Phosphinic peptide inhibitors of macrophage metalloelastase (MMP-12). Selectivity and mechanism of binding.
Pseudopeptide inhibitors of MMP-12 with a phosphinic dipeptide G psi[PO(2)H-CH(2)]L covering the P1-P1'- positions originating from a combinatorial solid phase library have been identified and kinetically analysed with respect to binding mechanism and selectivity towards MMP-7, MMP-9, MMP-13 and MMP-14. One compound with a low nanomolar dissociation constant for MMP-12 showed significantly lower affinity towards all other MMPs tested compared to MMP-12. Two compounds showed selectivity against MMP-9, MMP-13 and MMP-14. One additional compound showed selectivity against MMP-7. The selectivity of these compounds could partly be rationalized by analysis of homology models of the enzymes. Truncated versions of one inhibitor spanning P2 to P2', P3 to P2' or P2 to P3' showed that interactions on both the prime and the non-prime side are important for binding. A two-step binding mechanism, with a rate limiting second step, was shown for binding of a tryptophane containing inhibitor to MMP-12 by transient state analysis, using the tryptophane residue of the inhibitor as fluorescent probe.
Schiødt
C B
CB
OSTEOPRO A/S Herlev Hovedgade 207, Herlev, DK-2730, Denmark. cbs@osteopro.dk
Buchardt
J
J
Terp
G E
GE
Christensen
U
U
Brink
M
M
Berger Larsen
Y
Y
Meldal
M
M
Foged
N T
NT
D000818:Animals; D001665:Binding Sites; D004151:Dipeptides / Q000737:chemistry / Q000494:pharmacology*; D053508:Matrix Metalloproteinase 12; D020783:Matrix Metalloproteinase 7 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D061965:Matrix Metalloproteinase Inhibitors; D053504:Matrix Metalloproteinases, Membrane-Associated; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors* / Q000378:metabolism; D008958:Models, Molecular; D010721:Phosphinic Acids / Q000737:chemistry; D011487:Protein Conformation; D011817:Rabbits; D011994:Recombinant Proteins / Q000037:antagonists & inhibitors / Q000378:metabolism; D013329:Structure-Activity Relationship
D004151:Dipeptides; D061965:Matrix Metalloproteinase Inhibitors; D010721:Phosphinic Acids; D011994:Recombinant Proteins; D053504:Matrix Metalloproteinases, Membrane-Associated; D008666:Metalloendopeptidases; D020783:Matrix Metalloproteinase 7; D020780:Matrix Metalloproteinase 9; D053508:Matrix Metalloproteinase 12