title: Benzamidine derivatives inhibit the virulence of Porphyromonas gingivalis.
abstract: We have previously shown that benzamidine-type compounds can inhibit the activity of arginine-specific cysteine proteinases (gingipains HRgpA and RgpB); well-known virulence factors of Porphyromonas gingivalis. They also hinder in vitro growth of this important periodontopathogenic bacterium. Apparently growth arrest is not associated with their ability to inhibit these proteases, because pentamidine, which is a 20-fold less efficient inhibitor of gingipain than 2,6-bis-(4-amidinobenzyl)-cyclohexanone (ACH), blocked P. gingivalis growth far more effectively. To identify targets for benzamidine-derived compounds other than Arg-gingipains, and to explain their bacteriostatic effects, P. gingivalis ATCC 33277 and P. gingivalis M5-1-2 (clinical isolate) cell extracts were subjected to affinity chromatography using a benzamidine-Sepharose column to identify proteins interacting with benzamidine. In addition to HRgpA and RgpB the analysis revealed heat-shock protein GroEL as another ligand for benzamidine. To better understand the effect of benzamidine-derived compounds on P. gingivalis, bacteria were exposed to benzamidine, pentamidine, ACH and heat, and the expression of gingipains and GroEL was determined. Exposure to heat and benzamidine-derived compounds caused significant increases in GroEL, at both the mRNA and protein levels. Interestingly, despite the fact that gingipains were shown to be the main virulence factors in a fertilized egg model of infection, mortality rates were strongly reduced, not only by ACH, but also by pentamidine, a relatively weak gingipain inhibitor. This effect may depend not only on gingipain inhibition but also on interaction of benzamidine derivatives with GroEL. Therefore these compounds may find use in supportive periodontitis treatment.
authors.affiliation: Department of Experimental Anesthesiology, Clinic for Anesthesiology and Intensive Care Medicine, Jena University Hospital, Jena, Germany.
authors.forename: E
authors.initials: E
authors.lastname: Fröhlich
authors.forename: T
authors.initials: T
authors.lastname: Kantyka
authors.forename: K
authors.initials: K
authors.lastname: Plaza
authors.forename: K-H
authors.initials: KH
authors.lastname: Schmidt
authors.forename: W
authors.initials: W
authors.lastname: Pfister
authors.forename: J
authors.initials: J
authors.lastname: Potempa
authors.forename: S
authors.initials: S
authors.lastname: Eick
mesh_terms: D018829:Adhesins, Bacterial / Q000096:biosynthesis / Q000235:genetics; D000818:Animals; D001550:Benzamidines / Q000494:pharmacology*; D018834:Chaperonin 60 / Q000037:antagonists & inhibitors / Q000096:biosynthesis / Q000235:genetics; D002642:Chick Embryo; D002846:Chromatography, Affinity; D003512:Cyclohexanones / Q000494:pharmacology*; D003546:Cysteine Endopeptidases / Q000096:biosynthesis / Q000235:genetics; D000080867:Gingipain Cysteine Endopeptidases; D006358:Hot Temperature; D010419:Pentamidine / Q000494:pharmacology*; D016966:Porphyromonas gingivalis / Q000187:drug effects* / Q000472:pathogenicity*; D014774:Virulence / Q000187:drug effects; D037521:Virulence Factors / Q000037:antagonists & inhibitors / Q000096:biosynthesis
chemical_list: C012777:2,6-bis(4-amidinobenzyl)cyclohexanone; D018829:Adhesins, Bacterial; D001550:Benzamidines; D018834:Chaperonin 60; D003512:Cyclohexanones; D000080867:Gingipain Cysteine Endopeptidases; D037521:Virulence Factors; D010419:Pentamidine; D003546:Cysteine Endopeptidases; C032157:benzamidine