Patent GB-1497331-A (GB)
Title: Antibiotic substances and preparation thereof
IPC: C07G11/00
Filed: 1975-08-29
Abstract: 1497331 Antibiotic compositions AYERST MCKENNA & HARRISON Ltd 29 Aug 1975 35697/75 Heading A5B [Also in Division C3] A pharmaceutical composition, which inhibits the growth of pathogenic bacteria in a mammal, comprises a pharmaceutically-acceptable carrier and an antibacterial amount of an antibiotic selected from: (1) antibiotic complex 484 which (i) is soluble in water and methanol, and substantially insoluble in acetone, diethyl ether, benzene, ethyl acetate and chloroform (ii) forms acid addition salts (iii) gives two areas of antimicrobial activity 25 cm and 11 cm from the origin on descending paper chromatography at 24‹C for 48 hours on a fast medium paper which retains particles of 8 to 11 microns using a solvent system which consists of n-propanol, pyridine, acetic acid and water (30:20:6:20 by volume) as determined by bioautography on agar plates seed with Sarcina lutea (iv) does not exhibit any characteristic absorption between 220 nm and 350 nm in its ultraviolet spectrum (95% v/v, ethanol) (v) has a representative infrared absorption spectrum in KBr pellet given in Fig. 1 (not shown), showing bands (cm<SP>-1</SP>) at 3330, 2925, 2875 (sh), 1710, 1650, 1560 (sh), 1450, 1380, 1170, 1080, 1030, 970, 920, 820 (vi) inhibits the growth of Staphylococcus pyogenes (penicillin-sensitive), Staphylococcus pyogenes (penicillin-resistant), Streptococcus faecalis, Escherichia coli, Aerobacter aerogenes, Salmonella pullorum, Pseudomonas aeruginosa, Proteus mirabilis, Proteus vulgaris, Klebsiella pneumoniae, Serratia marcescens (vii) exhibits LD 50  (intraperitoneal, mice) of 350 to 450 mg of said antibiotic complex 484/kg of body weight and LD 50  (oral, mice) of greater than 1.0 of said antibiotic complex 484/kg of body weight (viii) has a specific rotation, [α]D<SP>25</SP> of -22 to -28‹ (1% w/v, methanol) (ix) gives a positive reaction with ninhydrin, a yellow colour with Elson-Morgan reagent (i.e. a negative reaction), a negative reaction with Sakuguchi reagent and a positive reaction with silver nitrate. (2) antibiotic component 484-A (i) is soluble in water and methanol, and substantially insoluble in acetone, diethyl ether, benzene, ethyl acetate and chloroform (ii) forms acid addition salts (iii) gives one zone of antimicrobial activity at 23 to 25 cm from origin on descending paper chromatography at 24‹C for 48 hours on a fast medium paper retaining particles of 8 to 11 particles using a solvent system which consists of npropanol, pyridine, acetic acid and water (30:20:6:20 by volume) as determined by bioautography on agar plates seed with Sarcina lutea and two spots 13 to 15cm and 23 to 25 cm from origin in the same system as revealed by ninhydrin spray (iv) forms a sulphate salt which gives two spots, Rf 0.15 to 0.16 and Rf 0.22 to 0.24 on ascending paper chromatography at 24‹C for 16 hours on a fast medium paper retaining particles of 8 to 11 microns) using a solvent system which consists of n-propanol, pyridine, acetic acid and water (30:20:6:24 by volume) as detected by ninhydrin (v) forms a sulphate salt giving two zones, as detected by ninhydrin spray, Rf 0.30 to 0.32 and Rf 0.40 to 0.42 on circular paper chromatography using a fast medium paper retaining particles of 8 to 11 microns and a solvent system which consists of n-propanol, pyridine, acetic acid and water (30:20:6:24 by volume) and in the same system gives one zone of antimicrobial activity at Rf 0.40 to 0.42 (vi) has a representative infrared absorption spectrum in KBr pellet given in Fig. 2 (not shown) showing bands (cm<SP>-1</SP>) at 3300, 3080 (sh), 2940 (sh), 1710 (sh), 1660, 1550, 1390, 1310, 1105, 955, 920, 850 (vii) inhibits growth of Staphylococcus pvogenes (penicillin-sensitive), Staphylococcus pyogenes (penicillin-resistant), Streptococcus faecalis, Escherichia coli, Aerobacter aerogenes, Salmonella pullorum, Pseudomonas aeruginosa, Proteus vulgaris, Klebsiella pneumoniae, Serratia marcescens, Proteus mirabilis (viii) exhibits LD 50  (intraperitoneal, mice) of about 1.0g of antibiotic component 484/kg of body weight (ix) has about 1 to 1 mole ratio of streptolidine and L-#-lysine moieties as indicated by amino acid analysis (x) has a specific rotation, [α]D<SP>25</SP> of -22 to -28‹ (1% w/v, water) (xi) gives positive reactions with ninhydrin and silver nitrate and negative reactions with Elson-Morgan and Sakaguchi reagents; (3) antibiotic component 484-B which (i) is soluble in water and methanol, and substantially insoluble in acetone, diethylether, benzene, ethyl acetate and chloroform (ii) forms acid addition salts (iii) gives a main area of antimicrobial activity at 17 cm and minor areas at 24cm and 8cm from the origin on descending paper chromatography at 24‹C for 48 hours on a fast medium paper retaining particles of 8 to 11 microns using a solvent system which consists of n-propanol, pyridine, acetic acid and water (6:4:1.5:4 by volume) as determined by bioautography on agar plates seeded with Sarcina lutea (iv) gives three zones, as detected by ninhydrin spray, Rf 0.16, 0.29 and 0.42, on circular paper chromatography using a fast-medium paper which retains particles of 8 to 11 microns and a solvent system which consists of n-propanol, pyridine, acetic acid and water (30:20:6:24 by volume) (v) has a representative infrared absorption spectrum in KBr pellet as given in Fig. 3 (not shown) showing bands (cm<SP>-1</SP>) at 3280, 3070, 2930, 1710, 1655, 1550, 1395, 1075, 920 (vi) inhibits the growth of Staphylococcus pyogenes (penicillin-sensitive), Staphylococcus pyogenes (penicillin-resistant), Streptococcus faecalis, Escherichia coli, Aerobacter aerogenes Salmonella pullorum, Pseudomonas aeruginosa, Proteus mirabilis, Proteus vulgaris, Klebsiella pneumoniae, Serratia marcescens (vii) exhibits LD 50  (intraperitoneal, mice) of about 100 mg of antibiotic component 484-B/kg of body weight (viii) has about 1 to 2 mole ratio of streptolidine and L-#-lysine moieties as indicated by amino acid analysis (ix) has a specific rotation, [α]D<SP>25</SP> of -10 to -14‹ (1% w/v, water).