Mechanism-specific assay design facilitates the discovery of Nav1.7-selective inhibitors

Chernov-Rogan, Tania; Li, Tianbo; Lü, Gang; Verschoof, Henry; Khakh, Kuldip; Jones, Steven W.; Beresini, Maureen H.; Liu, Chang; Ortwine, Daniel F.; McKerrall, Steven J.; Hackos, David H.; Sutherlin, Daniel P.; Cohen, Charles J.

doi:10.1073/pnas.1713701115

Cited by 27 publications

(30 citation statements)

References 47 publications

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“…These triggering agents can modify the pharmacological properties of channels and bias sensitivity to nonselective inhibitor types. [8][9][10] Automated electrophysiology platforms enable flexible and precise measurements of sodium channel function and pharmacology with the capacity to support HTS, 11,12 but display reduced throughput and higher cost when compared with fluorescent assays.…”

Section: Introductionmentioning

confidence: 99%

Correlation of Optical and Automated Patch Clamp Electrophysiology for Identification of NaV1.7 Inhibitors

Zhang

Moyer

et al. 2020

SLAS Discovery

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Section: Introductionmentioning

confidence: 99%

Correlation of Optical and Automated Patch Clamp Electrophysiology for Identification of NaV1.7 Inhibitors

Zhang

Moyer

et al. 2020

SLAS Discovery

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“…Veratridine is a Na v 1.7 activator 25, it was able to induce persistent Nav1.7 currents 26, and inhibited channel inactivation and generated enhanced window currents. PF-05089771 was previously identified as a state-dependent Na v 1.7 specific inhibitor interacting with Na v 1.7 voltage-sensor domain of domain IV 27, 28. We used flow cytometry analysis to investigate the consequences of veratridine and PF-05089771 on the cancer cell apoptosis.…”

Section: Resultsmentioning

confidence: 99%

The voltage-gated sodium channel Na_v1.7 associated with endometrial cancer

Liu¹,

Tan²,

Yang³

et al. 2019

J. Cancer

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Background: Endometrial cancer is the most common gynecologic malignancy in women in the developed countries. Despite recent progress in functional characterization of voltage-gated sodium channel (Na v) in multiple cancers, very little was known about the expression of Na v in human endometrial cancer. The present study sought to determine the role of Na v and molecular nature of this channel in the endometrial cancer. Methods: PCR approach was introduced to determine expression level of Na v subunits in endometrial cancer specimens. Pharmacological agents were used to investigate Na v function in endometrial cancer cells. Flow cytometry were used to test cancer apoptosis, and invasion assays were applied to test tumor metastasis. Results: Transcriptional levels of the all Na v α and β subunits were determined by real time-PCR in endometrial cancer with pair tissues of carcinoma and adjacent nonneoplastic tissue, Na v 1.7 was the most highly expressed Na v subtype in endometrial cancer tissues. Na v 1.7 level was closely associated with tumor size, local lymph node metastasis, and 5-year and 10-year survival ratio. Inhibition of this channel by Na v 1.7 blocker PF-05089771, promoted cancer apoptosis and attenuated cancer cell invasion. Conclusion: These results establish a relationship between voltage-gated sodium channel protein and endometrial cancer, and suggest that Na v 1.7 is a potential prognostic biomarker and could serve as a novel therapeutic target for endometrial cancer.

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“…geting toxins, isoform-specific compounds including G0766 (PF-771), G4936 (GX-936) 30 and A803467 31 have high specificities, as they do not bind to the common LA binding sites 30,31 . It's not surprising that they showed no effects on the NaChBac channel (Supplement Fig.…”

Section: Lack Of Effects Of Isoform-specific Compounds On Nachbac Chamentioning

confidence: 99%

Conservation and divergence in NaChBac and NaV1.7 pharmacology reveals novel drug interaction mechanisms

Zhu

Li²,

Silva

2020

Sci Rep

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Voltage-gated na + (na V) channels regulate homeostasis in bacteria and control membrane electrical excitability in mammals. Compared to their mammalian counterparts, bacterial Na V channels possess a simpler, fourfold symmetric structure and have facilitated studies of the structural basis of channel gating. However, the pharmacology of bacterial Na V remains largely unexplored. Here we systematically screened 39 Na V modulators on a bacterial channel (NaChBac) and characterized a selection of compounds on NaChBac and a mammalian channel (human Na V 1.7). We found that while many compounds interact with both channels, they exhibit distinct functional effects. For example, the local anesthetics ambroxol and lidocaine block both Na V 1.7 and NaChBac but affect activation and inactivation of the two channels to different extents. The voltage-sensing domain targeting toxin BDS-i increases na V 1.7 but decreases NaChBac peak currents. The pore binding toxins aconitine and veratridine block peak currents of Na V 1.7 and shift activation (aconitine) and inactivation (veratridine) respectively. In NaChBac, they block the peak current by binding to the pore residue F224. Nonetheless, aconitine has no effect on activation or inactivation, while veratridine only modulates activation of NaChBac. The conservation and divergence in the pharmacology of bacterial and mammalian na V channels provide insights into the molecular basis of channel gating and will facilitate organism-specific drug discovery. Electrical signaling is a highly conserved biological mechanism throughout the evolution of plants and animals 1,2. From prokaryotic organelles to vertebrates, ion channels are key regulators of cellular homeostasis, electrolyte balance and signaling 1. Single-celled eukaryotes first evolved voltage-gated calcium channels (Ca V), which are believed to have evolved subsequently into voltage-gated Na + (Na v) channels predating the origin of nervous systems in animals 3. In prokaryotes, Na V channels might have evolved independently to regulate cellular homeostasis, though their exact functions are not well understood. In humans, the malfunction of Na V channels, either through genetic mutations or off-target drug interactions, results in severe pathologies including cardiac arrhythmia and epilepsy 4. Given their critical physiological function, pathological relevance and utility as therapeutic targets, it is essential to understand the interaction of Na V channels with pharmacological agents. This task has been facilitated by the recent determination of prokaryotic 5-7 and eukaryotic 8-13 Na V channel structures. However, these structures do not provide a clear picture of the dynamic conformational changes that underlie state and voltage-dependent effects of pharmacological agents. Therefore, functional studies must be integrated with structural insights to elucidate the molecular mechanisms of compound-channel interaction 14,15. Eukaryotic Na V channels are large, complex membrane-bound proteins composed of four homologous domain...

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Mechanism-specific assay design facilitates the discovery of Nav1.7-selective inhibitors

Cited by 27 publications

References 47 publications

Correlation of Optical and Automated Patch Clamp Electrophysiology for Identification of NaV1.7 Inhibitors

Correlation of Optical and Automated Patch Clamp Electrophysiology for Identification of NaV1.7 Inhibitors

The voltage-gated sodium channel Na_v1.7 associated with endometrial cancer

Conservation and divergence in NaChBac and NaV1.7 pharmacology reveals novel drug interaction mechanisms

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Mechanism-specific assay design facilitates the discovery of Nav1.7-selective inhibitors

Cited by 27 publications

References 47 publications

Correlation of Optical and Automated Patch Clamp Electrophysiology for Identification of NaV1.7 Inhibitors

Correlation of Optical and Automated Patch Clamp Electrophysiology for Identification of NaV1.7 Inhibitors

The voltage-gated sodium channel Nav1.7 associated with endometrial cancer

Conservation and divergence in NaChBac and NaV1.7 pharmacology reveals novel drug interaction mechanisms

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The voltage-gated sodium channel Na_v1.7 associated with endometrial cancer