Allosteric Modulation of the CB1 Cannabinoid Receptor by Cannabidiol—A Molecular Modeling Study of the N-Terminal Domain and the Allosteric-Orthosteric Coupling

The CB₁ cannabinoid receptor (CB₁R) contains one of the longest N termini among class A G protein-coupled receptors. Mutagenesis studies suggest that the allosteric binding site of cannabidiol (CBD) involves residues from the N terminal domain. In order to study the allosteric binding of CBD to CB₁R we modeled the whole N-terminus of this receptor using the replica exchange molecular dynamics with solute tempering (REST2) approach. Then, the obtained structures of CB₁R with the N terminus were used for ligand docking. A natural cannabinoid receptor agonist, Δ⁹-THC, was docked to the orthosteric site and a negative allosteric modulator, CBD, to the allosteric site positioned between extracellular ends of helices TM1 and TM2. The molecular dynamics simulations were then performed for CB₁R with ligands: (i) CBD together with THC, and (ii) THC-only. Analyses of the differences in the residue-residue interaction patterns between those two cases allowed us to elucidate the allosteric network responsible for the modulation of the CB₁R by CBD. In addition, we identified the changes in the orthosteric binding mode of Δ⁹-THC, as well as the changes in its binding energy, caused by the CBD allosteric binding. We have also found that the presence of a complete N-terminal domain is essential for a stable binding of CBD in the allosteric site of CB₁R as well as for the allosteric-orthosteric coupling mechanism.     

Highly Selective,Amine-Derived Cannabinoid Receptor 2 Probes

The endocannabinoid (eCB) system is implied in various human diseases ranging from central nervous system to autoimmune disorders. Cannabinoid receptor 2 (CB₂R) is an integral component of the eCB system. Yet, the downstream effects elicited by this G protein-coupled receptor upon binding of endogenous or synthetic ligands are insufficiently understood—likely due to the limited arsenal of reliable biological and chemical tools. Herein, we report the design and synthesis of CB₂R-selective cannabinoids along with their in vitro pharmacological characterization (binding and functional studies). They combine structural features of HU-308 and AM841 to give chimeric ligands that emerge as potent CB₂R agonists with high selectivity over the closely related cannabinoid receptor 1 (CB₁R). The synthesis work includes convenient preparation of substituted resorcinols often found in cannabinoids. The utility of the synthetic cannabinoids in this study is showcased by preparation of the most selective high-affinity fluorescent probe for CB₂R to date.     

Virtual screening using docking and molecular dynamics of cannabinoid analogs against CB1 and CB2 receptors

BackgroundCannabis sativa has been attributed to different pharmacological properties. A number of secondary metabolites such as tetrahydrocannabinol (THC), cannabinol (CBD), and different analogs, with highly promising biological activity on CB₁ and CB₂ receptors, have been identified.MethodsThus, this study aimed was to evaluate the activity of THC, CBD, and their analogs using molecular docking and molecular dynamics simulations (MD) methods. Initially, the molecules (ligands) were selected by bioinformatics searches in databases. Subsequently, CB₁ and CB₂ receptors were retrieved from the protein data bank database. Afterward, each receptor and its ligands were optimized to perform molecular docking. Then, MD Simulation was performed with the most stable ligand-receptor complexes. Finally, the Molecular Mechanics-Generalized Born Surface Area (MM-PBSA) method was applied to analyze the binding free energy between ligands and cannabinoid receptors.ResultsThe results obtained showed that ligand LS-61176 presented the best affinity in the molecular docking analysis. Also, this analog could be a CB₁ negative allosteric modulator like CBD and probably an agonist in CB₂ like THC and CBD according to their dynamic behavior in silico. The possibility of having a THC and a CBD analog (LS-61176) as a promising molecule for experimental evaluation since it could have no central side-effects on CB₁ and have effects of CB₂ useful in pain, inflammation, and some immunological disorders. Docking results were validate using ROC curve for both cannabinoids receptor where AUC for CB1 receptor was 0.894±0.024, and for CB2 receptor AUC was 0.832±0032, indicating good affinity prediction.     

