Study of expression, activity, and inhibition of cathepsins B, L, and S by dipeptidyl nitriles using pancreatic adenocarcinoma cell lines

Abstract

Pancreatic Cancer has high mortality rate according to the Brazilian National Cancer Institute (INCA). Gemcitabine is a drug usually applied in this case, but it has many limitations. Therefore, novel therapeutic alternatives are needed. Cysteine proteases present enormous physiopathological relevance, being overexpressed in many Cancer types, including the pancreatic adenocarcinoma. Among them, cathepsins B, L, and S are involved in many processes, and they have been studied as therapeutic targets. Dipeptidyl nitrile derivatives are promising reversible covalent cathepsin inhibitors studied in a variety of cancer types. Here, in vitro studies using pancreatic adenocarcinoma cell lines (MIA PaCa-2 and BxPC-3) will be used to analyze the expression, activity, and inhibition of cathepsins B, L, and S with dipeptidyl nitrile derivatives. A myriad of assays and techniques will be performed in this study, including colorimetry, fluorimetry, epifluorescence microscopy, flow cytometry, Western blot, zymography and electron microscopy. Altogether, these studies will be done to correlate the enzymatic inhibition of the enzyme to the cell phenotypic pharmacological activity of these novel compounds. In the end, it is expected that the best substances will be used alone or in combination with known drugs to be developed to the next stage of the drug discovery pipeline. (AU)