BMS-345541 is a highly selective inhibitor of IκB kinase that binds at an allosteric site of the enzyme and blocks NF-κB-dependent transcription in mice
BMS-345541 is a highly selective inhibitor of IκB kinase that binds at an allosteric site of the enzyme and blocks NF-κB-dependent transcription in mice
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DOI：
10.1074/jbc.m209677200
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发表时间：
2003-01-17
影响因子：
4.8
通讯作者：
Zusi, FC
中科院分区：
文献类型：
--
作者：
Burke, JR;Pattoli, MA;Zusi, FC
IkappaBalpha丝氨酸32和36的信号诱导磷酸化在调节IkappaBalpha随后的泛素化和蛋白水解中至关重要，然后释放NF-kappaB促进基因转录。负责这种磷酸化的多亚基licB激酶包含两个催化亚基，称为licB激酶(IKK)-1和IKK-2。BMS-345541（4（2′-氨基乙基）氨基-1,8-二甲基咪唑（1,2-a）喹诺啉）是IKK催化亚基（IKK-2 IC50 = 0.3 muM, IKK-1 IC50 = 4 muM）的选择性抑制剂。该化合物不能抑制其他15种激酶，并选择性地抑制细胞中IkappaBalpha的受刺激磷酸化（IC50 = 4 muM），而不能影响细胞中c-Jun和STAT3的磷酸化，以及丝裂原活化蛋白激酶活化蛋白激酶2的活化。与IKK/NF-kappaB调控细胞因子转录的作用一致，BMS-345541抑制脂多糖刺激的THP-1细胞中的肿瘤坏死因子a、白细胞介素-1 β、白细胞介素-8和白细胞介素-6，IC50值在1 ~ 5-muM范围内。虽然BMS-345541对IKK-2抑制的Dixon图显示出非线性关系，表明非michaelis - menten动力学结合，但多重抑制分析表明，BMS-345541与IkappaBalpha的26-42氨基酸对应的肽抑制剂Ser-32和Ser-36以互斥的方式结合，与ADP的非互斥方式结合。在研究与IKK-1的结合时得到了相反的结果。提出了BMS-345541结合IKK-1和IKK-2上相似的变构位点的结合模型，从而对亚基的活性位点产生不同的影响。BMS-345541在小鼠体内也显示出良好的药代动力学，经口服表明该化合物在脂多糖腹腔刺激后能剂量依赖性地抑制血清肿瘤坏死因子a的产生。因此，该化合物对小鼠NF-kappaB活化有效，是研究IKK在疾病模型中的作用的重要工具。
The signal-inducible phosphorylation of serines 32 and 36 of IkappaBalpha is critical in regulating the subsequent ubiquitination and proteolysis of IkappaBalpha, which then releases NF-kappaB to promote gene transcription. The multi-subunit licB kinase responsible for this phosphorylation contains two catalytic subunits, termed licB kinase (IKK)-1 and IKK-2. BMS-345541 (4(2'-aminoethyl)amino-1,8-dimethylimidazo(1,2-a)quinoxaline) was identified as a selective inhibitor of the catalytic subunits of IKK (IKK-2 IC50 = 0.3 muM, IKK-1 IC50 = 4 muM). The compound failed to inhibit a panel of 15 other kinases and selectively inhibited the stimulated phosphorylation Of IkappaBalpha in cells (IC50 = 4 muM) while failing to affect c-Jun and STAT3 phosphorylation, as well as mitogen-activated protein kinase-activated protein kinase 2 activation in cells. Consistent with the role of IKK/NF-kappaB in the regulation of cytokine transcription, BMS-345541 inhibited lipopolysaccharide-stimulated tumor necrosis factor a, interleukin-1beta, interleukin-8, and interleukin-6 in THP-1 cells with IC50 values in the 1- to 5-muM range. Although a Dixon plot of the inhibition of IKK-2 by BMS-345541 showed a non-linear relationship indicating non-Michaelis-Menten kinetic binding, the use of multiple inhibition analyses indicated that BMS-345541 binds in a mutually exclusive manner with respect to a peptide inhibitor corresponding to amino acids 26-42 of IkappaBalpha with Ser-32 and Ser-36 changed to aspartates and in a non-mutually exclusive manner with respect to ADP. The opposite results were obtained when studying the binding to IKK-1. A binding model is proposed in which BMS-345541 binds to similar allosteric sites on IKK-1 and IKK-2, which then affects the active sites of the subunits differently. BMS-345541 was also shown to have excellent pharmacokinetics in mice, and peroral administration showed the compound to dose-dependently inhibit the production of serum tumor necrosis factor a following intraperitoneal challenge with lipopolysaccharide. Thus, the compound is effective against NF-kappaB activation in mice and represents an important tool for investigating the role of IKK in disease models.