Parameterization of Org27569: An allosteric modulator of the cannabinoid CB1 G protein‐coupled receptor

The cannabinoid CB₁ receptor is a class A G protein‐coupled receptor (GPCR) that is the most widely expressed GPCR in the brain. Many GPCRs contain allosteric binding sites for endogenous and/or synthetic ligands, which are topographically distinct from the agonist‐binding site that is known as the orthosteric site. While both endogenous and synthetic ligands that act at the CB₁ orthosteric site have been known for some time, compounds that act at a CB₁ allosteric site have only recently been discovered. The most studied of these is 5‐chloro‐3‐ethyl‐1H‐indole‐2‐carboxylic acid [2‐(4‐piperidin‐1‐ylphenyl)ethyl]amide (Org27569). Because allosteric ligands are thought to act through conformational changes in the receptor that are transmitted from the allosteric to the orthosteric site, computational studies of the structural and dynamic interactions of Org27569 with the CB₁ receptor are crucial to achieve a molecular level understanding of the basis of action of this important new class of compounds. To date, such computational studies have not been possible due to the lack of a complete set of molecular mechanics force field parameters for Org27569. Here, we present the development of missing CHARMM force field parameters for Org27569 using previously published methods and the validation and application of these new parameters using normal mode analysis and molecular dynamics simulations combined with experimental infrared measurements. © 2011 Wiley Periodicals, Inc. J Comput Chem, 2011     

Synthesis and evaluation of various heteroaromatic benzamides as analogues of –ylidene-benzamide cannabinoid type 2 receptor agonists

The CB₂ receptor is an attractive target for the treatment of a wide range of diseases and pathological conditions. Compounds that selectively activate the CB₂ receptor are desirable as this avoids CB₁-mediated psychoactive effects. Heteroarylidene-benzamides have demonstrated efficacy as selective CB₂ receptor agonists. We aimed to expand the structure-activity relationship studies of this series of compounds by investigating the heteroaromatic core via the synthesis and in vitro evaluation of a small library of various heteroaromatic benzamide analogues. As heteroaromatic amides are privileged scaffolds in drug design, methods to synthesise them are of interest. Concise and reliable synthetic strategies were developed to access these novel analogues. The –ylidene-benzamide moiety is shown to be essential for CB activity as all amide derivatives exhibit no functional activity at either CB₂ or CB₁ receptors.     

Cannabinoid Receptor 2 Alters Social Memory and Microglial Activity in an Age-Dependent Manner

Physiological brain aging is characterized by gradual, substantial changes in cognitive ability, accompanied by chronic activation of the neural immune system. This form of inflammation, termed inflammaging, in the central nervous system is primarily enacted through microglia, the resident immune cells. The endocannabinoid system, and particularly the cannabinoid receptor 2 (CB₂R), is a major regulator of the activity of microglia and is upregulated under inflammatory conditions. Here, we elucidated the role of the CB₂R in physiological brain aging. We used CB₂R^−/− mice of progressive ages in a behavioral test battery to assess social and spatial learning and memory. This was followed by detailed immunohistochemical analysis of microglial activity and morphology, and of the expression of pro-inflammatory cytokines in the hippocampus. CB₂R deletion decreased social memory in young mice, but did not affect spatial memory. In fact, old CB₂R^−/− mice had a slightly improved social memory, whereas in WT mice we detected an age-related cognitive decline. On a cellular level, CB₂R deletion increased lipofuscin accumulation in microglia, but not in neurons. CB₂R^−/− microglia showed an increase of activity markers Iba1 and CD68, and minor upregulation in tnfa and il6 expression and downregulation of ccl2 with age. This was accompanied by a change in morphology as CB₂R^−/− microglia had smaller somas and lower polarity, with increased branching, cell volume, and tree length. We present that CB₂Rs are involved in cognition and age-induced microglial activity, but may also be important for microglial activation itself.     

A [Fe(CB6)] platform for binding of small molecules: Insights from DFT calculations

The viability of making [Fe(CB₆)L] (L = H₂, N₂, O₂, nitric oxide [NO^?, NO, and NO⁺], CO₂, and hydrocarbons [CH₄, C₂H₆, C₂H₄, and C₆H₆]) has been investigated by density functional theory (DFT) calculations. The complexes 2 – 18 are thermodynamically stable and may be synthesized. The small molecules are activated to some extent after complexation. Molecular orbital and ΔG calculation revealed that the molecular hydrogen and hydrocarbons can be chemically adsorbed and desorbed on [Fe(CB₆)] without any significant chemical modification and therefore [Fe(CB₆)] may serve as a storage material. The N₂, O₂, and nitric oxide (NO^?, NO, and NO⁺) can be activated using [Fe(CB₆)]. Proton, carbon, boron, and nitrogen NMR chemical shift calculation predicts drastic chemical shift difference before and after the complexation of [Fe(CB₆)] with small molecules. This new findings suggest that the CB₆^2? ligand‐based complex may provide several applications in the future. © 2012 Wiley Periodicals, Inc.     

