Reaction equation
[NH2:1][C@H](C(O)=O)CCSC.[C:10](O)(=O)[CH:11]=O.[N:15]1[CH:20]=[CH:19][CH:18]=[CH:17][CH:16]=1.P([O-])([O-])([O-])=O>NC(N)=O.S([O-])([O-])(=O)=O.[Cu+2]>[CH3:16][C:17]1[CH:11]=[CH:10][C:20]([NH2:15])=[C:19]([NH2:1])[CH:18]=1Participants
Reactant 4
Product 1
Reagents, catalysts & solvents 2
Procedure
Forty mg of Met-NT-3 having methionine at its N-terminal obtained in Reference Example 3 was dissolved in 4 ml of 3 M urea solution. To the mixture was added a solution containing 50 mM copper sulfate 0.5 ml, glyoxylic acid 0.25 g and pyridine 0.5 ml and allowed to stand at 25° C. for 1 hour. The reaction solution was passed through Sephadex G-25 column (25 mmID×600 mmL) equilibrated with 2.5 M urea and 50 mM phosphate buffer solution (pH 6.0) and the column was washed with the same buffer solution at 6 ml/minute of flow rate to collect the fraction of diketone derivative of Met-NT-3. To this fraction was added the same volume of 4 M acetic acid-4 M sodium acetate solution and then added tolylene-3,4-diamine to give a final concentration of 40 mM, followed by vacuum and sealing with nitrogen gas, and the reaction was proceeded at 37° C. for 15 hours. The reaction solution was purified by the same method as described in Working Example 37 to obtain a purified NT-3.
Workup
- customobtained in Reference Example 3
- additionTo the mixture was added a solution
- washthe column was washed with the same buffer solution at 6 ml/minute of flow rate
- customto collect the fraction of diketone derivative of Met-NT-3
- additionTo this fraction was added the same volume of 4 M acetic acid-4 M sodium acetate solution
Sources & evidence
- ORD record
- ord-478491f1c905443fa2ca97113d974370
- Source dataset
- uspto-grants-2001_10
- Patent number
- US06309859B1
Linked notes
Loading…