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Abstract
Cannabinochromene (CBC, 1a) is the archetypal member of a class of more than twenty isoprenylated 5-hydroxy-7-alkyl(aralky)benzo[2H]pyranes first reported from Cannabis sativa L. but also occurring in unrelated plants (Rhododendron species) as well as liverworts and fungi. The chemistry, synthesis, and bioactivity of CBC (1a) is reviewed, highlighting its underexploited pharmacological potential and rich chemistry.
... To start with, CBC (4) is not a major constituent of cannabis. It was long confused with CBD (2) because the two compounds have the same molecular weight and very similar GC chromatographic retention time, but the actual concentration of CBC in cannabis is generally up to two orders of magnitude lower than the one of the dominant phytocannabinoid [6]. Furthermore, while it was possible to generate almost pure breeds of cannabis containing CBD (2), Δ 9 -THC (1) and CBG (3), this has proved much more difficult for CBC (4), whose production is associated with a juvenile trait orthogonal to the Mendelian allelic heritage associated with the production of CBD, Δ 9 -THC and CBG [7]. ...
... Furthermore, while it was possible to generate almost pure breeds of cannabis containing CBD (2), Δ 9 -THC (1) and CBG (3), this has proved much more difficult for CBC (4), whose production is associated with a juvenile trait orthogonal to the Mendelian allelic heritage associated with the production of CBD, Δ 9 -THC and CBG [7]. Therefore, the inclusion of CBC in the "Big Four" is basically only associated with its early isolation and easy availability via synthesis [6]. Next, the name "minor cannabinoids" seems out of place for acidic cannabinoids, the native form of phytocannabinoids, since the "Big Four" are derived from the decarboxylation of their corresponding acidic phytocannabinoids, whose concentration in the plant biomass is therefore at least equal to the one of their decarboxylated derivatives [5]. ...
... To start with, CBC (4) is not a major constituent of cannabis. It was long confused with CBD (2) because the two compounds have the same molecular weight and very similar GC chromatographic retention time, but the actual concentration of CBC in cannabis is generally up to two orders of magnitude lower than the one of the dominant phytocannabinoid [6]. Furthermore, while it was possible to generate almost pure breeds of cannabis containing CBD (2), ∆ 9 -THC (1) and CBG (3), this has proved much more difficult for CBC (4), whose production is associated with a juvenile trait orthogonal to the Mendelian allelic heritage associated with the production of CBD, ∆ 9 -THC and CBG [7]. ...
Despite the very large number of phytocannabinoids isolated from Cannabis (Cannabis sativa L.), bioactivity studies have long remained focused on the so called “Big Four” [Δ9-THC (1), CBD (2), CBG (3) and CBC (4)] because of their earlier characterization and relatively easy availability via isolation and/or synthesis. Bioactivity information on the chemical space associated with the remaining part of the cannabinome, a set of ca 150 compounds traditionally referred to as “minor phytocannabinoids”, is scarce and patchy, yet promising in terms of pharmacological potential. According to their advancement stage, we sorted the bioactivity data available on these compounds, better referred to as the “dark cannabinome”, into categories: discovery (in vitro phenotypical and biochemical assays), preclinical (animal models), and clinical. Strategies to overcome the availability issues associated with minor phytocannabinoids are discussed, as well as the still unmet challenges facing their development as mainstream drugs.
... The first phytocannabinoid was isolated from the Cannabis sativa family Cannabaceae, but it has a long controversial history of its use and abuse [1,2]. From C. sativa more than 113 phytocannabinoids were isolated and classified into several groups such as cannabidiols (CBDs), cannabigerols (CBGs), cannabicyclols (CBLs), cannabidiols (CBNDs), cannabinols (CBNs), cannabitriols (CBTs), cannabichromenes (CBCs), (−)-∆ 9 -trans-tetrahydrocannabinol (∆ 9 -THC) and miscellaneous cannabinoids [1,[3][4][5]. Compounds obtained from C. sativa predominately generate alkyl-type phytocannabinoids with a monoterpene isoprenyl and the pentyl side chain [4,6]. In C. sativa, CBD, CBG, CBC, cannabichromevarine (CBCV), and ∆ 9 -THC are the most abundant cannabinoids in their respective acidic form. ...
... Furthermore, these active phytocannabinoids undergo decarboxylation and spontaneous rearrangement reactions on exposure to heat, radiation, or during storage. Some phytocannabinoid having unknown C1-C4 alkyl side chain are synthesized from acetyl-CoA, propanoyl-CoA, or pentanoyl-CoA [1,5,31,35]. ...
... Like ∆ 9 -THCAS and CBDAS, DCAS is active enzymatically outside apoplastic spaces and dependent on O 2 . In Rh. dauricum, DCA decarboxylated forms produce confluentin; spontaneous decarboxylation occurs via heat, irradiation, and during storage, similar to the decarboxylation acidic to neutral phytocannabinoids in C. sativa trichomes [5,11,37]. ...
Phytocannabinoids are a structurally diverse class of bioactive naturally occurring compounds found in angiosperms, fungi, and liverworts and produced in several plant organs such as the flower and glandular trichrome of Cannabis sativa, the scales in Rhododendron, and oil bodies of liverworts such as Radula species; they show a diverse role in humans and plants. Moreover, phytocannabinoids are prenylated polyketides, i.e., terpenophenolics, which are derived from isoprenoid and fatty acid precursors. Additionally, targeted productions of active phytocannabinoids have beneficial properties via the genes involved and their expression in a heterologous host. Bioactive compounds show a remarkable non-hallucinogenic biological property that is determined by the variable nature of the side chain and prenyl group defined by the enzymes involved in their biosynthesis. Phytocannabinoids possess therapeutic, antibacterial, and antimicrobial properties; thus, they are used in treating several human diseases. This review gives the latest knowledge on their role in the amelioration of abiotic (heat, cold, and radiation) stress in plants. It also aims to provide synthetic and biotechnological approaches based on combinatorial biochemical and protein engineering to synthesize phytocannabinoids with enhanced properties.
... In an excellent review about the chemistry, synthesis and bioactivity of CBC, Pollastro et al. [52] summarized recent studies on the psychotropicity of CBC. Even though no narcotic effect was found in in vivo experiments, high doses of CBC may indeed exhibit responses typical for Δ 9 -THC (e.g., hypomotility, catalepsy, hypothermia and analgesia). ...
... Even though no narcotic effect was found in in vivo experiments, high doses of CBC may indeed exhibit responses typical for Δ 9 -THC (e.g., hypomotility, catalepsy, hypothermia and analgesia). The authors claim that the reason for this effect most likely derives from another than the typical mechanism, as CBC was found to show only marginal affinity for the cannabinoid receptors CB1 and CB2 [52]. Besides that, multiple other effects are related to CBC, among which the antibacterial and antifungal activity is the most noteworthy one as CBC outperforms other cannabinoids in this category. ...
... Besides that, multiple other effects are related to CBC, among which the antibacterial and antifungal activity is the most noteworthy one as CBC outperforms other cannabinoids in this category. According to the authors, little information exists regarding the biological profile of naturally occurring analogs of CBC [52]. ...
Cannabidiol (CBD) is a naturally occurring, non-psychotropic cannabinoid of the hemp plant Cannabis sativa L. and has been known to induce several physiological and pharmacological effects. While CBD is approved as a medicinal product subject to prescription, it is also widely sold over the counter (OTC) in the form of food supplements, cosmetics and electronic cigarette liquids. However, regulatory difficulties arise from its origin being a narcotic plant or its status as an unapproved novel food ingredient. Regarding the consumer safety of these OTC products, the question whether or not CBD might be degraded into psychotropic cannabinoids, most prominently tetrahydrocannabinol (THC), under in vivo conditions initiated an ongoing scientific debate. This feature review aims to summarize the current knowledge of CBD degradation processes, specifically the results of in vitro and in vivo studies. Additionally, the literature on psychotropic effects of cannabinoids was carefully studied with a focus on the degradants and metabolites of CBD, but data were found to be sparse. While the literature is contradictory, most studies suggest that CBD is not converted to psychotropic THC under in vivo conditions. Nevertheless, it is certain that CBD degrades to psychotropic products in acidic environments. Hence, the storage stability of commercial formulations requires more attention in the future.