Synthesis and Characterization of 2‐Mono‐ and 1,2‐Diaminocarba‐closo‐dodecaborates M[1‐R‐2‐H2N‐closo‐CB11H10] (R=H,Ph, H2N,CyHN)

The first primary 2‐aminocarba‐closo‐dodecaborates [1‐R‐2‐H₂N‐closo‐CB₁₁H₁₀]^? (R=H ( 1 ), Ph ( 2 )) have been synthesized by insertion reactions of (Me₃Si)₂NBCl₂ into the trianions [7‐R‐7‐nido‐CB₁₀H₁₀]^3?. The difunctionalized species [1,2‐(H₂N)₂‐closo‐CB₁₁H₁₀] ( 3 ) and 1‐CyHN‐2‐H₃N‐closo‐CB₁₁H₁₀ (H‐ 4 ) have been prepared analogously from (Me₃Si)₂NBCl₂ and 7‐H₃N‐7‐nido‐CB₁₀H₁₂. In addition, the preparation of [Et₄N][1‐H₂N‐2‐Ph‐closo‐CB₁₁H₁₀] ([Et₄N]‐ 5 ) starting from PhBCl₂ and 7‐H₃N‐7‐nido‐CB₁₀H₁₂ is described. Methylation of the [1‐Ph‐2‐H₂N‐closo‐CB₁₁H₁₀]^? ion ( 2 ) to produce 1‐Ph‐2‐Me₃N‐closo‐CB₁₁H₁₀ ( 6 ) is reported. The crystal structures of [Et₄N]‐ 2 , [Et₄N]‐ 5 , and 6 were determined and the geometric parameters were compared to theoretical values derived from DFT and ab initio calculations. All new compounds were studied by NMR, IR, and Raman spectroscopy, MALDI mass spectrometry, and elemental analysis. The discussion of the experimental NMR chemical shifts and of selected vibrational band positions is supported by theoretical data. The thermal properties were investigated by differential scanning calorimetry (DSC). The pK_a values of 2‐H₃N‐closo‐CB₁₁H₁₁ (H‐ 1 ), 1‐H₃N‐closo‐CB₁₁H₁₀ (H‐ 7 ), and 1,2‐(H₃N)₂‐closo‐CB₁₁H₁₀ (H₂‐ 3 ) were determined by potentiometric titration and by NMR studies. The experimental results are compared to theoretical data (DFT and ab initio). The basicities of the aminocarba‐closo‐dodecaborates agree well with the spectroscopic and structural properties.     

Visible-Light Photoswitchable Benzimidazole Azo-Arenes as β-Arrestin2-Biased Selective Cannabinoid 2 Receptor Agonists

The cannabinoid 2 receptor (CB₂R) has high therapeutic potential for multiple pathogenic processes, such as neuroinflammation. Pathway-selective ligands are needed to overcome the lack of clinical success and to elucidate correlations between pathways and their respective therapeutic effects. Herein, we report the design and synthesis of a photoswitchable scaffold based on the privileged structure of benzimidazole and its application as a functionally selective CB₂R “efficacy-switch”. Benzimidazole azo-arenes offer huge potential for the broad extension of photopharmacology to a wide range of optically addressable biological targets. We used this scaffold to develop compound 10 d , a “trans-on” agonist, which serves as a molecular probe to study the β-arrestin2 (βarr2) pathway at CB₂R. βΑrr2 bias was observed in CB₂R internalization and βarr2 recruitment, while no activation occurred when looking at Gα₁₆ or mini-Gα_i. Overall, compound 10 d is the first light-dependent functionally selective agonist to investigate the complex mechanisms of CB₂R-βarr2 dependent endocytosis.     

Tricyclic Pyrazole-Based Compounds as Useful Scaffolds for Cannabinoid CB1/CB2 Receptor Interaction

Cannabinoids comprise different classes of compounds, which aroused interest in recent years because of their several pharmacological properties. Such properties include analgesic activity, bodyweight reduction, the antiemetic effect, the reduction of intraocular pressure and many others, which appear correlated to the affinity of cannabinoids towards CB₁ and/or CB₂ receptors. Within the search aiming to identify novel chemical scaffolds for cannabinoid receptor interaction, the CB₁ antagonist/inverse agonist pyrazole-based derivative rimonabant has been modified, giving rise to several tricyclic pyrazole-based compounds, most of which endowed of high affinity and selectivity for CB₁ or CB₂ receptors. The aim of this review is to present the synthesis and summarize the SAR study of such tricyclic pyrazole-based compounds, evidencing, for some derivatives, their potential in the treatment of neuropathic pain, obesity or in the management of glaucoma.     

