DX-9065a inhibits proinflammatory events induced by gingipains and factor Xa.
OBJECTIVE
Arginine-specific cysteine proteases (Rgps) from Porphyromonas gingivalis are important virulent factors of periodontal diseases. However, there is no therapeutic drug that inhibits proinflammatory events induced by these enzymes. In this study, we investigated proinflammatory activities of Rgps and activated coagulation factor X (FXa) and examined the effect of DX-9065a, a new selective inhibitor of FXa, on proinflammatory events induced by these proteinases.


METHODS
Human gingival fibroblasts were stimulated with Rgps and FXa in the presence or absence of DX-9065a, and then interleukin-6 (IL-6) and matrix metalloproteinase-1 (MMP-1) release, their mRNA expression, and nuclear factor kappaB (NF-kappaB) activation were assessed using an enzyme-linked immunosorbent assay (ELISA), northern blotting, and a gel-mobility shift method, respectively.


RESULTS
Rgps and FXa activated IL-6 and MMP-1 release in human gingival fibroblasts through their amidolytic activities and in mitogen-activated protein kinase (MAPK) and NF-kappaB dependent manners. DX-9065a inhibited FXa-induced IL-6 mRNA expression and NF-kappaB activation. DX-9065a inhibited amidolytic activities of FXa and Rgps in vitro and ex vivo.


CONCLUSIONS
Rgps and FXa are potent inflammatory mediators and DX-9065a may be a useful therapeutic drug for periodontal disease.
Matsushita
Kenji
K
Department of Oral Disease Research, National Institute for Longevity Science, Aichi, Japan. kmatsu30@nils.go.jp
Imamura
Takahisa
T
Tomikawa
Munehiro
M
Tancharoen
Salunya
S
Tatsuyama
Syouko
S
Maruyama
Ikuro
I
D018829:Adhesins, Bacterial / Q000187:drug effects / Q000502:physiology*; D000925:Anticoagulants / Q000037:antagonists & inhibitors / Q000494:pharmacology*; D015152:Blotting, Northern; D002478:Cells, Cultured; D003546:Cysteine Endopeptidases / Q000187:drug effects / Q000502:physiology*; D015853:Cysteine Proteinase Inhibitors / Q000494:pharmacology; D004591:Electrophoresis, Polyacrylamide Gel; D015951:Factor Xa / Q000502:physiology*; D065427:Factor Xa Inhibitors; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism; D000080867:Gingipain Cysteine Endopeptidases; D005881:Gingiva / Q000166:cytology / Q000378:metabolism*; D005883:Gingival Crevicular Fluid / Q000201:enzymology; D006801:Humans; D018836:Inflammation Mediators / Q000037:antagonists & inhibitors*; D015850:Interleukin-6 / Q000096:biosynthesis; D020935:MAP Kinase Signaling System / Q000187:drug effects; D020781:Matrix Metalloproteinase 1 / Q000096:biosynthesis; D016328:NF-kappa B / Q000037:antagonists & inhibitors; D009281:Naphthalenes / Q000494:pharmacology*; D011422:Propionates / Q000494:pharmacology*; D015842:Serine Proteinase Inhibitors / Q000494:pharmacology*; D037521:Virulence Factors; D000093242:Interleukin-6 Inhibitors
D018829:Adhesins, Bacterial; D000925:Anticoagulants; D003546:Cysteine Endopeptidases; D015853:Cysteine Proteinase Inhibitors; D015951:Factor Xa; D065427:Factor Xa Inhibitors; D000080867:Gingipain Cysteine Endopeptidases; D018836:Inflammation Mediators; D015850:Interleukin-6; D020781:Matrix Metalloproteinase 1; D016328:NF-kappa B; D009281:Naphthalenes; D011422:Propionates; D015842:Serine Proteinase Inhibitors; D037521:Virulence Factors; D000093242:Interleukin-6 Inhibitors; C086910:(2S)-2-(4-(((3S)-1-acetimidoyl-3-pyrrolidinyl)oxy)phenyl)-3-(7-amidino-2-naphtyl)propanoic acid