A disintegrin derivative as a case study for PHIP labeling of disulfide bridged biomolecules.A specific labeling strategy for bioactive molecules is presented for eptifibatide (integrilin) an antiplatelet aggregation inhibitor, which derives from the disintegrin protein barbourin in the venom of certain rattlesnakes.  By specifically labeling the disulfide bridge this molecule becomes accessible for the nuclear spin hyperpolarization method of parahydrogen induced polarization (PHIP).  The PHIP-label was synthesized and inserted into the disulfide bridge of eptifibatide via reduction of the peptide and insertion by a double Michael addition under physiological conditions.  1H-PHIP-NMR experiments yield a factor of over 1000 as lower limit for the enhancement factor.  These results demonstrate the high potential of the labeling strategy for the introduction of site selective PHIP-labels into biomolecules' disulfide bonds.
PHIP
HPLC and MS spectra prove the successful insertion of the label. 
HPLC
HPLC and MS spectra prove the successful insertion of the label. 
MS