Role of Caryophyllane Sesquiterpenes in the Entourage Effect of Felina 32 Hemp Inflorescence Phytocomplex in Triple Negative MDA-MB-468 Breast Cancer Cells

Abstract

Cannabis sativa L. crops have been traditionally exploited as sources of fibers, nutrients, and bioactive phytochemicals of medical interest. In the present study, two terpene-rich organic extracts, namely FOJ and FOS, obtained from Felina 32 hemp inflorescences collected in June and September, respectively, have been studied for their in vitro anticancer properties. Particularly, their cytotoxicity was evaluated in different cancer cell lines, and the possible entourage effect between nonintoxicating phytocannabinoids (cannabidiol and cannabichromene) and caryophyllane sesquiterpenes (β-caryophyllene, β-caryophyllene oxide and α-humulene), as identified at GC/MS analysis, was characterized. Modulation of cannabinoid CB1 and CB2 receptors was studied as a mechanistic hypothesis. Results highlighted marked cytotoxic effects of FOJ, FOS, and pure compounds in triple negative breast cancer MDA-MB-468 cells, likely mediated by a CB2 receptor activation. Cannabidiol was the main cytotoxic constituent, although low levels of caryophyllane sesquiterpenes and cannabichromene induced potentiating effects; the presence in the extracts of unknown antagonistic compounds has been highlighted too. These results suggest an interest in Felina 32 hemp inflorescences as a source of bioactive phytocomplexes with anticancer properties and strengthen the importance of considering the possible involvement of minor terpenes, such as caryophyllane sesquiterpenes, in the entourage effect of hemp-based extracts.

Harpagophytum procumbens Root Extract Mediates Anti-Inflammatory Effects in Osteoarthritis Synoviocytes through CB2 Activation

“…The CB2 up-regulation induced by α-humulene, eugenol, and harpagoside is supported by the activation of CB2 induced by β-caryophyllene and by the action of other compounds present in the H. procumbens root extract, which may also affect other pathways. Potentiating properties for both eugenol and caryophyllane sesquiterpenes, such as β-caryophyllene and α-humulene, have been highlighted when these molecules were administered in combination with anticancer drugs or herbal phy-tocomplexes [42,43]. Particularly, the synergic effects of caryophyllane sesquiterpenes have been found to be associated with the inhibition of efflux pumps, which leads to the intracellular accumulation of other substances [44].…”

Novel Insights into the Immunomodulatory Effects of Caryophyllane Sesquiterpenes: A Systematic Review of Preclinical Studies

“…Similar effects on macrophage function have been found for β-caryophyllene oxide, albeit highlighted in just one study [33]. Considering the structural similarity with β-caryophyllene and the recently highlighted ability to modulate CB2 receptor expression [2], it is conceivable that it can act like the lead compound, thus suggesting the need for further investigation.…”

“…This is a critical methodologic feature as high impurity levels can dramatically affect the activity of the tested compounds, leading to unreliable results. Indeed, in our previous study, we highlighted that very low concentrations of caryophyllane sesquiterpenes were able to synergize cannabidiol cytotoxicity [2]; therefore, considering the possible modulating effects of minor compounds, the true mechanism of the tested substance is not ensured. As suggested by the best practices to be applied in phytopharmacological research, details about compound purity should be reported; in particular, the use of high purity (95-99%) compounds is recommended [66].…”

“…Syzygium aromaticum (L.) Merr. ), Hypericum perforatum L., Mentha suaveolens L., and Cannabis sativa L., along with herbal extracts from Piper nigrum L. and Harpagophytum procumbens DC are only some examples of the natural sources of these compounds [1][2][3][4][5][6]. Over the years, great attention has been devoted to their bioactivities, with β-caryophyllene being especially studied due to its analgesic, anti-inflammatory, antioxidant, neuroprotective, antidiabetic, and chemopreventive properties; moreover, its ability to modulate cannabinoid CB2 receptors has been highlighted too [1].…”

Sorafenib Chemosensitization by Caryophyllane Sesquiterpenes in Liver, Biliary, and Pancreatic Cancer Cells: The Role of STAT3/ABC Transporter Axis

“…Synergistic effects of β-caryophyllene oxide towards sorafenib were also confirmed in a xenograft model of liver cancer [20]. More recently, we reported the potentiating effects of caryophyllane sesquiterpenes towards cannabidiol and harpagoside in hemp and devil's claw extracts, respectively [18,22].…”