Synthesis and inhibitory properties of carboxylate-based inhibitors and their fluorescence-labeled analogues with high affinity for some Metzincins.

Matrix Metalloproteinases (MMPs) are a family of enzymes that are attracting growing interest as therapeutic targets. They are proteases whose fundamental role has been recognised in the degradation of the Extracellular Matrix (ECM) components. Their expression is finely regulated at many levels (transcription, activation, inhibition) while unregulated profiles have been found in many pathological conditions such as cancer, arthritis, atherosclerosis, and inflammatory diseases. Since the use of endogenous inhibitors is not easy to accomplish, the design and synthesis of new small molecules that could allow the regulation of these proteins is a vanguard in nowadays research of new anticancer drugs, as well as for other therapies. Some promising MMP inhibitors have been developed by Prof. Rossello’s group in recent years. The best results were achieved with an inhibitor particularly selective for MMP-9 and MMP-12 that was able to arrest human glioblastoma cancer cell invasiveness in models studied. On the basis of Prof. Rossello's group recent findings, in my Thesis project I was involved in the synthesis of a series of novel MMP inhibitors, derivatives of the previously discovered compounds. The newly synthesised entities have been tested on the principal targets, which are MMP-9 and MMP-12. The assays have been conducted in vitro on human recombinant MMPs by fluorometric HTS methods, using a fluorogenic peptide as substrate. All compounds are more active on MMP-12 than on MMP-9, with a lower inhibitory activity compared to previously achieved results. However, selectivity of the new inhibitors over other MMPs should be further investigated. In the second part of my Thesis, I carried out the synthesis of two derivatives of an ADAMTS-5(A Disintegrin And Metalloproteinase with Thrombospondin Motifs) selective inhibitor previously described by Prof. Rossello’s group(1). This specific enzyme inhibition has given promising results in the protection against osteoarthritis in mice models studied. The arylsulfonamidic structure of this inhibitor was slightly modified to improve potency and the synthesis of two new compounds was accomplished. The newly synthesised compounds have been tested for their inhibitory activity on ADAMTS-5 by Prof. Hideaki Nagase’s group (University of Oxford, UK). (1)Nuti, E.; Santamaria, S.; Casalini, F.; Yamamoto, K.; Marinelli, L.; La Pietra, V.; Novellino, E.; Orlandini, E.; Nencetti, S.; Marini, A. M.; Salerno, S.; Taliani, S.; Da Settimo, F.; Nagase, H.; Rossello, A., Arylsulfonamide inhibitors of aggrecanases as potential therapeutic agents for osteoarthritis: synthesis and biological evaluation, Eur. J. Med. Chem., (2013), 62: 379-394