| Lipid Signaling | Chemical Probes | Metabolomics |
Chemical proteomics (chemoproteomics) has greatly facilitated our ability to measure enzyme functional states in native biological systems using reporter-tagged, active-site directed small molecule probes. Standard activity-based probes contain broad-spectrum reactive groups that covalently label active sites of large number of enzymes based on conserved mechanistic or structural features (FP-Rh, Figure 1). In contrast to broad-spectrum probes, we have demonstrated the utility of developing activity-based probes with more tailored selectivity for enhanced detection of low-abundance enzymes. For example, first-generation tailored activity-based probes (HT-01, Figure 1) have transformed our ability to functionally interrogate the biosynthesis and signaling activity of endocannabinoids in living cells and animals. Future studies are aimed at expanding the coverage of tailored activity-based probes for highly focused analysis of enzymes not amenable to standard chemoproteomic approaches.
To facilitate our biological discovery efforts, we seek to develop selective chemical probes to study the function of individual enzymes in living systems. Our initial efforts will be directed at members of the serine hydrolase enzyme superfamily because of their diverse substrate specificity (lipids, small-molecule transmitters, peptides, and proteins) and widespread roles in mammalian biology and disease (e.g. inflammation). In previous studies, we have shown that 1,2,3-triazole ureas (1,2,3-TUs) serve as a versatile scaffold for developing selective and in vivo-active inhibitors as well as activity-based probes for members of this enzyme class (Figure 2). Integration of these pharmacological tools with genetic disruption mouse models has provided unique insights into the biological function of target enzymes that could not be achieved by either approach alone. Future studies are aimed at synthesis and optimization of selective inhibitors for functional studies of target enzymes of interest in vivo.
Representative publications
1. K.L. Hsu et al., DAGLbeta inhibition perturbs a lipid network involved in macrophage inflammatory responses. Nat Chem Biol 8, 999 (2012).
2. K.M. Nagano et al., Selective inhibitors and tailored activity probes for lipoprotein-associated phospholipase A(2). Bioorg Med Chem Lett 23, 839-843 (2013).
3. K.L. Hsu et al., Development and optimization of piperidyl-1,2,3-triazole ureas as selective chemical probes of endocannabinoid biosynthesis. J Med Chem 56, 8257 (2013).
4. K.L. Hsu et al., Discovery and optimization of piperidyl-1,2,3-triazole ureas as potent, selective, and in vivo-active inhibitors of alpha/beta-hydrolase domain containing 6 (ABHD6). J Med Chem 56, 8270-8279 (2013).
1. K.L. Hsu et al., DAGLbeta inhibition perturbs a lipid network involved in macrophage inflammatory responses. Nat Chem Biol 8, 999 (2012).
2. K.M. Nagano et al., Selective inhibitors and tailored activity probes for lipoprotein-associated phospholipase A(2). Bioorg Med Chem Lett 23, 839-843 (2013).
3. K.L. Hsu et al., Development and optimization of piperidyl-1,2,3-triazole ureas as selective chemical probes of endocannabinoid biosynthesis. J Med Chem 56, 8257 (2013).
4. K.L. Hsu et al., Discovery and optimization of piperidyl-1,2,3-triazole ureas as potent, selective, and in vivo-active inhibitors of alpha/beta-hydrolase domain containing 6 (ABHD6). J Med Chem 56, 8270-8279 (2013).