Benefits of VCE-003.2, a cannabigerol quinone derivative, against inflammation-driven neuronal deterioration in experimental Parkinson’s disease: possible involvement of different binding sites at the PPARγ receptor

Abstract

BackgroundNeuroprotection with cannabinoids in Parkinson’s disease (PD) has been afforded predominantly with antioxidant or anti-inflammatory cannabinoids. In the present study, we investigated the anti-inflammatory and neuroprotective properties of VCE-003.2, a quinone derivative of the non-psychotrophic phytocannabinoid cannabigerol (CBG), which may derive its activity at the peroxisome proliferator-activated receptor-γ (PPARγ). The compound is also an antioxidant.MethodsWe evaluated VCE-003.2 in an in vivo [mice subjected to unilateral intrastriatal injections of lipopolysaccharide (LPS)] model of PD, as well as in in vitro (LPS-exposed BV2 cells and M-213 cells treated with conditioned media generated from LPS-exposed BV2 cells) cellular models. The type of interaction of VCE-003.2 at the PPARγ receptor was furtherly investigated in bone marrow-derived human mesenchymal stem cells (MSCs) and sustained with transcriptional assays and in silico docking studies.ResultsVCE-003.2 has no activity at the cannabinoid receptors, a fact that we confirmed in this study using competition studies. The administration of VCE-003.2 to LPS-lesioned mice attenuated the loss of tyrosine hydroxylase (TH)-containing nigrostriatal neurons and, in particular, the intense microgliosis provoked by LPS in the substantia nigra, measured by Iba-1/Cd68 immunostaining. The analysis by qPCR of proinflammatory mediators such as tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and inducible nitric oxide synthase (iNOS) in the striatum showed they were markedly elevated by the LPS lesion and strongly reduced by the treatment with VCE-003.2. The effects of VCE-003.2 in LPS-lesioned mice implied the activation of PPARγ receptors, as they were attenuated when VCE-003.2 was co-administered with the PPARγ inhibitor T0070907. We then moved to some in vitro approaches, first to confirm the anti-inflammatory profile of VCE-003.2 in cultured BV2 cells exposed to LPS. VCE-003.2 was able to attenuate the synthesis and release of TNF-α and IL-1β, as well as the induction of iNOS and cyclooxygenase-2 (COX-2) elicited by LPS in these cells. However, we found such effects were not reversed by GW9662, another classic PPARγ antagonist. Next, we investigated the neuroprotective effects of VCE-003.2 in cultured M-213 neuronal cells exposed to conditioned media generated from LPS-exposed cultured BV2 cells. VCE-003.2 reduced M-213 cell death, but again, such effects were not reversed by T0070907. Using docking analysis, we detected that VCE-003.2 binds both the canonical and the alternative binding sites in the PPARγ ligand-binding pocket (LBP). Functional assays further showed that T0070907 almost abolished PPARγ transcriptional activity induced by rosiglitazone (RGZ), but it did not affect the activity of VCE-003.2 in a Gal4-Luc system. However, T0070907 inhibited the effects of RGZ and VCE-003.2 on the expression of PPARγ-dependent genes upregulated in MSCs.ConclusionsWe have demonstrated that VCE-003.2 is neuroprotective against inflammation-driven n...

Antioxidative and Anti-Inflammatory Properties of Cannabidiol

“…However, another CBD derivative, VCE-003, induces PPARγ-mediated antioxidant and anti-inflammatory activity that prevents neuronal damage caused by inflammation in the Parkinson's mouse model (intravascular LPS injection). The same effect was seen in the in vitro cellular model of neurological inflammation (BV2 cells exposed to LPS and M-213 cells treated with media prepared from BV2 cells exposed to LPS) [131].…”

A systematic review of minor phytocannabinoids with promising neuroprotective potential

“…VCE‐003.2 also attenuated TNF‐α and L‐1β secretion but from BV2 mouse microglial cells (5 μM) and astroglial cells (1 and 5 μM; Díaz‐Alonso et al, 2016; García et al, 2018; Rodríguez‐Cueto et al, 2018). Díaz‐Alonso et al (2016) and García et al (2018) deduced that VCE‐003.2 did not mediate its protective effects via CB 1 or CB 2 receptors due to poor binding affinity ( K i > 40 μM) and both groups found that VCE‐003.2 was an agonist at PPARγ (IC 50 of 1.2 μM).…”

“…Nine studies included in vitro data, and eight included in vivo data on CBG and its derivatives that are formed by the oxidation of CBG (Carrillo‐Salinas et al, 2014; Díaz‐Alonso et al, 2016; García et al, 2018). VCE‐003 and VCE‐003.2 have displayed increased affinity for PPARγ, thus maintaining their anti‐inflammatory properties while having little affinity for CB 1 and CB 2 receptors (VCE‐003 K i > 40 μM for CB 1 and K i > 1.76 μM CB 2 , Granja et al, 2012, and VCE‐003.2 K i > 40 μM for both CB 1 and CB 2 , García et al, 2018). All studies except one reported a positive effect of CBG, VCE‐003, or VCE‐002.3, compared with control in the disease model being studied.…”

“…and VCE‐003.2 (10 mg·kg −1 p.o./i.p.) successfully reduced immune cell activation in macrophages, microglia, and infiltrating neutrophils in models of EAE (to model MS) and Huntington's and LPS‐induced Parkinson's disease (PD; Aguareles et al, 2019; Carrillo‐Salinas et al, 2014; Díaz‐Alonso et al, 2016; García et al, 2018). In the in vivo Parkinson's disease model, García et al (2018) found that PPARγ antagonist T0070907 (5 mg·kg −1 ) blocked VCE‐003.2‐mediated decreases in TNF‐α, IL‐1β, and iNOS mRNA levels, but no other antagonists were investigated.…”

“…successfully reduced immune cell activation in macrophages, microglia, and infiltrating neutrophils in models of EAE (to model MS) and Huntington's and LPS‐induced Parkinson's disease (PD; Aguareles et al, 2019; Carrillo‐Salinas et al, 2014; Díaz‐Alonso et al, 2016; García et al, 2018). In the in vivo Parkinson's disease model, García et al (2018) found that PPARγ antagonist T0070907 (5 mg·kg −1 ) blocked VCE‐003.2‐mediated decreases in TNF‐α, IL‐1β, and iNOS mRNA levels, but no other antagonists were investigated. In a follow‐up study by the same group, 20 mg·kg −1 (but not 10 mg·kg −1 ) oral VCE‐003.2 promoted a trend towards recovery in the basal ganglia of LPS‐lesioned mice and was associated with decreases in IL‐1β gene expression, lysosomal‐associated membrane protein‐1 (LAMP‐1), and glial fibrillary acidic protein (GFAP) immunostaining (Burgaz et al, 2019).…”