2. Academic Validation
  3. Engineering of highly potent and selective HNTX-III mutant against hNav1.7 sodium channel for treatment of pain

Engineering of highly potent and selective HNTX-III mutant against hNav1.7 sodium channel for treatment of pain

  • J Biol Chem. 2021 Jan-Jun:296:100326. doi: 10.1016/j.jbc.2021.100326.
Yunxiao Zhang 1,  Li Wang 2,  Dezheng Peng 1,  Qingfeng Zhang 2,  Qiuchu Yang 2,  Jiayan Li 2,  Dan Li 2,  Dongfang Tang 2,  Minzhi Chen 2,  Songping Liang 2,  Yu Liu 3,  Sheng Wang 4,  Zhonghua Liu 5
Affiliations

Affiliations

  • 1 The National and Local Joint Engineering Laboratory of Animal Peptide Drug Development, College of Life Sciences, Hunan Normal University, Changsha, Hunan, China; Key Laboratory of Hunan Province for Advanced Carbon-based Functional Materials, School of Chemistry and Chemical Engineering, Hunan Institute of Science and Technology, Yueyang, Hunan, China.
  • 2 The National and Local Joint Engineering Laboratory of Animal Peptide Drug Development, College of Life Sciences, Hunan Normal University, Changsha, Hunan, China.
  • 3 Key Laboratory of Hunan Province for Advanced Carbon-based Functional Materials, School of Chemistry and Chemical Engineering, Hunan Institute of Science and Technology, Yueyang, Hunan, China. Electronic address: [email protected].
  • 4 Key Laboratory of Molecular Biophysics of the Ministry of Education, College of Life Science and Technology, Huazhong University of Science and Technology, Wuhan, Hubei, China. Electronic address: [email protected].
  • 5 The National and Local Joint Engineering Laboratory of Animal Peptide Drug Development, College of Life Sciences, Hunan Normal University, Changsha, Hunan, China. Electronic address: [email protected].
PMID: 33493520 DOI: 10.1016/j.jbc.2021.100326
Abstract

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.

Keywords

HNTX-III; analgesia; hNa(v)1.7; peptide; structure activity relationship.

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