## 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
Also, they brought attention to the fact that numerous U937 and Jurkat cells treated with the compound, especially at doses higher than 50 uM, halted predominantly in the S phase of the cell cycle. They then tested the effect of this HCAA on the epidermal growth factor receptor (EGFR), a tyrosine kinase that is overexpressed in certain cancer types. They observed a dose-dependent inhibition of EGFR by N-trans-coumaroyltyramine, with a 20% reduction in activity with 30 uM in the U937 cell line, and a 75% reduction in activity with 120 uM. Comparable results were produced for the Jurkat cell line. These results suggest that N-trans-coumaroyltyramine interferes with tyrosine kinase activity and its signaling process, necessary in cancerous cell replication, and could induce an apoptotic response. Its antiproliferative activity was tested on HeLa (cervical cancer) and A431 (epidermoid carcinoma) cell lines and it was found that N-p-trans-coumaroyltyramine significantly reduced cell viability in a dose- and time-dependent manner, at 10 uM for a 24 h treatment, and at 5 uM for 48 h of exposure [68]. N-p-trans-coumaroyltyramine was shown to enhance the sensitivity of indomethacin and diclofenac, two nonsteroidal anti-inflammatory drugs with antiproliferative activity against certain cancer types, in MCF-7 (breast cancer) and mitoxantrone-resistant MCF-7 (MCF-7/MX) cell lines. Exposure to 100 uM of N-p-trans-coumaroyltyramine for 48 h decreased MCF-7 viability by more than twofold when combined with 10 uM of both indomethacin and diclofenac [69]. For MCF-7/MX cells, 100 uM of N-p-trans-coumaroyltyramine decreased cell viability by twofold for 10 uM of indomethacin and 1.5-fold for diclofenac. Furthermore, 100 uM of N-p-trans-coumaroyltyramine increased the cytotoxic activity of endoplasmic reticulum stressors, such as thapsigargin (0.0125 uM), tunicamycin (0.3 uM), rotenone (1.0 uM), and hydrogen peroxide (10 uM) in both MCF-7 cell lines [69]. N-trans-feruoyltyramine extracted from laba garlic was reported to possess antiproliferative activity against HepG2 cells, with an IC\({}_{50}\) = 194 uM, whereas the positive control taxol had an IC\({}_{50}\) = 26 uM [70]. N-trans-feruoyltyramine and N-trans-caffeoyltyramine exhibited antiproliferative activity against A2780 (ovarian cancer) and Panc1 (pancreatic cancer) cell lines, with feruoyltyramine having an IC\({}_{50}\) = 9.1 uM (A2780) and IC\({}_{50}\) = 3.4 uM (Panc1), and caffeoyltyramine having an IC\({}_{50}\) = 11.2 uM (A2780) and IC\({}_{50}\) = 7.2 uM (Panc1) [71]. N-trans-coumaroyltyramine was reported to possess moderate acetylcholinesterase inhibitory activity, with an IC\({}_{50}\) of 48.8 uM [72]. The potential of lignanamides in Alzheimer's disease treatment was tested by verifying its effect on BACE 1, PPARy, and PGC-1\(\alpha\) expression in amyloid \(\beta\) precursor protein (APP)-producing N2a cells. The BACE 1 enzyme produces monomeric forms of amyloid \(\beta\) peptides, which accumulate in the brain, and is associated with a worsening of the symptoms of the disease. BACE 1 is activated by proinflammatory cytokines, namely IL-1\(\beta\), IL-6, and TNF-\(\alpha\). N-trans-caffeoyltyramine very significantly reduced the BACE 1 expression of N2a cells at 0.03 uM, more so at 24 h of exposure than 48 h [73]. Additionally, PPARy, a BACE 1 regulator whose activation inhibits BACE 1, was upregulated by more than twofold with 0.03 uM of N-trans-caffeoyltyramine after 24 h exposure, with a slight decrease after 48 h (just less than twofold), and with 0.08 uM of N-trans-caffeoyltyramine, PPARy was upregulated more than threefold after 24 h, also with a slight decrease after 48 h (just about threefold). PGC-1\(\alpha\) is a coactivator participating in PPARy transcription and decreased expression is thought to be linked to [PERSON]'s disease [73]. A total of 0.03 and 0.08 uM of N-trans-caffeoyltyramine had comparable effects on PGC-1\(\alpha\) expression, upregulating by approximately twofold after 24 h, again with a slight diminution after 48 h of exposure.