Glyoxylic Acid → 2,3-Diaminophenol

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 2,3-diaminophenol 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.

1. customobtained in Reference Example 3
2. additionTo the mixture was added a solution
3. washthe column was washed with the same buffer solution at 6 ml/minute of flow rate
4. customto collect the fraction of diketone derivative of Met-NT-3
5. additionTo this fraction was added the same volume of 4 M acetic acid-4 M sodium acetate solution

ORD record

ord-5f24ed609f86473bbf4104469f91d281

Source dataset

uspto-grants-2001_10

DOI

10.6084/m9.figshare.5104873.v1

Patent number

US06309859B1