Person:
Gómez Cañas, María

First Name

María

Last Name

Gómez Cañas

URI

https://hdl.handle.net/20.500.14352/85462

Affiliation

Universidad Complutense de Madrid

Faculty / Institute

Medicina

Department

Bioquímica y Biología Molecular

Identifiers

Full item page

Search Results

Now showing 1 - 10 of 27

Identification of Novel GPR55 Modulators Using Cell-Impedance-Based Label-Free Technology
(Journal of Medicinal Chemistry, 2016) Morales, Paula; Whyte, Lauren S.; Chicharro, Roberto; Gómez Cañas, María; Pazos Rodríguez, María Ruth; Goya, Pilar; Irving, Andrew J.; Fernández Ruiz, José Javier; Ross, Ruth A.; Jagerovic, Nadine
The orphan G protein-coupled receptor GPR55 has been proposed as a novel receptor of the endocannabinoid system. However, the validity of this categorization is still under debate mainly because of the lack of potent and selective agonists and antagonists of GPR55. Binding assays are not yet available for GPR55 screening, and discrepancies in GPR55 mediated signaling pathways have been reported. In this context, we have designed and synthesized novel GPR55 ligands based on a chromenopyrazole scaffold. Appraisal of GPR55 activity was accomplished using a label-free cell-impedance-based assay in hGPR55-HEK293 cells. The real-time impedance responses provided an integrative assessment of the cellular consequence to GPR55 stimulation taking into account the different possible signaling pathways. Potent GPR55 partial agonists (14b, 18b, 19b, 20b, and 21−24) have been identified; one of them (14b) being selective versus classical cannabinoid receptors. Upon antagonist treatment, chromenopyrazoles 21−24 inhibited lysophosphatidylinositol (LPI) effect. One of these GPR55 antagonists (21) is fully selective versus classic cannabinoid receptors. Compared to LPI, the predicted physicochemical parameters of the new compounds suggest a clear pharmacokinetic improvement
Δ9‐Tetrahydrocannabinolic acid alleviates collagen‐induced arthritis: Role of PPARγ and CB1 receptors
(British Journal of Pharmacology, 2020) Palomares, Belén; Gonzalo Consuegra, Claudia; Gómez Cañas, María; Fernández Ruiz, José Javier; Muñoz, Eduardo
Background and Purpose: Δ9 -Tetrahydrocannabinolic acid (Δ9 -THCA-A), the precur sor of Δ9 -THC, is a non-psychotropic phytocannabinoid that shows PPARγ agonist activity. Here, we investigated the ability of Δ9 -THCA-A to modulate the classic can nabinoid CB1 and CB2 receptors and evaluated its anti-arthritis activity in vitro and in vivo. Experimental Approach: Cannabinoid receptors binding and intrinsic activity, as well as their downstream signalling, were analysed in vitro and in silico. The anti-arthritis properties of Δ9 -THCA-A were studied in human chondrocytes and in the murine model of collagen-induced arthritis (CIA). Plasma disease biomarkers were identified by LC-MS/MS based on proteomic and ELISA assays. Key Results: Functional and docking analyses showed that Δ9 -THCA-A can act as an orthosteric CB1 receptor agonist and also as a positive allosteric modulator in the presence of CP-55,940. Also, Δ9 -THCA-A seemed to be an inverse agonist for CB2 receptors. In vivo, Δ9 -THCA-A reduced arthritis in CIA mice, preventing the infiltra tion of inflammatory cells, synovium hyperplasia, and cartilage damage. Furthermore, Δ9 -THCA-A inhibited expression of inflammatory and catabolic genes on knee joints. The anti-arthritic effect of Δ9 -THCA-A was blocked by either SR141716 or T0070907. Analysis of plasma biomarkers, and determination of cytokines and anti collagen antibodies confirmed that Δ9 -THCA-A mediated its activity mainly through PPARγ and CB1 receptor pathways. Conclusion and Implications: Δ9 -THCA-A modulates CB1 receptors through the orthosteric and allosteric binding sites. In addition, Δ9 -THCA-A exerts anti-arthritis activity through CB1 receptors and PPARγ pathways, highlighting its potential for the treatment of chronic inflammatory diseases such as rheumatoid arthritis.
Analogues of cannabinoids as multitarget drugs in the treatment of Alzheimer’s disease
(European Journal of Pharmacology, 2021) Fernandez, María; Villaro, Wilma; Gómez Cañas, María; García-Arencibia, Moisés; Egea, Javier; Fernández Ruiz, José Javier; Martín, María Isabel; Girón, Rocío; Sánchez Montero, José; Agis Torres, Ángel; Solano, David; Sollhuber Kretzer, Mónica; Sánchez Montero, José
