Development of a new therapy for periodontitis by regulation of proteinase cascades

通过调节蛋白酶级联开发治疗牙周炎的新疗法

基本信息

批准号：
12557193
负责人：
TORII Mitsuo
金额：
$ 8.13万
依托单位：
Kagoshima University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12557193/
关键词：
Periodontitis Porphyromonas gingivalis gingipain DX-9065a IL-6 MMP-1 Drug for periodontitis DX9065a プロテアーゼ阻害剤 DNAマイクロアレイ活性化血液凝固第X因子 cDNAマイクロアレイ KIAA 0263 炎症性サイトカイン P.gingivalis MMP

项目摘要

Proteolytic enzymes released from the periodontopathic bacteria and the host cells are considered to be important virulent factors of periodontal diseases. However, there is no therapeutic reagent of inhibit those enzyme activities and the enzyme-associated proinflammatory events. In this study, we investigated proinflammatory activities of arginine-specific cysteine protease (Rgp) from Porphyromonas gingivalis, a causative bacterium of adult periodontitis, and activated coagulation factor X (Fxa), a product by Rgps, and the effect of DX-9065a, a new selective inhibitor of Fxa, on the proinflammatory events induced by these proteinases. Rgps and Fxa increased of interleukin-6 (IL-6) and MMP-1 by human gingival fibroblasts (HGF) at 1 nM and above, which were completely inhibited by DX-9065a in a dose-dependent manner from a concentration of 0.18μM. DX-9065a also strongly inhibited Fxa-induced IL-6 mRNA expression and NF-kB activation. DX-9065a inhibited vascular permeability enhancing activity production from human plasma both by Rgps and whole cells of P. gingivalis almost completely at 100μM. DX-9065a selectively inhibited the growth of 3 strains of P. gingivalis and prevotella intermedia in a dose-dependent manner, and the effect on p. gingivalis was bactericidal. Trypsin-like proteinase activity was detected in P. gingivalis, and the activity was strongly by DX-9065a. DX-9065a even inhibited amidolytic activity of purified trypsin-like proteinase gingipain (RgpA and RgpB) from P. gingivalis. Furthermore, trypsin-like proteinase activity in gingival crevicular fluids was strongly inhibited by DX-9065a. These results suggest that Rgps and Fxa are potential inflammatory mediators at the periodontitis site and DX-9065a is a useful therapeutic drug for the periodontal disease.

从牙周病细菌和宿主细胞释放的蛋白水解酶被认为是牙周疾病的重要有毒因素。但是，没有抑制这些酶活性和与酶相关的促炎事件的治疗试剂。在这项研究中，我们研究了卟啉胞菌的精氨酸特异性半胱氨酸蛋白酶（RGP）的促炎活性蛋白酶。人牙龈成纤维细胞（HGF）在1 nm及以上增加了白介素-6（IL-6）和MMP-1的RGP和FXA增加，DX-9065A以剂量依赖性的方式完全抑制了DX-9065A的浓度。 DX-9065a还强烈抑制了FXA诱导的IL-6 mRNA表达和NF-KB激活。 DX-9065A抑制了由RGP和牙龈疟原虫的整个细胞从100μm的RGP和整个细胞中从人血浆中增强活性产生的血管通透性。 DX-9065a以剂量依赖性方式选择性地抑制了3株牙龈斑a和prevotella Intermedia的生长，对p的影响。牙龈是细菌。在牙龈疟原虫中检测到胰蛋白酶样蛋白酶活性，DX-9065A强烈抑制了活性。 DX-9065a甚至抑制了牙龈疟原虫纯化的胰蛋白酶样蛋白酶牙龈（RGPA和RGPB）的酰化活性。此外，DX-9065A强烈抑制牙龈裂纹液中胰蛋白酶样蛋白酶活性。这些结果表明，RGP和FXA是牙周炎部位的潜在炎症介质，DX-9065A是牙周疾病的有用治疗药物。