NaV1.5 sodium channel assays for preclinical safety studies

Why conduct a NaV1.5 sodium channel assay?

The voltage gated cardiac sodium ion channel NaV1.5, encoded by the SCN5a gene, is a key component for the initiation and transmission of this electrical signal throughout the heart. Its activation induces the depolarization of the cell membrane and initiates the rising phase of the action potential (dV/dtmax). When the NaV1.5 sodium channel cannot adequately conduct the electrical current across the cell membrane, this can result in a potentially fatal disorder called long QT syndrome 5LQT3) or Brugada syndrome.

The NaV1.5 sodium channel & preclinical studies

At a preclinical stage, NaV1.5 sodium channel assays aim to identify potential inhibitory effects early in drug discovery. A number of clinically successful drugs have had the tendency to inhibit NaV1.5 sodium channel and to create a concomitant risk of sudden death. As a result, a large number of drugs have been withdrawn from the market or during late stage clinical trials due to cardiotoxic effects. 

What we measure

A pharmacological reduction of the sodium current has been shown to delay the depolarization process and lead to a reduction of cardiac conduction. This makes the human NaV1.5 sodium channel a major target for the cardiac safety assessment.

Mutation LQT3: Mutations in the SCNA5 gene result in persistent depolarizing current throughout action potential duration causing the Long QT Syndrome 3 phenotype.

Why choose PhysioStim for your sodium channel assay ?

The experts at PhysioStim respect all ICH guidelines to perform screening and GLP-compliant studies. Moreover, our stimulation protocol is designed to make sure you get the highest value out of your assay:

Technique

  • Conventional manual Patch Clamp
  • 3 or 4 increasing concentrations of assay compound tested independently

Study model

  • Human Embryonic Kidney HEK-293 cells
  • 6 treated cells per concentration tested and 6 control cells (as basic design protocol but can be adapted)

Measured parameters

  • Amplitude of the peak current upon depolarization to –30mV (pA)
  • Amplitude of the base current at –110mV (pA)
  • Inhibition of NaV1.5 peak current amplitude (%)

Stimulation protocol

Nav1.5 sodium channel assay stimulation protocol and result curve

Typical effects of flecainide effects on NaV1.5 sodium currents recorded from HEK-293 cells

Results

  • ion current inhibition measurement
  • IC50 value

Ref­er­ence com­pounds or pos­i­tive con­trols

REFERENCE COM­POUNDS IC50
Fle­cainide 32.3 nM
Quini­dine 21.1 nM

Advice from our expert scientists

In order to confirm results during NaV1.5 sodium channel assays and the impact on cardiac conduction or ECG, PhysioStim recommends completing your NaV1.5 sodium channel assay with additional tests:

Need help with your preclinical safety study ?

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