Oedema and withdrawal latency were evaluated at 2 and 3?h after carrageenan injection, and nociceptive behaviour was observed for 60?min after formalin administration. Rimonabant and SR144528 were dissolved in a mixture of Tween80?:?DMSO?:?distilled water (1:2:7). To determine the ability of THCV to improve established signs of inflammation and pain, an additional series of experiments was performed in which this compound was administered to the mice at 0.3?mg��kg?1 i.p. 30?min after intraplantar injection of carrageenan, and for 3 consecutive days after the induction of inflammation. Behavioural evaluations were performed 2, 24, 48 and 72?h after carrageenan injection. No overt behavioural changes were observed in this study following administration of vehicle or of any of the drugs at the dosage used. The mice remained alert and generally active throughout these experiments. Values have been expressed as means and variability as SEM or as Afatinib datasheet
95% confidence intervals (CIs). The concentration of THCV and CP55940 that produced a 50% displacement of [3H]CP55940 from specific binding sites (IC50 value) was entered into the equation of Cheng and Prusoff (1973) to calculate the corresponding Ki value. Values obtained in vitro for EC50 and maximal effect (Emax) have been calculated by non-linear regression analysis using the equation for a sigmoid concentration�Cresponse curve. Statistical analysis of all in vivo data was performed by anova followed by the Newman�CKeuls multiple comparison test. All these CAL-101 clinical trial
statistical analyses were performed using GraphPAD software (San Diego, CA, USA). Differences were considered significant at P < 0.05. THCV, extracted from Cannabis, was a gift from GW Pharmaceuticals (Porton Down, Wiltshire, UK). It was 99.4% pure and contained no detectable ��9-tetrahydrocannabinol, cannabinol or cannabidiol. Rimonabant and SR144528 were kindly supplied by Sanofi-Aventis (Montpellier, France). Carrageenan and formalin were purchased from Sigma-Aldrich (Milan, Italy) Sitaxentan
and (�C)-cis-3-[2-hydroxy-4-(1,1-dimethylheptyl)phenyl]-trans-4-(3-hydroxypropyl)cyclohexanol (CP55940), from Tocris (Bristol, UK). Pertussis toxin, forskolin and rolipram were supplied by Sigma-Aldrich (Poole, Dorset, UK) and [3H]CP55940 (174.6?Ci��mmol?1) by PerkinElmer Life Sciences Inc (Boston, MA, USA). In our initial experiments, we investigated whether THCV shares the ability of established cannabinoid CB2 receptor agonists (Pertwee, 1999) to inhibit forskolin-induced stimulation of cyclic AMP production in hCB2-transfected CHO cells. As shown in Figure?2, we found that at concentrations in the nanomolar range THCV can indeed induce such inhibition in this bioassay, its EC50 and Emax values with 95% CIs shown in parentheses being 38?nM (12 and 124?nM) and 40% (32 and 48%) respectively. Corresponding values for the established CB1/CB2 receptor agonist, CP55940, which is thought to display full (high-efficacy) agonism at the cannabinoid CB2 receptor were 6.9?nM (3.