AM251

Alias: AM251; AM 251; AM-251

Cat No.:V1517 Purity: ≥98%

COA Product Data Instructions for use SDS

AM251 (AM-251; AM 251) is a novel, potent and selective cannabinoid (CB) receptor antagonist with potential anti-obesity effect.

AM251 Chemical Structure CAS No.: 183232-66-8
Product category: Cannabinoid Receptor

This product is for research use only, not for human use. We do not sell to patients.

Size	Price	Stock	Qty
5mg
10mg
25mg
50mg
100mg
250mg
500mg
1g
Other Sizes

ADD TO CART CHECKOUT BULK INQUIRY

1-708-310-1919 sales@invivochem.com

Official Supplier of:

Business Relationship with 5000+ Clients Globally
Major Universities, Research Institutions, Biotech & Pharma
Citations by Top Journals: Nature, Cell, Science, etc.

Top Publications Citing lnvivochem Products

Nature 2021, 597(7874):119-125.

Cell 2020,183:1202-1218.e25.

Science Adv 2022: 8, eabm9427.

Nature 597, 119–125 (2021)

Cell Stem Cell 2020,26(6):845-861.e12.

Science Trans Med 2023,15(701):eadd7872.

STTT 2023,8(1):91.

Cell Reports 2023,42(4):112313

Science Trans Med 2023, 15: eadd7872.

Cancer Cell 2021,39(4):529-547.e7.

Cancer Discov 2022,12(4):1106-1127.

Immunity 2023,56(3):620-634.e11.

J Am Chem Soc 2022,144:21831-21836.

Cell 2020,183:1202-1218.e25.

Science Adv 2022:8,eabm9427.

Nature 2021,597(7874):119-125.

Cell 2020,183:1202-1218.e25.

PNAS Nexus 2022,1(3):pgac063.

ACS Nano 2023,17(10):9110-9125.

Biomaterial 2023 Sep16:302:122333.

Purity & Quality Control Documentation

Current batch：

Purity: ≥98%

COA

MSDS

Instructions

Product Description
Bioactivity & Protocols
Physicochemical Properties
Solubility Data
Calculators
Biological Data

Product Description

AM251 (AM-251; AM 251) is a novel, potent and selective cannabinoid (CB) receptor antagonist with potential anti-obesity effect. It functions by inhibiting the brain's presynaptic cannabinoid 1 receptors, which binds to both endocannabinoids and synthetic cannabinoid agonists to suppress transmitter release. Rats' recognition memory is enhanced by AM251, and the ERK signaling pathway is activated, leading to nocifensive behavior. Furthermore, through the proteolytic degradation of ERRα, AM251 modifies mitochondrial physiology and reduces mechanical allodynia and thermal hyperalgesia following burn injuries.

Biological Activity I Assay Protocols (From Reference)

Targets

CB1 receptor ( IC50 = 8 nM )

ln Vitro

In vitro activity: AM251 is an inverse agonist and antagonist of the CB1 receptor. In HEK293 cells, AM251 elicits an agonist response akin to that observed in the yeast expression system[2]. AM-251 decreases the synthesis of cholesteryl ester in peritoneal macrophages that are CB2^+/+ and CB2^-/-, as well as in unstimulated and acetylated LDL-stimulated Raw 264.7 macrophages[3].

ln Vivo

AM251, a CB1 antagonist, reduces capsaicin-evoked nocifensive behavior (3 mg/kg, i.p.). This suppressive effect is genotype dependent, and there was a nearly significant interaction between AM251 and genotype effects. Compared to their respective vehicle controls, planned comparisons show that AM251 decreases nocifensive behaviors in fatty-acid amide hydrolase (FAAH) KO mice (p<0.01) but does not change nocifensive behavior in WT mice (p>0.2). The duration of heat hypersensitivity in FAAH KO mice is reduced by AM251 (3 mg/kg, i.p.) (F_1,9=21.43, p<0.01), but not in WT mice (p>0.3). In FAAH KO mice (F_5,9=4.349, p<0.01) but not in WT mice (p>0.3), AM251 reduces capsaicin-evoked heat hypersensitivity in a time-dependent manner. When compared to FAAH KO animals receiving AM251, post-hoc analysis shows that mice receiving vehicle (i.p.) exhibit increased thermal hypersensitivity at 30, 60, and 90 minutes after capsaicin injection (p<0.05, p<0.05, and p<0.001)[4].
A one-way ANOVA reveals that, in comparison to controls, AM251 (AM-251) injections considerably reduce the percentage of entries in the open arms and the amount of time spent in the open arms in the rats. The Tukey-Kramer test analysis shows that the amount of time that the rats spent in the open arms was significantly less for the doses of 1 mg/kg (P<0.05) and 5 mg/kg (P<0.01) when compared to the control rats. Additionally, at doses of 1 and 5 mg/kg (P<0.05), AM251 significantly reduces the percentage of entries in the open arms[5].

