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AP-18

Cat No.:V11418 Purity: ≥98%
AP-18 is a selective TRPA1 inhibitor.
AP-18
AP-18 Chemical Structure CAS No.: 55224-94-7
Product category: New1
This product is for research use only, not for human use. We do not sell to patients.
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10mg
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Product Description
AP-18 is a selective TRPA1 inhibitor. AP-18 blocks the activation of TRPA1 by 50 μM cinnamaldehyde, with IC50 of 3.1 μM and 4.5 μM in humans and mice respectively. AP-18 reverses CFA-induced mechanical hyperalgesia in mice. AP-18 attenuated Yo-Pro uptake induced by 30 μM AITC in a concentration-dependent fashion with IC50 of 10.3 μM.
Biological Activity I Assay Protocols (From Reference)
ln Vitro
Even at 50 μM, AP-18 did not substantially prevent TRPV1, TRPV2, TRPV3, TRPV4, or TRPM8 from activating. Ratiometric Ca2+ imaging in CHO cells demonstrates that AP-18 reversibly suppresses mouse TRPA1 responses to iodoacetamide, an irreversible cysteine alkylating agent. Additionally, AP-18 prevents mice from having their TRPA1 activated by cold oil and mustard oil. In ablated plaques of Xenopus oocytes, AP-18 inhibits TRPA1 currents generated by cinnamon aldehyde [1].
ln Vivo
AP18 (1 mM; injected into mice hind feet) effectively suppresses nociceptive events elicited by cinnamaldehyde dye but not capsaicin, exhibiting effectiveness and selectivity [1].
References

[1]. A role of TRPA1 in mechanical hyperalgesia is revealed by pharmacological inhibition. Mol Pain. 2007;3:40. Published 2007 Dec 17.

[2]. Pore dilation occurs in TRPA1 but not in TRPM8 channels. Mol Pain. 2009;5:3. Published 2009 Jan 21.

[3]. Nitrooleic acid, an endogenous product of nitrative stress, activates nociceptive sensory nerves via the direct activation of TRPA1. Mol Pharmacol. 2009;75(4):820-829.

These protocols are for reference only. InvivoChem does not independently validate these methods.
Physicochemical Properties
Molecular Formula
C11H12CLNO
Molecular Weight
209.67200
Exact Mass
209.06
CAS #
55224-94-7
PubChem CID
9584673
Appearance
White to off-white solid powder
Density
1.1±0.1 g/cm3
Boiling Point
348.4±34.0 °C at 760 mmHg
Flash Point
164.5±25.7 °C
Vapour Pressure
0.0±0.8 mmHg at 25°C
Index of Refraction
1.529
LogP
3.72
Hydrogen Bond Donor Count
1
Hydrogen Bond Acceptor Count
2
Rotatable Bond Count
2
Heavy Atom Count
14
Complexity
240
Defined Atom Stereocenter Count
0
SMILES
C/C(=C\C1=CC=C(C=C1)Cl)/C(=N/O)/C
InChi Key
MHTJEUOFLVQMCL-NJHPPEEMSA-N
InChi Code
InChI=1S/C11H12ClNO/c1-8(9(2)13-14)7-10-3-5-11(12)6-4-10/h3-7,14H,1-2H3/b8-7+,13-9+
Chemical Name
(NE)-N-[(E)-4-(4-chlorophenyl)-3-methylbut-3-en-2-ylidene]hydroxylamine
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

Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light.
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 : ≥ 100 mg/mL (~476.94 mM)
Solubility (In Vivo)
Solubility in Formulation 1: 2.5 mg/mL (11.92 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication.
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 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

Solubility in Formulation 2: ≥ 2.5 mg/mL (11.92 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 4.7694 mL 23.8470 mL 47.6940 mL
5 mM 0.9539 mL 4.7694 mL 9.5388 mL
10 mM 0.4769 mL 2.3847 mL 4.7694 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.