... Kenevir tohumu yağı, genellikle esansiyel yağ asitleri (EFA'lar) olarak bilinen PUFA'ların %80' den fazlasını içerir (7). Yağ; linoleik asit (%55, [41][42][43][44][45][46][47][48][49][50][51][52][53][54][55][56][57][58][59]64), α-linoleik asit (%16, 40), oleik asit (%11, 88), palmitik asit (%6,08-6,82) ve stearik asit (%2,34-2,67) içermektedir. Bazı türler ayrıca %4'e kadar γ-linoleik asit içerir. ...
... CBD, BACH1'in nükleer aktarımını ve bozulmasını indüklemekte olup oksidatif stres ve cilt yaşlanmasını azaltmaktadır (6,44,45). CBG, α2-adrenoseptörünün bir agonisti olup endokannabinoid membran taşıyıcısını inhibe etmektedir (6,(46)(47)(48)(49)(50). CBC, TRPA1 kanalının en güçlü agonistidir (6,46,51,52). Kannabidivarin, dopamin D2 benzeri reseptörlerin kısmi agonistidir (6,53). Son zamanlarda ciltte D2 benzeri reseptör agonizminin cilt bariyerinin iyileşmesini ve yara iyileşmesini desteklediği gösterilmiştir (6,(54)(55)(56). ...
... Despite this impressive variety, early studies in this field focused almost exclusively on the narcotic principle of marijuana, ∆ 9 -tetrahydrocannabinol (∆ 9 -THC, 1a, Figure 1), eventually expanding to the other related compounds, which are cannabidiol (CBD, 2a, Figure 1), cannabigerol (CBG, 3a, Figure 1), and cannabichromene (CBC, 4a, Figure 1), that together form a group of compounds often referred to as "the major cannabinoids" or "big four" [9]. As our understanding of the biological mechanisms underlying ∆ 9 -THC narcotic properties developed, the endocannabinoid system (ECS) was discovered [10], and its complexity, homeostatic role, and potential for drug discovery prompted a reconsideration Most of the studies on phytocannabinoids, both chemical and biological, have focused on the "big four" due to their high extraction yield from vegetable sources or easy accessibility through total synthesis [11][12][13][14][15][16][17]. However, cannabis plants are also capable of producing more than 150 other compounds referred to as "minor cannabinoids" [18], which have significant structural differences and specific biological properties [19,20]; among these compounds, one stands out in particular, namely cannabinol (CBN, 5a, Figure 1). ...
... CBN (5a) was probably considered a "minor" phytocannabioid owing to its unfortunate and confusing discovery: it was the first phytocannabinoid to be isolated from hash- Most of the studies on phytocannabinoids, both chemical and biological, have focused on the "big four" due to their high extraction yield from vegetable sources or easy accessibility through total synthesis [11][12][13][14][15][16][17]. However, cannabis plants are also capable of producing more than 150 other compounds referred to as "minor cannabinoids" [18], which have significant structural differences and specific biological properties [19,20]; among these compounds, one stands out in particular, namely cannabinol (CBN, 5a, Figure 1). ...
Cannabis (Cannabis sativa L.) is an outstanding source of bioactive natural products, with more than 150 different phytocannabinoids isolated throughout the decades; however, studies of their bioactivity have historically concentrated on the so-called “big four” [∆9-THC (1a), CBD (2a), CBG (3a) and CBC (4a)]. Among the remaining products, which have traditionally been referred to as “minor cannabinoids”, cannabinol (CBN, 5a) stands out for its important repercussions and implications on the global scientific landscape. Throughout this review, we will describe why CBN (5a) deserves a prominent place within the so-called “cannabinome”, providing an overview on its history, the syntheses developed, and its bioactivity, highlighting its promising pharmacological potential and the significant impact that the study of its chemistry had on the development of new synthetic methodologies.
... The anthoponoids E, G and H had also suppressed the LPS-induced inflammatory responses in RAW 264.7 macrophages. In addition, daurichromenic acid (DCA), the meroterpenoid consisting of orsellinic acid and sesquiterpene moieties, that analogued to the cannabinoid structure was also found [52]. ...
... R. dauricum L., widely spread throughout northeastern Asia, also produces unique secondary metabolites including DCAs (Table 2) [52]. The MeOH extract of the leaves and twigs had illustrated significant anti-HIV activity [53]. ...
Phytocannabinoids are isoprenylated resorcinyl polyketides produced mostly in glandular trichomes of Cannabis sativa L. These discoveries led to the identification of cannabinoid receptors, which modulate psychotropic and pharmacological reactions and are found primarily in the human central nervous system. As a result of the biogenetic process, aliphatic ketide phytocannabinoids are exclusively found in the cannabis species and have a limited natural distribution, whereas phenethyl-type phytocannabinoids are present in higher plants, liverworts, and fungi. The development of cannabinomics has uncovered evidence of new sources containing various phytocannabinoid derivatives. Phytocannabinoids have been isolated as artifacts from their carboxylated forms (pre-cannabinoids or acidic cannabinoids) from plant sources. In this review, the overview of the phytocannabinoid biosynthesis is presented. Different non-cannabis plant sources are described either from those belonging to the angiosperm species and bryophytes, together with their metabolomic structures. Lastly, we discuss the legal framework for the ingestion of these biological materials which currently receive the attention as a legal high.
... These compounds are produced by both male and female plants. However, their content, and those of the other phytocannabinoids present in cannabis, may differ significantly between plants, due to genetic variation (Pollastro et al. 2018a). Cannabigerol (C 21 H 32 O 2 , 2-[(2E)-3,7-dimethylocta-2,6-dienyl]-5-pentylbenzene-1,3-diol, MW = 316.5 g/mol, MP = 50-56°C and BP = 105°C) is the non-acidic form of the parent molecule cannabigerolic acid (CBGA) (Szaflarski and Bebin 2014). ...
... The relatively low content of compounds in nature warrants their synthetic preparation, an option frequently exploited to obtain drugs originating from natural sources. The synthesis of CBG was reported for the first time by Gaoni and Mechoulam (1964) Fig. 4 Biosynthesis of cannabigerol (CBG) and cannabigerolic acid (CBGA) (Pollastro et al. 2018a) by treating geraniol with olivetol in decalin for 36 h, in the absence of a catalyst. Heating probably favoured the condensation of the two starting alcohols, but mechanistic studies were not done. ...
Cannabigerol (CBG) is one of the major phytocannabinoids present in Cannabis sativa L. but is presumed to be an artefact or degradation product of cannabigerolic acid (CBGA), the principal precursor of several cannabinoids. The growing interest in CBG has been attributed to its non-psychotropic properties, low cannabinoid receptor potency and relative abundance in some commercial Cannabis varieties. A broad pharmacological profile has been described, where CBG is reported to exhibit anti-inflammatory, anticancer, anti-oxidant, antimicrobial, neuroprotective and appetite-enhancing properties, among others. Previous reviews on CBG have been limited to either the pharmacological potential of the molecule, with little detail on the chemistry, or to advances in the analytical tools used to explore CBG content in a wide range of biological samples. The current review seeks to highlight the chemistry, biosynthesis, pharmacology and safety aspects of the molecule. Additionally, we provide new insights into, and critical evaluation of, the pharmacological interactions of CBG via different receptor sites based on in silico predictions, using retrieved 3D structures of alpha-2 adrenergic receptors from the Protein Data Bank (PDB). For the first time, a bibliometric overview of the literature was performed, and with the aid of scientometric tools it was possible to present a visual overview of the research trends over the years, assess research performance of countries, and delineate the research output trajectory, hotspots and voids. In-depth analysis of the published literature revealed that clinical trials establishing the efficacy, safety and side-effects of CBG therapy in specific disease conditions are limited, as well as industry-friendly quality control procedures for CBG-enriched cannabis extracts.
... Since the late 1960s, several compounds known to be present in cannabis have been isolated and characterized, including the psychoactive cannabinoid Δ 9 -tetrahydrocannabinol (THC or Δ 9 -THC, 1, Figure 1) and the noneuphoric cannabidiol (CBD, 2, Figure 1) (Iversen, 2018;Sholler et al., 2020). These compounds are now referred to as "major cannabinoids", including other important ones like cannabigerol (CBG), cannabichromene (CBC), and cannabinol (CBN) (Pollastro et al., 2018;Anokwuru et al., 2022;Maioli et al., 2022). The two major phytocannabinoids THC and CBD already reached the approval by FDA for commercialization in different forms: Sativex ® , a combination of THC and CBD, is used to treat spasticity associated with multiple sclerosis, and CBD has also been developed as a single active pharmaceutical ingredient known as Epidiolex ® , drug of choice for the treatment of certain rare genetic forms of epilepsy. ...
