Synthesis and in vitro Evaluation of ADAM10 and ADAM17 Highly Selective Bioimaging Probes

摘要

Adisintegrin and metalloproteinase (ADAMs) are membrane- bound metalloproteases responsible forthe ectodomainshed- ding of varioustransmembrane proteins and play important roles in multiple relevant biological processes. Their altered ex- pression is involved in severalpathological conditions, and in particularADAM10 or ADAM17 overexpression is found in vari- ous forms of cancer. To better understandhow they are regu- lated in the cellular context, it is useful to visualize the specific ADAMs pathway by means of molecular imaging techniques. For this purpose, we synthesized bioactive fluorescent probes suitable for cell imaging and that are able to specifically target ADAM10 or ADAM17. Twopreviously developed ADAM17-and ADAM10-selective inhibitors werechosen for conjugation,re- spectively,toa Cy5.5 dye and to Cy5.5 and FITC dyes. Herein we also report the synthesis of agold-labeled compound as an additional bioimaging probe for ADAM10.The newly synthe- sized ligands were found to be active in vitro on human re- combinant ADAM10 and/or ADAM17, showing IC_(50) values in the nanomolar range anda good selectivity over matrix metal- loproteinases (MMPs).Finally,these newly developed probes were successfully used for ADAMs stainingondifferent lym- phoma cell lines and lymph node mesenchymal stromal cells.