反应详情

Streptomyces rimosus subsp. rimosus ATCC10970 maintained on a ¼ strength ATCC172 agar slope was inoculated as a loopful of spores into a 300 ml Erlenmeyer flask containing 50 ml of AS7-H 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 twenty 300 ml Erlenmeyer flask containing 50 ml of AP-5H production medium and incubated under the same conditions for a further 24 hours. At this point 5 mg of N-(4-(1,3-benzodioxol-5-yl)-3-{2-[(5-chloro-2-pyrimidinyl)oxy]ethoxy}-1-methyl-1H-pyrazol-5-yl)-5-isopropyl-2-pyridinesulfonamide (Example 103) dissolved in 0.5 ml of methanol was added to each of the flasks and the fermentation allowed to continue under the same conditions for a further 96 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 ¼ strength ATCC172 agar slope
2. additioncontaining 50 ml of AS7-H inoculum medium
3. additioncontaining 50 ml 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 96 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