxy-epi-17, is provided in such animals. Narasin, or a pharmaceutically acceptable salt thereof, is administered. 9. Feed for cattle consisting of fodder and 20-deoxynarasin, 20-deoxy-epi-17-narasin, deoxynara-sinibiotic complex or a pharmaceutically acceptable salt thereof. 10. A method of preventing or treating coccidiosis in poultry, characterized in that an effective anticocidal amount of 20-deoxynarasin, 20-deoxy-epi-17-narasin, deoxynarasin antibiotic complex or a pharmaceutically acceptable salt thereof is provided to poultry administered. Poultry feed, consisting of poultry feed and an anticoccide amount of 20-deoxynarasin, 20-deoxy-epi-17-narasin, deoxynarasin antibiotic complex or a pharmaceutically acceptable salt thereof. 12. A method of preventing or treating swine dysentery, characterized in that an effective amount of 20-deoxynarasin, 20-deoxy-epi-17-narasin, deoxynarasin-antibiotic complex or a pharmaceutically acceptable salt thereof is administered to pigs. 13. Pig feed consisting of pig feed and an effective amount for the control of pig dysentery of 20-deoxynarasin, 20-deoxy-epi-17-narasin, deoxynarasin antibiotic complex or a pharmaceutically acceptable salt thereof. 14. A method of increasing the contractile force of the heart muscle in a warm-blooded mammal, characterized in that an effective non-toxic dose of 20-deoxynarasin or a pharmaceutically acceptable salt thereof is administered parenterally. 15. A biologically pure culture of the microorganism Streptomyces aureofaciens identified as NRRL 11181, which culture has the ability to form 20-deoxy-narasin or 20-deoxy-epi-17-narasin after fermentation 7. ft O 2 1 7 t VVV tm '* in an aqueous nutrient medium containing assimilable sources of carbohydrate / nitrogen and inorganic salts. 16. NRRL 11181. 5 79 0 0 2 17