Acetonitrile → 4-Nitrobenzyl alcohol

HRID 1418803Notes

Reaction equation

HRID 1418803 reaction equation
[N+:1]([C:4]1[CH:32]=[CH:31][C:7]([CH2:8][O:9]C(=O)[C@H](CC2C=CC=CC=2)NC(OCC2C=CC=CC=2)=O)=[CH:6][CH:5]=1)([O-:3])=[O:2].C(#N)C>C(O)C(N)(CO)CO.CS(C)=O>[N+:1]([C:4]1[CH:5]=[CH:6][C:7]([CH2:8][OH:9])=[CH:31][CH:32]=1)([O-:3])=[O:2]

Participants

5

Reactant 2

HCID 6342AcetonitrileHCID 13188819HCID 13188819, compound

Product 1

HCID 692754-Nitrobenzyl alcohol

Reagents, catalysts & solvents 2

HCID 679Dimethyl SulfoxideSolventHCID 6503Tris(hydroxymethyl)aminomethaneSolvent

Procedure

To a solution of antibody 7G12 in 50 mM Tris-buffered solution, pH 8.0 (90 μL), was added a solution of a substrate compound, N-(benzyloxycarbonyl)-L-phenylalanine 4-nitrobenzyl ester (L-1) (R=PhCH2), synthesized in Preparation 3, in 2mM DMSO solution (10 μL) at 25° C., and they were mixed with stirring to obtain the reaction mixture containing 10.8 μM antibody and 200 μM substrate. By means of HPLC (YMCAM-303: C-18, 10 mm in diameter×250 mm in length, an aqueous solution of acetonitrile/0.1% trifluoroacetic acid=35/65 (0-7 minutes), 35/65-90/10 (7-15 minutes), 90/10 (15-17 minutes), 1.0 mL/minute, 278 nm), the amount of 4-nitrobenzyl alcohol (retention time: 5.9 minutes) produced in the reaction mixture was traced in time course for the purpose of determing initial rate. Next, the same procedure as mentioned above was repeated using N-(benzyloxycarbonyl)-D-phenylalanine 4-nitrobenzyl ester (D-1) (R=PhCH2) instead of N-(benzyloxycarbonyl)-L-phenylalanine 4-nitrobenzyl ester (L-1) (R=PhCH2) to determine the initial rate. Background hydrolytic reaction rate was also determined based on the amount of 4-nitrobenzyl alcohol produced by spontaneous degradation in the reaction solution free from antibody 7G12.

Workup

  1. additionthey were mixed
  2. customto obtain the reaction mixture
  3. additioncontaining 10.8 μM antibody and 200 μM substrate

Sources & evidence

ORD record
ord-edc715961eaf4579ba0c80fe7d31929b
Source dataset
uspto-grants-1999_03
Patent number
US05885816

Reaction notes

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