title: 4-alkylidene-azetidin-2-ones: novel inhibitors of leukocyte elastase and gelatinase.
abstract: In addition to their antibiotic potency, beta-lactams have recently been investigated as inhibitors of serine proteinase such as leukocyte elastase (LE), released by inflammatory cells. We describe the synthesis of a series of 4-alkylidene-beta-lactams, and investigate how substitutions on C-3, C-4, and N-1 of the beta-lactam ring affect the activity of human LE and gelatinases MMP-2 and MMP-9. LE activity was measured using a chromogenic substrate, while gelatin-zymography assay was used to evaluate gelatinase activity. We demonstrate that C-4 unsaturation on the beta-lactam ring determines the degree of biological activity, with a selectivity over LE by 3-[1-(tert-butyldimethylsilyloxy)-ethyl] derivatives (lowest IC(50) was 4 microM), and over gelatinase MMP-2 by C-3-unsubstituted 4-[1-ethoxycarbonyl]-ethylidene-beta-lactams (lowest IC(50) was 60 microM). (3S)-3-[(1R)-1-hydroxyethyl]-4-(1-ethoxycarbonyl)-ethylidene-azetidin-2-one inhibits gelatinase MMP-9. The compounds tested showed no cytotoxicity against NIH-3T3 murine fibroblasts. This is the first example of beta-lactams inhibiting metallo-proteinases instrumental in cancer invasion and angiogenesis. These molecules are good candidates for prototype drugs showing selective antibiotic, anti-inflammatory, and anti-invasion properties.
authors.affiliation: Department of Chemistry 'G Ciamician', University of Bologna, Via Selmi 2, I-40126 Bologna, Italy.
authors.forename: Gianfranco
authors.initials: G
authors.lastname: Cainelli
authors.forename: Paola
authors.initials: P
authors.lastname: Galletti
authors.forename: Spiridione
authors.initials: S
authors.lastname: Garbisa
authors.forename: Daria
authors.initials: D
authors.lastname: Giacomini
authors.forename: Luigi
authors.initials: L
authors.lastname: Sartor
authors.forename: Arianna
authors.initials: A
authors.lastname: Quintavalla
mesh_terms: D016475:3T3 Cells; D000818:Animals; D001384:Azetidines / Q000138:chemical synthesis / Q000494:pharmacology*; D005347:Fibroblasts / Q000187:drug effects; D018093:Gelatinases / Q000037:antagonists & inhibitors*; D006801:Humans; D020128:Inhibitory Concentration 50; D019272:Leukocyte Elastase / Q000037:antagonists & inhibitors*; D051379:Mice; D015394:Molecular Structure; D015842:Serine Proteinase Inhibitors / Q000138:chemical synthesis / Q000494:pharmacology*; D013329:Structure-Activity Relationship
chemical_list: D001384:Azetidines; D015842:Serine Proteinase Inhibitors; D019272:Leukocyte Elastase; D018093:Gelatinases