Prodipine hydrochloride

Kinase experiment:

Dipeptidyl peptidase IV activity is determined using the fluorogenic substrate Gly-Pro-4-methoxy-2-naphthylamide. Hydrolysis of L-Ala-4-methoxy-2-naphthylamide in 60 mM phosphate buffer pH 7.4 is chosen for the determination of aminopeptidase M activity. The intramolecularly quenched fluorogenic substrate Lys (2,4 dinitrophenyl)-Pro-Pro-NH-CH2-CH,-NH,-aminobenzoyl-2HCl is used to assay aminopeptidase P activity. Prolyl oligopeptidase (PO) is measured with N-benzyloxycarbonyl-Gly-Pro-7-amino-4-methylcoumarin. One unit of enzymatic activity is defined as the amount of enzyme catalyzing the formation of 1 μmol of assay product per minute under the conditions used[1].

Animal experiment:

Rabbits[1] Male New Zealand white rabbits (3-4 kg) are used. EXPERIMENT 1 The rabbits (n=1 for each condition) receive a single iv. injection of 10 mg Prodipine or saline. Blood samples are taken 5 min before injection and 30 min, 1 hr, 2 hr, and 24 hr after the treatment. EXPERIMENT 2 The rabbits (n=1 for each condition) receive a single i.v. bolus injection of saline or are treated with 1, 5, or 10 mg Prodipine (also in a single i.v. bolus) at time 0. The animals are kept under observation for 40 days. At several time points (-5 min, 4 hr, and at days 1, 2, 3, 6, 12, 21, and 40), blood samples are taken and body weight determined. Rabbits are killed with pentobarbital after termination of the observation period.

References:

[1]. De Meester I, et al. In vivo inhibition of dipeptidyl peptidase IV activity by pro-pro-diphenyl-phosphonate (Prodipine). Biochem Pharmacol. 1997 Jul 1;54(1):173-9.