Given that neuronal degeneration in Alzheimer's disease (AD) is caused by the combination of multiple neurotoxic insults, current directions in the research of novel therapies to treat this disease attempts to design multitarget strategies that could be more effective than the simply use of acetylcholinesterase inhibitors; currently, the most used therapy for AD. One option, explored recently, is the synthesis of new analogues of cannabinoids that could competitively inhibit the acetylcholinesterase (AChE) enzyme and showing the classic neuroprotective profile of cannabinoid compounds. In this work, molecular docking has been used to design some cannabinoid analogues with such multitarget properties, based on the similarities of donepezil and Δ9-tetrahydrocannabinol. The analogues synthesized, compounds 1 and 2, demonstrated to have two interesting characteristics in different in vitro assays: competitive inhibition of AChE and competitive antagonism at the CB1/CB2 receptors. They are highly lipophilic, highlighting that they could easily reach the CNS, and apparently presented a low toxicity. These results open the door to the synthesis of new compounds for a more effective treatment of AD.
The cannabinoid quinol VCE-004.8 alleviates bleomycin-induced scleroderma and exerts potent antifibrotic effects through peroxisome proliferator-activated receptor-γ and CB2 pathways
(Scientific Reports, 2016) Río, Carmen del; Gómez Cañas, María; Pazos Rodríguez, María Ruth; Fernández Ruiz, José Javier; Muñoz, Eduardo
Scleroderma is a group of rare diseases associated with early and transient inflammation and vascular injury, followed by fibrosis affecting the skin and multiple internal organs. Fibroblast activation is the hallmark of scleroderma, and disrupting the intracellular TGFβ signaling may provide a novel approach to controlling fibrosis. Because of its potential role in modulating inflammatory and fibrotic responses, both PPARγ and CB2 receptors represent attractive targets for the development of cannabinoid-based therapies. We have developed a non-thiophilic and chemically stable derivative of the CBD quinol (VCE 004.8) that behaves as a dual agonist of PPARγ and CB2 receptors, VCE-004.8 inhibited TGFβ-induced Col1A2 gene transcription and collagen synthesis. Moreover, VCE-004.8 inhibited TGFβ–mediated myofibroblast differentiation and impaired wound-healing activity. The anti-fibrotic efficacy in vivo was investigated in a murine model of dermal fibrosis induced by bleomycin. VCE-004.8 reduced dermal thickness, blood vessels collagen accumulation and prevented mast cell degranulation and macrophage infiltration in the skin. These effects were impaired by the PPARγ antagonist T0070907 and the CB2 antagonist AM630. In addition, VCE-004.8 downregulated the expression of several key genes associated with fibrosis, qualifying this semi-synthetic cannabinoid as a novel compound for the management of scleroderma and, potentially, other fibrotic diseases
Tricyclic pyrazoles. Part 8. Synthesis, biological evaluation and modelling of tricyclic pyrazole carboxamides as potential CB2 receptor ligands with antagonist/inverse agonist properties
(European Journal of Medicinal Chemistry, 2016) Deiana, Valeria; Gómez Cañas, María; Pazos Rodríguez, María Ruth; Fernández Ruiz, José Javier; García Arencibia, Moisés; Pinna, Gerard A.
Previous studies have investigated the relevance and structure-activity relationships (SARs) of pyrazole derivatives in relation with cannabinoid receptors, and the series of tricyclic 1,4-dihydroindeno[1,2-c] pyrazoles emerged as potent CB2 receptor ligands. In the present study, novel 1,4-dihydroindeno[1,2-c] pyrazole and 1H-benzo[g]indazole carboxamides containing a cyclopropyl or a cyclohexyl substituent were designed and synthesized to evaluate the influence of these structural modifications towards CB1 and CB2 receptor affinities. Among these derivatives, compound 15 (6-cyclopropyl-1-(2,4- dichlorophenyl)-N-(adamantan-1-yl)-1,4-dihydroindeno[1,2-c]pyrazole-3-carboxamide) showed the highest CB2 receptor affinity (Ki ¼ 4 nM) and remarkable selectivity (KiCB1/KiCB2 ¼ 2232), whereas a similar affinity, within the nM range, was seen for the fenchyl