In vivo inhibition of dipeptidyl peptidase IV activity by pro-pro-diphenyl-phosphonate (Prodipine).
Dipeptidyl peptidase IV (DPP IV, EC 3.4.14.5), also known as CD26, is a membrane-bound serine protease that cleaves off aminoterminal dipeptides from peptides with a penultimate proline (or, at a much slower rate, a penultimate alanine). Recently, we synthesized and characterized a number of dipeptide-derived diphenylphosphonates. Out of the resulting series of slow-binding irreversible inhibitors of DPP IV, diphenyl 1-(S)-prolylpyrrolidine-2(R,S)-phosphonate hydrochloride (Pro-Pro-diphenylphosphonate or Prodipine) was selected for further study. We investigated the in vivo applicability of Prodipine. Male rabbits weighing 3-4 kg received a single intravenous injection with 10 mg Prodipine or saline. After 1 hr, plasma DPP IV activity had decreased to less than 20% of the preinjection value and remained unchanged during a 24-hr observation period. In a next step, we aimed to study (i) the dose dependency and (ii) the duration of the effect after a single intravenous dose of Prodipine. A profound and long-lasting inhibition of plasma DPP IV activity was observed in the treated animals (1, 5 or 10 mg). It took 5 to 8 days to reach half of the pretreatment DPP IV activity and generally more than 20 days for a complete recovery. Systemic treatment with Prodipine not only led to inhibition of plasma DPP IV activity but also decreased tissue DPP IV activity in circulating mononuclear cells, kidney cortex, thymus, spleen, lung, and liver. No differences in activities of the related peptidases aminopeptidase P (APP, EC 3.4.11.9), prolyl oligopeptidase (PO, EC 3.4.21.26), or aminopeptidase M (mAAP, EC 3.4.11.2) were detected between Prodipine-treated and control rabbits. The in vivo applicability of this chemically stable, irreversible inhibitor of DPP IV opens new possibilities, not only to further unravel the biological functions of this intriguing ectopeptidase, but also to explore this enzyme as a new target in various fields of pharmacological research.
De Meester
I
I
Laboratory of Medical Biochemistry, University of Antwerp, Wilrijk, Belgium. idmeest@uia.ua.ac.be
Belyaev
A
A
Lambeir
A M
AM
De Meyer
G R
GR
Van Osselaer
N
N
Haemers
A
A
Scharpé
S
S
D000626:Aminopeptidases / Q000378:metabolism; D000818:Animals; D018819:Dipeptidyl Peptidase 4 / Q000097:blood / Q000302:isolation & purification / Q000378:metabolism*; D007672:Kidney Cortex / Q000201:enzymology; D008099:Liver / Q000201:enzymology; D008168:Lung / Q000201:enzymology; D008297:Male; D063208:Methionyl Aminopeptidases; D009000:Monocytes / Q000201:enzymology; D010880:Piperidines / Q000494:pharmacology*; D000085822:Prolyl Oligopeptidases; D011817:Rabbits; D012697:Serine Endopeptidases / Q000378:metabolism; D013154:Spleen / Q000201:enzymology; D013950:Thymus Gland / Q000201:enzymology
D010880:Piperidines; C016695:prodipin; D000626:Aminopeptidases; D063208:Methionyl Aminopeptidases; C022750:X-Pro aminopeptidase; D018819:Dipeptidyl Peptidase 4; D012697:Serine Endopeptidases; D000085822:Prolyl Oligopeptidases