Kataoka Shinsuke, Baba Atsuyo, Suda Yoshimitsu, Takii Ryosuke, Hashimoto Munetaka, Kawakubo Tomoyo, Asao Tetsuji, Kadowaki Tomoko, Yamamoto Kenji
Life Science Research Laboratories, Lion Corporation, Kanagawa, Japan;
Department of Pharmacology, Graduate School of Dental Science, and.
FASEB J. 2014 Aug;28(8):3564-78. doi: 10.1096/fj.14-252130. Epub 2014 Apr 28.
The periodontal pathogen Porphyromonas gingivalis produces a unique class of cysteine proteinases termed gingipains that comprises Arg-gingipain (Rgp) and Lys-gingipain (Kgp). Growing evidence indicates that these 2 types of gingipains synergistically contribute to the entire virulence of the organism and increase the risk of periodontal disease (PD) by disrupting the host immune system and degrading the host tissue and plasma proteins. Therefore, a dual inhibitor of both gingipains would have attractive clinical potential for PD therapy. In this study, a novel, potent, dual inhibitor of Rgp and Kgp was developed through structure-based drug design, and its biological potency was evaluated in vitro and in vivo. This inhibitor had low nanomolar inhibitory potency (Ki=40 nM for Rgp, Ki=0.27 nM for Kgp) and good selectivity for host proteases and exhibited potent antibacterial activity against P. gingivalis by abrogating its manifold pathophysiological functions. The therapeutic potential of this inhibitor in vivo was also verified by suppressing the vascular permeability that was enhanced in guinea pigs by the organism and the gingival inflammation in beagle dog PD models. These findings suggest that a dual inhibitor of Rgp and Kgp would exhibit noteworthy anti-inflammatory activity in the treatment of PD.
牙周病原体牙龈卟啉单胞菌产生一类独特的半胱氨酸蛋白酶，称为牙龈蛋白酶，它包括精氨酸牙龈蛋白酶（Rgp）和赖氨酸牙龈蛋白酶（Kgp）。越来越多的证据表明，这两种类型的牙龈蛋白酶协同作用，对该生物体的整体毒力有贡献，并通过破坏宿主免疫系统以及降解宿主组织和血浆蛋白，增加患牙周病（PD）的风险。因此，一种针对两种牙龈蛋白酶的双重抑制剂在PD治疗方面具有诱人的临床潜力。在本研究中，通过基于结构的药物设计开发了一种新型、高效的Rgp和Kgp双重抑制剂，并在体外和体内评估了其生物学活性。该抑制剂具有低纳摩尔抑制活性（对Rgp的Ki = 40 nM，对Kgp的Ki = 0.27 nM），对宿主蛋白酶具有良好的选择性，并且通过消除牙龈卟啉单胞菌的多种病理生理功能，对其表现出强大的抗菌活性。通过抑制该生物体在豚鼠中增强的血管通透性以及比格犬PD模型中的牙龈炎症，也验证了该抑制剂在体内的治疗潜力。这些发现表明，Rgp和Kgp双重抑制剂在PD治疗中会表现出显著的抗炎活性。
