Cannabinoid receptor 2 (CB2); Peroxisome proliferator-activated receptor-gamma (PPARγ) [1]

β-Caryophyllene has been shown to selectively bind to the CB2 receptor and act as a full agonist. It can inhibit pathways that typically lead to the expression of proinflammatory cytokines (IL-1β, IL-6, IL-8, and TNF-α). [1]
The anti-inflammatory effect of β-caryophyllene involves both CB2 receptor activation and the PPARγ pathway. [1]
β-Caryophyllene is a lipophilic molecule that can easily cross cell membranes. [1]

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For the tested cancer cell lines, β-Caryophyllene showed a selective anti-proliferative effect against HCT 116 (colon cancer, IC50=19 μM), PANC-1 (pancreatic cancer, IC50=27 μM), and HT29 (colon cancer, IC50=63 μM). Conversely, β-Caryophyllene showed either moderate or poor cytotoxic effects against ME-180, PC3, K562, and MCF-7. Based on the results, it can be observed that β-Caryophyllene has a higher selectivity towards colorectal cancer cells (HCT 116), with a selectivity index (SI) of 27.9. PANC-1 and HT 29 cells follow with SIs of 19.6 and 8, respectively. After 24 hours of treatment with β-Caryophyllene, the estimated apoptotic index for HCT 116 cells is 64± 0.04. After 6 hours of treatment, 10 μM concentration of β-Caryophyllene results in considerable nuclei condensation. HCT 116 cell motility is inhibited by β-caryophyllene in a time- and dose-dependent manner[2].

In APP/PS1 transgenic Alzheimer's disease mice, oral administration of β-caryophyllene (16, 48, or 144 mg/kg, once daily for 10 weeks) prevented cognitive impairment in a dose-dependent manner, as measured by the Morris water maze test (escape latency and swimming distance). The two higher doses (48 and 144 mg/kg) were more effective than the lowest dose, with no significant difference between them. [1]
β-Caryophyllene treatment reduced β-amyloid burden in both the hippocampus and cerebral cortex of APP/PS1 mice, as measured by immunohistochemistry. [1]
β-Caryophyllene reduced astrogliosis in the cerebral cortex (but not hippocampus) of APP/PS1 mice, as shown by GFAP immunohistochemistry and Western blotting. [1]
β-Caryophyllene (48 mg/kg) reduced microglial activation (Iba-1 protein levels by 51.47% compared to vehicle-treated APP/PS1 mice), decreased COX-2 protein levels, and reduced mRNA levels of proinflammatory cytokines TNF-α and IL-1β in the cerebral cortex of APP/PS1 mice. IL-10 mRNA levels were unchanged. [1]
The CB2-selective antagonist AM630 (10 mg/kg, i.p., given 30 min before β-caryophyllene) significantly reversed the protective effects of β-caryophyllene on behavioral tests, β-amyloid burden, astrogliosis, microglial activation, and mRNA expression of TNF-α and IL-1β. [1]
The PPARγ-selective antagonist GW9662 (1 mg/kg, i.p., given 30 min before β-caryophyllene) significantly blocked the beneficial effects of β-caryophyllene by partially reversing the improvement in behavioral tests, β-amyloid burden, astrogliosis, microglial activation, and mRNA expression of TNF-α and IL-1β. [1]
In C57BL/6 mice, β-caryophyllene (100 mg/kg, i.p., 60 min before PTZ) increased the latency to PTZ-induced myoclonic jerks by 167% on average compared to vehicle-treated mice. There was a significant positive correlation between the doses of β-caryophyllene (10, 30, 100 mg/kg) and the latency to PTZ-induced myoclonic seizures (rs = 0.4237, P < 0.005). Onset latency and duration of the first PTZ-induced generalized tonic-clonic seizure did not significantly change. Mortality rate was zero. [3]
EEG recordings confirmed that β-caryophyllene (100 mg/kg) delayed the appearance of PTZ-induced myoclonic jerks but did not alter the duration or generalized seizure-associated wave patterns. [3]
β-Caryophyllene (100 mg/kg) did not significantly alter performance in the open-field test (crossings, rearings, time spent in center), rotarod test (latency to fall), or forced swim test (immobility time) in mice. [3]
β-Caryophyllene (100 mg/kg) significantly improved the object recognition index in mice compared to vehicle-treated controls. Total time spent in object exploration was not significantly different. [3]
β-Caryophyllene (100 mg/kg) did not prevent PTZ-induced oxidative stress: TBARS levels increased in the cerebral cortex (but not hippocampus), and NPSH content decreased in the hippocampus (but not cerebral cortex). [3]

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There are no impacts on swimming speed during the test when β-Caryophyllene treatment is administered at different doses. β-Caryophyllene given orally to transgenic mice reduces the rise in β-amyloid deposition in a roughly dose-dependent manner, with the two higher dosages having almost comparable effects on β-amyloid burden modification. In mouse brains treated with vehicle, there are more activated astrocyte cells than in brains treated with varying dosages of β-Caryophyllene. When APP/PS1 mice are given with a vehicle, the increase of the COX-2 protein level can be effectively reduced by β-caryophyllene[1]. The object recognition index is higher in animals treated with β-Caryophyllene than in those treated with vehicles [t(14)=4.204, P<0.05]. There is no significant difference observed in the total amount of time spent object exploring throughout the test trial between animals treated with β-Caryophyllene and those treated with vehicle (t(14)=0.5874, P>0.05). β-Caryophyllene treatment has no discernible effect on these neurochemical alterations brought on by seizures[3].