Difunctionalized {closo‐1‐CB11} Clusters: 1‐ and 2‐Amino‐12‐ethynylcarba‐closo‐dodecaborates

Carba‐closo‐dodecaborate anions with two functional groups have been synthesized via a simple two‐step procedure starting from monoamino‐functionalized {closo‐1‐CB₁₁} clusters. Iodination at the antipodal boron atom provided access to [1‐H₂N‐12‐I‐closo‐1‐CB₁₁H₁₀]^? ( 1 a ) and [2‐H₂N‐12‐I‐closo‐1‐CB₁₁H₁₀]^? ( 2 a ), which have been transformed into the anions [1‐H₂N‐12‐RC?C‐closo‐1‐CB₁₁H₁₀]^? (R=H ( 1 b ), Ph ( 1 c ), Et₃Si ( 1 d )) and [2‐H₂N‐12‐RC?C‐closo‐1‐CB₁₁H₁₀]^? (R=H ( 2 b ), Ph ( 2 c ), Et₃Si ( 2 d )) by microwave‐assisted Kumada‐type cross‐coupling reactions. The syntheses of the inner salts 1‐Me₃N‐12‐RC?C‐closo‐1‐CB₁₁H₁₀ (R=H ( 1 e ), Et₃Si ( 1 f )) and 2‐Me₃N‐12‐RC?C‐closo‐1‐CB₁₁H₁₀ (R=H ( 2 e ), Et₃Si ( 2 f )) are the first examples for a further derivatization of the new anions. All {closo‐1‐CB₁₁} clusters have been characterized by multinuclear NMR and vibrational spectroscopy as well as by mass spectrometry. The crystal structures of Cs 1 a , [Et₄N] 2 a , K 1 b , [Et₄N] 1 c , [Et₄N] 2 c , 1 e , and [Et₄N][1‐H₂N‐2‐F‐12‐I‐closo‐1‐CB₁₁H₉]?0.5 H₂O ([Et₄N ]4 a ?0.5 H₂O) have been determined. Experimental spectroscopic data and especially spectroscopic data and bond properties derived from DFT calculations provide some information on the importance of inductive and resonance‐type effects for the transfer of electronic effects through the {closo‐1‐CB₁₁} cage.     

1‐Amino‐2‐fluoromonocarba‐closo‐dodecaborates: K[1‐H2N‐2‐F‐closo‐1‐CB11H10] and [Et4N][1‐H2N‐2‐F‐closo‐1‐CB11I10]

The potassium salt of the [1‐H₂N‐2‐F‐closo‐1‐CB₁₁H₁₀]^– anion ( 1 ) was obtained from an insertion reaction of Li₃[7‐H₂N‐nido‐7‐CB₁₀H₁₀] with BF₃ · OEt₂. Anion 1 was protonated to the neutral species 1‐H₃N‐2‐F‐closo‐1‐CB₁₁H₁₀ (H 1 ) and it was iodinated with ICl to the [1‐H₂N‐2‐F‐closo‐1‐CB₁₁I₁₀]^– anion ( 2 ). All species were characterized by multinuclear NMR, IR, and Raman spectroscopy as well as by elemental analysis. The structure of H 1· (CH₃)₂CO was studied by single‐crystal X‐ray diffraction and the experimentally determined bond lengths are compared to values derived from density functional calculations.     

Group 4 complexes derived from o-carborane: synthesis, structures and ethylene polymerization properties

Nine thermally stable complexes (η⁵-Cp*)[η⁵-(C₅H₄)CMe₂CB₁₀H₁₀CR]MCl₂ (R=H and Me) and (η⁵-Cp*)[η⁵; η¹-(C₅H₄)CMe₂(CB₁₀H₁₀C)]MCl have been prepared via metathesis reactions of Cp*MCl₃ (M=Ti, Zr and Hf, Cp*=pentamethylcyclopentadienyl) with monolithium salts of (C₅H₅)CMe₂(CB₁₀H₁₀CR) (R=H and Me) and with dilithium salt of (C₅H₅)CMe₂(CB₁₀H₁₀CH), respectively. These compounds have been fully characterized by various spectroscopic methods and elemental analyses. All of the compounds except (η⁵-Cp*)[η⁵-(C₅H₄)CMe₂CB₁₀H₁₀CMe]HfCl₂ were additionally characterized by a single crystal X-ray diffraction study, establishing their monomeric bent metallocene structural feature with carborane acting as a substituent or an ancillary ligand. The titanium and zirconium complexes produce high-density polyethylenes with the activity range of about 10³-10⁴ g PE per mol of M bar h in the presence of modified methylaluminoxane cocatalyst.     

