## 4 Biological Activity of Cannabis Metabolites
### Biological Activity of Cannabis Alkaloids and Other Nitrogen-Containing Compounds
#### 4.2.1 In Vitro Activity of Cannabis Nitrogen-Containing Compounds
Furthermore, N-trans-caffeoyltyramine and N-trans-feruoyltyramine extracted from _Bassia indica_ and _Agathophora alopecuroides_ exhibited the good inhibition of factors implicated in [PERSON]'s disease, namely BACE1 and MAO-B activity, A\(\beta\) aggregation, and Tau-protein phosphorylation. N-trans-caffeoyltyramine had an IC\({}_{50}\) of 10.2 ug/mL (BACE1), 2.6 ug/mL (MAO-B), 6.0 ug/mL (A\(\beta\) aggregation), and 6.1 ug/mL (Tau protein), whilst N-trans-feruoyltyramine had, respectively, 11.6 ug/mL (BACE1), 2.2 ug/mL (MAO-B), 1.2 ug/mL (A\(\beta\) aggregation), and 2.1 ug/mL (Tau protein) [74]. N-trans-caffeoyltyramine extracted from _Lycium chinese_ was reportedly effective in protecting PC12 cells from apoptotic cell death induced by H\({}_{2}\)O\({}_{2}\)[75]. Where control PC12 cells had an apoptotic rate of 2.7%, the H\({}_{2}\)O\({}_{2}\) control had a rate of 23.8%. Cells pretreated with 20 uM and 40 uM of N-trans-caffeoyltyramine had an apoptotic rate of 11.7% and 7.3%, respectively, suggesting a neuroprotective function. In addition to the neuroprotective effects, HCAAs possess antioxidant activity, which seems to be related with the groups in position 3 and 4 of the benzene ring of the cinnamic acid moiety. Free hydroxyl groups in those positions are thought to be key to the enhanced antioxidant effect. Indeed, N-caffeoyltyramine has the highest antioxidant capacity among them in a DPPH-radical scavenging assay, followed by N-feruloyltyramine, N-sinapoyltyramine, and N-p-coumaroyltyramine [76]. Another DPPH-radical assay performed on a Celtis occidentalis extract by [PERSON] et al. in 2020 corroborates these findings. They reported the antioxidant activity of HCAAs as such: N-trans-caffeoyltyramine (IC\({}_{50}=31.6\) uM) \(>\) N-trans-feruloyltyramine (IC\({}_{50}=47.9\) uM) \(>\) N-\(p\)-trans-coumaroyltyramine (IC\({}_{50}=1378\) uM) [77]. Although it is worth considering that the literature data seem to trend in the same general direction regarding HCAA antioxidant activity, published results sometimes have discrepancies in them, so much so that it brought some to hypothesize that plant sources might have an influence on their antioxidant capabilities [78]. N-trans-caffeoyltyramine exhibited anti-inflammatory activity through the inhibition of TNF-\(\alpha\)-induced NF-\(\kappa\)B production in HEK293/NF-\(\kappa\)B-luciferase cells [79]. At 30 uM, N-trans-caffeoyltyramine inhibited NF-\(\kappa\)B production by 74% (IC\({}_{50}=18.4\) uM). N-trans-feruloyltyramine, and to a lesser extent N-p-trans-coumaroyltyramine, extracted from Allium hookeri, showed senomorphic activity, inhibiting a senescence-associated secretory phenotype (SASP) [80]. Additionally, they exhibited anti-inflammatory activity, as they decreased IL-1\(\alpha\) and IL-8 mRNA in replicative senescent HDF cells at 10 uM. This may suggest that N-trans-feruloyltyramine is a good candidate for fighting age- and senescence-related illnesses. N-trans-feruloyltyramine and N-trans-caffeoyltyramine possess affinity for hepatic nuclear factor 4\(\alpha\) (HNF-4\(\alpha\)), especially N-trans-caffeoyltyramine, acting as agonists [81]. The activation of HNF-4\(\alpha\) induces lipophagy, which results in a release of lipids in hepatic tissue, which can alleviate non-alcoholic fatty liver disease (NAFLD). Also, [PERSON] et al. (2021) brought evidence that 20 uM of N-trans-caffeoyltyramine increased mRNA levels of insulin and HNF-4\(\alpha\) by twentyfold and fivefold, respectively; as for N-trans-feruloyltyramine (20 uM), mRNA levels increased more than tenfold for insulin and twofold for HNF-4\(\alpha\). These results suggest that both HCAAs could be beneficial prospects in type II diabetes and hepatic fat regulation. The antifungal activity of N-trans-caffeoyltyramine extracted from _Lycium dlinense_ was evaluated by [PERSON] et al. (2004).