Cannabinoid subtype 1 receptors (CB 1 Rs) are an important class of G protein-coupled receptors (GPCRs) belonging to the endocannabinoid system. CB 1 Rs play a crucial modulatory role in the functioning of other neurotransmitter systems and are involved in a wide range of physiological functions and dysfunctions; thus, they are considered one of the most important targets for drug development, as well as diagnostic purposes. Despite this, only a few molecules targeting this receptor are available on the pharmaceutical market, thus emphasizing the need to gain a deeper understanding of the complex activation pathways of CB 1 Rs and how they regulate diseases. As part of this review, we provide an overview of pharmacological and imaging tools useful for detecting CB 1 Rs. Herein, we summarize the derivations of cannabinoids and terpenoids with fluorescent compounds, radiotracers, or photochromic motifs. CB 1 Rs’ molecular probes may be used in vitro and, in some cases, in vivo for investigating and exploring the roles of CB 1 Rs together with the starting point for the development of CB 1 R-targeted drugs.
... CBG is an agonist of α2-adrenoceptor [73,98], it inhibits the endocannabinoid membrane transporter [50,79,82]. CBC is the most potent agonist of the TRPA1 channel [81,82,99]. CBDV is a partial agonist of dopamine D2-like receptors [100]. ...
Cannabis sativa L. plant is currently attracting increasing interest in cosmetics and dermatology. In this review, the biologically active compounds of hemp are discussed. Particularly the complex interactions of cannabinoids with the endocannabinoid system of the skin to treat various conditions (such as acne, allergic contact dermatitis, melanoma, and psoriasis) with clinical data. Moreover, the properties of some cannabinoids make them candidates as cosmetic actives for certain skin types. Hemp seed oil and its minor bioactive compounds such as terpenes, flavonoids, carotenoids, and phytosterols are also discussed for their added value in cosmetic formulation.
... Terpenylation was then carried out mutuating chemistry developed for the synthesis of the corresponding natural n-pentyl phytocannabinoids. C-Menthylation with (4R)-2,9-p-menthadien-1-ol (11) afforded either DMH-CBD (1b) or DMH-∆ 8 -THC (6b) depending on the reaction conditions [14], while geranylation with BF 3 on alumina gave DMH-CBG (7b) [15], and chromenyation with citral (13) under basic conditions generated DMH-CBC (8b) [16], next aromatized to DMH-CBN (9b) by treatment with iodine (Scheme 1) [17]. ...
The affinity of cannabinoids for their CB1 and CB2 metabotropic receptors is dramatically affected by a combination of α-branching and elongation of their alkyl substituent, a maneuver exemplified by the n-pentyl -> α,α-dimethylheptyl (DMH) swap. The effect of this change on other cannabinoid end-points is still unknown, an observation surprising since thermo-TRPs are targeted by phytocannabinoids with often sub-micromolar affinity. To fill this gap, the α,α-dimethylheptyl analogues of the five major phytocannabinoids [CBD (1a), Δ8-THC (6a), CBG (7a), CBC (8a) and CBN (9a)] were prepared by total synthesis, and their activity on thermo-TRPs (TRPV1-4, TRPM8, and TRPA1) was compared with that of one of their natural analogues. Surprisingly, the DMH chain promoted a shift in the selectivity toward TRPA1, a target involved in pain and inflammatory diseases, in all investigated compounds. A comparative study of the putative binding modes at TRPA1 between DMH-CBC (8b), the most active compound within the series, and CBC (8a) was carried out by molecular docking, allowing the rationalization of their activity in terms of structure–activity relationships. Taken together, these observations qualify DMH-CBC (8b) as a non-covalent TRPA1-selective cannabinoid lead that is worthy of additional investigation as an analgesic and anti-inflammatory agent.
... 15,23 CBC is highly scalemic or even racemic and is not present in significant amounts in cannabis flower heads, being produced mostly in the early stages of development of the plant. 23 Given also the very low yield and harsh conditions required for the chemical conversion, derivation of Δ 9 -cis-THC from CBC seems unlikely. However, it is possible that Δ 9 -cis-THC (3) and CBC (10) are derived from alternative pericyclic processes from cannabigerolic acid (11). ...
The cis-stereoisomers of Δ⁹-THC [(−)-3 and (+)-3] were identified and quantified in a series of low-THC-containing varieties of Cannabis sativa registered in Europe as fiber hemp and in research accessions of cannabis. While Δ⁹-cis-THC (3) occurs in cannabis fiber hemp in the concentration range of (−)-Δ⁹-trans-THC [(−)-1], it was undetectable in a sample of high-THC-containing medicinal cannabis. Natural Δ⁹-cis-THC (3) is scalemic (ca. 80–90% enantiomeric purity), and the absolute configuration of the major enantiomer was established as 6aS,10aR [(−)-3] by chiral chromatographic comparison with a sample available by asymmetric synthesis. The major enantiomer, (−)-Δ⁹-cis-THC [(−)-3], was characterized as a partial cannabinoid agonist in vitro and elicited a full tetrad response in mice at 50 mg/kg doses. The current legal discrimination between narcotic and non-narcotic cannabis varieties centers on the contents of “Δ⁹-THC and isomers” and needs therefore revision, or at least a more specific wording, to account for the presence of Δ⁹-cis-THCs [(+)-3 and (−)-3] in cannabis fiber hemp varieties.
... In the crude IGB strain extract, CBD was abundant in comparison to CBC. Indeed, CBC concentrations in most cannabis strains rarely exceed 0.2-0.3% on a dry weight basis, much lower than the other "major" phytocannabinoids [21]. Yet, CBC + THC (the latter in minute amounts) acts similarly to CBD in terms of cytotoxicity (i.e., comparable concentrations are needed for significant cytotoxicity). ...
Cannabis sativa contains more than 500 constituents, yet the anticancer properties of the vast majority of cannabis compounds remains unknown. We aimed to identify cannabis compounds and their combinations presenting cytotoxicity against bladder urothelial carcinoma (UC), the most common urinary system cancer. An XTT assay was used to determine cytotoxic activity of C. sativa extracts on T24 and HBT-9 cell lines. Extract chemical content was identified by high-performance liquid chromatography (HPLC). Fluorescence-activated cell sorting (FACS) was used to determine apoptosis and cell cycle, using stained F-actin and nuclei. Scratch and transwell assays were used to determine cell migration and invasion, respectively. Gene expression was determined by quantitative Polymerase chain reaction (PCR). The most active decarboxylated extract fraction (F7) of high-cannabidiol (CBD) C. sativa was found to contain cannabichromene (CBC) and Δ9-tetrahydrocannabinol (THC). Synergistic interaction was demonstrated between CBC + THC whereas cannabinoid receptor (CB) type 1 and type 2 inverse agonists reduced cytotoxic activity. Treatments with CBC + THC or CBD led to cell cycle arrest and cell apoptosis. CBC + THC or CBD treatments inhibited cell migration and affected F-actin integrity. Identification of active plant ingredients (API) from cannabis that induce apoptosis and affect cell migration in UC cell lines forms a basis for pre-clinical trials for UC treatment.
... The hidden Bronsted acidity of iodine, that is, its capacity to generate HI by interaction with hydroxyl groups from the substrate or from traces of protic solvents, 25 was ruled out as a possible mechanism since treatment of CBC with Bronsted acids affords compounds resulting from the formation of a benzyl cation and not by cycloreversion. 7 A soft polarizability/polarization of the I−X bond seems, conversely, critical to avoid electrophilic attack to the electron-rich aromatic ring and the dihydropyrane double bond. Electroreversion could then be promoted by halogen bonding to the chromene oxygen and/or to its aromatic ring, 25 while the final aromatization eventually funnels the various equilibria toward the generation of dibenzochromenes. ...
