PloS one, 2018-05, Vol.13 (5), p.e0196791-e0196791
2018
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- Autor(en) / Beteiligte
- Titel
- Pharmacological characterization of potent and selective NaV1.7 inhibitors engineered from Chilobrachys jingzhao tarantula venom peptide JzTx-V
- Ist Teil von
- PloS one, 2018-05, Vol.13 (5), p.e0196791-e0196791
- Ort / Verlag
- United States: Public Library of Science
- Erscheinungsjahr
- 2018
- Quelle
- Free E-Journal (出版社公開部分のみ）
- Beschreibungen/Notizen
- Identification of voltage-gated sodium channel NaV1.7 inhibitors for chronic pain therapeutic development is an area of vigorous pursuit. In an effort to identify more potent leads compared to our previously reported GpTx-1 peptide series, electrophysiology screening of fractionated tarantula venom discovered the NaV1.7 inhibitory peptide JzTx-V from the Chinese earth tiger tarantula Chilobrachys jingzhao. The parent peptide displayed nominal selectivity over the skeletal muscle NaV1.4 channel. Attribute-based positional scan analoging identified a key Ile28Glu mutation that improved NaV1.4 selectivity over 100-fold, and further optimization yielded the potent and selective peptide leads AM-8145 and AM-0422. NMR analyses revealed that the Ile28Glu substitution changed peptide conformation, pointing to a structural rationale for the selectivity gains. AM-8145 and AM-0422 as well as GpTx-1 and HwTx-IV competed for ProTx-II binding in HEK293 cells expressing human NaV1.7, suggesting that these NaV1.7 inhibitory peptides interact with a similar binding site. AM-8145 potently blocked native tetrodotoxin-sensitive (TTX-S) channels in mouse dorsal root ganglia (DRG) neurons, exhibited 30- to 120-fold selectivity over other human TTX-S channels and exhibited over 1,000-fold selectivity over other human tetrodotoxin-resistant (TTX-R) channels. Leveraging NaV1.7-NaV1.5 chimeras containing various voltage-sensor and pore regions, AM-8145 mapped to the second voltage-sensor domain of NaV1.7. AM-0422, but not the inactive peptide analog AM-8374, dose-dependently blocked capsaicin-induced DRG neuron action potential firing using a multi-electrode array readout and mechanically-induced C-fiber spiking in a saphenous skin-nerve preparation. Collectively, AM-8145 and AM-0422 represent potent, new engineered NaV1.7 inhibitory peptides derived from the JzTx-V scaffold with improved NaV selectivity and biological activity in blocking action potential firing in both DRG neurons and C-fibers.
- Sprache
- Englisch
- Identifikatoren
- ISSN: 1932-6203eISSN: 1932-6203DOI: 10.1371/journal.pone.0196791Titel-ID: cdi_plos_journals_2034348513
- Format
- –
- Schlagworte
- Action potential, Action Potentials - drug effects, Amino Acid Substitution, Analgesics - isolation & purification, Analgesics - pharmacology, Animals, Binding sites, Biological activity, Biology and Life Sciences, Capsaicin, Capsaicin - pharmacology, Cell Line, Chilobrachys jingzhao, Chimeras, Chronic pain, Conformation, Dorsal root ganglia, Drug Evaluation, Preclinical, Electric potential, Electrophysiology, Engineering and Technology, Ganglia, Ganglia, Spinal - drug effects, Humans, Inhibitors, Male, Medicine and Health Sciences, Mice, Inbred C57BL, Muscles, Mutagenesis, Site-Directed, Mutation, NAV1.7 Voltage-Gated Sodium Channel - drug effects, Nerve Fibers, Unmyelinated - drug effects, Neurons, NMR, Nuclear magnetic resonance, Nuclear Magnetic Resonance, Biomolecular, Optimization, Pain, Patch-Clamp Techniques, People and Places, Peptides, Peptides - chemistry, Pharmacology, Physical Stimulation, Protein Engineering, Recombinant Proteins - drug effects, Science Policy, Selectivity, Skeletal muscle, Skin, Skin preparations, Sodium Channel Blockers - isolation & purification, Sodium Channel Blockers - pharmacology, Sodium channels (voltage-gated), Spider Venoms - chemistry, Structure-Activity Relationship, Tetrodotoxin, Tetrodotoxin - pharmacology, Venom