The chemical composition of PipeNig® is characterized by the presence of sesquiterpene (E)-β-caryophyllene, a sequiterpene hydrocarbon (Figure 1).

La composizione chimica di PipeNig® è caratterizzata dalla presenza del (E)-β-cariofillene, un sequiterpene idrocarburico (Figura 1).

Figure 1: Structure formula of (E)-β-caryophyllene

PipeNig® is analyzed by gas chromatography (GC) coupled to mass spectrometry (MS) for qualitative analysis of the compounds and by GC coupled to flame ionization detection (FID) for quantitative analysis. GC-FID quantitative analyses revealed a total percentage of 80% (E)-β-caryophyllene in the volatile fraction of PipeNig®-FL and 30% in PipeNig®-PWD. The precise quantification of (E)-β-caryophyllene and its standardization in PipeNig® allows the preparation and accurate dosage of products based on PipeNig®. For instance, in a recent pilot study (see below) we tested the effect of capsules contaning 100 mg PipeNig®-PWD, corresponding to 30 mg (E)-β-caryophyllene. Figure 2 shows a typical GC-MS profile of PipeNig®.

Figure 2: GC-MS chromatogram of PipeNig®


The endogenous cannabinoid system (ECS) plays an important role in the immune response to an infection. At present, two cannabinoid (CB) receptors are described: cannabinoid type 1 receptor (CB1) and cannabinoid type 2 receptor (CB2), both G-protein coupled receptors. CB2 receptor represents the peripheral CB, due to its expression on circulating immune cells. However, studies have also found CB2 expression in the brain, such as cerebellum and microglial cells. CB2 receptor is involved in the attenuation of inflammatory immune responses. CB2 receptor ligands have been shown to inhibit inflammation and edema formation, exhibit analgesic effects, play a protective role in hepatic ischemia-reperfusion injury, and prevent experimental colitis by reducing inflammation.

CB2 receptor pathway activation entails the suppression of cytokine release from immune cells and thereby dampening of the inflammatory response (immunosuppression). CB2 receptor-selective agonists that are devoid of the psychoactive side effects typically associated with CB1 receptor activation are potential candidates as a valid support of a range of different diseases. (E)-β-caryophyllene (BCP, Figure 1) is a sesquiterpene hydrocarbon that selectively binds to the CB2 receptor and is a functional CB2 agonist. BCP exerts toxicity at doses higher than 2000 mg/kg body weight and also has an anxiolytic-like and anti-depressant effects. The possibility that BCP may ameliorate mood disorders offers exciting prospects for future studies

A consistent number of in vitro and in vivo studies on BCP indicates that its biological effects include anti-inflammatory, antimicrobial, antioxidant and analgesic activities. These biological effects derive from the ability of BCP to activate the CB2 endocannabinoid receptor, thus behaving like an endo-cannabinoid. The specificity of BCP for the CB2 receptor, mainly expressed in peripheral tissues, and its inability to bind CB1 which is predominantly expressed at the level of the central nervous system, implies that its action is devoid of the known psychoactive effects associated with the activation of CB1 and suggests its potential use. Thus, BCP offers the possibility of obtaining a pain-killer and/or anti-inflammatory action as an interesting alternative to the use of Cannabis.
Recent findings have shown significant implications for clinical research and strongly supports the effectiveness of BCP as a novel molecule to target in the development of effective therapeutic agents for multiple sclerosis. In a pilot study performed by the Farmacia Centrale (Cambiano, Italy), a balanced sample of 31 volunteers (19 females and 12 males, with ages ranging from 36 to older than 76 years) exhibiting either acute (48%) or chronic (52%) pain pathologies, including headache (8%), cervical pain (20%), joint pain (32%), muscle aches (16%), lower back pain (12%) and other pain (12%), was studied for two months and compared to different NSAIDs. The volunteers received 2 capsules/day containing 100 mg of PipeNig® (corresponding to 30 mg of bioactive BCP) for 10 days or 2 capsules/day until pain relief. A score was recorded for all volunteers based on a questionnaire reporting their direct experience with PipeNig®. In general, volunteers reported a 60% reduction of pain between the 3rd and the 4th day after administration (Fig. 2A) and the perceived effect was similar or slightly lower than the NSAID used for 38% and 31% of the volunteers, respectively (Fig. 2B). Finally, when volunteers were asked whether they would use regularly PipeNig®, more than 70% responded affirmatively (Fig. 2C).

Figure 3 Result of the pilot study performed by the Farmacia Centrale (Cambiano, Italy) on a balanced sample of 31 volunteers exhibiting either acute or chronic pain pathologies after administration of PipeNig® as compared to different NSAIDs. A, timing of pain reduction after administration of PipeNig®. B, perception of the effect of PipeNig® as compared to the NSAID usually taken to reduce pain. C, response of volunteers to the question whether they would use again PipeNig® as a pain-killer. Values are expressed as percentage of responses.

The careful authentication the bioactive BCP by GC-MS and the quantification and standardization by GC-FID are necessary to prepare effective doses for analgesic activity. Our pilot study indicates PipeNig®-PWD and PipeNig®-FL as attractive candidates for the development of novel natural painkiller preparations for the reduction of the most distressing pain pathologies.

The recommended dosage of PipeNig®-PWD is 100 mg/dose. This dosage has been demonstrated to be effective when administered once a day for at least 10 days.

PipeNig®-FL is an alcohol-free liquid source with the highest content of (800 g/kg standardized BCP) available on the market and is particularly suitable for all liquid applications, including softgels. PipeNig®-FL suggested dosage is 30-50 mg/dose.

For further reading on PipeNig® refer to the publication:

Maffei, M. (2018)
PipeNig®, a black pepper (Piper nigrum) extract with a standardized content of (E)-β-caryophyllene. A natural source of an endo-cannabinoid agonist of the CB2 receptor.
Nutrafoods (2018) 17:N15-N17 – (Download pdf).


BCP has also been demonstrated to directly activate peroxisome proliferator-activated receptor-α (PPARα)2, involved in liver lipid metabolism, and to trigger the activation of PPAR3, involved in adipogenesis (Machado et al. Phytother. Res. 2018;32:2376-88).

Giving the growing scientific interest in BCP, we investigated the metabolic effects of PipeNig® (Endophyllene®), containing 80% β-caryophyllene. In particular, we focused on its potential antiobesogenic and antidiabetic activities in three in vitro cell models: 3T3-L1 preadipocytes, C2C12 myotubes and HepG2 hepatocytes.

PipeNig® reduces 3T3-L1 adipocyte differentiation and lipid accumulation. Moreover, acute exposure of C2C12 myotubes with different concentrations of PipeNig® improves glucose uptake activity.

The HepG2 hepatocyte steatosis model showed the ability of PipeNig® to interfere with free fatty acids-induced steatosis.

Altogether, our study indicates that 3T3-L1, C2C12 and HepG2 can be useful in vitro models to study adipogenesis and glucose uptake, in physiological conditions, and obesity, diabetes and steatosis in pathological contexts. Moreover, it highlights novel and interesting properties of β-caryophyllene, suggesting potential applications as a natural support to alleviate metabolic diseases (Scandiffio et al., Eur. J. Histochem. 2019, 63:30). Further studies are under way to assess the mechanism of action of BCP.