The thermal degradation of cannabichromene (CBC, 3) is dominated by cationic reactions and not by the pericyclic rearrangements observed in model compounds. The rationalization of these differences inspired the development of a process that coupled, in an aromatization-driven single operational step, the condensation of citral and alkylresorciniols to homoprenylchromenes and their in situ deconstructive annulation to benzo[c]chromenes. This process was applied to a total synthesis of cannabinol (CBN, 5) and to its molecular editing.
Cannabichromene (CBC, 1a) occurs in Cannabis (Cannabis sativa) as a scalemate having a composition that is strain-dependent in terms of both enantiomeric excess and enantiomeric dominance. In the present work, the chirality of CBC (1a), a noncrystalline compound, was shown not to be significantly affected by standard conditions of isolation and purification, and enantiomeric self-disproportionation effects were minimized by carrying out the chiral analysis on crude fractions rather than on purified products. A genetic basis for the different enantiomeric state of CBC in Cannabis therefore seems to exist, implying that the chirality status of natural CBC (1a) in the plant is associated with the differential expression of CBCA-synthase isoforms and/or of associated directing proteins with antipodal enantiospecificity. The biological profile of both enantiomers of CBC should therefore be investigated independently to assess the contribution of this compound to the activity of Cannabis preparations.
Cannabis belongs to the family Cannabaceae, and phytocannabinoids are produced by the Cannabis sativa L. plant. A long-standing debate regarding the plant is whether it contains one or more species. Phytocannabinoids are bioactive natural products found in flowers, seeds, and fruits. They can be beneficial for treating human diseases (such as multiple sclerosis, neurodegenerative diseases, epilepsy, and pain), the cellular metabolic process, and regulating biological function systems. In addition, several phytocannabinoids are used in various therapeutic and pharmaceutical applications. This study provides an overview of the different sources of phytocannabinoids; further, the biosynthesis of bioactive compounds involving various pathways is elucidated. The structural classification of phytocannabinoids is based on their decorated resorcinol core and the bioactivities of naturally occurring cannabinoids. Furthermore, phytocannabinoids have been studied in terms of their role in animal models and antimicrobial activity against bacteria and fungi; further, they show potential for therapeutic applications and are used in treating various human diseases. Overall, this review can help deepen the current understanding of the role of biotechnological approaches and the importance of phytocannabinoids in different industrial applications.
Cannabinoids are bioactive meroterpenoids comprising prenylated polyketide molecules that can modulate a wide range of physiological processes. Cannabinoids have been shown to possess various medical/therapeutic effects, such as anti-convulsive, anti-anxiety, anti-psychotic, antinausea, and anti-microbial properties. The increasing interest in their beneficial effects and application as clinically useful drugs has promoted the development of heterologous biosynthetic platforms for the industrial production of these compounds. This approach can help circumvent the drawbacks associated with extraction from naturally occurring plants or chemical synthesis. In this review, we provide an overview of the fungal platforms developed by genetic engineering for the biosynthetic production of cannabinoids. Different yeast species, such as Komagataella phaffii (formerly P. pastoris) and Saccharomyces cerevisiae, have been genetically modified to include the cannabinoid biosynthetic pathway and to improve metabolic fluxes in order to increase cannabinoid titers. In addition, we engineered the filamentous fungus Penicillium chrysogenum for the first time as a host microorganism for the production of Δ9-tetrahydrocannabinolic acid from intermediates (cannabigerolic acid and olivetolic acid), thereby showing the potential of filamentous fungi as alternative platforms for cannabinoid biosynthesis upon optimization.
The number of people diagnosed with diabetes mellitus and its complications is markedly increasing worldwide, leading to a worldwide epidemic across all age groups, from children to older adults. Diabetes is associated with premature aging. In recent years, it has been found that peripheral overactivation of the endocannabinoid system (ECS), and in particular cannabinoid receptor 1 (CB1R) signaling, plays a crucial role in the progression of insulin resistance, diabetes (especially type 2), and its aging-related comorbidities such as atherosclerosis, nephropathy, neuropathy, and retinopathy. Therefore, it is suggested that peripheral blockade of CB1R may ameliorate diabetes and diabetes-related comorbidities. The use of synthetic CB1R antagonists such as rimonabant has been prohibited because of their psychiatric side effects. In contrast, phytocannabinoids such as cannabidiol (CBD) and tetrahydrocannabivarin (THCV), produced by cannabis, exhibit antagonistic activity on CB1R signaling and do not show any adverse side effects such as psychoactive effects, depression, or anxiety, thereby serving as potential candidates for the treatment of diabetes and its complications. In addition to these phytocannabinoids, cannabis also produces a substantial number of other phytocannabinoids, terpenes, and flavonoids with therapeutic potential against insulin resistance, diabetes, and its complications. In this review, the pathogenesis of diabetes, its complications, and the potential to use cannabinoids, terpenes, and flavonoids for its treatment are discussed.
Analysis of hemp collection samples based on the content of minor (rare) non-psychotropic cannabinoids, such as cannabichromene (CBC), cannabidivarin (CBDV), and cannabinol (CBN); determination of correlation relationships between them and common compounds; selection of valuable breeding genotypes. Methods. Field, biochemical (gas chromatography of cannabinoid compounds), and statistical (pair, partial, and multiple linear correlations). Results. Quantitative analysis of 210 samples of various ecological-geographical and genetic origin (local and wild forms, self-filing lines, hybrids, varieties, synthetic populations, polyploids) with a tetrahydrocannabinol (THC) content of less than 0.08% in dried plants showed the level of manifestation of the trait from its absence within the sensitivity of the gas chromatograph up to 0.6838% CBC, 0.1719% CBC and 0.3274% CBN. In the studied hemp samples, a medium negative relationship was found between the signs of the CBC and cannabidiol (CBD) contents (r = –0.53), a weak negative relationship between CBC and CBDV contents (r = –0.35), medium positive relationships between the signs of CBC and THC contents (r = 0.57) and CBC and CBN contents (r = 0.59). A medium positive correlation (r = 0.57) was found between the signs of CBDV and CBD contents, while CBN had a strong positive relationship with THC (r = 0.82). There is almost no correlation between cannabigerol (CBG) and the minor cannabinoids under study. The biosynthesis of minor cannabinoid compounds is quite complex. Signs manifestation is affected by many genetic and external factors. Partial correlation coefficients (given that one of the three signs is eliminated) and multiple correlation coefficients (given that the relationship of one sign is determined and two other signs are combined) give grounds to state that the gene for CBCA-synthase affects the production of CBD and, in particular THC. Conclusions. The closeness of the linear relationships between minor cannabinoids and common components allows selecting valuable hemp samples with a high content of one or several compounds under the absence or low content of psychotropic THC.
Cannabis sativa is an extraordinarily versatile species. Hemp and its cousin marijuana, both C. sativa, have been used for millennia as a source of fibre, oil, and for medicinal, spiritual, and recreational purposes. Because the consumption of Cannabis can have psychoactive effects, the plant has been widely banned throughout the last century. In the past decade, evidence of its medicinal properties did lead to the relaxation of legislation in many countries around the world. Consequently, the genetics and development of Cannabis as well as Cannabis-derived products are the subject of renewed attention.
Here, we review the biology of C. sativa, including recent insights from taxonomy, morphology, and genomics, with an emphasis on the genetics of cannabinoid synthesis. Because the female Cannabis flower is of special interest as the site of cannabinoid synthesis, we explore flower development, flowering time well as the species' unique sex determination system in detail.
Furthermore, we outline the tremendous medicinal, engineering, and environmental opportunities that Cannabis bears. Together, the picture emerges that our understanding of Cannabis biology currently progresses at an unusual speed. A future challenge will be to preserve the multi-purpose nature of Cannabis, and to harness its medicinal properties and sustainability advantages simultaneously.
With the advent of medical cannabis usage globally, there has been a renewed interest in exploring the chemical diversity of this unique plant. Cannabis produces hundreds of unique phytocannabinoids, which not only have diverse chemical structures but also a range of cellular and molecular actions, interesting pharmacological properties, and biological actions. In addition, it produces other flavonoids, stilbenoids, and terpenes that have been variably described as conferring additional or so-called entourage effects to whole-plant extracts when used in therapeutic settings. This review explores this phytochemical diversity in relation to specific bioactivity ascribed to phytocannabinoids as neuroprotective agents. It outlines emergent evidence for the potential for selected phytocannabinoids and other cannabis phytochemicals to mitigate factors such as inflammation and oxidative stress as drivers of neurotoxicity, in addition to focusing on specific interactions with pathological misfolding proteins, such as amyloid β, associated with major forms of neurodegenerative diseases such as Alzheimer’s disease.