Enzyme Assay

In 12-well culture plates, macrophages are seeded at a density of 2×10⁸/well. Seven-ketocholesterol (7KC) from a 2 mg/mL ethanol stock solution is added one hour before AM-251 or SR144528 are added from 4 mM stock solutions made in DMSO. To give controls the same amounts of ethanol and DMSO, adjustments are made. The activity of caspase-3 is measured after 16 hours. The data is displayed as the mean RFLU/mg protein±SD for each treatment, which is carried out in triplicate[3].

Cell Assay

In A375 human melanoma cells, treatment with AM251 (5 μmol/l) caused apoptosis, G2/M cell cycle arrest, and an increase in cAMP. Furthermore, AM-251 prevented Raw 264.7 macrophages from undergoing apoptosis caused by 7-ketocholesterol.

Animal Protocol

Mice: In total, 246 mice weighing between 17 and 48 g are employed in these studies. Mice are given a single intraperitoneal injection (5 mL/kg) of either vehicle (n = 6 per group), AM251 (3 mg/kg, n = 5 per group), or AMG9810 (3 mg/kg) after baseline responding has been determined. I.p. injections are done half an hour before administering vehicle or capsaicin intraperitoneally. Before and 10, 30, 60, 90, and 120 minutes following an intradermal injection of capsaicin or a vehicle, paw withdrawal latencies are measured. Paw withdrawal latencies are recorded as the average of the two duplicate measurements made by each animal, averaged over subjects, and are measured in duplicate in each paw at each time point.
Rats: We use male Wistar rats weighing between 250 and 350 grams.In this study, the following agents are used: endocannabinoid breakdown inhibitor, URB-597 (0.03, 0.1, and 0.3 mg/kg, i.p.); CB1 receptor antagonist, Win-55212 (0.3, 1 and 5 mg/kg, i.p.); and CB1 receptor antagonist, AM251 (0.3, 1 and 5 mg/kg, i.p.). The vehicle is physiological saline, which contains 0.9% sodium chloride. Every medication is made fresh and given intraperitoneally (i.p.) to rats in a volume of 0.1 mL per 10 g of body weight. Each drug is administered 30 minutes prior to the elevated plus-maze test after being dissolved in physiological saline.

References

[1]. Beyond radio-displacement techniques for identification of CB1 ligands: the first application of afluorescence-quenching assay. Sci Rep. 2014 Jan 20;4:3757.

[2]. Pharmacological characterization of GPR55, a putative cannabinoid receptor. Pharmacol Ther. 2010 Jun;126(3):301-13

[3]. AM-251 and SR144528 are acyl CoA:cholesterol acyltransferase inhibitors. Biochem Biophys Res Commun. 2009 Apr 3;381(2):181-6.

[4]. A pro-nociceptive phenotype unmasked in mice lacking fatty-acid amide hydrolase. Mol Pain. 2016 May 13;12. pii: 1744806916649192.

[5]. Study the Effect of Endocannabinoid System on Rat Behavior in Elevated Plus-Maze. Basic Clin Neurosci. 2015 Jul;6(3):147-53.

[6]. Role of cannabinoid receptor type 1 in tibial and pudendal neuromodulation of bladder overactivity in cats. Am J Physiol Renal Physiol. 2017 Mar 1;312(3):F482-F488.

[7]. Endocannabinoid activation of CB1 receptors contributes to long-lasting reversal of neuropathic pain by repetitive spinal cord stimulation. Eur J Pain. 2017 May;21(5):804-814.

These protocols are for reference only. InvivoChem does not independently validate these methods.

