反应详情

Streptomyces rimosus subsp. rimosus ATCC10970 maintained on a quarter strength ATCC172 agar slope was inoculated as a loopful of spores into a 300 mm Erlenmeyer flask containing 50 mm of AS-7H inoculum medium. This was allowed to incubate for 2 days at 28° C., 200 rpm on an Infors Multitron™ Shaker with 1″ throw. 2 mls of this inoculum medium was then transferred to each of six 300 mm Erlenmeyer flask containing 50 mm of AP-5H production medium and incubated under the same conditions for a further 24 hours. At this point 5 mg of N-[6-{2-[(5-bromo-2-pyrimidinyl)oxy]ethoxy}-5-(4-methylphenyl)-4-pyrimidinyl]-4-(tert-butyl)benzenesulfonamide (EP 0658548B1) dissolved in 0.5 mm of dimethylsulfoxide was added to each of the flasks and the fermentation allowed to continue under the same conditions for a further 3 hours. Each flask was then extracted with 200 mls of ethyl acetate and the combined ethyl acetate solubles concentrated to dryness to give a gum solid.

1. temperaturerimosus ATCC10970 maintained on a quarter strength ATCC172 agar slope
2. additioncontaining 50 mm of AS-7H inoculum medium
3. additioncontaining 50 mm of AP-5H production medium
4. waitincubated under the same conditions for a further 24 hours
5. additionwas added to each of the flasks
6. waitto continue under the same conditions for a further 3 hours
7. extractionEach flask was then extracted with 200 mls of ethyl acetate
8. concentrationthe combined ethyl acetate solubles concentrated to dryness
9. customto give a gum solid