Tags

Type your tag names separated by a space and hit enter

Seven novel modulators of the analgesic target NaV 1.7 uncovered using a high-throughput venom-based discovery approach.
Br J Pharmacol. 2015 May; 172(10):2445-58.BJ

Abstract

BACKGROUND AND PURPOSE

Chronic pain is a serious worldwide health issue, with current analgesics having limited efficacy and dose-limiting side effects. Humans with loss-of-function mutations in the voltage-gated sodium channel NaV 1.7 (hNaV 1.7) are indifferent to pain, making hNaV 1.7 a promising target for analgesic development. Since spider venoms are replete with NaV channel modulators, we examined their potential as a source of hNaV 1.7 inhibitors.

EXPERIMENTAL APPROACH

We developed a high-throughput fluorescent-based assay to screen spider venoms against hNaV 1.7 and isolate 'hit' peptides. To examine the binding site of these peptides, we constructed a panel of chimeric channels in which the S3b-S4 paddle motif from each voltage sensor domain of hNaV 1.7 was transplanted into the homotetrameric KV 2.1 channel.

KEY RESULTS

We screened 205 spider venoms and found that 40% contain at least one inhibitor of hNaV 1.7. By deconvoluting 'hit' venoms, we discovered seven novel members of the NaSpTx family 1. One of these peptides, Hd1a (peptide μ-TRTX-Hd1a from venom of the spider Haplopelma doriae), inhibited hNaV 1.7 with a high level of selectivity over all other subtypes, except hNaV 1.1. We showed that Hd1a is a gating modifier that inhibits hNaV 1.7 by interacting with the S3b-S4 paddle motif in channel domain II. The structure of Hd1a, determined using heteronuclear NMR, contains an inhibitor cystine knot motif that is likely to confer high levels of chemical, thermal and biological stability.

CONCLUSION AND IMPLICATIONS

Our data indicate that spider venoms are a rich natural source of hNaV 1.7 inhibitors that might be useful leads for the development of novel analgesics.

Authors+Show Affiliations

Centre for Pain Research, Institute for Molecular Bioscience, St. Lucia, Qld, Australia.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article
Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

25754331

Citation

Klint, Julie K., et al. "Seven Novel Modulators of the Analgesic Target NaV 1.7 Uncovered Using a High-throughput Venom-based Discovery Approach." British Journal of Pharmacology, vol. 172, no. 10, 2015, pp. 2445-58.
Klint JK, Smith JJ, Vetter I, et al. Seven novel modulators of the analgesic target NaV 1.7 uncovered using a high-throughput venom-based discovery approach. Br J Pharmacol. 2015;172(10):2445-58.
Klint, J. K., Smith, J. J., Vetter, I., Rupasinghe, D. B., Er, S. Y., Senff, S., Herzig, V., Mobli, M., Lewis, R. J., Bosmans, F., & King, G. F. (2015). Seven novel modulators of the analgesic target NaV 1.7 uncovered using a high-throughput venom-based discovery approach. British Journal of Pharmacology, 172(10), 2445-58. https://doi.org/10.1111/bph.13081
Klint JK, et al. Seven Novel Modulators of the Analgesic Target NaV 1.7 Uncovered Using a High-throughput Venom-based Discovery Approach. Br J Pharmacol. 2015;172(10):2445-58. PubMed PMID: 25754331.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Seven novel modulators of the analgesic target NaV 1.7 uncovered using a high-throughput venom-based discovery approach. AU - Klint,Julie K, AU - Smith,Jennifer J, AU - Vetter,Irina, AU - Rupasinghe,Darshani B, AU - Er,Sing Yan, AU - Senff,Sebastian, AU - Herzig,Volker, AU - Mobli,Mehdi, AU - Lewis,Richard J, AU - Bosmans,Frank, AU - King,Glenn F, Y1 - 2015/03/04/ PY - 2014/07/03/received PY - 2014/11/08/revised PY - 2014/12/08/accepted PY - 2015/3/11/entrez PY - 2015/3/11/pubmed PY - 2016/5/25/medline SP - 2445 EP - 58 JF - British journal of pharmacology JO - Br. J. Pharmacol. VL - 172 IS - 10 N2 - BACKGROUND AND PURPOSE: Chronic pain is a serious worldwide health issue, with current analgesics having limited efficacy and dose-limiting side effects. Humans with loss-of-function mutations in the voltage-gated sodium channel NaV 1.7 (hNaV 1.7) are indifferent to pain, making hNaV 1.7 a promising target for analgesic development. Since spider venoms are replete with NaV channel modulators, we examined their potential as a source of hNaV 1.7 inhibitors. EXPERIMENTAL APPROACH: We developed a high-throughput fluorescent-based assay to screen spider venoms against hNaV 1.7 and isolate 'hit' peptides. To examine the binding site of these peptides, we constructed a panel of chimeric channels in which the S3b-S4 paddle motif from each voltage sensor domain of hNaV 1.7 was transplanted into the homotetrameric KV 2.1 channel. KEY RESULTS: We screened 205 spider venoms and found that 40% contain at least one inhibitor of hNaV 1.7. By deconvoluting 'hit' venoms, we discovered seven novel members of the NaSpTx family 1. One of these peptides, Hd1a (peptide μ-TRTX-Hd1a from venom of the spider Haplopelma doriae), inhibited hNaV 1.7 with a high level of selectivity over all other subtypes, except hNaV 1.1. We showed that Hd1a is a gating modifier that inhibits hNaV 1.7 by interacting with the S3b-S4 paddle motif in channel domain II. The structure of Hd1a, determined using heteronuclear NMR, contains an inhibitor cystine knot motif that is likely to confer high levels of chemical, thermal and biological stability. CONCLUSION AND IMPLICATIONS: Our data indicate that spider venoms are a rich natural source of hNaV 1.7 inhibitors that might be useful leads for the development of novel analgesics. SN - 1476-5381 UR - https://www.unboundmedicine.com/medline/citation/25754331/Seven_novel_modulators_of_the_analgesic_target_NaV_1_7_uncovered_using_a_high_throughput_venom_based_discovery_approach_ L2 - https://doi.org/10.1111/bph.13081 DB - PRIME DP - Unbound Medicine ER -