Joo Jung Hee, Huh Jeong-Eun, Lee Jee Hyun, Park Doo Ri, Lee Yoonji, Lee Seul Gee, Choi Sun, Lee Hwa Jeong, Song Seong-Won, Jeong Yongmi, Goo Ja-Il, Choi Yongseok, Baek Hye Kyung, Yi Sun Shin, Park Soo Jin, Lee Ji Eun, Ku Sae Kwang, Lee Won Jae, Lee Kee-In, Lee Soo Young, Bae Yun Soo
Department of Life Science, Ewha Womans University, Seoul, Republic of Korea.
College of Pharmacy and Graduate School of Pharmaceutical Sciences, Ewha Womans University, Seoul 120-750, Korea.
Sci Rep. 2016 Mar 15;6:22389. doi: 10.1038/srep22389.
Osteoclast cells (OCs) are differentiated from bone marrow-derived macrophages (BMMs) by activation of receptor activator of nuclear factor κB (NF-κB) ligand (RANKL). Activation of NADPH oxidase (Nox) isozymes is involved in RANKL-dependent OC differentiation, implicating Nox isozymes as therapeutic targets for treatment of osteoporosis. Here, we show that a novel pyrazole derivative, Ewha-18278 has high inhibitory potency on Nox isozymes. Blocking the activity of Nox with Ewha-18278 inhibited the responses of BMMs to RANKL, including reactive oxygen species (ROS) generation, activation of mitogen-activated protein (MAP) kinases and NF-κB, and OC differentiation. To evaluate the anti-osteoporotic function of Ewha-18278, the derivative was applied to estrogen-deficient ovariectomized (OVX) ddY mice. Oral administration of Ewha-18278 (10 mg/kg/daily, 4 weeks) into the mice recovered bone mineral density, trabecular bone volume, trabecular bone length, number and thickness, compared to control OVX ddY mice. Moreover, treatment of OVX ddY mice with Ewha-18278 increased bone strength by increasing cortical bone thickness. We provide that Ewha-18278 displayed Nox inhibition and blocked the RANKL-dependent cell signaling cascade leading to reduced differentiation of OCs. Our results implicate Ewha-18278 as a novel therapeutic agent for the treatment of osteoporosis.
破骨细胞（OCs）通过核因子κB（NF-κB）受体激活剂配体（RANKL）的激活，从骨髓来源的巨噬细胞（BMMs）分化而来。NADPH氧化酶（Nox）同工酶的激活参与RANKL依赖的OC分化，这意味着Nox同工酶是治疗骨质疏松症的治疗靶点。在此，我们表明一种新型吡唑衍生物Ewha-18278对Nox同工酶具有高抑制效力。用Ewha-18278阻断Nox的活性可抑制BMMs对RANKL的反应，包括活性氧（ROS）生成、丝裂原活化蛋白（MAP）激酶和NF-κB的激活以及OC分化。为了评估Ewha-18278的抗骨质疏松功能，将该衍生物应用于雌激素缺乏的去卵巢（OVX）ddY小鼠。与对照OVX ddY小鼠相比，对小鼠口服给予Ewha-18278（10 mg/kg/每日，4周）可恢复骨矿物质密度、小梁骨体积、小梁骨长度、数量和厚度。此外，用Ewha-18278治疗OVX ddY小鼠可通过增加皮质骨厚度来提高骨强度。我们证明Ewha-18278表现出Nox抑制作用，并阻断了RANKL依赖的细胞信号级联反应，导致OC分化减少。我们的结果表明Ewha-18278是一种治疗骨质疏松症的新型治疗剂。
