Antibiotic nebramycin and preparation thereof

Claim 2

An antibiotic substance selected from the class consisting of nebramycin V and the acid addition salts thereof; said nebramycin V being characterized as follows: a crystalline white compound having titratable groups of pK''a values of about 5.5, 7.0, 8.0, and 9.1 as determined by electrometric titration in water; has an apparent molecular weight of about 424 as determined by titration; has a specific rotation, ( Alpha )25D, of +118*, (C 1, weight per volume); has the approximate percentage composition of C, 41.05; H, 7.60; N, 13.62; O, 36.76; and has infrared spectral absorption maxima as a mineral oil mull of 3.05, 3.19, 6.32, 7.45, 8.75, 9.7, 11.15, and 12.25 microns.

Claim 3

An antibiotic substance selected from the class consisting of nebramycin V'' and the acid addition salts thereof; said nebramycin V'' being characterized as follows: a crystalline white substance which is soluble in ethanol, pyridine, dimethylformamide, and dimethylsulfoxide and substantially insoluble in the common ketone and ester solvents; has titratable groups of pK''a values of about 5.6, 6.9, 7.7, 8.6, and 9.9 as determined by electrometric titration; has an apparent molecular weight of about 510; has a specific rotation, ( Alpha )25D, of +125*, (C 1, weight per volume); has the approximate percentage compositions of C, 41.6; H, 7.49; N, 14.9; O, 36.0; and has infrared spectral absorption maxima as a mineral oil mull of 2.87, 3.00, 3.18, 3.66, 5.75, 5.82, 6.1, 6.23, 6.28, 6.86, 7.03, 7.28, 7.37, 7.42, 7.61, 7.84, 7.95, 8.17, 8.47, 8.55, 8.77, 8.94, 9.23, 9.40, 9.63, 9.87, 10.08, 10.50, 11.36, 11.53, 11.61, 11.8, 12.5, 12.95, 13.46, and 14.00 microns.

Claim 4

An antibiotic substance selected from the class consisting of nebramycin VI and the acid addition salts thereof; said nebramycin VI being characterized as follows: a crystalline white substance having titratable groups of pK''a values of about 5.8, 6.8, 7.1, 7.9, and 9.3 as determined by electrometric titration in water; has a specific rotation, ( Alpha )25D, of +131*, (C 1, weight per volume); has an apparent molecular weight of about 467 as determined by titration; has the approximate percentage composition of C, 44.3; H, 8.1; N, 14.3; O, 32.8; and has infrared spectral absorption maxima as a mineral oil mull of 3.05, 3.19, 6.35, 7.47, 8.75, 9.8, 12.3 and 12.95 microns.

Claim 5

An antibiotic complex designated as nebramycin which when adsorbed upon a cation exchange resin and subjected to a stepwise elution procedure employing 0.1 N ammonium hydroxide and 0.3 N ammonium hydroxide yields from about 35 to about 50 percent by weight of nebramycin II as defined in claim 1, about 15 to 20 percent by weight of nebramycin IV as defined in claim 2, about 30 percent by weight of nebramycin V as defined in claim 3, and a combined total of about 10 percent by weight of nebramycin V'' as defined in claim 4, and nebramycin VI as defined in claim 5 said antibiotic complex being prepared by cultivating Streptomyces tenebrarius, ATCC 17920, in a culture medium containing assimilable sources of carbon, nitrogen, and inorganic salts under submerged aerobic conditions until a substantial amount of nebramycin is produced by said organism in said culture medium.

Claim 6

A method of producing nebramycin as defined in claim 6 which comprises cultivating Streptomyces tenebrarius, ATCC 17920, in a culture medium containing assimilable sources of carbon, nitrogen, and inorganic salts under submerged aerobic conditions until a substantial amount of antibiotic activity is imparted by said organism to said culture medium.