derivative (compound 10: Ki ¼ 6 nM), for the bornyl analogue (compound 14: Ki ¼ 38 nM) and, to a lesser extent, for the aminopiperidine de rivative (compound 6: Ki ¼ 69 nM). Compounds 10 and 14 were also highly selective for the CB2 receptor (KiCB1/KiCB2 > 1000), whereas compound 6 was relatively selective (KiCB1/KiCB2 ¼ 27). The four com pounds were also subjected to GTPgS binding analysis showing antagonist/inverse agonist properties (IC50 for compound 14 ¼ 27 nM, for 15 ¼ 51 nM, for 10 ¼ 80 nM and for 6 ¼ 294 nM), and this activity was confirmed for the three more active compounds in a CB2 receptor-specific in vitro bioassay con sisting in the quantification of prostaglandin E2 release by LPS-stimulated BV2 cells, in the presence and absence of WIN55,212-2 and/or the investigated compounds. Modelling studies were also conducted with the four compounds, which conformed with the structural requirements stated for the binding of antagonist compounds to the human CB2 receptor.
VCE-004.3, a cannabidiol aminoquinone derivative, prevents bleomycin-induced skin fibrosis and inflammation through PPARγ- and CB2 receptor-dependent pathways
(British Journal of Pharmacology, 2018) Río, Carmen del; Gómez Cañas, María; Fernández Ruiz, José Javier; Muñoz, Eduardo
BACKGROUND AND PURPOSE The endocannabinoid system and PPARγ are important targets for the development of novel compounds against fibrotic diseases such as systemic sclerosis (SSc), also called scleroderma. The aim of this study was to characterize VCE-004.3, a novel cannabidiol derivative, and study its anti-inflammatory and anti-fibrotic activities. EXPERIMENTAL APPROACH The binding of VCE-004.3 to CB1 and CB2 receptors and PPARγ and its effect on their functional activities were studied in vitro and in silico. Anti-fibrotic effects of VCE-004.3 were investigated in NIH-3T3 fibroblasts and human dermal fibroblasts. To assess its anti-inflammatory and anti-fibrotic efficacy in vivo, we used two complementary models of bleomycin-induced fibrosis. Its effect on ERK1/2 phosphorylation induced by IgG from SSc patients and PDGF was also investigated. KEY RESULTS VCE-004.3 bound to and activated PPARγ and CB2 receptors and antagonized CB1 receptors. VCE-004.3 bound to an alternative site at the PPARγ ligand binding pocket. VCE-004.3 inhibited collagen gene transcription and synthesis and prevented TGFβ induced fibroblast migration and differentiation to myofibroblasts. It prevented skin fibrosis, myofibroblast differentiation and ERK1/2 phosphorylation in bleomycin-induced skin fibrosis. Furthermore, it reduced mast cell degranulation, macrophage acti vation, T-lymphocyte infiltration, and the expression of inflammatory and profibrotic factors. Topical application of VCE-004.3 also alleviated skin fibrosis. Finally, VCE-004.3 inhibited PDGF-BB- and SSc IgG-induced ERK1/2 activation in fibroblasts. CONCLUSIONS AND IMPLICATIONS VCE-004.3 is a novel semisynthetic cannabidiol derivative that behaves as a dual PPARγ/CB2 agonist and CB1 receptor modulator that could be considered for the development of novel therapies against different forms of scleroderma.
A Cannabigerol Quinone Alleviates Neuroinflammation in a Chronic Model of Multiple Sclerosis
(Journal of Neuroimmune Pharmacology, 2012) Granja, Aitor G.; Carrillo-Salinas, Francisco; Pagani, Alberto; Negri, Roberto; Navarrete, Carmen; Mecha, Miriam; Mestre, Leyre; Fiebich, Bend L.; Cantarero, Irene; Calzado, Marco A.; Bellido, Maria L.; Appendino, Giovanni; Guaza, Carmen; Muñoz, Eduardo; Gómez Cañas, María; Fernández Ruiz, José Javier
Phytocannabinoids like ∆9-tetrahydrocannabinol (THC) and cannabidiol (CBD) show a beneficial effect on neuroinflammatory and neurodegenerative processes through cell membrane cannabinoid receptor (CBr)-dependent and -independent mechanisms. Natural and synthetic cannabinoids also target the nuclear receptor peroxisome proliferator-activated receptor-gamma (PPARγ), an attractive molecular target for the treatment of neuroinflammation. As part of a study on the SAR of phytocannabinoids, we have investigated the effect of the oxidation modification in the resorcinol moiety of cannabigerol (CBG) on CB1, CB2 and PPARγ binding affinities, identifying cannabigerol quinone (VCE-003) as a potent anti-inflammatory