A-803467

Injected i.p. 30 minutes before behavioral testing

5-(4-chlorophenyl)-N-(3, 5-dimethoxyphenyl)furan-2-carboxamide

ca.100 mg/kg

Dissolved in 5% DMSO/95% polyethylene glycol (PEG 400)

A-803467 potently blocks recombinant human or rat NaV1.8 channels with IC50 of 8 nM and 45 nM, respectively, at a holding potential of -40 mV. At a resting state, A-803467 also potently blocks human NaV1.8 channels with IC50 of 79 nM. A-803467 blocks tetrodotoxin-resistant (TTX-R) currents in rat dorsal root ganglion neurons in a concentration-dependent manner with IC50 of 140 nM, more potently compared with both mexiletine and lamotrigine with IC50 of >30 uM. A-803467 displays 300- to 1, 000-fold higher selectivity for hNaV1.8 over hNaV1.2, hNaV1.3, hNaV1.5, and hNaV1.7 channels with IC50 of 7.38 uM, 2.45 uM, 7.34 uM, and 6.74 uM, respectively. A-803467 shows no significant activity against other channels and receptors expressed in peripheral sensory neurons including TRPV1, P2X2/3, CaV2.2 and KCNQ2/3 channels with IC50 >10 uM. A-803467 also shows no or weak activity against a broad screening panel of cell-surface receptors, ion channels, and enzymes with IC50 of >2 uM. A-803467 at 0.3 uM but not 0.1 uM significantly inhibits the generation of spontaneous and electrically evoked action potentials. [1]

Recombinant human sodium channels, HEK-293 cells expressing recombinant human NaV1.8 channels are grown in DMEM/high-glucose Dulbecco's modified, 10% FBS, 2 mM sodium pyruvate, and G418. For whole-cell voltage-clamp recordings, patch pipettes are pulled from borosilicate glass on a Flaming-Brown micropipette puller. Pipettes have a tip resistance of 0.8-2.5Momega with the internal solutions 135 mM CsF, 10 mM CsCl, 5 mM EGTA, 5 mM NaCl, 10 mM Hepes, pH to 7.3, with 5 M CsOH, and voltage offset is zeroed before seal formation. The external buffer consists of 132 mM NaCl, 5.4 mM KCl, 0.8 mM MgCl2, 1.8 mM CaCl2, 5 mM Glucose, and 10 mM Hepes-free acid, pH to 7.3, with 6 M NaOH. After establishment of a whole-cell recording, cellular capacitance is minimized by using the analog compensation available on the recording amplifier. Series resistance is 85% in all experiments, resulting in a final series resistance no greater than 0.75 Momega. Signals are low-pass filtered at 5-10 kHz, digitized at 20-50 kHz, and stored on a computer for later analysis. Voltage protocols are generated, and data acquisition and analysis are performed by using pCLAMP software. All experiments are performed at room temperature. Liquid junction potentials are <10mV and are not corrected. The concentration-response curve is generated after application of different concentrations of A-803467 and IC50 value of A-803467 is assessed at a holding potential of -40 mV.

A-803467 Chemical Structure

2 years -20CPowder, 2 weeks4Cin DMSO, 2 months-80Cin DMSO