Stability of B12(CN)122−: Implications for Lithium and Magnesium Ion Batteries

Multiply charged negative ions are seldom stable in the gas phase. Electrostatic repulsion leads either to autodetachment of electrons or fragmentation of the parent ion. With a binding energy of the second electron at 0.9 eV, B₁₂H₁₂^2? is a classic example of a stable dianion. It is shown here that ligand substitution can lead to unusually stable multiply charged anions. For example, dodecacyanododecaborate, B₁₂(CN)₁₂^2?, created by substituting H by CN is found to be highly stable with the second electron bound by 5.3 eV, which is six times larger than that in the B₁₂H₁₂^2?. Equally important is the observation that CB₁₁(CN)₁₂^2?, which contains one electron more than needed to satisfy the Wade‐Mingos rule, is also stable with its second electron bound by 1.1 eV, while CB₁₁H₁₂^2? is unstable. The ability to stabilize multiply charged anions in the gas phase by ligand manipulation opens a new door for multiply charged species with potential applications as halogen‐free electrolytes in ion batteries.     

Cannabinoid CB1 receptor antagonists in therapeutic and structural perspectives

The observed antiobesity effect of rimonabant ( 1 ) in a pharmacological rodent model 10 years ago has led to a surge in the search for novel cannabinoid CB₁ antagonists as a new therapeutic target for the treatment of obesity. Rimonabant showed clinical efficacy in the treatment of obesity and also improved cardiovascular and metabolic risk factors. Cannabinoid CB₁ receptor antagonists have also good prospects in other therapeutic areas, including smoking and alcohol addiction as well as cognitive impairment. Solvay's research achievements in this fast‐moving field are reported in relation with the current state of the art. Several medicinal chemistry strategies have been pursued. The application of the concept of conformational constraint led to the discovery of more rigid analogs of the prototypic CB₁ receptor antagonist rimonabant. Replacement of the central heterocyclic pyrazole ring in rimonabant yielded imidazoles, triazoles, and thiazoles as selective CB₁ receptor antagonists. Dedicated medium‐throughput screening efforts delivered one 3,4‐diarylpyrazoline hit. Its poor pharmacokinetic properties were successfully optimized which led to the discovery of orally active and highly CB₁/CB₂ receptor selective analogs in this series. Regioisomeric 1,5‐diarylpyrazolines, 1,2‐diarylimidazolines, and water‐soluble imidazoles have been designed as novel CB₁ receptor antagonist structure classes. © 2008 The Japan Chemical Journal Forum and Wiley Periodicals, Inc. Chem Rec 8: 156–168; 2008: Published online in Wiley InterScience ( www.interscience.wiley.com ) DOI 10.1002/tcr.20147     

Temperature dependent product distribution in photolysis of o-alkylphenacyl benzoates

Temperature effect on photochemical reactions of ortho-alkylphenacyl benzoates, one of the recently developed photoremovable protecting groups (PPG), giving indanones (IN) and benzocyclobutenols (CB) was examined. As temperature was lowered, CB was formed preferentially over IN and the amount of IN increased as temperature was elevated. Arrhenius plot of ln(IN/CB) versus 1/T gave a straight line from which Ea_IN–Ea_CB and A_IN/A_CB were obtained. The least sterically hindered 2-methylphenacyl benzoate (1) gave the highest Ea_IN–Ea_CB and A_IN/A_CB among the ortho-alkylphenacyl benzoates tested in this research including 2,4,6-trimethylphenacyl benzoate (2) and 2,4,6-triisopropylphenacyl benzoate (3).     

A study of 4-carba-arachno-nonaborane(14) and the derived tridecahydro-4-carba-arachno-nonaborane(−1) anion: Structure and CH bond activation as shown by 1- and 2-D NMR

The ¹¹B and ¹H NMR spectra of the carbaborane CB₈H₁₄, and its anion CB₈H₁₃^?, have been recorded and assigned unambiguously by decoupling and COSY techniques. The results indicate the presence of unsymmetrical bridging hydrogen atoms in both the neutral and anionic species. More significantly the endo-hydrogen attached to the carbon atom is involved in exchange with all the three bridging hydrogen atoms in the CB₈H₁₃^? anion, whereas no such exchange is observed in the neutral CB₈H₁₄.     