Calculator

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An example of molarity calculation using the molarity calculator is shown below:
What is the mass of compound required to make a 10 mM stock solution in 5 ml of DMSO given that the molecular weight of the compound is 350.26 g/mol?
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  • The answer of 17.513 mg appears in the Mass box. In a similar way, you may calculate the volume and concentration.

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What volume of a given 10 mM stock solution is required to make 25 ml of a 25 μM solution?
Using the equation C1V1 = C2V2, where C1=10 mM, C2=25 μM, V2=25 ml and V1 is the unknown:
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  • The answer of 62.5 μL (0.1 ml) appears in the Volume (Start) box
g/mol

Molecular Weight Calculator allows you to calculate the molar mass and elemental composition of a compound, as detailed below:

Note: Chemical formula is case sensitive: C12H18N3O4  c12h18n3o4
Instructions to calculate molar mass (molecular weight) of a chemical compound:
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Definitions of molecular mass, molecular weight, molar mass and molar weight:
  • Molecular mass (or molecular weight) is the mass of one molecule of a substance and is expressed in the unified atomic mass units (u). (1 u is equal to 1/12 the mass of one atom of carbon-12)
  • Molar mass (molar weight) is the mass of one mole of a substance and is expressed in g/mol.
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In vivo Formulation Calculator (Clear solution)
Step 1: Enter information below (Recommended: An additional animal to make allowance for loss during the experiment)
Step 2: Enter in vivo formulation (This is only a calculator, not the exact formulation for a specific product. Please contact us first if there is no in vivo formulation in the solubility section.)
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Calculation results

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.
             (2) Be sure to add the solvent(s) in order.

Biological Data
  • AP18 blocks TRPA1 activation. (A) Chemical structures of the AP18 (upper) and cinnamaldehyde (lower). (B) Dose-response relationships for block of the calcium influx by AP18 into CHO cells expressing mouse and human TRPA1 elicited by 50 μM cinnamaldehyde (left panel). Calcium influx was measured using a standard FLIPR assay, data points are the average of four wells (~8,000 cells/well) and error bars show standard error. Values are normalized to the maximal response (observed in the absence of AP18). (C) Ratiometric Ca2+ imaging of average of ~50 mTRPA1-expressing CHO cells in response to 1 mM Iodoacetamide (IA) and 100 μM of AP18 (black trace). Grey trace represents cells that were treated with IA alone. (D) Current-voltage relationship of TRPA1. Outward rectifying currents elicited by cinnamaldehyde (left panel) in inside-out macropatches derived from TRPA1-expressing Xenopus oocytes were suppressed by AP18 coapplications (right panel).[1]. A role of TRPA1 in mechanical hyperalgesia is revealed by pharmacological inhibition. Mol Pain. 2007;3:40. Published 2007 Dec 17.
  • AP18 specifically blocks TRPA1. Calcium influx (FLIPR) in HEK cells transiently transfected with the indicated cDNAs in response to 200 nM capsaicin (TRPV1), 150 μM 2-APB (TRPV2), 30 μM 2-APB (TRPV3), 20 μM 4alpha-PDD (TRPV4), 20 μM menthol (TRPM8), and 80 μM cinnamaldehyde (TRPA1). Values are given as a percentage of agonist response in the absence of AP18.[1]. A role of TRPA1 in mechanical hyperalgesia is revealed by pharmacological inhibition. Mol Pain. 2007;3:40. Published 2007 Dec 17.
  • AP18 suppresses acute nociceptive behaviours caused by cinnamaldehyde but not capsaicin. Time spent licking and flicking hindpaws injected with cinnamaldehyde (16.4 mM) or capsaicin (0.328 mM) was measured for five minutes and compared with animals co-injected with AP18 (1 mM). Numbers of cases for each experiment from the left is 8, 8, 6 and 6, respectively (*p < 0.05, two-tailed Student's t-test).[1]. A role of TRPA1 in mechanical hyperalgesia is revealed by pharmacological inhibition. Mol Pain. 2007;3:40. Published 2007 Dec 17.
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