TBARS content measurement for lipid peroxidation: Tissue homogenate (100 μL) was mixed with 15 μL of 8.1% SDS, 60 μL of acetic acid buffer (2.5 M, pH 3.4), and 115 μL of 0.81% thiobarbituric acid. The mixture was heated at 95°C for 120 minutes in a water bath. After cooling to room temperature, absorbance was measured in the supernatant at 532 nm. Results were calculated as nmol MDA/mg protein. [3]
NPSH content measurement: Homogenates were precipitated with 10% TCA and centrifuged at 3000 × g at 4°C for 10 minutes. The supernatant (100 μL) was added to a reaction medium containing potassium phosphate buffer (1 M, pH 7.4) and 10 mM DTNB. NPSH levels were measured spectrophotometrically at 412 nm. Results were calculated using a standard curve constructed with reduced glutathione and expressed as nmol NPSH/mg protein. [3]

APP/PS1 mouse model: Male double transgenic APP/PS1 mice and wild-type littermates were used. Animals were group-housed (3-5 animals/cage) with a 12-hour light/dark cycle and ad libitum access to food and water. β-Caryophyllene was orally administered by gavage at 16, 48, or 144 mg/kg every morning for 10 weeks starting at 7 months of age. β-Caryophyllene was initially dissolved in DMSO and further diluted (2%) in sterile PBS with 10% Cremophor EL. For antagonist studies, mice received intraperitoneal injections of AM630 (10 mg/kg) or GW9662 (1 mg/kg) every morning for 10 weeks, given 30 minutes before β-caryophyllene (48 mg/kg). [1]
\nMorris water maze test: A steel circular pool (diameter 100 cm, height 35 cm) was filled with non-toxic white-dyed water at 22-25°C. A transparent platform (diameter 8 cm, height 10 cm) was submerged 1 cm below the water surface. Mice underwent daily acquisition tests (3 times/day) for 5 consecutive days. Maximum trial length was 90 seconds. A probe trial was conducted 24 hours after the last acquisition test for 60 seconds. [1]
\nImmunohistochemistry: Brain slices (4 μm) were processed with antigen retrieval, blocked with 3% hydrogen peroxide, and incubated with normal goat serum for 90 minutes. Sections were incubated with anti-β-amyloid (1:100) or anti-GFAP (1:200) antibodies overnight at 4°C. Development was conducted using biotin-streptavidin IHC detection kits, visualized by DAB oxidation, counterstained with hematoxylin for 60 seconds, and mounted. [1]
\nPTZ seizure model in C57BL/6 mice: Adult C57BL/6 mice (25-35 g, 60-90 days old) of both genders were used. β-Caryophyllene (10, 30, or 100 mg/kg) or vehicle (0.9% NaCl containing 0.05% Tween 80) was injected intraperitoneally. Sixty minutes later, PTZ (60 mg/kg, i.p.) was injected, and animals were observed for 15 minutes. Latency to myoclonic jerks, latency to generalized seizure, and duration of the first generalized seizure were recorded. [3]
\nEEG recordings: Mice were anesthetized with ketamine (80 mg/kg) and xylazine (10 mg/kg) and placed in a stereotaxic apparatus. Two stainless steel screw electrodes were placed over the parietal cortex with a ground lead over the nasal sinus. Six days after surgery, mice were habituated for 20 minutes before EEG recording. A 30-minute baseline recording was obtained. Mice were injected with β-caryophyllene (100 mg/kg) or vehicle, and 60 minutes later PTZ was injected. EEG signals were recorded for 15 minutes, amplified, filtered (0.1 to 50.0 Hz bandpass), and digitized (sampling rate 1024 Hz). [3]
\nOpen-field test: Mice were placed in the center of a round open field (56 cm diameter) with floor divided into 10 equal areas. β-Caryophyllene (100 mg/kg) or vehicle was injected 60 minutes before the test. Number of crossings and rearing responses were recorded for 5 minutes. [3]
\nObject recognition test: The test consisted of habituation #1 (first training session, 10 min with two identical objects), habituation #2 (second training session, 4 hours later, one object replaced with a new object, 10 min), and test session (24 hours after habituation #1, the new object replaced with another new object, 10 min). Recognition index = time spent in new object / (time spent in new object + time spent in familiar object). β-Caryophyllene (100 mg/kg) or vehicle was injected 60 minutes before the test session. [3]
\nRotarod test: Fine motor coordination was assessed using a rotarod apparatus (3.7 cm rod diameter, 8 rpm constant speed). The task consisted of one training session and one testing session 24 hours apart. Cutoff time was 60 seconds. β-Caryophyllene (100 mg/kg) or vehicle was injected 60 minutes before the test session. [3]
\nForced swim test: Mice were placed in individual PVC cylinders (30 cm tall × 10 cm diameter) containing 23-25°C water (20 cm deep). Immobility time during the 5-minute test was recorded. β-Caryophyllene (100 mg/kg) or vehicle was injected 60 minutes before the test session. [3]\n

