selective cytotoxicity of amidinopiperidine based compounds towards burkitt’s lymphoma cells involves proteasome inhibition基于amidinopiperidine化合物的选择性细胞毒性对伯基特淋巴瘤细胞包括蛋白酶体抑制.pdfVIP

Selective Cytotoxicity of Amidinopiperidine Based Compounds Towards Burkitt’s Lymphoma Cells Involves Proteasome Inhibition Martina Gobec, Ales Obreza, Matevz Prijatelj, Boris Brus, Stanislav Gobec, Irena Mlinaric-Rascan* University of Ljubljana, Faculty of Pharmacy, Ljubljana, Slovenia Abstract Serine proteases have proven to be promising pharmacological targets in contemporary drug discovery for cancer treatment. Since azaphenylalanine-based compounds manifest cytotoxic activity, we have selected serine protease inhibitors designed and synthesized in-house with large hydrophobic naphthalene moiety for screening. The cytotoxic potential of screened molecules was correlated to modifications of R1 residues. The most cytotoxic were compounds with greater basicity; amidinopiperidines, piperidines and benzamidines. Amidinopiperidine-based compounds exert cytotoxicity in low mM range, with IC50 18 mM and 22 mM for inhibitors 15 and 16 respectively. These compounds exhibited selective cytotoxicity towards the Burkitt’s lymphoma cells Ramos and Daudi, and proved nontoxic to PMBC, Jurkat and U937. They induce caspase-dependent apoptotic cell death, as demonstrated by the use of a pan-caspase inihibitor, zVADfmk, which was able to rescue Ramos cells from compound(s)-induced apoptosis. We confirm a disruption of the pro-survival pathway in Burkitt’s lymphoma through NFkB inhibition. The accumulation of phosphorylated precursor (p105) and inhibitory (IkB) molecules with no subsequent release of active NFkB implicated the involvement of proteasome. Indeed, we show that the amidinopiperidine-based compounds inhibit all three proteolytical activities of the human 20S proteasome, with the most prominent effect being on the trypsin-like activity. Consistently, treatment of Ramos cells with these