title: Metalloproteinase inhibitors for the disintegrin-like metalloproteinases ADAM10 and ADAM17 that differentially block constitutive and phorbol ester-inducible shedding of cell surface molecules.
abstract: The transmembrane metzinkin-proteases of the ADAM (a disintegrin and a metalloproteinase)-family ADAM10 and ADAM 17 are both implicated in the ectodomain shedding of various cell surface molecules including the IL6-receptor and the transmembrane chemokines CX3CL1 and CXCL16. These molecules are constitutively released from cultured cells, a process that can be rapidly enhanced by cell stimulation with phorbol esters such as PMA. Recent research supports the view that the constitutive cleavage predominantly involves ADAM10 while the inducible one is mediated to a large extent by ADAM17. We here describe the discovery of hydroxamate compounds with different potency against ADAM10 and ADAM17 and different ability to block constitutive and inducible cleavage of IL6R, CX3CL1 and CXCL16 by the two proteases. By screening a number of hydroxamate inhibitors for the inhibition of recombinant metalloproteinases, a compound was found inhibiting ADAM10 with more than 100-fold higher potency than ADAM17, which may be explained by an improved fit of the compound to the S1' specificity pocket of ADAM10 as compared to that of ADAM17. In cell-based cleavage experiments this compound (GI254023X) potently blocked the constitutive release of IL6R, CX3CL1 and CXCL16, which was in line with the reported involvement of ADAM10 but not ADAM17 in this process. By contrast, the compound did not affect the PMA-induced shedding, which was only blocked by GW280264X, a potent inhibitor of ADAM17. As expected, GI254023X did not further decrease the residual release of CX3CL1 and CXCL16 in ADAM10-deficient cells verifying that the compound's effect on the constitutive shedding of these molecules was exclusively due to the inhibition of ADAM10. Thus, GI254023X may by of use as a preferential inhibitor of constitutive shedding events without effecting the inducible shedding in response to agonists acting similar to PMA.
authors.affiliation: Biochemical Institute, Christian-Albrechts-University, Kiel, Germany. aludwig@biochem.uni-kiel.de
authors.forename: Andreas
authors.initials: A
authors.lastname: Ludwig
authors.forename: Christian
authors.initials: C
authors.lastname: Hundhausen
authors.forename: Millard H
authors.initials: MH
authors.lastname: Lambert
authors.forename: Neil
authors.initials: N
authors.lastname: Broadway
authors.forename: Robert C
authors.initials: RC
authors.lastname: Andrews
authors.forename: D Mark
authors.initials: DM
authors.lastname: Bickett
authors.forename: M Anthony
authors.initials: MA
authors.lastname: Leesnitzer
authors.forename: J David
authors.initials: JD
authors.lastname: Becherer
mesh_terms: D051722:ADAM Proteins; D000072198:ADAM17 Protein; D053829:Amyloid Precursor Protein Secretases; D002478:Cells, Cultured; D054428:Chemokine CX3CL1; D018925:Chemokines / Q000378:metabolism; D020523:Chemokines, CX3C / Q000378:metabolism; D019743:Chemokines, CXC / Q000378:metabolism; D004347:Drug Interactions; D010450:Endopeptidases / Q000187:drug effects / Q000378:metabolism*; D006877:Hydroxamic Acids / Q000737:chemistry / Q000494:pharmacology*; D015850:Interleukin-6 / Q000378:metabolism; D008565:Membrane Proteins / Q000187:drug effects / Q000378:metabolism*; D008666:Metalloendopeptidases / Q000037:antagonists & inhibitors / Q000378:metabolism*; D010703:Phorbol Esters / Q000378:metabolism / Q000494:pharmacology*; D011480:Protease Inhibitors / Q000494:pharmacology*
chemical_list: D054428:Chemokine CX3CL1; D018925:Chemokines; D020523:Chemokines, CX3C; D019743:Chemokines, CXC; D006877:Hydroxamic Acids; D015850:Interleukin-6; D008565:Membrane Proteins; D010703:Phorbol Esters; D011480:Protease Inhibitors; D053829:Amyloid Precursor Protein Secretases; D010450:Endopeptidases; D051722:ADAM Proteins; D008666:Metalloendopeptidases; D000072198:ADAM17 Protein