Hempseed Lignanamides Rich-Fraction: Chemical Investigation and Cytotoxicity towards U-87 Glioblastoma Cells

Abstract

The weak but noteworthy presence of (poly)phenols in hemp seeds has been long overshadowed by the essential polyunsaturated fatty acids and digestible proteins, considered responsible for their high nutritional benefits. Instead, lignanamides and their biosynthetic precursors, phenylamides, seem to display interesting and diverse biological activities only partially clarified in the last decades. Herein, negative mode HR-MS/MS techniques were applied to the chemical investigation of a (poly)phenol-rich fraction, obtained from hemp seeds after extraction/fractionation steps. This extract contained phenylpropanoid amides and their random oxidative coupling derivatives, lignanamides, which were the most abundant compounds and showed a high chemical diversity, deeply unraveled through high resolution tandem mass spectrometry (HR-MS/MS) tools. The effect of different doses of the lignanamides-rich extract (LnHS) on U-87 glioblastoma cell line and non-tumorigenic human fibroblasts was evaluated. Thus, cell proliferation, genomic DNA damage, colony forming and wound repair capabilities were assessed, as well as LnHS outcome on the expression levels of pro-inflammatory cytokines. LnHS significantly inhibited U-87 cancer cell proliferation, but not that of fibroblasts, and was able to reduce U-87 cell migration, inducing further DNA damage. No modification in cytokines’ expression level was found. Data acquired suggested that LnHS acted in U-87 cells by inducing the apoptosis machinery and suppressing the autophagic cell death.

Chemical Analysis of Minor Bioactive Components and Cannabidiolic Acid in Commercial Hemp Seed Oil

“…Recently, hydroxycinnamoyl amides, lignanamides, and flavonol glycosides were identified as minor constituents of hemp seed [18,27], and their presence is favorably revealed in hemp seed meal, whereas their content is highly variable in hemp seed oil, being strongly dependent on the extraction method applied, and/or cultivar considered for oil obtainment [28]. Smeriglio et al [29] also characterized the polyphenolic compounds of cold-pressed seed oil from Finola cultivar, finding a Total Phenol Content (TPC) equal to 267.5 mg Gallic Acid Equivalents (GAE)/100 g. The cultivar identity could also affect TPC content, as a value 10-fold lower (2.1 mg GAE/100 g) was estimated for hemp seed oil from Fedora cultivar by Siano et al [26], who monitored the distribution of hemp seed components in the flour and oil after fractionation by cold-pressing.…”

A Cup of Hemp Coffee by Moka Pot from Southern Italy: An UHPLC-HRMS Investigation

“…The chemical composition and the nutraceutical nature of these fruits have been recently reviewed, pointing out that its functional benefits are still far from being fully understood [4]. In fact, besides essential polyunsaturated fatty acids with a ratio nearly equal to 3:1 (ω-6: ω-3), which characterize also the oil together with vitamins, minerals, amino acids and phytosterols, the presence of antioxidant (poly)phenols is worth mentioning, not only for their preservation of oil stability and quality but also for their numerous health benefits for consumers [5][6][7].…”

Cancer Initiation, Progression and Resistance: Are Phytocannabinoids from Cannabis sativa L. Promising Compounds?

“…These data have been confirmed in the U-87 glioblastoma cell line in which CBD led to a concentration-related inhibition of the U87 cell viability [ 92 ]. In accordance, Nigro et al analyzed, in the same glioblastoma cell line, the effects of a heterogenous extract from Cannabis Sativa L., finding the inhibition of both proliferation and migration from a dose of 25 µg/mL [ 32 ]. The underlying molecular mechanism is not completely clear but the authors evidenced DNA damage [ 32 ].…”

“…In accordance, Nigro et al analyzed, in the same glioblastoma cell line, the effects of a heterogenous extract from Cannabis Sativa L., finding the inhibition of both proliferation and migration from a dose of 25 µg/mL [ 32 ]. The underlying molecular mechanism is not completely clear but the authors evidenced DNA damage [ 32 ]. Recently, Singer et al demonstrated that apoptosis together with ROS production are two additional mechanisms involved in CBD inhibitory activity of 3832 and 387 glioma primary stem cell lines (GSC) with an IC50 value of 3.5 μM [ 93 ].…”

“…Methylated prenylated flavones, cannflavins A, B, and C, as well as flavonols glycosides, such as kaempferol 3- O -sophoroside and quercetin 3- O -sophoroside, were also isolated from hemp pollen [ 31 ]. Furthermore, from Cannabis sativa L. fruits, beyond to a source to produce an oil rich in tocopherols and mono-, di-, and polyunsaturated fatty acids (PUFA) such as α-linolenic (ALA), γ-linolenic acid (GLA), and stearidonic acid (SDA), contained different phenylamides, tyramine and octopamine derivatives of hydroxycinnamic acids, and their lignanamides, also called cannabisins [ 32 , 33 ].…”