This dataset comprises the findings obtained in the study aimed at investigating the anti-migraine effects of two URB597 analogs with improved solubility and bioavailability – ARN14633 ([4-fluoro-3-[3-(methylcarbamoyl)phenyl]phenyl] N-cyclohexylcarbamate) and ARN14280 ([3-(3-carbamoylphenyl)-4-(difluoromethoxy)phenyl] N-cyclohexylcarbamate), in the migraine rat model based by nitroglycerin (NTG) administration. Two compounds significantly attenuated the nocifensive behavior induced by NTG compared with original URB597 administered in the same conditions. Additionally, both inhibitors reverted the neurobiological changes induced by NTG in specific peripheral and central areas. Although the precise mechanism underlying these effects remains to be elucidated, our results support further investigational studies of FAAH blockade as a potential therapeutic strategy to treat migraine conditions. ARN14633 (1 mg/kg, i.p.), ARN14280 (3 mg/kg, i.p.) and URB597 (2mg/kg, i.p.) were administered to adult male Sprague-Dawley rats 3 hours after NTG injection. One hour after the administration of either compound, rats were subjected to the orofacial formalin test and compared. In addition, the anti-migraine effects of two URB597 analogs were evaluated by neuronal nitric oxide synthase, pain mediators peptides, pro-inflammatory cytokines gene expression and lipid amides levels in specific areas. The in vivo and ex vivo assessments were: 1) mRNA expression levels: CGRP, SP, interleukin-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha) and IL-1beta in cervical spinal cord, medulla-pons and trigeminal ganglia. mRNA levels were measured by rt-PCR. All samples were assayed in triplicate and gene expression levels were calculated according to 2−∆∆Ct = 2− (∆Ct gene − ∆Ct housekeeping gene) formula by using Ct (cycle threshold) values. 2) Central and peripheral Endocannabinoids levels: Lipid amides levels were measured using a Xevo TQ UPLC-MS/MS system equipped with a reversed-phase BEH C18 column (2.1 × 50 mm, 1.7 μm particle size) (Waters, Milford, USA). The mobile phase consisted of 0.1% formic acid in water as solvent A and 0.1% formic acid in acetonitrile as solvent B. A linear gradient was used: 0.0–0.5 min 20% B; 0.5–2.5 min 20 to 100% B; and 2.5–3.0 min maintained at 100% B. The column was reconditioned to 20% B for 1 min. Analysis time was 4 min and the injection volume was 5 μL. Results in brief ARN14633 and ARN14280 attenuated NTG-induced nocifensive behavior compared with URB597. Additionally, the new FAAH inhibitors reduced transcription of genes encoding neuronal nitric oxide synthase, pain mediators peptides (calcitonin gene-related peptide, substance P) and pro-inflammatory cytokines (tumor necrosis factor-alpha, interleukin-1beta and 6) in the trigeminal ganglion, cervical spinal cord and medulla. Finally, both compounds strongly elevated levels of endocannabinoids and/or other FAAH substrates in cervical spinal cord and medulla, and, to a lesser extent, in the trigeminal ganglia.

Funding: Ricerca Finalizzata 13A, Ministry of Health (Italy)