Dimethyl Sulfoxide → 3,4-Methylenedioxymethamphetamine

HRID 1828966Notes

Reaction equation

HRID 1828966 reaction equation
CS(C)=O.O=C1CCC(=O)N1OC(=O)CCC(NC[CH:20]1[O:24][C:23]2[CH:25]=[CH:26][C:27]([CH2:29][CH:30]([N:32](C)[C:33](=O)C(F)(F)F)[CH3:31])=[CH:28][C:22]=2[O:21]1)=O.N[C@H](C(O)=O)CCCCN.[N+](C1C=C([N+]([O-])=O)C=C([N+]([O-])=O)C=1S([O-])(=O)=O)([O-])=O>P([O-])([O-])([O-])=O.[K+].[K+].[K+].CN(C=O)C>[CH3:31][CH:30]([NH:32][CH3:33])[CH2:29][C:27]1[CH:26]=[CH:25][C:23]2[O:24][CH2:20][O:21][C:22]=2[CH:28]=1

Participants

7

Reactant 4

HCID 679Dimethyl SulfoxideHCID 5962L-LysineHCID 1151422,4,6-Trinitrobenzene SulfonateHCID 87616818N-(5-{2-[methyl-(2,2,2-trifluoro-acetyl)-amino]-propyl}-benzo[1,3]dioxol-2-yl methyl)-succinamic Acid 2,5-dioxo-pyrrolin-1-yl Ester

Product 1

HCID 16153,4-Methylenedioxymethamphetamine

Reagents, catalysts & solvents 2

HCID 6228DimethylformamideSolventHCID 62657Potassium Phosphate, TribasicSolvent

Procedure

A solution of 173 mg of keyhole limpet hemocyanin in 7 ml of 50 mM potassium phosphate (pH 7.5) was cooled in an ice-bath. To the solution were added 10.5 mL of dimethylsulfoxide (DMSO) dropwise, and the reaction temperature was maintained below room temperature. To the protein solution was added a solution of 40.2 mg of 1L in 1.5 mL of DMF dropwise. The mixture was allowed to stir at room temperature 18 hours. The resulting conjugate was placed in a dialysis tube (10,000 MW cut-off) and dialyzed in 1 L of 70% DMSO in 50 mM potassium phosphate (pH 7.5, 3 changes, at least 3 hours each), 1 L of 50% DMSO in 50 mM potassium phosphate (at least 3 hours), 1 L of 30% DMSO in 50 mM potassium phosphate (at least 3 hours), 1 L of 10% DMSO in 50 mM potassium phosphate (at least 3 hours) at room temperature. The trifluoroacetamido group of the conjugate was deprotected by dialysis of the resulting conjugate against 10% ammonium hydroxide for 3 days (1 L each for approximately 24 hours each), followed by 6 changes with 50 mM potassium phosphate (pH 7.5) at 4° C. (1 L each for at least 6 hours each). The protein concentration was determined to be 2.9 mg/mL using BioRad Coomassie blue protein assay (Bradford, M., Anal. Biochem. 72:248, 1976). A total of 34 mL of the conjugate was obtained. The extent of available lysine modification was determined to be 72% by the 2,4,6-trinitrobenzene sulfonate (TNBS) method (Habeeb AFSA, Anal. Biochem. 14:328-34, 1988).

Workup

  1. temperaturethe reaction temperature was maintained below room temperature
  2. customThe resulting conjugate was placed in a dialysis tube
  3. custom(10,000 MW cut-off) and dialyzed in 1 L of 70% DMSO in 50 mM potassium phosphate (pH 7.5, 3 changes, at least 3 hours each), 1 L of 50% DMSO in 50 mM potassium phosphate (at least 3 hours), 1 L of 30% DMSO in 50 mM potassium phosphate (at least 3 hours)
  4. custom1 L of 10% DMSO in 50 mM potassium phosphate (at least 3 hours)
  5. customat room temperature
  6. customThe trifluoroacetamido group of the conjugate was deprotected by dialysis of the resulting conjugate against 10% ammonium hydroxide for 3 days (1 L each for approximately 24 hours each),
  7. customfollowed by 6 changes with 50 mM potassium phosphate (pH 7.5) at 4° C.
  8. customfor at least 6 hours
  9. concentrationThe protein concentration
  10. customA total of 34 mL of the conjugate was obtained

Sources & evidence

ORD record
ord-acbaa32e167145c59171232444cb6a81
Source dataset
uspto-grants-2006_06
Patent number
US07060847B2

Reaction notes

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