In the last few years research into Cannabis and its constituent phytocannabinoids has burgeoned, particularly in the potential application of novel cannabis phytochemicals for the treatment of diverse illnesses related to neurodegeneration and dementia, including Alzheimer’s (AD), Parkinson’s (PD) and Huntington’s disease (HD). To date, these neurological diseases have mostly relied on symptomatological management. However, with an aging population globally, the search for more efficient and disease-modifying treatments that could delay or mitigate disease progression is imperative. In this context, this review aims to present a state of art in the research with cannabinoids and novel cannabinoid-based drug candidates that have been emerged as novel promising alternatives for drug development and innovation in the therapeutics of a number of diseases, especially those related to CNS-disturbance and impairment.
Marijuana (Cannabis sativa) has long been known to contain antibacterial cannabinoids, whose potential to address antibiotic resistance has not yet been investigated. All five major cannabinoids (cannabidiol (1b), cannabichromene (2), cannabigerol (3b), Delta (9)-tetrahydrocannabinol (4b), and cannabinol (5)) showed potent activity against a variety of methicillin-resistant Staphylococcus aureus (MRSA) strains of current clinical relevance. Activity was remarkably tolerant to the nature of the prenyl moiety, to its relative position compared to the n-pentyl moiety (abnormal cannabinoids), and to carboxylation of the resorcinyl moiety (pre-cannabinoids). Conversely, methylation and acetylation of the phenolic hydroxyls, esterification of the carboxylic group of pre-cannabinoids, and introduction of a second prenyl moiety were all detrimental for antibacterial activity. Taken together, these observations suggest that the prenyl moiety of cannabinoids serves mainly as a modulator of lipid affinity for the olivetol core, a per se poorly active antibacterial pharmacophore, while their high potency definitely suggests a specific, but yet elusive, mechanism of activity.
The Transient Receptor Potential Ankyrin 1 channel (TRPA1), is a member of the large TRP family of ion channels, and functions as a Ca2+ permeable non-selective cation channel in many different cell processes, ranging from sensory to homeostatic tasks. TRPA1 is highly conserved across the animal kingdom. The only mammalian TRPA subfamily member, TRPA1, is widely expressed in neuronal (e.g. sensory dorsal root and trigeminal ganglia neurons)- and in non-neuronal cells (e.g. epithelial cells, hair cells). It exhibits 14–19 amino-(N-)terminal ankyrin repeats, an unusual structural feature. The TRPA1 channel is activated by noxious cold (<17 °C) as well as by a plethora of chemical compounds that includes not only electrophilic compounds and oxidants that can modify, in an alkylative or oxidative fashion, nucleophilic cysteine residues in the channel’s N-terminus, but also compounds that do not covalently bind to the channel proteins (e.g. menthol, nifedipin). Based on localization and functional properties, TRPA1 is considered a key player in acute and chronic (neuropathic) pain and inflammation. Moreover, its role in the (patho)physiology of nearly all organ systems is anticipated, and will be discussed along with the potential of TRPA1 as a drug target for the management of various pathological conditions.
Thermal isomerization of CBC(an) to THC(an) [nonaromatic analogues of plant cannabinoids cannabichromene (CBC) and Delta(1)-tetrahydrocannabinol (THC), respectively] is predicted in silico and demonstrated experimentally. Density functional theory calculations support a similar isomerization mechanism for the corresponding plant cannabinoids. Docking studies suggest that THC(an), although nonaromatic, has a CB(1) receptor binding affinity similar to that of natural THC.
Delta(9)-Tetrahydrocannabinol (THC) exhibits antitumor effects on various cancer cell types, but its use in chemotherapy is limited by its psychotropic activity. We investigated the antitumor activities of other plant cannabinoids, i.e., cannabidiol, cannabigerol, cannabichromene, cannabidiol acid and THC acid, and assessed whether there is any advantage in using Cannabis extracts (enriched in either cannabidiol or THC) over pure cannabinoids. Results obtained in a panel of tumor cell lines clearly indicate that, of the five natural compounds tested, cannabidiol is the most potent inhibitor of cancer cell growth (IC(50) between 6.0 and 10.6 microM), with significantly lower potency in noncancer cells. The cannabidiol-rich extract was equipotent to cannabidiol, whereas cannabigerol and cannabichromene followed in the rank of potency. Both cannabidiol and the cannabidiol-rich extract inhibited the growth of xenograft tumors obtained by s.c. injection into athymic mice of human MDA-MB-231 breast carcinoma or rat v-K-ras-transformed thyroid epithelial cells and reduced lung metastases deriving from intrapaw injection of MDA-MB-231 cells. Judging from several experiments on its possible cellular and molecular mechanisms of action, we propose that cannabidiol lacks a unique mode of action in the cell lines investigated. At least for MDA-MB-231 cells, however, our experiments indicate that cannabidiol effect is due to its capability of inducing apoptosis via: direct or indirect activation of cannabinoid CB(2) and vanilloid transient receptor potential vanilloid type-1 receptors and cannabinoid/vanilloid receptor-independent elevation of intracellular Ca(2+) and reactive oxygen species. Our data support the further testing of cannabidiol and cannabidiol-rich extracts for the potential treatment of cancer.
A liquid chromatography-tandem mass spectrometry single-laboratory validation was performed for the detection and quantification of the 10 major cannabinoids of cannabis, namely, (−)-trans-Δ9-tetrahydrocannabinol, cannabidiol, cannabigerol, cannabichromene, tetrahydrocannabivarian, cannabinol, (−)-trans-Δ8-tetrahydrocannabinol, cannabidiolic acid, cannabigerolic acid, and Δ9-tetrahydrocannabinolic acid-A, in the root extract of Cannabis sativa. Acetonitrile : methanol (80 : 20, v/v) was used for extraction; d3-cannabidiol and d3- tetrahydrocannabinol were used as the internal standards. All 10 cannabinoids showed a good regression relationship with r 2 > 0.99. The validated method is simple, sensitive, and reproducible and is therefore suitable for the detection and quantification of these cannabinoids in extracts of cannabis roots. To our knowledge, this is the first report for the quantification of cannabinoids in cannabis roots.
Six new pairs of bibenzyl-based meroterpenoid enantiomers, (±)-rasumatranin A–D (1–4) and (±)-radulanin M and N (5 and 6), and six known compounds were isolated from the adnascent Chinese liverwort, Radula sumatrana. Their structures were elucidated based on spectroscopic data and chiral phase HPLC-ECD analyses. The structures of 1 and 7 were also confirmed by single-crystal X-ray diffraction analysis. Cytotoxicity tests of the isolated compounds showed that 6-hydroxy-3-methyl-8-phenylethylbenzo[b]oxepin-5-one (8) showed activity against the human cancer cell lines MCF-7, PC-3, and SMMC-7721, with IC50 values of 3.86, 6.60, and 3.58 μM, respectively, and induced MCF-7 cell death through a mitochondria-mediated apoptosis pathway.
By using the Inverted Chirality Columns Approach (ICCA) we have developed an enantioselective UHPSFC method to determine the enantiomeric excess (e.e.) for (–)-Δ ⁹ -THC in medicinal marijuana (Bedrocan ® ). The e.e. was...
Daurichromenic acid (DCA) synthase catalyzes the oxidative cyclization of grifolic acid to produce DCA, an anti-HIV meroterpenoid isolated from Rhododendron dauricum. We identified a novel cDNA encoding DCA synthase by transcriptome-based screening from young leaves of R. dauricum. The gene coded for a 533-amino acid polypeptide with moderate homologies to FAD oxidases from other plants. The primary structure contained an N-terminal signal peptide, and conserved amino acid residues to form bicovalent linkage to FAD isoalloxazine ring at His112 and Cys175. In addition, the recombinant DCA synthase, purified from the culture supernatant of transgenic Pichia pastoris, exhibited structural and functional properties as a flavoprotein. The reaction mechanism of DCA synthase characterized herein partly shares a similarity with those of cannabinoid synthases from Cannabis sativa, whereas DCA synthase catalyzes a novel cyclization reaction of farnesyl moiety of a meroterpenoid natural product of plant origin. Moreover, in this study, we present evidence that DCA is biosynthesized and accumulated specifically in the glandular scales, on the surface of R. dauricum plants, based on various analytical studies at chemical, biochemical and molecular levels. Extracellular localization of DCA was also confirmed by a confocal microscopic analysis of its autofluorescence. These data highlight the unique feature of DCA; the final step of biosynthesis is completed in apoplastic space, and it is highly accumulated outside the scale cells.
