Development of a Rapid Throughput Assay for Identification of hNav1.7 Antagonist Using Unique Efficacious Sodium Channel Agonist, Antillatoxin
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- Fang Zhao
- State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing 211198, China
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- Xichun Li
- State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing 211198, China
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- Liang Jin
- State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing 211198, China
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- Fan Zhang
- Jiangsu Provincial Key laboratory for TCM Evaluation and Translational Development, China Pharmaceutical University, Nanjing 211198, China
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- Masayuki Inoue
- Graduate School of Pharmaceutical Sciences, The University of Tokyo, Tokyo 113-0033, Japan
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- Boyang Yu
- State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing 211198, China
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- Zhengyu Cao
- State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing 211198, China
書誌事項
- 公開日
- 2016-02-16
- 資源種別
- journal article
- 権利情報
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- https://creativecommons.org/licenses/by/4.0/
- DOI
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- 10.3390/md14020036
- 公開者
- MDPI AG
説明
<jats:p>Voltage-gated sodium channels (VGSCs) are responsible for the generation of the action potential. Among nine classified VGSC subtypes (Nav1.1–Nav1.9), Nav1.7 is primarily expressed in the sensory neurons, contributing to the nociception transmission. Therefore Nav1.7 becomes a promising target for analgesic drug development. In this study, we compared the influence of an array of VGSC agonists including veratridine, BmK NT1, brevetoxin-2, deltamethrin and antillatoxin (ATX) on membrane depolarization which was detected by Fluorescence Imaging Plate Reader (FLIPR) membrane potential (FMP) blue dye. In HEK-293 cells heterologously expressing hNav1.7 α-subunit, ATX produced a robust membrane depolarization with an EC50 value of 7.8 ± 2.9 nM whereas veratridine, BmK NT1, and deltamethrin produced marginal response. Brevetoxin-2 was without effect on membrane potential change. The ATX response was completely inhibited by tetrodotoxin suggesting that the ATX response was solely derived from hNav1.7 activation, which was consistent with the results where ATX produced a negligible response in null HEK-293 cells. Six VGSC antagonists including lidocaine, lamotrigine, phenytoin, carbamazepine, riluzole, and 2-amino-6-trifluoromethylthiobenzothiazole all concentration-dependently inhibited ATX response with IC50 values comparable to that reported from patch-clamp experiments. Considered together, we demonstrate that ATX is a unique efficacious hNav1.7 activator which offers a useful probe to develop a rapid throughput screening assay to identify hNav1.7 antagonists.</jats:p>
収録刊行物
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- Marine Drugs
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Marine Drugs 14 (2), 36-, 2016-02-16
MDPI AG
- Tweet
キーワード
- antillatoxin; FMPblue; membrane potential; hNa<sub>v</sub>1.7; rapid throughput
- Patch-Clamp Techniques
- QH301-705.5
- Voltage-Gated Sodium Channels
- Peptides, Cyclic
- Article
- antillatoxin
- Inhibitory Concentration 50
- Lipopeptides
- Drug Discovery
- hNav1.7
- Humans
- FMPblue
- Biology (General)
- Voltage-Gated Sodium Channel Blockers
- Analgesics
- Dose-Response Relationship, Drug
- 3002 Drug Discovery
- NAV1.7 Voltage-Gated Sodium Channel
- 500
- Voltage-Gated Sodium Channel Agonists
- High-Throughput Screening Assays
- HEK293 Cells
- Drug Design
- rapid throughput
- membrane potential
- Sodium Channel Blockers
詳細情報 詳細情報について
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- CRID
- 1360004239503347072
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- ISSN
- 16603397
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- PubMed
- 26891306
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- 資料種別
- journal article
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- データソース種別
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- Crossref
- KAKEN
- OpenAIRE
