We designed AM1241, a selective CB2 cannabinoid receptor agonist, and used it to test the hypothesis that CB2 receptor activation would reverse the sensory hypersensitivity observed in neuropathic pain states. AM1241 exhibits high affinity and selectivity for CB 2 receptors. It also exhibits high potency in vivo. AM1241 dose-dependently reversed tactile and thermal hypersensitivity produced by ligation of the L5 and L6 spinal nerves in rats. These effects were selectively antagonized by a CB 2 but not by a CB1 receptor antagonist, suggesting that they were produced by actions of AM1241 at CB 2 receptors. AM1241 was also active in blocking spinal nerve ligation-induced tactile and thermal hypersensitivity in mice lacking CB 1 receptors (CB1 ؊/؊ mice), confirming that AM1241 reverses sensory hypersensitivity independent of actions at CB 1 receptors. These findings demonstrate a mechanism leading to the inhibition of pain, one that targets receptors localized exclusively outside the CNS. Further, they suggest the potential use of CB 2 receptor-selective agonists for treatment of human neuropathic pain, a condition currently without consistently effective therapies. CB 2 receptor-selective agonist medications are predicted to be without the CNS side effects that limit the effectiveness of currently available medications. N europathic pain is defined as pain initiated or caused by a primary lesion or dysfunction in the nervous system (1). It affects Ϸ1% of the population and results from a variety of etiologies including trauma, infection, diabetes, immune deficiencies, ischemic disorders, and toxic neuropathies (1, 2). It can be excruciating, and some patients are unable to work or to perform normal daily activities. Neuropathic pain often responds poorly to medical therapy (3,4). This may be due, in part, to adverse side effects of available medications that limit drug dosage (5). Medications currently used for the treatment of neuropathic pain act on neurotransmitter systems or ion channels and typically produce significant CNS side effects. For example, gabapentin, a drug commonly used to treat neuropathic pain because of its modest side effect profile compared with other therapeutic options, produces somnolence in 19% of patients and dizziness in 17% (Neurontin prescribing information, Parke-Davis). A therapy directed at targets not found in the CNS would avoid these problems. CB 2 cannabinoid receptors are one such potential target.CB 2 receptor mRNA is not detected in brain (6, 7). In addition, the CB 2 receptor-selective antagonist SR144528 did not displace the nonselective cannabinoid ligand [ 3 H]CP55,940 from binding to rat brain (7). Finally, binding of [ 3 H]CP55,940 to mouse brain was eliminated by disruption of the CB 1 receptor gene (8) but was not affected by disruption of the CB 2 receptor gene (9). These studies suggest that CB 2 receptors are not found in the normal CNS, although they do not fully exclude the possibility that CB 2 receptors are expressed in the CNS in small, but functionally signi...

SUMMARY The extensive physiological influence of transmission through the CB2 cannabinoid receptor makes this G protein-coupled receptor (GPCR) a promising therapeutic target for treating neuropathic pain, inflammation, and immune disorders. However, there is little direct structural information pertaining to either GPCR or CB2-receptor ligand recognition and activation. The present work helps characterize experimentally the ligand-binding interactions of the human CB2 (hCB2) receptor. This study illustrates how our overall experimental approach, “ligand-assisted protein structure” (LAPS), affords direct determination of the requirements for ligand binding to the hCB2 receptor and discrimination among the binding motifs for ligands that activate therapeutically relevant GPCRs.

mann. Activation of cannabinoid CB 2 receptors suppresses C-fiber responses and windup in spinal wide dynamic range neurons in the absence and presence of inflammation. J Neurophysiol 92: 3562-3574, 2004. First published August 18, 2004 doi:10.1152/jn.00886.2003. Effects of the CB 2 -selective cannabinoid agonist AM1241 on activity evoked in spinal wide dynamic range (WDR) neurons by transcutaneous electrical stimulation were evaluated in urethane-anesthetized rats. Recordings were obtained in both the absence and the presence of carrageenan inflammation. AM1241, administered intravenously or locally in the paw, suppressed activity evoked by transcutaneous electrical stimulation during the development of inflammation. Decreases in WDR responses resulted from a suppression of C-fiber-mediated activity and windup. A␤-and A␦-fiber-mediated responses were not reliably altered. The AM1241-induced suppression of electrically evoked responses was blocked by the CB 2 antagonist SR144528 but not by the CB 1 antagonist SR141716A. AM1241 (33 g/kg intraplantar [ipl]), administered to the carrageenan-injected paw, suppressed activity evoked in WDR neurons relative to groups receiving vehicle in the same paw or AM1241 in the opposite (noninflamed) paw. The electrophysiological effects of AM1241 (330 g/kg intravenous [iv]) were greater in rats receiving ipl carrageenan compared with noninflamed rats receiving an ipl injection of vehicle. AM1241 failed to alter the activity of purely nonnociceptive neurons recorded in the lumbar dorsal horn. Additionally, AM1241 (330 g/kg iv and ipl; 33 g/kg ipl) reduced the diameter of the carrageenaninjected paw. The AM1241-induced decrease in peripheral edema was blocked by the CB 2 but not by the CB 1 antagonist. These data demonstrate that activation of cannabinoid CB 2 receptors is sufficient to suppress neuronal activity at central levels of processing in the spinal dorsal horn. Our findings are consistent with the ability of AM1241 to normalize nociceptive thresholds and produce antinociception in inflammatory pain states. I N T R O D U C T I O NCannabinoids attenuate nociceptive responses in behavioral

Activation of cannabinoid CB 2 receptors suppresses neuropathic pain induced by traumatic nerve injury. The present studies were conducted to evaluate the efficacy of cannabinoid CB 2 receptor activation in suppressing painful peripheral neuropathy evoked by chemotherapeutic treatment with the antitumor agent paclitaxel. Rats received paclitaxel (2 mg/kg i.p./day) on 4 alternate days to induce mechanical hypersensitivity (mechanical allodynia). Mechanical allodynia was defined as a lowering of the threshold for paw withdrawal to stimulation of the plantar hind paw surface with an electronic von Frey stimulator. Mechanical allodynia developed in paclitaxel-treated animals relative to groups receiving the Cremophor EL/ethanol/saline vehicle at the same times. Two structurally distinct cannabinoid CB 2 agonists, the Administration of either the CB 1 or CB 2 antagonist alone failed to alter paclitaxel-evoked mechanical allodynia. Moreover, (R,S)-AM1241 did not alter paw withdrawal thresholds in rats that received the Cremophor EL vehicle in lieu of paclitaxel, whereas AM1714 induced a modest antinociceptive effect. Our data suggest that cannabinoid CB 2 receptors may be important therapeutic targets for the treatment of chemotherapy-evoked neuropathy.Painful peripheral neuropathy is a well documented side effect of chemotherapeutic treatment (for review, see Aley and Levine, 2002). The major classes of antineoplastic agents, the vinca alkaloids (e.g., vincristine), taxane (e.g., paclitaxel), and platinum-derived (e.g., cisplatin) compounds, are associated with the development of dose-limiting neuropathic pain. The chemotherapeutic agent used, dosing schedule, form of cancer, and presence of additional medical complications can affect the occurrence and severity of chemotherapy-induced neuropathy (for review, see Cata et al., 2006).Paclitaxel is commonly used for the treatment of solid tumors and ovarian and breast cancers. Paclitaxel induces