agent. VCE-003 protected neuronal cells from excitotoxicity, activated PPARγ transcriptional activity and inhibited the release of pro-inflammatory mediators in LPS-stimulated microglial cells. Theiler’s murine encephalomyelitis virus (TMEV) model of multiple sclerosis (MS) was used to investigate the anti-inflammatory activity of this compound in vivo. Motor function performance was evaluated and the neuroinflammatory response and gene expression pattern in brain and spinal cord were studied by immunostaining and qRT-PCR. We found that VCE-003 ameliorated the symptoms associated to TMEV infection, decreased microglia reactivity and modulated the expression of genes involved in MS pathophysiology. These data lead us to consider VCE-003 to have high potential for drug development against MS and perhaps other neuroinflammatory diseases.s.
Caracterización de nuevos ligandos cannabinoides con potencial neuroprotector
(2017) Gómez Cañas, María; Pazos Rodríguez, Mª Ruth; García Arencibia, Moisés; Fernández Ruiz, Javier
Desde que se descubrió el principal componente psicoactivo de la planta Cannabis sativa (Δ9-THC) y sus efectos farmacológicos sobre el organismo, se investigó la posible existencia de los receptores sobre los que podría actuar esta molécula, así como la de sus ligandos endógenos cannabimiméticos y la maquinaria enzimática responsable de la síntesis y degradación de eCBs. Al conjunto de estos elementos se le denominó SCE. En la actualidad se conocen dos receptores cannabinoides, ambos de la familia de GPCRs, denominados CB1 y CB2, cuya homología es del 44%, la cual asciende al 68% si se atiende a los segmentos transmembrana. El primero se localiza fundamentalmente en SNC, aunque también se ha observado su presencia en varios órganos periféricos; mientras que la distribución del segundo se ha considerado clásicamente periférica, principalmente en células del sistema inmune, aunque también se ha observado su presencia en distintos tipos de células del SNC, especialmente en células gliales activadas. Los efectos psicoactivos provocados por los cannabinoides, se deben a la activación del receptor CB1, al estar ubicado en neuronas de determinadas áreas del cerebro relacionadas con procesos cognitivos y emocionales. Existen además, nuevas dianas para los cannabinoides como el receptor ionotrópico TRPV1, los receptores nucleares PPAR y otros receptores metabotrópicos, sobre los que aún existe controversia sobre su ligando endógeno, como los receptores GPR55 o GPR18. En cuanto a las moléculas con capacidad para activar los receptores cannabinoides, se encuentran los fitocannabinoides (cannabinoides presentes en la planta), los eCBs (moléculas endógenas de origen animal) y cannabinoides de nueva síntesis, que se han desarrollado para mejorar las propiedades de los cannabinoides naturales y ser utilizados como terapia o como herramienta en el laboratorio. Los principales eCBs son la AEA y el 2-AG, ambos derivados del AA, los cuales se comportan como agonistas de los receptores CB1 y CB2. También se conocen más moléculas cannabimiméticas endógenas que tienen capacidad de activar los receptores, o bien de potenciar la señalización endocannabinoide, impidiendo la degradación de los principales eCBs. Se han estudiado los mecanismos de síntesis y degradación fundamentalmente de la AEA y el 2-AG, los cuales son sintetizados a demanda y degradados tras su acción. En cuanto a la AEA, su síntesis es fundamentalmente llevada a cabo por la acción consecutiva de dos enzimas dependientes de calcio: la NAT y la NAPE-PLD, aunque existen otras rutas alternativas, y su degradación tiene lugar principalmente por la acción de una serín-hidrolasa llamada FAAH. En cuanto al 2-AG, su síntesis es llevada a cabo a partir de DAG mediante la acción de una DAGL, aunque al igual que para la AEA, también hay otras rutas alternativas para su síntesis, y su degradación tiene lugar por la acción de la enzima MAGL, aunque existen otras con capacidad semejante. Aunque no se consideran mecanismos de degradación por la cantidad de metabolitos secundarios que se originan, los eCBs también pueden ser sometidos a rutas de metabolismo oxidativo, originando EETs y HETEs por la acción de LOXs y COXs, respectivamente...
Structure–Effect Relationships of Novel Semi-Synthetic Cannabinoid Derivatives
