Cannabinoids are gaining attention in many sectors, including the pharmaceutical, nutraceutical, and cosmetic sectors. To date, several conventional and alternative techniques have been applied for Cannabis sativa L (C. sativa L.) extraction at the industrial scale. The conventional methods are liquid solvent extraction, including polar and non-polar solvents such as ethanol, hexane, petroleum ether, and other solvent extraction. Pressurized gas extraction is another conventional method and comprises gaseous hydrocarbon extraction such as n-butane and n-propane, supercritical or subcritical carbon dioxide extraction, and extraction using a refrigerant gas such as 1,1,1,2-tetrafluoroethane (HFC 134a). Alternative extraction methods include microwave-assisted extraction (MAE), ultrasound-assisted extraction (UAE), hydrodynamic cavitation, and pulsed electric fields (PEF). This review thoroughly analyzes and compares the main extraction techniques and technologies at the industrial scale found in the patent literature. In addition, several aspects of the pretreatment of cannabis plant material and its influence on extraction are also discussed.

Minor cannabinoid and non-cannabinoid molecules have been proposed to significantly contribute to the pharmacological profile of cannabis extracts. Phytoplant Research has developed highly productive cannabis cultivars with defined chemotypes, as well as proprietary methods for the extraction and purification of cannabinoids. Here, we investigate the effect of solvent selection and decarboxylation on the composition and pharmacological activity of cannabis extracts. A library of forty cannabis extracts was generated from ten different cannabis cultivars registered by Phytoplant Research at the EU Community Plant Variety Office. Plant material was extracted using two different solvents, ethanol and hexane, and crude extracts were subsequently decarboxylated or not. Cannabinoid content in the resulting extracts was quantified, and biological activity was screened in vitro at three molecular targets involved in hypoxia and inflammation (NF-κB, HIF-1α and STAT3). Changes in transcriptional activation were strongly associated to solvent selection and decarboxylation. Two decarboxylated extracts prepared with hexane were the most potent at inhibiting NF-κB transcription, while HIF-1α activation was preferentially inhibited by ethanolic extracts, and decarboxylated extracts were generally more potent at inhibiting STAT3 induction. Our results indicate that solvent selection and proper decarboxylation represent key aspects of the standardized production of cannabis extracts with reproducible pharmacological activity.

Background: Cannabidiol (CBD) is a phytocannabinoid with potential in one of the most prevalent syndromes occurring at birth, the hypoxia of the neonate. CBD targets a variety of proteins, cannabinoid CB₂ and serotonin 5HT_1A receptors included. These two receptors may interact to form heteromers (CB₂–5HT_1A-Hets) that are also a target of CBD. Aims: We aimed to assess whether the expression and function of CB₂–5HT_1A-Hets is affected by CBD in animal models of hypoxia of the neonate and in glucose- and oxygen-deprived neurons. Methods: We developed a quantitation of signal transduction events in a heterologous system and in glucose/oxygen-deprived neurons. The expression of receptors was assessed by immuno-cyto and -histochemistry and, also, by using the only existing technique to visualize CB₂–5HT_1A-Hets fixed cultured cells and tissue sections (in situ proximity ligation PLA assay). Results: CBD and cannabigerol, which were used for comparative purposes, affected the structure of the heteromer, but in a qualitatively different way; CBD but not CBG increased the affinity of the CB₂ and 5HT_1A receptor–receptor interaction. Both cannabinoids regulated the effects of CB₂ and 5HT_1A receptor agonists. CBD was able to revert the upregulation of heteromers occurring when neurons were deprived of oxygen and glucose. CBD significantly reduced the increased expression of the CB₂–5HT_1A-Het in glucose/oxygen-deprived neurons. Importantly, in brain sections of a hypoxia/ischemia animal model, administration of CBD led to a significant reduction in the expression of CB₂–5HT_1A-Hets. Conclusions: Benefits of CBD in the hypoxia of the neonate are mediated by acting on CB₂–5HT_1A-Hets and by reducing the aberrant expression of the receptor–receptor complex in hypoxic-ischemic conditions. These results reinforce the potential of CBD for the therapy of the hypoxia of the neonate.

Cannabidiol (CBD) is a phytocannabinoid with potential as a therapy for a variety of diseases. CBD may act via cannabinoid receptors but also via other G-protein-coupled receptors (GPCRs), including the adenosine A_2A receptor. Homogenous binding and signaling assays in Chinese hamster ovary (CHO) cells expressing the human version of the A_2A receptor were performed to address the effect of CBD on receptor functionality. CBD was not able to compete for the binding of a SCH 442416 derivative labeled with a red emitting fluorescent probe that is a selective antagonist that binds to the orthosteric site of the receptor. However, CBD reduced the effect of the selective A_2A receptor agonist, CGS 21680, on G_s-coupling and on the activation of the mitogen activated kinase signaling pathway. It is suggested that CBD is a negative allosteric modulator of the A_2A receptor.

Cannabis sativa L. is an ancient cultivar that has found applications in various fields, e.g., medicine, due to its beneficial effects. However, due to its psychotropic effects, the regulation of this cultivar has increased throughout the decades. In this context, the need for rapid and reliable analytical methods to ensure the quality control of Cannabis cultivars has become of extreme importance. NIRS has arisen as a powerful tool in this field due to its multiple advantages, e.g., non-destructive, rapid, and cost-effective. In this article, the chemometric techniques commonly employed in NIRS method development are described, along with their application for the analysis of Cannabis samples. Regarding qualitative methods, different mathematical treatments and classification models are explained. As for quantitative methods, the representative linear and non-linear modelling techniques applied for the development of prediction equations are described, alongside their application in the Cannabis field. To the best of our knowledge, this is the first time this type of review is written, since there are several articles which address cannabinoid determination, but the main purpose of this review is to enhance the potential of NIRS over the traditional techniques employed for the analysis of Cannabis samples.

Adenosine regulates neurotransmission, immunity, inflammation, and tissue protection via four G protein-coupled receptors, among which the adenosine A₃ receptor (A₃R) remains comparatively understudied despite its therapeutic promise. This study examined whether cannabidiol (CBD) and cannabigerol (CBG) modulate human A₃R signalling, alone or with added adenosine, using cAMP assays, molecular docking, binding-energy calculations, and molecular dynamics simulations. Both cannabinoids inhibited forskolin-stimulated cAMP accumulation in A₃R-expressing cells, an effect absent in non-transfected cells and blocked by the selective A₃R antagonist PSB-10 but not by the selective A_2A receptor antagonist SCH 58261, supporting A₃R-mediated activity under these conditions. CBG was more potent than CBD (IC₅₀ = 0.130 vs. 1.05 µM), whereas their maximal inhibitory responses were comparable. In the presence of 100 nM CBD, the apparent IC₅₀ of adenosine decreased from 138 to 42 nM, whereas 100 nM CBG reduced it to 1.0 nM, corresponding to an approximately 138-fold increase in apparent adenosine potency; CBG also increased the maximal inhibitory response. These results identify CBG as an effective enhancer of adenosine potency at human A₃R, suggesting it stabilizes an occupancy-dependent secondary pose that fine-tunes the orthosteric agonist-receptor interaction network. Further work, encompassing direct binding studies, proximal signalling assays, receptor-subtype profiling, and structural analyses, is needed to validate this mechanism.