Physicochemical Properties

Molecular Formula

C22H21CL2IN4O

Molecular Weight

555.24

Exact Mass

554.01

Elemental Analysis

C, 47.59; H, 3.81; Cl, 12.77; I, 22.86; N, 10.09; O, 2.88

CAS #

183232-66-8

Related CAS #

183232-66-8

Appearance

Solid powder

SMILES

CC1=C(N(N=C1C(=O)NN2CCCCC2)C3=C(C=C(C=C3)Cl)Cl)C4=CC=C(C=C4)I

InChi Key

BUZAJRPLUGXRAB-UHFFFAOYSA-N

InChi Code

InChI=1S/C22H21Cl2IN4O/c1-14-20(22(30)27-28-11-3-2-4-12-28)26-29(19-10-7-16(23)13-18(19)24)21(14)15-5-8-17(25)9-6-15/h5-10,13H,2-4,11-12H2,1H3,(H,27,30)

Chemical Name

1-(2,4-dichlorophenyl)-5-(4-iodophenyl)-4-methyl-N-piperidin-1-ylpyrazole-3-carboxamide

Synonyms

AM251; AM 251; AM-251

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: 25~40 mg/mL (45.0~72.0 mM)

Water: <1 mg/mL

Ethanol: <1 mg/mL

Solubility (In Vivo)

1%DMSO+30% polyethylene glycol+1%Tween 80: 8 mg/mL

(Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	1.8010 mL	9.0051 mL	18.0102 mL
	5 mM	0.3602 mL	1.8010 mL	3.6020 mL
	10 mM	0.1801 mL	0.9005 mL	1.8010 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

Mass

Concentration

Volume

Molecular Weight*

Calculate Reset

Molarity Calculator allows you to calculate the mass, volume, and/or concentration required for a solution, as detailed below:

Calculate the Mass of a compound required to prepare a solution of known volume and concentration
Calculate the Volume of solution required to dissolve a compound of known mass to a desired concentration
Calculate the Concentration of a solution resulting from a known mass of compound in a specific volume

See Example

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?

Enter 350.26 in the Molecular Weight (MW) box
Enter 10 in the Concentration box and choose the correct unit (mM)
Enter 5 in the Volume box and choose the correct unit (mL)
Click the “Calculate” button
The answer of 17.513 mg appears in the Mass box. In a similar way, you may calculate the volume and concentration.

Concentration (Start)

Volume (Start)

Concentration (End)

Volume (End)

Calculate Reset

Dilution Calculator allows you to calculate how to dilute a stock solution of known concentrations. For example, you may Enter C1, C2 & V2 to calculate V1, as detailed below:

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:

Enter 10 into the Concentration (Start) box and choose the correct unit (mM)
Enter 25 into the Concentration (End) box and select the correct unit (mM)
Enter 25 into the Volume (End) box and choose the correct unit (mL)
Click the “Calculate” button
The answer of 62.5 μL (0.1 ml) appears in the Volume (Start) box

Molecular formula

Total molecular weight

g/mol

Calculate Reset

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:

To calculate molar mass of a chemical compound, please enter the chemical/molecular formula and click the “Calculate’ button.

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.

Volume (to add to vial)

Mass (in vial)

Desired Reconstitution Concentration

Calculate Reset

Reconstitution Calculator allows you to calculate the volume of solvent required to reconstitute your vial.

Enter the mass of the reagent and the desired reconstitution concentration as well as the correct units
Click the “Calculate” button
The answer appears in the Volume (to add to vial) box

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal to make allowance for loss during the experiment)

Dosage mg/kg

Average weight of animals g

Dosing volume per animal μL

Number of animals

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.)

% DMSO

% Tween 80

% ddH₂O

Calculate Reset

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.

Note： (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

Theoretical Binding modes of AM251 and T1117 within CB₁ receptor. Sci Rep . 2014 Jan 20:4:3757. doi: 10.1038/srep03757.
AM-251 and SR144528 inhibit the stimulation of cholesterol esterification by acLDL independent of CB2 expression. Biochem Biophys Res Commun . 2009 Apr 3;381(2):181-6.
AM251 attenuates capsaicin-evoked nocifensive behavior in FAAH KO but not WT mice whereas AMG9810 increases it in WT but not FAAH KO mice. Mol Pain . 2016 May 13:12:1744806916649192.

Effect of AM251 (0.3, 1 and 5 mg/kg) administration on the elevated plus-maze performance: total distance covered by rats (A), the percentage of entries in open arms (B), time spent in open arms (C) and number of closed arms entry (D) during the 10 min test session in EPM. Basic Clin Neurosci . 2015 Jul;6(3):147-53.