Ethylmorpholine → HCID 86610815, compound

HRID 494901Notes

Reaction equation

HRID 494901 reaction equation
[S:1]([NH:17][CH2:18][CH2:19][CH2:20][CH2:21][C@@H:22]([C:24]([OH:26])=[O:25])[NH2:23])([C:4]1[C:16]2[CH:15]=[CH:14][CH:13]=[C:9]([N:10]([CH3:12])[CH3:11])[C:8]=2[CH:7]=[CH:6][CH:5]=1)(=[O:3])=[O:2].C(N1[CH2:34][CH2:33][O:32]CC1)C.C[O:36][CH:37](OC)[C:38]1[CH:43]=C[C:41](C(O)=O)=[CH:40][CH:39]=1.S(N[C@H](C(O)=O)CCCCN)(C1C2C=CC=C(N(C)C)C=2C=CC=1)(=O)=O>CN(C)C=O.C(O)(=O)C>[S:1]([NH:17][CH2:18][CH2:19][CH2:20][CH2:21][C@@H:22]([C:24]([OH:26])=[O:25])[NH:23][C:33](=[O:32])[C:34]1[CH:41]=[CH:40][CH:39]=[C:38]([CH:37]=[O:36])[CH:43]=1)([C:4]1[C:16]2[CH:15]=[CH:14][CH:13]=[C:9]([N:10]([CH3:12])[CH3:11])[C:8]=2[CH:7]=[CH:6][CH:5]=1)(=[O:2])=[O:3]

Participants

7

Reactant 4

HCID 7525Ethylmorpholine2×HCID 121945DansyllysineHCID 10691138α-Monodansyl-L-lysineHCID 10888839HCID 10888839, compound

Product 1

HCID 86610815HCID 86610815, compound

Reagents, catalysts & solvents 2

HCID 176Acetic AcidSolventHCID 6228DimethylformamideSolvent2×

Procedure

Nε-dansyl-L-lysine (Sigma, 9.5 mg) was dissolved in 100 μl dimethylformamide under gentle warming. N-ethyl-morpholine (3 μl) was added, so that the apparent pH, as judged by spotting onto moist pH paper, was between 8 and 8.5. This solution was mixed with a solution of 7.3 mg of the hydroxysuccinimido ester of 4-carboxybenzaldehyde dimethylacetal in 50 μl dimethylformamide. The apparent pH was checked in the same way as before, and adjusted with N-ethylmorpholine (1 μl at a time) if necessary. After 4 hours and 30 minutes (the apparent pH having been checked from time to time and adjusted if necessary), a similar solution of 7.3 mg of the active ester was added and the reaction allowed to continue for a further 4 hours. The reaction, as judged by t.l.c., was then essentially complete. The t.l.c. system was that described in Example 1(c) and the Rf values were approx. 0.3 and 0.6 for dansyl-lysine and the product, respectively. The mixture was then diluted to 1 ml with acetic acid (1%. v/v), centrifuged, and the supernatant subjected to HPLC (system of Example 4(c)). The major fluorescent peak eluting on the gradient (around 40% acetonitrile) was pooled, and the acetonitrile was driven off in a current of air. The product was characterized by measurement of its electrophoretic mobility at pH 6.5 (predicted and observed, 0.5 [mAsp=1.0]). To this stock solution (600 μl) were added 6 μl M HCl, and the solution was diluted ten-fold with 10−3M HCl. This brought the concentration to 0.7 mM, as judged by direct comparison of the u.v. absorption spectrum with that of a solution of dansyl-lysine of known concentration. It was assumed that the optical density of the dansyl chromophore would be approximately the same in the two substances.

Workup

  1. custom30 minutes
  2. additionwas added
  3. waitto continue for a further 4 hours
  4. customThe reaction
  5. washThe major fluorescent peak eluting on the gradient (around 40% acetonitrile)
  6. additionTo this stock solution (600 μl) were added 6 μl M HCl
  7. additionthe solution was diluted ten-fold with 10−3M HCl
  8. concentrationthe concentration to 0.7 mM
  9. customabsorption spectrum with that
  10. concentrationof a solution of dansyl-lysine of known concentration

Sources & evidence

ORD record
ord-d3592f6f392943c388185dd14d66e33c
Source dataset
uspto-grants-2009_05
Patent number
US07538182B2

Reaction notes

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