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 ml Erlenmeyer flask containing 50 ml 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 one 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 1 mg of 4-tert butyl-N-[4-(2-hydroxyethoxy)-5-(2-methoxyphenoxy)-2-(2-pyrimidinyl)pyrimidin-6-yl]benzenesulphonamide (CA 2071193) dissolved in 0.5 ml of dimethylsulfoxide was added to each of the flasks and the fermentation allowed to continue under the same conditions for a further 24 hours. The 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 ml of AS-7H 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 24 hours
7. extractionThe 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