Engineering of highly potent and selective HNTX-III mutant against hNa(v)1.7 sodium channel for treatment of pain.
Engineering of highly potent and selective HNTX-III mutant against hNa(v)1.7 sodium channel for treatment of pain.
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针对 hNav1.7 钠通道的高效选择性 HNTX-III 突变体工程用于治疗疼痛
DOI：
10.1016/j.jbc.2021.100326
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发表时间：
2021-01
期刊：
影响因子：
--
通讯作者：
Liu Z
中科院分区：
文献类型：
--
作者：
Zhang Y;Wang L;Peng D;Zhang Q;Yang Q;Li J;Li D;Tang D;Chen M;Liang S;Liu Y;Wang S;Liu Z
人电压门控钠通道Nav1.7（hNav1.7）参与神经性和伤害性疼痛信号的产生和传导。令人信服的遗传和临床前研究已经证实，hNav1.7是治疗疼痛的治疗靶点;然而，目前缺乏能够以高效力和特异性靶向hNav1.7的化合物。海南毒素-III（HNTX-III）是来自海南鸟蛛（Ornithoctonus hainana）毒液的33个残基的多肽。它是神经元河豚毒素敏感性电压门控钠通道的选择性拮抗剂。在这里，我们报告了源自HNTX-III支架的hNav1.7抑制肽的改进的效力和Nav选择性的工程化。丙氨酸扫描诱变显示HNTX-III与hNav1.7相互作用的关键残基。定点突变分析表明hNav1.7上的关键残基与HNTX-III相互作用。进行分子对接以阐明HNTX-III与Nav1.7之间的结合界面并指导分子工程过程。最终，我们基于HNTX-III和hNav1.7的分子对接获得了H4 [K 0 G1-P18 K-A21 L-V]，其效价提高了30倍（IC 50 0.007 ± 0.001 μM），对Nav1.4和Nav1.5的选择性提高了>1000倍。H4在急性和慢性炎性疼痛模型和神经性疼痛模型中也显示出稳健的镇痛作用。因此，我们的研究结果提供了对肽毒素的进一步了解，这些肽毒素可能有助于指导开发具有改善的效力和选择性的抑制剂，用于具有强大镇痛作用的Nav亚型。
Human voltage-gated sodium channel Nav1.7 (hNav1.7) is involved in the generation and conduction of neuropathic and nociceptive pain signals. Compelling genetic and preclinical studies have validated that hNav1.7 is a therapeutic target for the treatment of pain; however, there is a dearth of currently available compounds capable of targeting hNav1.7 with high potency and specificity. Hainantoxin-III (HNTX-III) is a 33-residue polypeptide from the venom of the spider Ornithoctonus hainana. It is a selective antagonist of neuronal tetrodotoxin-sensitive voltage-gated sodium channels. Here, we report the engineering of improved potency and Nav selectivity of hNav1.7 inhibition peptides derived from the HNTX-III scaffold. Alanine scanning mutagenesis showed key residues for HNTX-III interacting with hNav1.7. Site-directed mutagenesis analysis indicated key residues on hNav1.7 interacting with HNTX-III. Molecular docking was conducted to clarify the binding interface between HNTX-III and Nav1.7 and guide the molecular engineering process. Ultimately, we obtained H4 [K0G1-P18K-A21L-V] based on molecular docking of HNTX-III and hNav1.7 with a 30-fold improved potency (IC50 0.007 ± 0.001 μM) and >1000-fold selectivity against Nav1.4 and Nav1.5. H4 also showed robust analgesia in the acute and chronic inflammatory pain model and neuropathic pain model. Thus, our results provide further insight into peptide toxins that may prove useful in guiding the development of inhibitors with improved potency and selectivity for Nav subtypes with robust analgesia.
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