Neuroprotective Effect of N-Cyclohexylethyl-[A/G]-[D/E]-X-V Peptides on Ischemic Stroke by Blocking nNOS-CAPON Interaction
Neuroprotective Effect of N-Cyclohexylethyl-[A/G]-[D/E]-X-V Peptides on Ischemic Stroke by Blocking nNOS-CAPON Interaction
复制标题
N-环己基乙基-[A/G]-[D/E]-X-V 肽通过阻断 nNOS-CAPON 相互作用对缺血性中风的神经保护作用
DOI：
10.1021/acschemneuro.0c00739
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发表时间：
2021
影响因子：
5
通讯作者：
Li Tingyou
中科院分区：
文献类型：
--
作者：
Qin Yajuan;Feng Lingling;Fan Xin;Zheng Liping;Zhang Yu;Chang Lei;Li Tingyou
The protein–protein interaction between neuronal nitric oxide syntheses (nNOS) and the carboxy-terminal PDZ ligand of nNOS (CAPON) is a potential target for the treatment of ischemic stroke. Our previous study had identified ZLc-002 as a promising lead compound for inhibiting nNOS–CAPON coupling. To find better neuroprotective agents disrupting the ischemia-induced nNOS–CAPON interaction, a series ofN-cyclohexylethyl-[A/G]-[D/E]-X-V peptides based on the carboxy-terminal tetrapeptide of CAPON was designed, synthesized, and evaluated in this study. Herein, we reported an affinity-based fluorescence polarization (FP) method using 5-carboxyfluorescein (5-FAM) labeled CAPON (496–506) peptide as the probe for high-throughput screening of the small-molecule inhibitors of the PDZ domain of nNOS.N-Cyclohexylethyl-ADAV displayed the most potent affinity for the nNOS PDZ domain in the FP and isothermal titration calorimetry (ITC) (ΔH= −1670 ± 151.0 cal/mol) assays. To improve bioavailability, lipophilicity, and membrane permeability, the Asp methylation was employed to getN-cyclohexylethyl-AD(OMe)AV, which possesses good blood–brain barrier (BBB) permeabilityin vitroparallel artificial membrane permeability assay (PAMPA)-BBB (Pe = 6.07 cm/s) andin vivoassays. In addition,N-cyclohexylethyl-AD(OMe)AV (10 mg/kg body weight, i.v., immediately after reperfusion) substantially reduced infarct size in rats, which was measured 24 h after reperfusion and subjected to 120 min of middle cerebral artery occlusion (MCAO).
