β-Caryophyllene ((-)-(E)-Caryophyllene) acts as an CB2 receptor agonist.

β-Caryophyllene shows selective anti-proliferative effects against three tested cancer cell lines: HCT 116 (colon cancer, IC50=19 μM), PANC-1 (pancreatic cancer, IC50=27 μM), and HT29 (colon cancer, IC50=63 μM), but exhibits moderate or poor cytotoxicity against ME-180, PC3, K562, and MCF-7. It demonstrates higher selectivity for colorectal cancer cells, particularly HCT 116 (SI=27.9), followed by PANC-1 (SI=19.6) and HT 29 (SI=8). The apoptotic index for HCT 116 cells after 24 hours of β-Caryophyllene treatment is 64±0.04. At 10 μM, it causes significant nuclear condensation after 6 hours and exhibits dose- and time-dependent inhibitory effects on HCT 116 cell motility.

During the examination period, administering varying doses of β-Caryophyllene had no impact on swimming speed. However, oral administration of β-Caryophyllene significantly reduced β-amyloid deposition in transgenic mice in a nearly dose-dependent manner, with the two highest doses showing comparable efficacy in decreasing β-amyloid accumulation. Notably, vehicle-treated mice displayed an increased number of activated astroglial cells compared to those treated with any dose of β-Caryophyllene. Furthermore, β-Caryophyllene effectively lowered the elevated COX-2 protein levels observed in vehicle-treated APP/PS1 mice. Additionally, animals receiving β-Caryophyllene treatment demonstrated a higher object recognition index than their vehicle-treated counterparts, indicating improved memory retention [t(14)=4.204, P<0.05]. There was no significant difference in the total time spent exploring objects between the β-Caryophyllene and vehicle-treated groups during the test trial (t(14)=0.5874, P>0.05). β-Caryophyllene treatment did not significantly affect seizure-induced neurochemical alterations.

Stock solution (10 mM) of β-Caryophyllene is prepared using DMSO. Further, various concentrations (3 to 100 μM) of β-Caryophyllene are prepared by serially diluting the stock with respective culture medium.Panel of human cancer cells such as, pancreatic (PANC-1), colorectal (HCT-116 and HT-29), invasive squamous cell carcinoma (ME-180), leukemia (K562), hormone sensitive and invasive breast cancer cell line (MCF-7), and prostatic (PC3) adenocarcinoma cell lines are used. Cells are incubated in a humidified CO2 incubator at 37°C supplied with 5% CO2. Inhibitory effect of β-Caryophyllene on proliferation of the cell lines is tested using the MTT assay. The selectivity index (SI) for the cytotoxicity of β-Caryophyllene is calculated using the ratio of IC50 of β-Caryophyllene on a normal cell line (NIH-3T3) to the IC50 of β-Caryophyllene on cancer cell lines. They are for reference only.

β-Caryophyllene is initially dissolved in dimethyl sulfoxide and further diluted (2%) in sterile phosphate-buffered saline (PBS) with 10% Cremophor EL (poluoxyl-35 hydrogenated castor oil).Male double transgenic APP/PS1 mice and wild-type littermates are used. The mice are group housed (3 to 5 animals/cage) with a 12:12-hour light/dark cycle and ad libitum access to food and water. In this experiment, animals are orally treated by gavage with 16, 48, or 144 mg/kg of β-Caryophyllene every morning for 10 weeks starting at the age of 7 months. All vehicle solutions are used for the respective control animal treatments and the Morris water maze test is performed. They are for reference only.