The Peripheral Cannabinoid Receptor Type 1 (CB1) as a Molecular Target for Modulating Body Weight in Man

The cannabinoid 1 (CB₁) receptor regulates appetite and body weight; however, unwanted central side effects of both agonists (in wasting disorders) or antagonists (in obesity and diabetes) have limited their therapeutic utility. At the peripheral level, CB₁ receptor activation impacts the energy balance of mammals in a number of different ways: inhibiting satiety and emesis, increasing food intake, altering adipokine and satiety hormone levels, altering taste sensation, decreasing lipolysis (fat break down), and increasing lipogenesis (fat generation). The CB₁ receptor also plays an important role in the gut–brain axis control of appetite and satiety. The combined effect of peripheral CB₁ activation is to promote appetite, energy storage, and energy preservation (and the opposite is true for CB₁ antagonists). Therefore, the next generation of CB₁ receptor medicines (agonists and antagonists, and indirect modulators of the endocannabinoid system) have been peripherally restricted to mitigate these issues, and some of these are already in clinical stage development. These compounds also have demonstrated potential in other conditions such as alcoholic steatohepatitis and diabetic nephropathy (peripherally restricted CB₁ antagonists) and pain conditions (peripherally restricted CB₁ agonists and FAAH inhibitors). This review will discuss the mechanisms by which peripheral CB₁ receptors regulate body weight, and the therapeutic utility of peripherally restricted drugs in the management of body weight and beyond.     

Non-isothermal kinetic for lyophilized leachate from sanitary landfill and composting usine

Leachate samples from a sanitary landfill of Araraquara city and composting usine of Vila Leopoldina, São Paulo, Brazil were lyophilized to remove the water content. TG/DTG curves at different heating rates were recorded. The second step of the thermal decomposition of leachate from the Araraquara landfill (CB₁), from the composting usine from Vila Leopoldina (CB₂) from the organic phase extracted (FO) and aqueous phase (FA) were all kinetically evaluated using the non-isothermal method.By Flynn-Wall isoconversional method the following values were obtained: E=234±3.65 kJ mol^?1 and logA=29.7±0.58 min^?1 for CB₁; E=129±1.66 kJ mol^?1 and logA=11.8±0.10 min^?1 for CB₂; E=51.6±1.35 kJ mol^?1 and logA=6.09±0.09 min^?1 for FO and E=76.91±6.33 kJ mol^?1 and logA=8.88±0.7 min^?1 for FA with 95% confidence level. Applying the procedures of Málek and Koga, SB kinetic model (?esták-Berggren) is the most appropriate to describe the decomposition of CB₁, CB₂, FO and FA.     

Zerumbone Ameliorates Neuropathic Pain Symptoms via Cannabinoid and PPAR Receptors Using In Vivo and In Silico Models

Neuropathic pain is a chronic pain condition persisting past the presence of any noxious stimulus or inflammation. Zerumbone, of the Zingiber zerumbet ginger plant, has exhibited anti-allodynic and antihyperalgesic effects in a neuropathic pain animal model, amongst other pharmacological properties. This study was conducted to further elucidate the mechanisms underlying zerumbone’s antineuropathic actions. Research on therapeutic agents involving cannabinoid (CB) and peroxisome proliferator-activated receptors (PPARs) is rising. These receptor systems have shown importance in causing a synergistic effect in suppressing nociceptive processing. Behavioural responses were assessed using the von Frey filament test (mechanical allodynia) and Hargreaves plantar test (thermal hyperalgesia), in chronic constriction injury (CCI) neuropathic pain mice. Antagonists SR141716 (CB₁ receptor), SR144528 (CB₂ receptor), GW6471 (PPARα receptor) and GW9662 (PPARγ receptor) were pre-administered before the zerumbone treatment. Our findings indicated the involvement of CB₁, PPARα and PPARγ in zerumbone’s action against mechanical allodynia, whereas only CB₁ and PPARα were involved against thermal hyperalgesia. Molecular docking studies also suggest that zerumbone has a comparable and favourable binding affinity against the respective agonist on the CB and PPAR receptors studied. This finding will contribute to advance our knowledge on zerumbone and its significance in treating neuropathic pain.