反应详情

17-Allyl-1,14-di-hydroxy-12-[2-(4-hydroxy-3-methoxy-cyclohexyl)-1-methyl-vinyl]-23,25-dimethoxy-13,19,21,27-tetramethyl-11,28-dioxa-4-aza-tricyclo[22.3.1.04,9]octacos-18-ene-2,3,10,16-tetraone (FK506; 8.04 mg, 0.01 mmol) was dissolved in acetonitrile (CH3CN; 100 μl), a solution of phosgene (COCl2) in toluene (1.24 mmol/ml; 8 μl, 0.01 mmol) was added at room temperature, and this was followed by stirring for 1.5 hours. After the reaction solution was concentrated under reduced pressure, TOYO-Pearl resin (TSKgel AF-amino, 100 μl, free amino group (available amino group) 0.01 mmol; manufactured by Tosoh Corporation) and acetonitrile (CH3CN; 500 μl) were added, and this was followed by stirring at room temperature for 1.5 hours. After completion of the reaction, the resin was washed with DMF two times and with water two times, a saturated aqueous solution of sodium hydrogen carbonate (1 ml) was added, and this was followed by stirring for 1 minute. After the resin was washed with water two times and with DMF two times, acetic anhydride (100 μl) and DMF (400 μl) were added, and this was followed by stirring at room temperature for 5 minutes. Subsequently, the resin was washed with DMF three times and with 20% aqueous ethanol two times, and the TOYO Pearl resin with FK506 obtained was used in the binding experiment described below.

1. concentrationAfter the reaction solution was concentrated under reduced pressure, TOYO-Pearl resin (TSKgel AF-amino, 100 μl, free amino group (available amino group) 0.01 mmol; manufactured by Tosoh Corporation) and acetonitrile (CH3CN; 500 μl)
2. additionwere added
3. stirringby stirring at room temperature for 1.5 hours
4. customAfter completion of the reaction
5. washthe resin was washed with DMF two times and with water two times
6. additiona saturated aqueous solution of sodium hydrogen carbonate (1 ml) was added
7. stirringby stirring for 1 minute
8. washAfter the resin was washed with water two times and with DMF two times, acetic anhydride (100 μl) and DMF (400 μl)
9. additionwere added
10. stirringby stirring at room temperature for 5 minutes
11. washSubsequently, the resin was washed with DMF three times and with 20% aqueous ethanol two times