Design, synthesis and evaluation of small molecules as modulators of ion channels and enzymes.

The work reported in this thesis spaces between two different main projects, carried out at the University of Florence, and a third one at the Moffitt Cancer Center and Research Institute in the United States. Since these three projects are completely different, they have been discussed separately. The first project discusses the design and synthesis of isoform-selective HCN channel inhibitors through suitable modification on the structure of EC18, a known isoform-preferring HCN channel inhibitor. The second project discuss the design, synthesis and enzymatic evaluation of carbonic anhydrase's (CA) modulators. The first part discusses the preparation and enzymatic evaluation of a series of histidine (or histamine) piperazine-containing compounds as CA activators. The compounds prepared showed remarkable selectivity toward isoform XIII. The second part discusses the preparation and enzymatic evaluation of a new series of di-substituted piperazine carbonic anhydrase inhibitors. These compounds showed an interesting selectivity toward CA IX and XII. The third project discussed was carried out in the United States, at the Moffitt Cancer Center and Research Institute, in Tampa, Florida. The project had the aim to provide not only the practical synthesis and the late-stage installation on a drug scaffold of a new covalent warhead prepared previously by the group of Prof. Justin Lopchuk, but also to provide a vast quantity of information on fine-tuning reactivity, solvent stability, DMPK properties both in vitro and in vivo, efficacy against the target protein, proteome-wide selectivity analysis, and ultimately efficacy against the conventional drug. New modalities for the installation of these novel warheads on complex drug scaffolds have been explored through a series of novel transfer reagents developed in the Lopchuk laboratory.