The systemic administration of the anandamide transport inhibitor ARN272 ([(4-(5-(4-hydroxy-phenyl)-3,4-diaza-bicyclo[4.4.0]deca-1(6),2,4,7,9-pentaen-2-ylamino)-phenyl)-phenylamino-methanone]) was used to evaluate the prevention of LiCl-induced nausea-induced behaviour (conditioned gaping) in rats, and LiCl-induced emesis in shrews (Suncus murinus). The mechanism of how prolonging anandamide availability acts to regulate nausea in rats was explored by the antagonism of cannabinoid 1 (CB1) receptors with the systemic co-administration of SR141716.The systemic administration of ARN272 produced a dose-dependent suppression of nausea-induced conditioned gaping in rats, and produced a dose-dependent reduction of vomiting in shrews. The systemic co-administration of SR141716 with ARN272 (at 3.0 mg·kg(-1)) in rats produced a complete reversal of ARN272-suppressed gaping at 1.0 mg·kg(-1). SR141716 alone did not differ from the vehicle solution.These results suggest that anandamide transport inhibition by the compound ARN272 tonically activates CB1 receptors and as such produces a type of indirect agonism to regulate toxin-induced nausea and vomiting. The results also provide behavioural evidence in support of a facilitated transport mechanism used in the cellular reuptake of anandamide.[1]

A total of 58 naïve male Sprague-Dawley rats were used for assessment of anti-nausea-induced behaviour. Rats were single-housed in 48 × 26 × 20 cm shoebox cages in a colony room at an ambient temperature of 21°C. All animals were maintained on a reverse light/dark cycle (0700 h lights off; 1900 h lights on) with free access to food (Iams rodent chow, 18% protein) and tap water, except during testing, which occurred during the dark cycle. All animals were provided with environmental enrichment from two clean paper towels (replenished weekly during cage changes) and a soft plastic container 14 cm long and 12 cm in diameter.
A total of 21 S. murinus, house musk shrews, were bred and raised in a colony at the University of Guelph. Shrews were single-housed in cages at an ambient temperature of 21°C on a 14/10 light dark schedule (lights off at 2100 h). Shrews were tested during their light cycle, between 0900 and 1400 h. Both males (42.9–53.0 g) and females (26.1–32.9 g) were used and equally distributed among the groups, with subjects ranging from 98 to 814 days of age. The sexes did not significantly differ in vomiting frequency in any analysis; therefore, males and females were pooled in all reported analyses. The shrews had previous emetic experience with the limitation of a minimum of 3 weeks recovery between treatments.
The anandamide transport inhibitor, ARN272 , was prepared in a vehicle solution (VEH) of 1:1:8 PEG400, Tween 80 and physiological saline, respectively, for all experiments. All systemic injections were administered i.p. In experiment 1, ARN272 was delivered to rats at concentrations of 0.1 mg·mL−1 (0.1 mg·kg−1 dose), 1 mg·mL−1 (1 mg·kg−1 dose) and 3 mg·mL−1 (3 mg·kg−1 dose), and at a volume of 1 mL·kg−1, chosen on the basis of previous experiments performed by Fu et al. (2012) where 1 mg·kg−1 increased plasma anandamide levels 2 h post-administration. In experiment 2, ARN272 was delivered to shrews at concentrations of 3.0 mg·mL−1 (9.0 mg·kg−1 dose) at a volume of 3 and 3.0 mg·mL−1 (18 mg·kg−1 dose) at a volume of 6 mL·kg−1, chosen on the basis of previous experiments where S. murinus required a dose increase by at least a factor of three times an effective dose in rats . The concentration of SR141716 in experiment 1 was delivered at 1.0 mg·mL−1 (1.0 mg·kg−1 dose) at a volume of 1 mL·kg−1. The dose of 1.0 mg·kg−1 SR141716 was chosen based on prior effectiveness in reversing the breakpoint and reinstatement of nicotine self-administration , while also being found to not to potentiate the effects of emetic agents unlike doses of SR141716 at 2.5 mg·kg−1 or higher .
The LiCl drug treatment 0.15 M used in all experiments was prepared in sterile water and administered at volumes of 20 mL·kg−1 (127 mg·kg−1) in rats in experiment 1, and 60 mL kg−1 (390 mg·kg−1) in shrews in experiment 2.

Br J Pharmacol.2013 Nov;170(5):1130-6.

C27H20N4O2

432.473305702209

432.158

488793-85-7

1386351

Typically exists as solid at room temperature

1.4±0.1 g/cm3

629.1±55.0 °C at 760 mmHg

334.3±31.5 °C

0.0±1.9 mmHg at 25°C

1.757

4.58

3

5

5

33

624

0

OC(C=C1)=CC=C1C2=NN=C(C3=C2C=CC=C3)NC4=CC=C(C(NC5=CC=CC=C5)=O)C=C4

UPKNGUQNXSMHBE-UHFFFAOYSA-N

InChI=1S/C27H20N4O2/c32-22-16-12-18(13-17-22)25-23-8-4-5-9-24(23)26(31-30-25)28-21-14-10-19(11-15-21)27(33)29-20-6-2-1-3-7-20/h1-17,32H,(H,28,31)(H,29,33)

4-[[4-(4-hydroxyphenyl)phthalazin-1-yl]amino]-N-phenylbenzamide

ARN272; ARN-272; ARN 272

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

DMSO : ~125 mg/mL (~289.04 mM)

Solubility in Formulation 1: ≥ 2.08 mg/mL (4.81 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (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 20.8 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.

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Solubility in Formulation 2: ≥ 2.08 mg/mL (4.81 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 20.8 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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