Carbonic anhydrases are enzymes responsible for the reversible hydration of carbon dioxide in the body, and they have been linked to various diseases like glaucoma, edema, altitude sickness, epilepsy, and cancer. Low molecular weight compounds are utilized to inhibit these enzymes and regulate their activity. However, existing drugs affecting carbonic anhydrases are non-selective, inhibiting the target enzymes and other isoenzymes. Consequently, efforts are underway to develop new compounds more selective for the target isoenzymes. Still, progress has been slow, with the last carbonic anhydrase inhibitor registered 30 years ago.
The house mouse (Mus musculus) is a commonly used model organism in pre-clinical studies; conducting control studies on the selectivity of test compounds with recombinant mouse carbonic anhydrase proteins and characterizing these proteins would enable an assessment of whether the observed effects of potential drugs in the mouse model can be attributed to the selectivity of the test compounds for the chosen human carbonic anhydrase isoenzyme.
Hence, the primary goal of this project is to characterize recombinant mouse carbonic anhydrase isoenzymes and explore their interactions with inhibitors using various biochemical and biophysical methods. The outcomes of these studies would enhance our understanding of the mechanisms involved in carbonic anhydrase-inhibitor interactions, facilitating more precise planning of pre-clinical studies.
The project will involve expressing and purifying mouse carbonic anhydrases, enzymatic activity and inhibition studies, employing thermal shift and isothermal calorimetry techniques for inhibitor binding studies, and determining the crystal structures of these proteins and their inhibitor complexes.