Konda Veera Reddy, Desai Anuradha, Darland Gary, Bland Jeffrey S, Tripp Matthew L
MetaProteomics, LLC, Gig Harbor, Washington 98332, USA.
Arthritis Rheum. 2010 Jun;62(6):1683-92. doi: 10.1002/art.27441.
The multikinase inhibitor META060 has been shown to inhibit NF-kappaB activation and expression of markers of inflammation. This study was undertaken to investigate the effect of META060 on biomarkers associated with bone and cartilage degradation in vitro and its antiinflammatory efficacy in vivo in both acute and chronic inflammation models.
Glycogen synthase kinase 3beta (GSK3beta)-dependent beta-catenin phosphorylation was evaluated in RAW 264.7 macrophages to assess kinase inhibition. The inhibition of osteoclastogenesis and tartrate-resistant acid phosphatase (TRAP) activity was evaluated in RANKL-treated RAW 264.7 cells. The inhibition of interleukin-1beta (IL-1beta)-mediated markers of inflammation was analyzed in human rheumatoid arthritis synovial fibroblasts (RASFs). Mice with carrageenan-induced acute inflammation and collagen-induced arthritis (CIA) were used to assess efficacy.
META060 inhibited the activity of kinases (spleen tyrosine kinase [Syk], Bruton's tyrosine kinase [Btk], phosphatidylinositol 3-kinase [PI 3-kinase], and GSK3) associated with RA and inhibited beta-catenin phosphorylation. META060 inhibited osteoclastogenesis, as indicated by decreased transformation of RAW 264.7 cells to osteoclasts and reduced TRAP activity, and inhibited IL-1beta-activated prostaglandin E(2), matrix metalloproteinase 3, IL-6, IL-8, and monocyte chemotactic protein 1 in RASFs. In mice with acute inflammation, oral administration of META060 reduced paw swelling similar to the effect of aspirin. In mice with CIA, META060 significantly reduced the arthritis index and decreased bone, joint, and cartilage degradation. Serum IL-6 concentrations in these mice were inhibited in a dose-dependent manner.
Our findings indicate that META060 reduces swelling in a model of acute inflammation and inhibits bone and cartilage destruction in a model of chronic inflammation. Its efficacy is associated with the inhibition of multiple protein kinases, including Syk, Btk, PI 3-kinase, and GSK3. These results warrant further clinical testing of META060 for its therapeutic potential in the treatment of inflammatory diseases.
多激酶抑制剂META060已被证明可抑制核因子κB的激活及炎症标志物的表达。本研究旨在探讨META060对体外与骨和软骨降解相关生物标志物的影响及其在急性和慢性炎症模型中的体内抗炎疗效。
在RAW 264.7巨噬细胞中评估糖原合酶激酶3β（GSK3β）依赖性β-连环蛋白磷酸化，以评估激酶抑制情况。在经RANKL处理的RAW 264.7细胞中评估破骨细胞生成及抗酒石酸酸性磷酸酶（TRAP）活性的抑制情况。在人类风湿关节炎滑膜成纤维细胞（RASFs）中分析白细胞介素-1β（IL-1β）介导的炎症标志物的抑制情况。使用角叉菜胶诱导的急性炎症小鼠和胶原诱导的关节炎（CIA）小鼠评估疗效。
META060抑制了与类风湿关节炎相关的激酶（脾酪氨酸激酶[Syk]、布鲁顿酪氨酸激酶[Btk]、磷脂酰肌醇3激酶[PI 3激酶]和GSK3）的活性，并抑制β-连环蛋白磷酸化。META060抑制破骨细胞生成，表现为RAW 264.7细胞向破骨细胞的转化减少及TRAP活性降低，并抑制RASFs中IL-1β激活的前列腺素E2、基质金属蛋白酶3、IL-6、IL-8和单核细胞趋化蛋白1。在急性炎症小鼠中，口服META060可减轻爪肿胀，效果与阿司匹林相似。在CIA小鼠中，META060显著降低关节炎指数，并减少骨、关节和软骨破坏。这些小鼠血清IL-6浓度呈剂量依赖性受到抑制。
我们的研究结果表明，META060可减轻急性炎症模型中的肿胀，并抑制慢性炎症模型中的骨和软骨破坏。其疗效与抑制包括Syk、Btk、PI 3激酶和GSK3在内的多种蛋白激酶有关。这些结果为进一步临床测试META060在治疗炎症性疾病方面的治疗潜力提供了依据。
