Réaction #531852

ord-df53c23fcbed4461a3f88914925f5408

Équation de réaction

CC(C)(COP(=O)(O)OP(=O)(O)OC[C@H]1O[C@@H](n2cnc3c(N)ncnc32)[C@H](O)[C@@H]1OP(=O)(O)O)[C@@H](O)C(=O)NCCC(=O)NCCS.[Na].[Na]
coenzyme A disodium salt
CCN(CC)CC
Et3N
CC(C)(COP(=O)(O)OP(=O)(O)OC[C@H]1O[C@@H](n2cnc3c(N)ncnc32)[C@H](O)[C@@H]1OP(=O)(O)O)[C@@H](O)C(=O)NCCC(=O)NCCS
desired product
CC(C)(COP(=O)(O)OP(=O)(O)OC[C@H]1O[C@@H](n2cnc3c(N)ncnc32)[C@H](O)[C@@H]1OP(=O)(O)O)[C@@H](O)C(=O)NCCC(=O)NCCS
CoA

Solvants

Conditions de réaction

Conditions détaillées
See reaction.notes.procedure_details.

Traitement

  1. 1
    Autrethe completion of the reaction

Mode opératoire

To a solution of N-(ε-maleimidocaproic acid) hydrazide 2 (1 mg, 0.0044 mmol) in 50 μl DMF, a solution of coenzyme A disodium salt (3.6 mg, 0.0044 mmol, 1 eq.) in a DMF/buffer mixture (70 μl DMF/30 μl 50 mM Tris Cl, pH 7.5) is added. The reaction is followed by analytical HPLC (detection at 260 nm) to verify the completion of the reaction. Subsequently, a solution of 3-amino-3-deoxydigoxigenin hemisuccinamide succinimidyl ester (2.6 mg, 0.0044 mmol, 1 eq.) dissolved in 50 μl DMF and 10 μl Et3N is added, and the reaction mixture stirred for 4 hours at room temperature. The reaction is submitted to preparative HPLC and fractions containing the desired product are concentrated in vacuo, dissolved in DMSO, and analyzed by analytical HPLC for purity. Fractions containing pure product are combined. The concentration of CoA-Dg is determined using the extinction coefficient of adenine (ε(adenine, 260 nm)=15′300 [M−1 cm−1]). The yield of CoA-Dg is 2.37 mg (37%).

Source

DOI: 10.6084/m9.figshare.5104873.v1Brevet: US08476031B2uspto-grants-2013_07