HCID 19867502, compound → Indoline-2-carboxylic acid

HRID 1198866Notes

Reaction equation

HRID 1198866 reaction equation
C([O:6][C:7]([CH:9]1[CH2:17][C:16]2[C:11](=[CH:12][CH:13]=[CH:14][CH:15]=2)[NH:10]1)=[O:8])CCCC>P([O-])([O-])([O-])=O>[NH:10]1[C:11]2[C:16](=[CH:15][CH:14]=[CH:13][CH:12]=2)[CH2:17][CH:9]1[C:7]([OH:8])=[O:6]

Participants

3

Reactant 1

HCID 19867502HCID 19867502, compound

Product 1

HCID 86074Indoline-2-carboxylic acid

Reagents, catalysts & solvents 1

HCID 1061PhosphateSolvent2×

Procedure

After adding 10 g of lipase (Steapsin made by Wako Purechemical Industries, Ltd.) having (R)-selective esterase activity to 100 ml of 0.1M phosphate buffer at pH 7.0, a mixture was stirred and the insoluble material was removed. To the filtrate was added 60 g wet weight (containing 71% of water) of the methacrylate porous adsorbent Amberlite XAD-7 (made by Rohm & Haas Co.), which was washed with methanol and then with water, and a mixture was slowly stirred in a cold room (4° C.) for a night to immobilize the enzyme on the support by adsorption. The suspension containing the immobilized enzyme was filtered with suction by means of glass filter and the the filtrate was washed with 0.1M phosphate buffer at pH 7.0 three times (100 ml×3), followed by filtration with suction to give the wet immobilized enzyme. The obtained immobilized lipase was filled in the column (inner diameter: 2.2 cm, length: 15 cm), which was kept at 33° C. On the column was loaded 5 g of racemic amyl-indoline-2-carboxylate [(R,S)-Ia] and 0.1M phosphate buffer at pH 7.0 was passed through the column at a flow rate of 6 ml/h. Each 12 ml of the eluate collected from the column by a fraction collector was subjected to a liquid chromatography analysis. Only hydrophilic indoline-2-carboxylic acid produced by the asymmetric hydrolysis was contained. 180 ml of the phosphate buffer fraction was saturated with ammonium sulfate and the pH of the mixture was adjusted to 5.0, followed by extraction of indoline-2-carboxylic acid with an equivalent amount of ethyl acetate three times. The obtained ethyl acetate layer was dehydrated and concentrated under reduced pressure, followed by recrystallization in acetone-hexane (5 ml-1 ml). After driness in vacuum, 1.26 g of (R)-indoline2-carboxylic acid [(R)-II] having a specific rotatory power [α]D25 -32.5° (c=1.0, DMFA) (the value described in the literature J. Med. Chem. 26, 394 (1983) by D. H. Kim et al. is [α]D25 +34.5° (c=0.91, DMFA) was obtained as a white powder.

Workup

  1. additionAfter adding 10 g of lipase (Steapsin made by Wako Purechemical Industries, Ltd.)
  2. customthe insoluble material was removed
  3. additionTo the filtrate was added 60 g wet weight (containing 71% of water) of the methacrylate porous adsorbent Amberlite XAD-7 (made by Rohm & Haas Co.), which
  4. washwas washed with methanol
  5. stirringwith water, and a mixture was slowly stirred in a cold room (4° C.) for a night
  6. additionThe suspension containing the immobilized enzyme
  7. filtrationwas filtered with suction by means of glass
  8. filtrationfilter
  9. washthe the filtrate was washed with 0.1M phosphate buffer at pH 7.0 three times (100 ml×3)
  10. filtrationfollowed by filtration with suction
  11. customto give the wet immobilized enzyme
  12. additionThe obtained immobilized lipase was filled in the column (inner diameter: 2.2 cm, length: 15 cm), which
  13. customwas kept at 33° C
  14. customEach 12 ml of the eluate collected from the column by a fraction collector

Sources & evidence

ORD record
ord-abe19ee41268459db31b8447369e27ad
Source dataset
uspto-grants-1990_02
Patent number
US04898822

Reaction notes

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