Introduction To The Effects of Terpenes 

Cahn.. CBN was the first marijuana cannabinoid for which a structure had been identified. Cannabidiol (CBD) was identified a few years later (Adams et al., 1940a), and the same research group later came close to identifying the structure of tetrahydrocannabinols, in a study involving isomerization of CBD (Adams et al., 1940b). In the time after, tetrahydrocannabinols were isolated from marijuana resin (Wollner et al., 1942)–though it was more than twenty years before chemical analytical and scientifical methods were adequate for deducting the final structure of the primary psychoactive component of cannabis, (-)D9-tetrahydrocannabinol (D9-THC; Gaoni and Mechoulam, 1964).

Loewe was also among the first to distinguish between cannabinoids ( Loewe, 1946). In a study, Loewe distinguished D 9–THC and a synthetic analogue from CBD. Both the former and the latter were able to cause central excitation as well as catalepsy. Since then, at the most, 489 compounds (″>Loewe, 1946) have been identified as cannabinoids. 9CBD and Aizpurua ­Olaizola (2016) are the most prevalent. Scherma (2018) is also a popular compound. 9THC acts partly as an antagonist at type 1 and 2 cannabinoid receptors. These receptors are found mainly in the central nervous system and peripheral nervous system. They mediate the effects of cannabis. It also acts upon type 2 cannabinoid receptors (CB2), the ones most expressed in immune cell (reviewed in Turcotte (2016)). It is generally believed that CBD has non-cannabinoid effects ( Turner. However, CBD has been shown not to bind CB2 at high concentrations (micromolar).

Recent research has focused on the biological activities and “entourage effects” of less-abundant, “minor,” phytocannabinoids. This phenomenon was first described for endogenous glycerol esters (Ben-Shabat et al., 1998), when 2-linoleoylglycerol and 2-palmitoylglycerol were found to increase the on-target affinity and efficacy of the endogenous cannabinoid 2-arachidonoylglycerol (2-AG), with which they can  co-occur, yet they cause no detectable direct combination with the cannabinoid receptors themselves (even though in this study these data points were not resolved). Similar observations have been described for N-palmitoylethanolamide and N-oleoylethanolamide (which are co-synthesized with anandamide) and may potentiate anandamide-induced relaxation of arteries (Ho et al., 2008)

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Ben-Shabbat et al. study, the term “entourage effect” has been co-opted to refer to the idea that complete cannabis contains a higher therapeutic benefit than its separate components , with a greater number of informational cannabis websites alluding that terpenes can modify the high produced by D9-THC. Dephles to Dudieva in 2007. 120 different terpenes have been discovered in the  cannabis Sativa plant . Of these, some of the most popular are linalool (myrcene), limonene, and a– and b–pinene. Although previous work suggests that b-caryophyllene acts as a CB2 receptor antagonist ( al. (2008)), subsequent research has questioned this ( al., 2019).

The evidence for entourage activity by cannabis-derived compounds is also lacking. A recent study looked at the six terpenoids discussed above in order to find out if they have potential entourage activity on cannabinoid receptors. Use of terpenes in combination or as a single agent to stimulate AtT-20 cells with CB1 or B2 expressing CB1 or D -THC-induced hyperpolarization was undetectable ( Santiago. et al. 2019). Furthermore, no GIRK channel related modulatory effects were observed in this molecular research for any of the six terpenes. Receptor desensitization was not affected by the same GIRK assay paradigm ( Santiago. et. al., 2019).

The current study sought to clarify five terpenoids of particular interest that act specifically (on target) through CB1/CB2, in an activity pathway (CAMP) that can capture receptor effects with high sensitivities. In the study design were also included effects on orthostatic interaction.


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Materials and Methods Of Terpenes 


True Terpenes in Portland, OR sold all terpenes. Terpene molarities have been calculated using the information provided by the supplier ( ). The terpenes were diluted with DMSO to 10 mM (Sigma Aldrich St Louis MO USA). The DMSO was kept constant at 1:1,000. Terpenes were assessed in assays separately, and in three different mixtures (Table 2): commercial analysis of multiple cannabis variants indicate huge variability in terpenoid formulations between strains (e.g.,, and these mixtures were therefore intended to capture some of this variability.


The entourage phenomenon was suggested as an explanation for biological findings that endocannabinoid binding activities can also be modified by other fats released from cells. There has been an increase in reports of plant activity, as well as interest, which led to research into volatile terpenoids (myrcene), a- and/or b-pinenes, b–caryophyllene and limonene. However, it has yet to be identified a clear interaction. These test were performed with Terpenes purchased from True Terpenes from Portland Oregon. The current study was designed to determine whether terpenes in the cannabis plant have detectable receptor-mediated activity, or modify the activity of D9-tetrahydrocannabinol, cannabidiol, or the endocannabinoid 2-arachidonylglycerol at the cannabinoid receptors. The standard radioligand binding paradigm was used to detect allosteric, orthostatic interactions. With the possible exception of a weak interaction of b-caryophyllene with CB2, no data were produced to support the hypothesis that any of the five terpenes tested (either alone or in mixtures) have direct interactions with CB1 or CB2, as the binding of radioligand ([3H]-CP55,940), D9-tetrahydrocannabinol, and cannabidiol were unaltered by the presence of terpenes. Similarly, terpene functional effects were also not detected, either alone or in combination with D9-tetrahydrocannabinol, cannabidiol, or 2-arachidonoylglycerol. This study reinforces the fact that the putative entourage effect is not explained by CB1 effects.

Keywords: cannabis, cannabinoid, terpenoid, terpene, entourage effect, signaling, binding