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Eact

Cat No.:V9860 Purity: ≥98%
Eact is a selective activator of TMEM16A that directly activates TRPV1 channels in sensory nociceptors, producing itch, acute nociception, and thermosensitivity responses.
Eact
Eact Chemical Structure CAS No.: 461000-66-8
Product category: New1
This product is for research use only, not for human use. We do not sell to patients.
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Product Description
Eact is a selective activator of TMEM16A that directly activates TRPV1 channels in sensory nociceptors, producing itch, acute nociception, and thermosensitivity responses.
Biological Activity I Assay Protocols (From Reference)
ln Vitro
Eact Eact depends on TRPV1 to elicit itch and associated pain behaviors. With an EC50 of 11.6±2.5 μM, Eact stimulates membrane currents in HEK293T cells expressing mtrpv1 in a concentration-dependent manner. TRPV1 channels are damaged and rearranged when activated by the TMEM16A activator Eact [1].
References

[1]. Eact, a small molecule activator of TMEM16A, activates TRPV1 and elicits pain- and itch-related behaviours. Br J Pharmacol. 2016 Apr;173(7):1208-18.

These protocols are for reference only. InvivoChem does not independently validate these methods.
Physicochemical Properties
Molecular Formula
C22H24N2O5S
Molecular Weight
428.501364707947
Exact Mass
428.141
CAS #
461000-66-8
PubChem CID
3173542
Appearance
White to off-white solid powder
Density
1.237±0.06 g/cm3(Predicted)
Boiling Point
595.9±60.0 ℃(Predicted)
LogP
4.129
Hydrogen Bond Donor Count
0
Hydrogen Bond Acceptor Count
7
Rotatable Bond Count
9
Heavy Atom Count
30
Complexity
520
Defined Atom Stereocenter Count
0
InChi Key
ZUXNHFFVQWADJL-UHFFFAOYSA-N
InChi Code
InChI=1S/C22H24N2O5S/c1-26-11-10-24(22-23-17(14-30-22)15-8-6-5-7-9-15)21(25)16-12-18(27-2)20(29-4)19(13-16)28-3/h5-9,12-14H,10-11H2,1-4H3
Chemical Name
3,4,5-trimethoxy-N-(2-methoxyethyl)-N-(4-phenyl-1,3-thiazol-2-yl)benzamide
HS Tariff Code
2934.99.9001
Storage

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Shipping Condition
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
Solubility Data
Solubility (In Vitro)
DMSO : ~50 mg/mL (~116.69 mM)
Solubility (In Vivo)
Solubility in Formulation 1: ≥ 2.5 mg/mL (5.83 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

 (Please use freshly prepared in vivo formulations for optimal results.)
Preparing Stock Solutions 1 mg 5 mg 10 mg
1 mM 2.3337 mL 11.6686 mL 23.3372 mL
5 mM 0.4667 mL 2.3337 mL 4.6674 mL
10 mM 0.2334 mL 1.1669 mL 2.3337 mL

*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.

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Note: Chemical formula is case sensitive: C12H18N3O4  c12h18n3o4
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Step 1: Enter information below (Recommended: An additional animal to make allowance for loss during the experiment)
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Working concentration mg/mL;

Method for preparing DMSO stock solution mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.

Method for preparing in vivo formulation:Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O,mix and clarify.

