## 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
They reported a MIC of 5 ug/mL against _Candidia albincans_, 10 ug/mL against _Saccharomyces cerevisiae_, and 5-10 ug/mL against _Trichospor beigelli_, compared to the positive control amphotericin B, which had an MIC of 0.63 ug/mL, 5 ug/mL, and 1.25 ug/mL, with respect to each fungus. N-p-trans-coumaroyltyramine exhibited antimicrobial activity against _Escherichia coli_, where the reported IC\({}_{50}\) was 0.8 ug/mL, compared to the positive control ciprofloxacin (IC\({}_{50}=0.01\) ug/mL) [82]. N-\(p\)-trans-coumaroyltyramine's antimicrobial activity could possibly extend to urease-positive bacteria like _Helicobacter pylori_, because of its reported anti-urease activity (IC\({}_{50}=45.2\) uM) against the _Canvalia ensiformis_ (jack bean) urease enzyme [83]. N-\(p\)-trans-coumaroyltyramine and N-trans-feruloyltyramine were reportedly produced in response to _Chilo suppressalis_ exposure in rice (_Orza sativa_, cv. Oochikara), suggesting a role in herbivore insect management [84]. The presence of these compounds in the feed given to _C. suppressalis_ resulted in a weight loss at concentrations of 100 ug/g, suggesting anti-insect and antifeedant activity. N-\(p\)-trans-coumaroyltyramine, N-trans-feruloyltyramine, and N-trans-caffeoyltyramine exhibited anti-melanogenesis activity at 50 uM, reducing the amount of melanin produced by alpha melanocyte stimulating hormone (\(\alpha\)-MSH) activation in B16F10 melanoma cells [85]. Anti-melanogenesis activity is ranked as follows: N-trans-caffeoyltyramine (IC\({}_{50}=0.8\) uM), N-\(p\)-trans-comamaroyltyramine (IC\({}_{50}=6.3\) uM), and N-trans-feruloyltyramine (IC\({}_{50}=20.2\) uM). It is worth noting that N-\(p\)-trans-coumaroyltyramine at 50 uM decreased B16F10 cell viability by more than 50% [85]. All three cinnamamides also exhibited anti-tyrosinase activity. Lignanamide Activity In Vitro  
Cannabisin G extracted from _Berberis vulgaris_ was reported to possess strong antioxidant activity, with an IC\({}_{50}\) = 2.7 ug/mL in a DPPH radical-scavenging assay [86]. Cannabisin B extracted from _C. sativa_ hemp seed hulls would also possess decent antioxidant properties, with a reported IC\({}_{50}\) = 11.2 ug/mL in a DPPH radical-scavenging assay, slightly lower than N-\(p\)-trans-caffeoyltyramine (IC\({}_{50}\) = 9.42 ug/mL) [87]. In another study concerning hemp seed hulls, cannabis B demonstrates antiproliferative activity in HepG2 (liver cancer) cells through the inhibition of the AKT/mTOR pathway and autophagy, at concentrations exceeding 167 uM. Furthermore, HepG2 cells treated with cannabis B tended to arrest the cell cycle in the S phase in a dose-dependent manner [88]. Lignanamides extracted from hemp seeds were tested by [PERSON] et al. (2015) for their antioxidant and anticholinesterase abilities. They found that cannabis A and D were among the most effective lignanamides tested in DPPH (IC\({}_{50}\) = 32.9 uM and 23.9 uM), ABTS (IC\({}_{50}\) = 6.6 uM and 0.5 uM), and ORAC (IC\({}_{50}\) = 7.3 uM and 73.0 uM) assays, against the positive control quercetin (IC\({}_{50}\) = 25.5 uM, 0.4 uM, and 9.2 uM) [41]. C. _sativa_ lignanamides presenting anticholinesterase activity are 3,3\({}^{\prime}\)-demethyl-heliotropamide (IC\({}_{50}\) = 46.2 uM) and 3,3\({}^{\prime}\) dimethyl-grossamide (IC\({}_{50}\) = 38.7 uM), against the positive control galanthamine (IC\({}_{50}\) = 2.8 uM). Cannabisin F reportedly exhibited anti-inflammatory activity against BV2 microglia cells. The pretreatment of BV2 cells with doses of 15 uM of cannabis F significantly reduced IL-6 and TNF-\(\alpha\) production induced by LPS (100 ng/mL), at both protein and mRNA levels, and in a dose-dependent manner [89]. [PERSON] and colleagues also provided evidence that the anti-inflammatory activity of cannabis F is SIRT1-dependent, and acts by inhibiting NF-\(\kappa\)B signaling. Grossamide also reduced pro-inflammatory cytokines IL-6 and TNF-\(\alpha\) levels of BV2 cells treated with LPS (100 ng/mL, 24 h), with concentrations as low as 10 uM [90].