Daurichromenic acid is a meroterpenoid with various pharmacological activities that is biosynthesized from grifolic acid in Rhododendron dauricum. Heterologous expression of grifolic acid synthases from Stachybotrys bisbyi and a daurichromenic acid synthase from R. dauricum in Aspergillus oryzae mediated three-step combinatorial biosynthesis of (+)-daurichromenic acid through enantioselective 6-endo-trig cyclization. Additional introduction of a halogenase from Fusarium sp. into the strain resulted in the biosynthesis of (+)-5-chlorodaurichromenic acid, which exceeds the antibacterial activity of the original compounds.
Total synthesis of biologically interesting (?)-cannabichromene, (?)-cannabichromenic acid, and (?)-daurichromenic acid is described. The key step in the synthetic strategy involves the formation of benzopyrans by ethylenediamine diacetate-catalyzed reactions of resorcinols with \alpha,\beta-unsaturated aldehydes.
Cannabis sativa L. is a prolific, but not exclusive, producer of a diverse group of isoprenylated resorcinyl polyketides collectively known as phytocannabinoids. The modular nature of the pathways that merge into the phytocannabinoid chemotype translates in differences in the nature of the resorcinyl side-chain and the degree of oligomerization of the isoprenyl residue, making the definition of phytocannabinoid elusive from a structural standpoint. A biogenetic definition is therefore proposed, splitting the phytocannabinoid chemotype into an alkyl-and a b-aralklyl version, and discussing the relationships between phytocannabinoids from different sources (higher plants, liverworts, fungi). The startling diversity of cannabis phytocannabinoids might be, at least in part, the result of non-enzymatic transformations induced by heat, light, and atmospheric oxygen on a limited set of major constituents (CBG, CBD, D 9-THC and CBC and their corresponding acidic versions), whose degradation is detailed to emphasize this possibility. The diversity of metabotropic (cannabinoid receptors), ionotropic (thermos-TRPs), and transcription factors (PPARs) targeted by phytocannabinoids is discussed. The integrated inventory of these compounds and their biological macromolecular end-points highlights the opportunities that phytocannabinoids offer to access desirable drug-like space beyond the one associated to the narcotic target CB 1 .
Acne is a common skin disease characterized by elevated sebum production and inflammation of the sebaceous glands. We have previously shown that a non-psychotropic phytocannabinoid ((–)-cannabidiol [CBD]) exerted complex anti-acne effects by normalizing ‘pro-acne agents’-induced excessive sebaceous lipid production, reducing proliferation and alleviating inflammation in human SZ95 sebocytes. Therefore, in this study we aimed to explore the putative anti-acne effects of further non-psychotropic phytocannabinoids ((–)-cannabichromene [CBC], (–)-cannabidivarin [CBDV], (–)-cannabigerol [CBG], (–)-cannabigerovarin [CBGV] and (–)-Δ9-tetrahydrocannabivarin [THCV]). Viability and proliferation of human SZ95 sebocytes were investigated by MTT and CyQUANT assays; cell death and lipid synthesis were monitored by DilC1(5)-SYTOX Green labelling and Nile Red staining, respectively. Inflammatory responses were investigated by monitoring expressions of selected cytokines upon lipopolysaccharide treatment (RT-qPCR, ELISA). Up to 10 μm, the phytocannabinoids only negligibly altered the viability of the sebocytes, whereas high doses (≥50 μm) induced apoptosis. Interestingly, basal sebaceous lipid synthesis was differentially modulated by the substances: CBC and THCV suppressed it, and CBDV had only minor effects, whereas CBG and CBGV increased it. Importantly, CBC, CBDV and THCV significantly reduced arachidonic acid (AA)-induced ‘acne-like’ lipogenesis. Moreover, THCV suppressed proliferation, and all phytocannabinoids exerted remarkable anti-inflammatory actions. Our data suggest that CBG and CBGV may have potential in the treatment of dry-skin syndrome, whereas CBC, CBDV and especially THCV show promise to become highly efficient, novel anti-acne agents. Moreover, based on their remarkable anti-inflammatory actions, phytocannabinoids could be efficient, yet safe novel tools in the management of cutaneous inflammations.
Development of an oxa-[3+3] annulation of vinyliminium salts with resorcinols as a 1,3-diketo equivalent is described. This annulation constitutes a cascade of Knoevenagel condensation-oxa-electrocyclization leading to a direct access to chromenes. A series of attempts was made to demonstrate its synthetic utility in natural product synthesis, culminating in a total synthesis of (±)-rhododaurichromanic acid A that also featured an intramolecular Gassman-type cationic [2+2] cycloaddition.
Simple and facile synthetic routes for the preparation of biologically interesting cyclol bearing polycycles were developed using FeCl3-promoted [2 + 2] cycloaddition from readily available benzopyrans possessing a variety of substituents. As examples of this methodology, one-step syntheses of cannabicyclol, cannabicyclovarin, and ranhuadujuanine A were accomplished in good yield.
Isolation and structure elucidation of cannabischromene is reported. Cannabichromen is the first compound isolated from hashish which contains the skeleton of chromene.
Citral condenses with olivetol in the presence of pyridine (1 mol) to give citrylidene-cannabis(III)(26%), cannabi-chromen (IV)(15%), its isomer (V)(3%) the bischromen (II)(6%), cannabicyclol (IX)(1%), its isomer (XV)(3%), and the bis-compound (XVI)(1·3%). Cannabichromen when heated with pyridine gives citrylidene-cannabis and cannabicyclol: the latter is also formed from cannabichromen under acid catalysis, and photochemically, in preparatively acceptable yields. Reasons for revising to (IX) the earlier structural proposal (VII) for cannabicyclo, are presented. Cannabichromen and cannabicycol are extractives of Cannabis(hashish).
Two novel chromane derivatives (1 and 2) and the known chromene (3) were isolated from the leaves and twigs of Rhododendron dauricum. The absolute stereostructure of 1 was established by spectroscopic examination and X-ray crystallographic analysis. The absolute stereostructures of 2 and 3 were also confirmed by photochemical conversion of 3 into 1 and 2. Daurichromenic acid (3) demonstrated potent anti-HIV activity with an EC50 value of 0.00567μg/mL and therapeutic index (TI) of 3,710. Rhododaurichromanic acid A (1) also showed relatively potent anti-HIV activity with an EC50 value of 0.37μg/mL, and a TI of 91.9, whereas rhododaurichromanic acid B (1) displayed no anti-HIV activity.
Total syntheses of a series of chromane natural products that contain a cyclobutane ring are described. A unified theme in the strategy employed for all these syntheses is an oxa-[3 + 3] annulation for constructing the chromane nucleus and a stepwise cationic [2 + 2] cycloaddition for the cyclobutane formation. More importantly, the two reactions could be rendered in tandem, thereby providing an expeditious approach to this family of natural products.
The natural chromenes D,L-cannabichromene, franklinone, alloevodionol, evodionol methyl ether and the trimethy ether of flemingin C were synthesized by cyclodehydrogenation of the corresponding isoprenylphenols with 2,3-dichloro-5,6-dicyanobenzoquinone (DDQ) or via the chromanes and dehydrogenation with DDQ. The biogenetical interest of this route is emphasized
Background and purpose:
The non-psychotropic cannabinoid cannabichromene is known to activate the transient receptor potential ankyrin-type1 (TRPA1) and to inhibit endocannabinoid inactivation, both of which are involved in inflammatory processes. We examined here the effects of this phytocannabinoid on peritoneal macrophages and its efficacy in an experimental model of colitis.