(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
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Clinical Trial Information
NCT Number Recruitment interventions Conditions Sponsor/Collaborators Start Date Phases
NCT03817229 COMPLETED Behavioral: Education microlearning sessions
Behavioral: Automated digital reminders
Behavioral: Problem-solving mHealth module
Behavioral: Individualized Adherence Feedback Report
Epilepsy Children's Hospital Medical Center, Cincinnati 2019-04-15 Not Applicable
Biological Data
  • Genetic ablation or pharmacological blockade of TRPV1 function severely attenuates or abolishes Eact‐induced itch‐ and pain‐related behaviours. (A) I.d. injection of 50 μL Eact (4.67 mM) into the rostral back induced a scratching response that was attenuated by pretreatment with AMG9810 (50 mg kg−1, i.p.) and was completely absent in the Trpv1 −/− mice. (B) Intraplantar injection of 20 μL of Eact (4.67 mM) produced flinching and licking behaviours that were significantly reduced by i.p. injection of AMG9810 (50 mg kg−1) 30 min before paw injection of Eact. Genetic ablation of TRPV1 function abolished the nocifensive responses evoked by Eact. (C) Time course of thermal hypersensitivity in animals treated with Eact. Intraplantar injection of 20 μL of Eact (4.67 mM) induced thermal hypersensitivity in wt. mice. AMG9810 (50 mg kg−1; i.p.) significantly inhibited the effect of Eact, and Eact‐elicited thermal hypersensitivity was absent in the Trpv1 −/− mice. **P < 0.01, ***P < 0.001 Eact WT group compared with vehicle WT group; ++P < 0.01, +++P < 0.001 Eact + AMG WT group compared with Eact WT group; #P < 0.05, ###P < 0.001 Eact Trpv1 −/− group compared with Eact WT group; n = 7–8 mice per group. AMG = AMG9810.[1].Liu S, et al. Eact, a small molecule activator of TMEM16A, activates TRPV1 and elicits pain- and itch-related behaviours. Br J Pharmacol. 2016 Apr;173(7):1208-18.
  • Eact activates recombinant TRPV1. (A) The representative trace on top shows that Eact (100 μM) evoked an increase in [Ca2 +]i in a HEK293T cell transiently transfected with mTRPV1, which also responded to capsaicin (Cap: 500 nM). The second to forth representative traces from the top illustrate that Eact (100 μM) had no effect on [Ca2 +]i in hTRPA1‐, hTRPM8‐ or rTRPV4‐expressing HEK293T cells, which were activated by their respective agonists allyl isothiocyanate (AITC, 100 μM), menthol ((−)‐Men, 100 μM) or GSK1016790A (GSK) (0.3 μM) respectively. (B) Representative current traces show that Eact activated an outward (at +60 mV) and an inward (at −60 mV) current in a concentration‐dependent manner in a mTRPV1‐expressing HEK293T cell. (C) Representative current–voltage (I–V) curves taken at the specified time points from the trace on (B) illustrate that an outwardly rectifying whole‐cell current was evoked by Eact at 10 μM (b), whereas 100 μM (c) Eact activated a current with a linear I–V relationship in a mTRPV1‐expressing HEK293T cell (a refers to the baseline response). (D) The concentration‐response curve of Eact‐activated outward currents at +60 mV. The inset graph illustrates the maximal current densities evoked by saturating concentrations of Eact (100 μM) and capsaicin (3 μM). (E) The single‐channel current traces on top show that Eact increased single channel opening in an inside‐out membrane patch isolated from a mTRPV1‐expressing HEK293T cell. The graph at the bottom illustrates that Eact significantly increased single channel open probability (nPo) in inside‐out patches excised from mTRPV1‐expressing HEK293T cells (**P < 0.0001 vs. baseline response).[1].Liu S, et al. Eact, a small molecule activator of TMEM16A, activates TRPV1 and elicits pain- and itch-related behaviours. Br J Pharmacol. 2016 Apr;173(7):1208-18.
  • Structural requirements for Eact activation of TRPV1. (A) Concentration‐response curves of Eact‐activated outward currents at +60 mV in WT and TRPV1 mutants with disrupted domains for proton or heat activation of TRPV1. (B) Concentration‐response curves of Eact‐activated outward currents at +60 mV in WT and TRPV1 mutants carrying single or double point mutations in the ‘vanilloid‐binding pocket’. (C) Schematic diagram illustrates structural elements required for activation/modulation of TRPV1 channels by capsaicin, protons and heat.[1].Liu S, et al. Eact, a small molecule activator of TMEM16A, activates TRPV1 and elicits pain- and itch-related behaviours. Br J Pharmacol. 2016 Apr;173(7):1208-18.
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