A drug development study for a periodontal disease : Biological properties of synthetic lipid A of Porphyromonas gingivalis lipopolysaccharide

牙周病药物开发研究：牙龈卟啉单胞菌脂多糖合成脂质A的生物学特性

• 批准号: 17592170
• 负责人: KUMADA Hidefumi
• 金额: $ 2.3万
• 依托单位: Kanagawa Dental College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2005
• 资助国家: 日本
• 起止时间: 2005 至 2006
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-17592170/
• 关键词: periodontal disease; lipopolysaccharide; synthetic lipid A; biological properties; Porphyromanas gingivalis; 化学合成

A pentaacyl and diphosphoryl lipid A molecule (J Bacteriol 1995: 177: 2098) found in the lipid A isolated from P. gingivalis LPS was chemically synthesized, and its characteristics were evaluated to reconfirm its interesting bioactivities including low endotoxicity and activity against LPS-unresponsive C3H/HeJ mouse cells. The synthesized P. gingivalis lipid A (synthetic Pg-LA) exhibited strong activities almost equivalent to those of Escherichia coli-type synthetic lipid A (compound 506) in all assays on LPS-responsive mice, and cells. LPS and native lipid A of P. gingivalis displayed overall endotoxic activities, but its potency was reduced in comparison to the synthetic analogs. In the assays using C3H/HeJ mouse cells, the LPS and native lipid A significantly stimulated splenocytes to cause mitosis, and peritoneal macrophages to induce TNF-D and IL-6 production. However, synthetic Pg-LA and compound 506 showed no activity on the LPS-unresponsive cells. Inhibition assays using some i … More nhibitors including anti-human TLR 2 and TLR4/MD-2 complex monoclonal antibodies showed that the biological activity of synthetic Pg-LA was mediated only through the TLR4 signaling pathway that might act as a receptor for LPS, whereas TLR2, possibly together with CD14, was associated with the signaling cascade for LPS and native lipid A of P. gingivalis, in addition to the TLR4 pathway. These results suggested that the moderated and reduced biological activity of P. gingivalis LPS and native lipid A, including the activity on C3H/HeJ mouse cells by TLR2-mediated pathway, may be mediated by bioactive contaminants or low acylated molecules present in the native preparations having multiple lipid A moieties.In conclusion, these findings suggested that the moderated and reduced biological activity of P. gingivalis LPS and native lipid A, including the activity on C3H/HeJ mouse cells by TLR2-mediated pathway, may be mediated by bioactive contaminants or low acylated molecules present in the native preparations having high heterogeneity in lipid A moiety. To elucidate these problems, we are now attempting to evaluate the biological characterizations of tetra-acylated monophosphorylated or diphosphorylated species with the predominant molecules found in P. gingivalis native lipid A, using each chemically synthesized analog. Less

A pentaacyl and diphosphoryl lipid A molecule (J Bacteriol 1995: 177: 2098) found in the lipid A isolated from P. gingivalis LPS was chemically synthesized, and its characteristics were evaluated to reconfirm its interesting bioactivities including low endotoxicity and activity against LPS-unresponsive C3H/HeJ mouse cells.在LPS响应性小鼠和细胞上的所有测定中，合成的牙龈疟原虫脂质A（合成PG-LA）几乎表现出强大的活性，几乎相当于coli-type合成脂质A（化合物506）的活性。牙龈疟原虫的LPS和天然脂质A显示出总体内毒性活性，但与合成类似物相比，其效力降低了。在使用C3H/HEJ小鼠细胞的测定中，LPS和天然脂质有明显刺激的脾细胞引起有丝分裂，以及腹膜巨噬细胞诱导TNF-D和IL-6产生。然而，合成PG-LA和化合物506在LPS无反发细胞上没有活性。使用某些I…更多的核糖体（包括抗人TLR 2和TLR4/MD-2复杂的单克隆抗体）的抑制作用调查表明，合成PG-LA的生物学活性仅通过TLR4信号传导途径介导，这些信号传导途径可能与LPS的接收器以及CASC相关，而CASC则与CD14相关的接收器，而CD14则可能与CASC相关。除TLR4途径外，牙龈。这些结果表明，在TLR2介导的途径上，通过TLR2介导的途径在C3H/HEJ小鼠细胞上的适度和减少的生物学活性可能是通过生物活性污染物或低酰基化的分子介导的，或在生物学制剂中存在于Moieties and Moieties。 Gingivalis LPS和天然脂质A，包括通过TLR2介导的途径对C3H/HEJ小鼠细胞的活性，可以通过生物活性污染物或在脂质A部分中具有高异质性的天然制剂中存在的生物活性污染物或低酰基化的分子介导。为了阐明这些问题，我们使用每种化学合成的类似物，试图评估四酰化单磷酸化或二磷酸化的物种的生物学特性，其主要分子在牙龈疟原虫天然脂质A中发现的主要分子。较少的