Metalloproteinase inhibitors for the disintegrin-like metalloproteinases ADAM10 and ADAM17 that differentially block constitutive and phorbol ester-inducible shedding of cell surface molecules
Metalloproteinase inhibitors for the disintegrin-like metalloproteinases ADAM10 and ADAM17 that differentially block constitutive and phorbol ester-inducible shedding of cell surface molecules
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DOI：
10.2174/1386207053258488
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发表时间：
2005-03-01
影响因子：
1.8
通讯作者：
Becherer, JD
中科院分区：
文献类型：
--
作者：
Ludwig, A;Hundhausen, C;Becherer, JD
ADAM（一种崩解素和金属蛋白酶）家族ADAM10和adam17的跨膜metzinkin- proteasic都涉及各种细胞表面分子的外胞域脱落，包括il6受体和跨膜趋化因子CX3CL1和CXCL16。这些分子组成性地从培养的细胞中释放出来，这一过程可以通过phorbol酯（如PMA）刺激细胞而迅速增强。最近的研究支持了这样一种观点，即组成性切割主要涉及ADAM10，而诱导性切割在很大程度上是由ADAM17介导的。我们在这里描述了对ADAM10和ADAM17具有不同效价的羟酸酯化合物的发现，以及这两种蛋白酶阻断IL6R、CX3CL1和CXCL16的组成性和诱导性切割的不同能力。通过筛选多种羟肟酸抑制剂抑制重组金属蛋白酶，发现一种化合物抑制ADAM10的效价比ADAM17高100倍以上，这可能是由于该化合物比ADAM17更适合ADAM10的S1'特异性囊。在基于细胞的切割实验中，该化合物（G1254023X）能有效阻断IL6R、CX3CL1和CXCL16的组成性释放。这与ADAM10而非ADAM17参与这一过程的报道一致。相比之下，该化合物不影响pma诱导的脱落，仅被ADAM17的强效抑制剂GW280264X阻断。正如预期的那样，G1254023X并没有进一步减少ADAM10缺陷细胞中CX3CL1和CXCL16的残留释放，这证明该化合物对这些分子的组成性脱落的影响完全是由于抑制ADAM10。因此，G1254023X可以作为本构脱落事件的优先抑制剂，而不影响诱导脱落，以响应类似于PMA的激动剂。
The transmembrane metzinkin-proteascs 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 (G1254023X) 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, G1254023X 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, G1254023X 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.