Experimental approach:
Murine peritoneal macrophages were activated in vitro by LPS. Nitrite levels were measured using a fluorescent assay; inducible nitric oxide (iNOS), cyclooxygenase-2 (COX-2) and cannabinoid (CB1 and CB2 ) receptors were analysed by RT-PCR (and/or Western blot analysis); colitis was induced by dinitrobenzene sulphonic acid (DNBS). Endocannabinoid (anandamide and 2-arachidonoylglycerol), palmitoylethanolamide and oleoylethanolamide levels were measured by liquid chromatography-mass spectrometry. Colonic inflammation was assessed by evaluating the myeloperoxidase activity as well as by histology and immunohistochemistry.
Key results:
LPS caused a significant production of nitrites, associated to up-regulation of anandamide, iNOS, COX-2, CB1 receptors and down-regulation of CB2 receptors mRNA expression. Cannabichromene significantly reduced LPS-stimulated nitrite levels, and its effect was mimicked by cannabinoid receptor and TRPA1 agonists (carvacrol and cinnamaldehyde) and enhanced by CB1 receptor antagonists. LPS-induced anandamide, iNOS, COX-2 and cannabinoid receptor changes were not significantly modified by cannabichromene, which, however, increased oleoylethanolamide levels. In vivo, cannabichromene ameliorated DNBS-induced colonic inflammation, as revealed by histology, immunohistochemistry and myeloperoxidase activity.
Conclusion and implications:
Cannabichromene exerts anti-inflammatory actions in activated macrophages - with tonic CB1 cannabinoid signalling being negatively coupled to this effect - and ameliorates experimental murine colitis.
The first asymmetric synthesis of daurichromenic acid and confluentin is described. The key step of the sequence leading to both natural products is a highly enantioselective domino aldol/oxa Michael reaction (97% ee) of farnesal and 2-methoxy-4-methylsalicylaldehyde.
The mechanism that controls the proportion of cannabichromene (CBC), a potential pharmaceutical, in the cannabinoid fraction
of Cannabis sativa L. is explored. As with tetrahydrocannabinol (THC) and cannabidiol (CBD), CBC is an enzymatic conversion product of the precursor
cannabigerol (CBG). CBC is reported to dominate the cannabinoid fraction of juveniles and to decline with maturation. This
ontogeny was confirmed in inbred lines with different mature chemotypes. A consistent CBC presence was found in early leaves
from a diverse clone collection, suggesting that CBC synthase is encoded by a fixed locus. Morphological variants possessing
a ‘prolonged juvenile chemotype’ (PJC), a substantial proportion of CBC persisting up to maturity, are presented. PJC is associated
with a reduced presence of floral bracts, bracteoles, and capitate-stalked trichomes. Genetic factors causing these features
were independent of the allelic chemotype locus B that was previously postulated and regulates THC and CBD synthesis and CBG accumulation. In contrast to previously described
Cannabis chemotypes, the cannabinoid composition of PJCs showed plasticity in that reduced light levels increased the CBC proportion.
The ability of PJC plants to enable the production of pharmaceutical raw material with high CBC purity is demonstrated.
Cannabichromene (CBC) is a major non-psychotropic phytocannabinoid that inhibits endocannabinoid inactivation and activates the transient receptor potential ankyrin-1 (TRPA1). Both endocannabinoids and TRPA1 may modulate gastrointestinal motility. Here, we investigated the effect of CBC on mouse intestinal motility in physiological and pathological states.
Inflammation was induced in the mouse small intestine by croton oil. Endocannabinoid (anandamide and 2-arachidonoyl glycerol), palmitoylethanolamide and oleoylethanolamide levels were measured by liquid chromatography-mass spectrometry; TRPA1 and cannabinoid receptors were analysed by quantitative RT-PCR; upper gastrointestinal transit, colonic propulsion and whole gut transit were evaluated in vivo; contractility was evaluated in vitro by stimulating the isolated ileum, in an organ bath, with ACh or electrical field stimulation (EFS).
Croton oil administration was associated with decreased levels of anandamide (but not 2-arachidonoyl glycerol) and palmitoylethanolamide, up-regulation of TRPA1 and CB₁ receptors and down-regulation of CB₂ receptors. Ex vivo CBC did not change endocannabinoid levels, but it altered the mRNA expression of TRPA1 and cannabinoid receptors. In vivo, CBC did not affect motility in control mice, but normalized croton oil-induced hypermotility. In vitro, CBC reduced preferentially EFS- versus ACh-induced contractions. Both in vitro and in vivo, the inhibitory effect of CBC was not modified by cannabinoid or TRPA1 receptor antagonists.
CBC selectively reduces inflammation-induced hypermotility in vivo in a manner that is not dependent on cannabinoid receptors or TRPA1.
A liquid chromatography-tandem mass spectrometry (LC-MS-MS)method was developed for the analysis of marijuana cannabinoids in mouse brain tissue using an Applied Biosystems 3200 Q trap with a turbo V source for TurbolonSpray attached to a Shimadzu SCL HPLC system. The method included cannabichromene (CBC),cannabidiol (CBD), Δ⁹-tetrahydrocannabinol (THC), 11-hydroxytetrahydrocannabinol (11-OH-THC), and 11-nor-Δ⁹-tetrahydrocannabinol-9-carboxylic acid (THC-COOH). These compounds were isolated by liquid-liquid extraction using cold acetonitrile. The following transition ions were monitored by multiple reaction monitoring (MRM): m/z 315>193, 315>259 for THC/CBD/CBC; m/z 331>193, 331>105 for 11-OH-THC; m/z 345>299, 345>193 for THC-COOH; m/z 318>196 for THC-d₃; m/z 334>196 for 11-OH-THC-d₃, and m/z 348>302 for THC-COOH-d₃. Linearity for THC, 1-OH-THC, and THC-COOH was 1-200 ng/g; for CBC and CBD, it was 0.5-20 ng/g. Within-run and between-run precisions for all the analytes yielded coefficients of variation of < 20%. Four C57BL6 mice were sacrificed 20 min after nose-only exposure to the smoke of 200 mg of marijuana containing 0.44 mg CBC, 0.93 mg CBD, and 8.81 mg THC. The mean brain concentrations were 3.9 ± 1.5 ng/g CBC, 21 ± 3.9 ng/g CBD, 364 ± 74 ng/g THC, and 28 ± 5.9 ng/g 11-OH-THC. THC-COOH was not detected. The relative mean brain cannabinoid concentrations correlated to the amounts of the cannabinoids in the inhaled marijuana.
Cannabidiol (CBD) and Δ(9) -tetrahydrocannabinol (THC) interact with transient receptor potential (TRP) channels and enzymes of the endocannabinoid system.
The effects of 11 pure cannabinoids and botanical extracts [botanical drug substance (BDS)] from Cannabis varieties selected to contain a more abundant cannabinoid, on TRPV1, TRPV2, TRPM8, TRPA1, human recombinant diacylglycerol lipase α (DAGLα), rat brain fatty acid amide hydrolase (FAAH), COS cell monoacylglycerol lipase (MAGL), human recombinant N-acylethanolamine acid amide hydrolase (NAAA) and anandamide cellular uptake (ACU) by RBL-2H3 cells, were studied using fluorescence-based calcium assays in transfected cells and radiolabelled substrate-based enzymatic assays. Cannabinol (CBN), cannabichromene (CBC), the acids (CBDA, CBGA, THCA) and propyl homologues (CBDV, CBGV, THCV) of CBD, cannabigerol (CBG) and THC, and tetrahydrocannabivarin acid (THCVA) were also tested.
CBD, CBG, CBGV and THCV stimulated and desensitized human TRPV1. CBC, CBD and CBN were potent rat TRPA1 agonists and desensitizers, but THCV-BDS was the most potent compound at this target. CBG-BDS and THCV-BDS were the most potent rat TRPM8 antagonists. All non-acid cannabinoids, except CBC and CBN, potently activated and desensitized rat TRPV2. CBDV and all the acids inhibited DAGLα. Some BDS, but not the pure compounds, inhibited MAGL. CBD was the only compound to inhibit FAAH, whereas the BDS of CBC > CBG > CBGV inhibited NAAA. CBC = CBG > CBD inhibited ACU, as did the BDS of THCVA, CBGV, CBDA and THCA, but the latter extracts were more potent inhibitors.
These results are relevant to the analgesic, anti-inflammatory and anti-cancer effects of cannabinoids and Cannabis extracts.
