title: Azaphenylalanine-based serine protease inhibitors induce caspase-mediated apoptosis.
abstract: Molecules regulating cell death constitute prominent therapeutic targets. The pro-apoptotic role of serine protease inhibitors prompted us to search for novel modulators of this process. We have tested some recently synthesized antithrombotic compounds for their potential to induce apoptotic cell death. Cell based analyses revealed that inhibitors built on the azaphenylalanine scaffold are, for B-cell lymphoma cells, severely cytotoxic, while other compounds tested were moderate or non-cytotoxic. These inhibitors induced the time and concentration dependent biochemical and morphological characteristics of apoptosis, such as DEVDase activation, loss of mitochondrial membrane potential, nuclear degradation and genomic DNA fragmentation. Most of the inhibitors proved to be selective for thrombin, with inhibition constants (K(i)) in the nanomolar range. However, they could also inhibit at least one additional serine protease (trypsin, chymotrypsin and/or coagulation factor X) with K(i) values in the nanomolar or low micromolar range. These serine protease inhibitors constitute novel apoptosis inducing compounds in B-cell lymphoma cells.
authors.affiliation: University of Ljubljana, Faculty of Pharmacy, Ljubljana, Slovenia.
authors.forename: Teja
authors.initials: T
authors.lastname: Celhar
authors.forename: Tina
authors.initials: T
authors.lastname: Batista Napotnik
authors.forename: Ales
authors.initials: A
authors.lastname: Obreza
authors.forename: Anamarija
authors.initials: A
authors.lastname: Zega
authors.forename: Petra Stefanic
authors.initials: PS
authors.lastname: Anderluh
authors.forename: Danijel
authors.initials: D
authors.lastname: Kikelj
authors.forename: Irena
authors.initials: I
authors.lastname: Mlinaric-Rascan
mesh_terms: D017209:Apoptosis / Q000187:drug effects*; D020169:Caspases / Q000378:metabolism*; D045744:Cell Line, Tumor; D006801:Humans; D016393:Lymphoma, B-Cell / Q000378:metabolism / Q000473:pathology; D010649:Phenylalanine / Q000031:analogs & derivatives* / Q000494:pharmacology; D015842:Serine Proteinase Inhibitors / Q000737:chemistry* / Q000494:pharmacology*; D013997:Time Factors
chemical_list: D015842:Serine Proteinase Inhibitors; C493947:azaphenylalanine; D010649:Phenylalanine; D020169:Caspases