The provided texts indicate that β-caryophyllene is a lipophilic molecule that can pass the blood-brain barrier and act on the central nervous system. It can exert an anxiolytic-like effect on mice, supporting that it reaches the brain. However, no detailed pharmacokinetic data (e.g., half-life, bioavailability, clearance) are reported. [1]

Acute administration of β-caryophyllene at doses up to 5 g/kg in mice did not cause mortality or signs of toxicity. The LD50 in rats or rabbits exceeded 5 g/kg. [3]
β-Caryophyllene was not mutagenic in Salmonella typhimurium strains or in an unscheduled DNA synthesis assay at concentrations up to 150 mg/plate and 10 mg/mL. [3]
In C57BL/6 mice, β-caryophyllene (100 mg/kg, i.p.) did not cause significant adverse effects on spontaneous locomotor activity (open-field test), forced locomotor activity (forced swim test), or motor coordination (rotarod test). [3]

[1]. β-Caryophyllene ameliorates the Alzheimer-like phenotype in APP/PS1 Mice through CB2 receptor activation and the PPARγ pathway. Pharmacology. 2014;94(1-2):1-12.

[2]. The Anticancer, Antioxidant and Antimicrobial Properties of the Sesquiterpene β-Caryophyllenefrom the Essential Oil of Aquilaria crassna. Molecules. 2015 Jun 26;20(7):11808-29.

[3]. Anticonvulsant activity of β-caryophyllene against pentylenetetrazol-induced seizures. Epilepsy Behav. 2016 Mar;56:26-31.

β-Caryophyllene is a pale yellow, oily liquid with an odor between clove and turpentine. (NTP, 1992)
(-)-β-Caryophyllene is a β-caryophyllene with an S-configuration at the stereocenter adjacent to the outer ring double bond and an R-configuration at the remaining stereocenters. It is the most common form of β-caryophyllene and is found in many essential oils, especially clove oil. It is used as a nonsteroidal anti-inflammatory drug, fragrance, metabolite, and insect attractant. It is the enantiomer of (+)-β-caryophyllene.
Caryophyllene has been reported in tea (Camellia sinensis), Trichogonia grazielae, and several other organisms with relevant data.
See also: isocaryophyllene (related); humulene (related). (+)-β-caryophyllene (is the enantiomer of…)…see more…

C15H24

204.35

204.187

C, 88.16; H, 11.84

87-44-5

5281515

Colorless to light yellow liquid

0.9±0.1 g/cm3

268.4±10.0 °C at 760 mmHg

< 25 °C

104.9±13.8 °C

0.0±0.3 mmHg at 25°C

1.495

6.78

0

0

0

15

293

2

C1(C([H])([H])[H])(C([H])([H])[H])C([H])([H])[C@]2([H])C(=C([H])[H])C([H])([H])C([H])([H])C([H])=C(C([H])([H])[H])C([H])([H])C([H])([H])[C@@]12[H] |c:25|

NPNUFJAVOOONJE-GFUGXAQUSA-N

InChI=1S/C15H24/c1-11-6-5-7-12(2)13-10-15(3,4)14(13)9-8-11/h6,13-14H,2,5,7-10H2,1,3-4H3/b11-6+/t13-,14-/m1/s1

(1R,4E,9S)-4,11,11-trimethyl-8-methylidenebicyclo[7.2.0]undec-4-ene

CCRIS 8094; AI3-36121; BETA-CARYOPHYLLENE; Caryophyllene; (-)-trans-Caryophyllene; 87-44-5; L-Caryophyllene; Caryophyllene

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Ethanol :≥ 176.67 mg/mL (~864.55 mM)
DMSO : ~50 mg/mL (~244.68 mM)
H2O : ~0.67 mg/mL (~3.28 mM)

Solubility in Formulation 1: ≥ 13.25 mg/mL (64.84 mM) (saturation unknown) in 10% EtOH + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 132.5 mg/mL clear EtOH stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: 13.25 mg/mL (64.84 mM) (saturation unknown) in 10% EtOH + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 132.5 mg/mL clear EtOH stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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Solubility in Formulation 3: ≥ 13.25 mg/mL (64.84 mM) (saturation unknown) in 10% EtOH + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 132.5 mg/mL clear EtOH stock solution to 900 μL of corn oil and mix evenly.

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Solubility in Formulation 4: ≥ 2.5 mg/mL (12.23 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 5: ≥ 2.5 mg/mL (12.23 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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Solubility in Formulation 6: ≥ 2.5 mg/mL (12.23 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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Solubility in Formulation 7: 100 mg/mL (489.36 mM) in PBS (add these co-solvents sequentially from left to right, and one by one), clear solution; with ultrasonication.

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 (Please use freshly prepared in vivo formulations for optimal results.)