Novel, Self-Assembling Dimeric Inhibitors of Human β Tryptase

β-tryptase, a homotetrameric serine protease, has four identical active sites facing a central pore, presenting an optimized setting for the rational design of bivalent inhibitors that bridge two adjacent sites. Using diol, hydroxymethyl phenols or benzoyl methyl hydroxamates and boronic acid chemistries to reversibly join two [3-(1-acylpiperidin-4-yl)phenyl]methanamine core ligands, we have successfully produced a series of self-assembling heterodimeric inhibitors. These heterodimeric tryptase inhibitors demonstrate superior activity compared to monomeric modes of inhibition. X-ray crystallography validated the dimeric mechanism of inhibition, and compounds demonstrated high selectivity against related proteases, good target engagement and tryptase inhibition in HMC1 xenograft models. Screening 3,872 possible combinations from 44 boronic acid and 88 diol derivatives, revealed several combinations that produced nano-molar inhibition and 7 unique pairs produced greater than 100-fold improvement in potency ...