A967079

SR144528—a CB2 receptor antagonist, was purchased from Abcam, Cambridge, UK. A967079—a TRPA1 Receptor antagonist and BCTC—a TRPV1 Receptor antagonist, were purchased from Alomone Labs, Jerusalem, Israel. GSK2193874—a TRPV4 antagonist, was purchased from SIGMA-ALDRICH, Rehovot, Israel. CID16020046—a GPR55 antagonist, was purchased from Cayman, MI, USA and GW9662—a PPARγ antagonist, was purchased from Enzo Life Sciences, New York, NY, USA.

For the acute zymosan studies, the same vehicle (10% DMSO, 10% Tween-80 in saline) was used for both A967079 (Alomone Labs, Israel) ((1E,3E)-1-(4-Fluorophenyl)-2-methyl-1-pentene-3-one oxime), a selective TRPA1 antagonist [40 (link)], or MCC950 (Insight Biosciences, UK) (N-[[(1,2,3,5,6,7-hexahydro-s-indacen-4-yl)amino]carbonyl]-4-(1-hydroxy-1-methylethyl)-2-furansulfonamide), a selective NLRP3 antagonist [41 (link)]. The drugs were administered (i.p., 100 mg/kg) 30 min prior to the injection of zymosan. For longer-term CFA studies, A967079 or MCC950 was administered i.p. at 100 mg/kg or 10 mg/kg, respectively, 30 min prior to CFA injection at day 0. A967079 was then administered once daily at 60 mg/kg i.p. thereafter, while MCC950 was administered i.p. once daily (10 mg/kg) on days 1, 2, then every 2 days thereafter until the end of the study period. These respective treatment regimens were chosen as they previously showed to be effective in various pain and inflammatory murine models [40 (link),41 (link)].

Cinnamaldehyde (CA) (#W228613, Sigma-Aldrich, Gillingham, UK, >95% purity), capsaicin (#M2028, Sigma-Aldrich, Gillingham, UK >95%), and menthol (#2772, Sigma-Aldrich, Gillingham, UK >99%) was prepared as a 20 μL solution with a final topical dose of 10%. Cinnamaldehyde, capsaicin, and menthol were dissolved in an ethanol vehicle solution which contained 10% DMSO as a solvent. The vehicle solutions that were used were 10% DMSO in ethanol. A967079 (#A-225, Alomone, Jerusalem, Israel >99% purity) powder was dissolved in 10% DMSO + 10% Tween-80 in saline and given at a final dose of 100 mg/kg. The vehicle for A967079 was 10% DMSO + 10% Tween-80 that was dissolved in saline. N-(3-Aminopropyl)-2-[(3-methylphenyl)methoxy]-N-(2-thienylmethyl)benzamidehydrochloride (AMTB) (#A-305, Alomone, Jerusalem, Israel >97% purity) was dissolved with 10% DMSO in saline to produce a final dose of 10 mg/kg. The A967079, AMTB, and vehicle solution were then given orally via a mixture with 200 µL chocolate-spread Nutella (containing 13% hazelnuts) Ferrero SpA, Alba, Italy), which the mice readily consumed.