Sodium Hydroxide → HCID 10178874, compound

HRID 1743647Notes

Reaction equation

HRID 1743647 reaction equation
C[O:2][C:3](=[O:28])[C@H:4]([CH2:13][S:14][C:15]1[CH:20]=[CH:19][C:18]([C:21]([O:23][CH2:24][CH:25]=[CH2:26])=[O:22])=[CH:17][C:16]=1[NH2:27])[NH:5][C:6]([O:8][C:9]([CH3:12])([CH3:11])[CH3:10])=[O:7].[OH-].[Na+]>CC(OC)(C)C>[C:9]([O:8][C:6]([NH:5][C@H:4]([C:3]([OH:28])=[O:2])[CH2:13][S:14][C:15]1[CH:20]=[CH:19][C:18]([C:21]([O:23][CH2:24][CH:25]=[CH2:26])=[O:22])=[CH:17][C:16]=1[NH2:27])=[O:7])([CH3:10])([CH3:11])[CH3:12]

Participants

4

Reactant 2

HCID 14798Sodium HydroxideHCID 58604712HCID 58604712, compound

Product 1

HCID 10178874HCID 10178874, compoundYield 94.2%

Reagents, catalysts & solvents 1

HCID 15413tert-Butyl methyl etherSolvent

Procedure

6.4 g (15.658 mmol) of N-tert-butoxycarbonyl-3-(4-alloxycarbonyl-2-aminophenylthio)-L-alanine methyl ester (99%) was dissolved in 480 ml TBME and emulsified in 1.5 L buffer solution (0.1M sodium chloride; 160 mM sodium phosphate pH 7.5) under vigorous stirring. 12.0 ml Alcalase 2.4 L [a subtilisin Carlsberg from Novo Nordisk] was added and the pH maintained at 7.5 under vigorous stirring by the controlled addition (pH-static) of 1.0N sodium hydroxide solution. After 48.5 h, 16.6 ml of 1.0N sodium hydroxide solution was consumed and the conversion degree was >97% (HPLC analysis). After phase separation, the aqueous phase was extracted once with 1 L TBME. The combined TBME phases were extracted with 2×0.4 L 0.1 M potassium phosphate buffer pH 7.6. The combined aqueous phases were acidified to pH 2 with 32% hydrochloric acid and extracted with 3×0.5 L ethyl acetate. In case a stabile emulsion was formed, the phase separation was achieved by filtration on Dicalite. The combined ethyl acetate phases were dried on anhydrous sodium sulfate and evaporated. The residue was dissolved in dichloromethane, evaporated and dried at HV to give 5.85 g N-tert-butoxycarbonyl-3-(4-alloxycarbonyl-2-aminophenylthio)-L-alanine as a yellow highly viscous oil (yield: 94.6%). Analytical data: 1H-NMR (DMSO, 400 MHz) 1.338 (s, 9H, OC(CH3)); 3.077 (dd, 1H, SCH2, J=12.8 Hz, J=6.8 Hz); 3.238 (dd, 1H, SCH2, J=13.8 Hz, J=4.6 Hz); 3.867 (m, 1H, COCHNH); 4.753 (d, 2H, COOCH2, J=5.2 Hz); 5.526 (d, 1H, CH═CH2, J=10 Hz); 5.538 (d, 1H, CH=CH2, J=17.6 Hz); 5.595 (m, 2H, NH2); 6.018(ddtr, 1H, CH2CH═CH2, J=16.6 Hz, J=10.6 Hz, J=5.6 Hz); 6.519 (m, 1H, CONHCH); 6.876 (d, 1H, arom. CSCH, J=8.4 Hz); 7.330 (d, 1H, arom. CSCHCH, J=8.4 Hz); 7.338 (s, 1H, arom. CHCNH2). IR (ATR-IR) 3420 and 3355 cm−1 (—NH, —NH2); 2980 and 2935 cm−1 broad (—COOH); 1705 cm−1 (COOH —C═O, carbamate —C═O); 1510 cm−1 (amide —CO—NH); 1486 cm−1 aromate; 1229 and 1160 cm−1 (COOH); 982 and 932 cm−1 (vinyl, —C═CH2). MS (ESI-positive ionisation) m/z=397.1 [M+H+]; m/z=419.4 [M+Na+]. OR [α]D=+16.16° (CHCl3; c=1.0). HPLC analysis: column: ABZ+plus; mobile phase: A: 0.1% TFA in H20; B: MeCN; gradient B: 30–80% 0–15 min, 80–30% 15–16 min, 30% 16–19.5 min; flow; 1 ml/min; pressure:50–80 bar; detection: UV, 300 nm; retention times: 11.2 min (product-acid); 12.7 min (substrate-ester).

Workup

  1. addition12.0 ml Alcalase 2.4 L [a subtilisin Carlsberg from Novo Nordisk] was added
  2. temperaturethe pH maintained at 7.5
  3. custom16.6 ml of 1.0N sodium hydroxide solution was consumed
  4. customAfter phase separation
  5. extractionthe aqueous phase was extracted once with 1 L TBME
  6. extractionThe combined TBME phases were extracted with 2×0.4 L 0.1 M potassium phosphate buffer pH 7.6
  7. extractionextracted with 3×0.5 L ethyl acetate
  8. customIn case a stabile emulsion was formed
  9. customthe phase separation
  10. filtrationwas achieved by filtration on Dicalite
  11. dry with materialThe combined ethyl acetate phases were dried on anhydrous sodium sulfate
  12. customevaporated
  13. dissolutionThe residue was dissolved in dichloromethane
  14. customevaporated
  15. customdried at HV

Sources & evidence

ORD record
ord-c1a4940f9cd74fc1ba39f1f6d18ce702
Source dataset
uspto-grants-2006_03
Patent number
US07011960B2

Reaction notes

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