Data Sheet 1_v3_Modulation of NMDA receptor activity by CR4056, an imidazoline-2 receptor ligand with analgesic properties.pdf

CR4056 is an imidazoline-2 receptor ligand having potent analgesic activity and synergistic effect with opioids. Very recently it has been found that CR4056 can revert the cognitive impairment in animal models of Alzheimer's disease (AD). Since several lines of evidence highlight the importance of NMDAR modulators in nociceptive signaling and in AD progression, we considered as important to investigate the effects of CR4056 on NMDAR activity. In primary culture of cortical neurons, application of NMDA and glycine elicits a current that is decreased in a dose-dependent fashion by CR4056 (IC50 5.3 ± 0.1 µM). CR4056 antagonism is reversible, not competitive and voltage-independent and it is not blocked by pertussis toxin. CR4056 interacts with the co-agonist glycine site in a competitive way, indeed high glycine concentrations diminish its effect. Fibroblasts expressing different recombinant NMDA receptors are differently modulated by CR4056: the potency and the efficacy of the compound are higher in GluN1- GluN2B than in GluN1-GluN2A containing receptors. In lamina II neurons of spinal cord slices, single stimulation of afferent fibers evokes an NMDA-mediated current that is inhibited by 10 µM CR4056. Repetitive stimulation of the dorsal root at high frequency and high intensity produces a firing activity that is significatively depressed by CR4056. Taken together, our results broad the understanding of the molecular mechanisms of CR4056 analgesic activity, involving the modulation of NMDAR activity. Therefore, we propose that the analgesic action of CR4056 and the neuroprotective effects in AD models may be mediated also by NMDAR inhibition.</p

Exploring Multitarget Interactions to Reduce Opiate Withdrawal Syndrome and Psychiatric Comorbidity

Opioid addiction is often characterized as a chronic relapsing condition due to the severe somatic and behavioral signs, associated with depressive disorders, triggered by opiate withdrawal. Since prolonged abstinence remains a major challenge, our interest has been addressed to such objective. Exploring multitarget interactions, the present investigation suggests that <b>3</b> or its (<i>S</i>)-enantiomer and <b>4</b>, endowed with effective α_2C-AR agonism/α_2A-AR antagonism/5-HT_1A-R agonism, or <b>7</b> and <b>9</b>–<b>11</b> producing efficacious α_2C-AR agonism/α_2A-AR antagonism/I₂–IBS interaction might represent novel multifunctional tools potentially useful for reducing withdrawal syndrome and associated depression. Such agents, lacking in sedative side effects due to their α_2A-AR antagonism, might afford an improvement over current therapies with clonidine-like drugs

Additional file 4: of Pharmacological characterisation of CR6086, a potent prostaglandin E2 receptor 4 antagonist, as a new potential disease-modifying anti-rheumatic drug

Data table showing the effect of repeated administration of CR6086, MTX and their combination in arthritic mice (CIA model). Arthritic CIA mice, recruited upon arthritis onset, were treated with test drugs for 16 days. CR6086 was administered orally once daily, whereas MTX was administered intraperitoneally every third day. Clinical score (a) and paw swelling in millimetres (b) were reported as median (IQR) and mean (SD), respectively. (DOCX 45 kb

Additional file 3: of Pharmacological characterisation of CR6086, a potent prostaglandin E2 receptor 4 antagonist, as a new potential disease-modifying anti-rheumatic drug

Data table showing cytokine serum concentrations (pg/ml) of samples determined from 3-PLEX (MSD) in CIA mice. Arthritis was induced in mice by intradermal injection of bovine type II collagen. Upon onset, animals were recruited and randomised into experimental groups. Oral treatments with drugs were administered daily and lasted 10 days. At the end of the study, sera were isolated for determination of indicated cytokines by multiplex analysis on the MSD platform (Artialis, Liège, Belgium). Data represent mean ¹ SEM of the number of animals per group: N = 8 (sham, vehicle, 30 mg/kg CR6086), 6 (60 mg/kg CR6086) and 7 (60 mg/kg naproxen). (DOCX 40 kb