(Frontiers in Pharmacology, 2019) Götz, Marcus R.; Fernández Ruiz, José Javier; García Toscano, Laura; Gómez Cañas, María; Pazos Rodríguez, María Ruth; Holzgrabe, Ulrike
Background: As a library of cannabinoid (CB) derivatives with (‑)-trans-cannabidiol (CBD) or (‑)-trans-cannabidivarin (CBDV) scaffold, we synthesized nine novel cannabinoids: 2-hydroxyethyl cannabidiolate (2-HEC), 2-hydroxypentyl cannabidiolate (2-HPC), 2,3-dihydroxypropyl cannabidiolate (GCBD), cyclohexyl cannabidiolate (CHC), n-hexyl cannabidiolate (HC), 2-(methylsulfonamido)ethyl cannabidiolate (NMSC), 2-hydroxyethyl cannabidivarinolate (2-HECBDV), cyclohexyl cannabidivarinolate (CHCBDV), and n-hexyl cannabidivarinolate (HCBDV). Their binding and intrinsic effects at the CB1- and CB2 receptors and the effects on inflammatory signaling cascades were investigated in in vitro and ex vivo cell models. Materials and Methods: Binding affinity was studied in membranes isolated from CB-receptor-transfected HEK293EBNA cells, intrinsic functional activity in Chinese hamster ovary (CHO) cells, and activation of nuclear factor κB (NF-κB) and nuclear factor of activated T-cells (NFAT) in phorbol 12-myristate 13-acetate (PMA)/ionomycin (IO) treated Jurkat T-cells. Inhibition of interleukin (IL)-17-induced pro-inflammatory cytokines and chemokines [IL-6, IL-1β, CC-chemokine ligand 2 (CCL2), and tumor necrosis factor (TNF)-α] was studied in RAW264.7 macrophages at the RNA level. Pro-inflammatory cytokine (IL-1β, IL-6, IL-8, and TNF-α) expression and prostaglandin E2 (PGE2) expression were investigated at the protein level in lipopolysaccharide (LPS)-treated primary human monocytes. Results: Derivatives with long aliphatic side chains at the ester position at R1 [HC (5)] as well as the ones with polar side chains [2-HECBDV (7), NMSC (6), and 2-HEC (1)] can be selective for CB2-receptors. The CBDV-derivatives HCBDV and CHCBDV demonstrated specific binding at CB1- and CB2-receptors at nanomolar concentrations. 2-HEC, 2-HPC, GCBD, and NMSC were agonists at CB2-receptor and antagonists at CB1-receptor. CHC bound both receptors at submicromolar ranges and was an agonist for these receptors. 2-HECBDV was an agonist at CB2-receptor and an antagonist at the CB1-receptor despite its modest affinity at this receptor (micromolar range). NMSC inhibited NF-κB and NFAT activity, and 2-HEC, 2-HPC, and GCBD dose-dependently inhibited PMA/IO-stimulated NFAT activation. CHC and HC dose-dependently reduced IL-1β and CCL2 messenger RNA (mRNA) expression. NMSC inhibited IL-1β, CCL2, and TNF-α at lower doses. At higher doses, it induced a pronounced increase in IL-6 mRNA. 2-HEC, 2-HPC, and GCBD dose-dependently inhibited LPS-induced IL-1β, TNF-α, and IL-6 synthesis. NMSC further increased LPS-stimulated IL-1β release but inhibited IL-8, TNF-α, and PGE2. Conclusion: The CBD- and CBDV-derivatives studied are suitable for targeting CB-receptors. Some may be used as selective CB2 agonists. The length of the aliphatic rest at R2 of CBD (pentyl) and CBDV (propyl) did not correlate with the binding affinity. Higher polarity at R1 appeared to favor the agonistic activity at CB2-receptors.
Targeting CB2-GPR55 Receptor Heteromers Modulates Cancer Cell Signaling
(THE JOURNAL OF BIOLOGICAL CHEMISTRY, 2014) Moreno, Estefanía; Andradas, Clara; Medrano, Mireia; Caffarel, María M.; Blasco-Benito, Sandra; Pazos, M. Ruth; Irving, Andrew J.; Lluís, Carme; Canela, Enric I.; McCormick, Peter J.; Gómez Cañas, María; Pérez Gómez, Eduardo; Fernández Ruiz, José Javier; Guzmán Pastor, Manuel; Sánchez García, María Cristina
The G protein-coupled receptors CB2 (CB2R) and GPR55 are overexpressed in cancer cells and human tumors. Because a modulation of GPR55 activity by cannabinoids has been suggested, we analyzed whether this receptor participates in cannabinoid effects on cancer cells. Here we show that CB2R and GPR55 form heteromers in cancer cells, that these structures possess unique signaling properties, and that modulation of these heteromers can modify the antitumoral activity of cannabinoids in vivo. These findings unveil the existence of previously unknown signaling platforms that help explain the complex behavior of cannabinoids and may constitute new targets for therapeutic intervention in oncology.