In contrast to the numerous reports on the pharmacological effects of Δ(9)-tetrahydrocannabinol (THC), the pharmacological activity of another substituent of Cannabis sativa, cannabichromene (CBC) remains comparatively unknown. In the present study, we investigated whether CBC elicits cannabinoid activity in the tetrad assay, which consists of the following four endpoints: hypomotility, antinociception, catalepsy, and hypothermia. Because cannabinoids are well documented to possess anti-inflammatory properties, we examined CBC, THC, and combination of both phytocannabinoids in the lipopolysaccharide (LPS) paw edema assay. CBC elicited activity in the tetrad that was not blocked by the CB(1) receptor antagonist, rimonabant. Moreover, a behaviorally inactive dose of THC augmented the effects of CBC in the tetrad that was associated with an increase in THC brain concentrations. Both CBC and THC elicited dose-dependent anti-inflammatory effects in the LPS-induced paw edema model. The CB(2) receptor, SR144528 blocked the anti-edematous actions of THC, but not those produced by CBC. Isobolographic analysis revealed that the anti-edematous effects of these cannabinoids in combination were additive. Although CBC produced pharmacological effects, unlike THC, its underlying mechanism of action did not involve CB(1) or CB(2) receptors. In addition, there was evidence of a possible pharmacokinetic component in which CBC dose-dependently increased THC brain levels following an i.v. injection of 0.3mg/kg THC. In conclusion, CBC produced a subset of behavioral activity in the tetrad assay and reduced LPS-induced paw edema through a noncannabinoid receptor mechanism of action. These effects were augmented when CBC and THC were co-administered.
Synthetic and naturally occurring cannabidiol and cannabichromene were distinctly separated without derivation by GLC using a 6% OV-1 column; an artifact of cannabichromene, cannabicyclol, was separated from (minus)-delta9-trans-tetrahydrocanna-bivarin. This procedure is versatile and applicable for the quantitation of Cannabis containing both cannabidiol and cannabichromene. Biological interaction among (minus)-delta9-trans-tetrhydrocanabinol, cannabichromene, and other cannabinoids in natural Cannabis preparations can now be studied. In the phenyl methyl silicone polymer series, cannabidiol precedes ca-nabichromene on columns containing below a 50% phenyl-to-methyl ratio. Columns containing below a 50% phenyl-to-methyl ratio. Columns containing a 50:50 or greater ratio of phenyl to methyl reverse the separation order with cannabichromene preceding cannabidiol.
The metabolism of delta-9-tetrahydrocannabinol (delta-9-THC), delta-8-THC, delta-11-THC, cannabidiol (CBD), cannabinol (CBN), cannabichromene (CBC), cannabigerol (CBG) and the equatorial-isomer of hexahydrocannabinol (HHC) was studied in microsomal preparations obtained from rats, mice, guinea pigs, rabbits, hamsters, gerbils and a cat. Identification of metabolites was by GC/MS and quantification by gas chromatography. Major metabolites were monohydroxylated compounds but the pattern of hydroxylation varied considerably between the species, no doubt reflecting the variable nature of the cytochrome P-450 mixed-function oxidases. Although the primary carbon allylic to the endocyclic double bond of tricyclic cannabinoids was usually the major site of attack, the 4' (side-chain, omega-1 position) and the terpene ring were usually favoured by the cat and hamster respectively. The guinea pig generally produced more metabolites hydroxylated in the side-chain (all positions) than did the other species. The results from HHC were very similar to those from THC, namely hydroxylation at C-11 in most species, and the production of high concentrations of 8 alpha-hydroxy-HHC in the mouse and 8 beta-hydroxy-HHC in the hamster. As this molecule lacks the double bond of the THCs and, hence, the allylic nature of C-11 and C-8, the results suggest that it is the orientation of the molecule to the active site of the cytochrome P-450 mixed-function oxidase rather than the reactivity of the C-H bond that governs the position of hydroxylation.
Metabolites of cannabichromene (CBC) produced by hepatic microsomal incubates from rabbits and mice were examined by gas chromatography/mass spectrometry (GC/MS) as trimethylsilyl (TMS) and (2H9)TMS derivatives. Most metabolites were hydroxylated compounds whose mass spectra gave very little information on metabolite structure as fragmentation was dominated by formation of the substituted chromenyl ion. This prevented charge localization and diagnostic fragmentation at the site of metabolic attack. This paper describes the identification of these metabolites by GC/MS techniques using both deuterium-exchange reactions and hydrogenation of the metabolites to tetrahydro derivatives; the latter method was used to suppress chromenyl ion formation and to enhance the relative abundance of diagnostic fragment ions. Twenty-one metabolites were identified. Metabolites were found hydroxylated in all positions of both aliphatic chains, with additional compounds formed by epoxidation and reduction of the aliphatic double bond in the methylpentenyl chain. Dihydroxy metabolites were hydoxylated in both the pentyl and methylpentenyl chains in positions common to those hydroxylated in the monohydroxy metabolites.
Metabolism of cannabichromene (CBC) was studied in hepatic microsomal incubates from mouse, rat, rabbit, guinea pig, cat, hamster, and gerbil. Metabolites were extracted with ethyl acetate, concentrated by chromatography on Sephadex LH-20, and identified by GC/MS as trimethylsilyl derivatives of both the metabolites themselves and their hydrogenated analogues. Thirteen metabolites were identified. The major metabolites were monohydroxy compounds with hydroxylation at all positions of the pentyl and methylpentenyl chains. An epoxide and its derived dihydrodiol were formed from the double bond in the methylpentenyl chains. Several unidentified decomposition products were found in the extracts from mouse, gerbil, and cat; these appeared to have been produced by the opening of the dihydropyran ring. Metabolism varied considerably between the species, although the trans-hydroxy metabolite 5'-hydroxy-CBC was the major metabolite in most cases. Metabolites hydroxylated in the pentyl chain were more abundant in mouse, rabbit, and cat; the hamster, gerbil, and cat produced the most epoxide-derived material.
1. Neither cannabichromene (CBC) nor delta 9-tetrahydrocannabinol (THC) protected mice from electroshock-induced seizures, although THC inhibited postictal mortality. Minor effects were produced on seizure latency and duration. 2. CBC had a weak analgetic action in mice; THC had a moderate and lengthy effect, which was potentiated at 2 hr by concurrent CBC. 3. Both CBC (10-75 mg/kg, i.p.) and THC (20 mg/kg) reduced motility of mice, the THC equalling the highest dose of CBC. 4. Performance of a conditioned avoidance response was strongly impaired by THC, but not by CBC, nor did CBC combined with THC have influence on the effects of THC.
Cannabichromene homologs, analogs, and isomers as well as the C1-homolog and isomer of cannabigerol were prepared and tested for their antimicrobial and antifungal properties. Spectral data of all compounds synthesized are presented.
Cannabichromene (CBC) is one of four major cannabinoids in Cannabis sativa L. and is the second most abundant cannabinoid in drug-type cannabis. Cannabichromene and some of its homologs, analogs, and isomers were evaluated for antiinflammatory, antibacterial, and antifungal activity. Antiinflammatory activity was evaluated by the carrageenan-induced rat paw edema and the erythrocyte membrane stabilization method. In both tests, CBC was superior to phenylbutazone. Antibacterial activity of CBC and its isomers and homologs was evaluated using gram-positive, gram-negative, and acid-fast bacteria. Antifungal activity was evaluated using yeast-like and filamentous fungi and a dermatophyte. Antibacterial activity was strong, and the antifungal activity was mild to moderate.
Cannabiorcichromenic acid and 8-chlorocannabiorcichromenic acid [8-chloro-5-hydroxy-2,7-dimethyl-2-(4-methyl-3-pentenyl)-2H-1-benzopyran -6- carboxylic acid] were identified as active components in cultures of Cylindrocarpon olidum which antagonized various other fungi. Experiments performed with the purified acids confirmed the antifungal activity; in addition, they revealed that the acids had antibiotic properties towards gram-positive bacteria and were toxic to nematodes.
Cannabichromenic acid synthase was purified to apparent homogeneity by sequential column chromatography including DEAE-cellulose, phenyl-Sepharose CL-4B, and hydroxylapatite. The enzyme catalysed the oxidocyclization of cannabigerolic acid and cannabinerolic acid to cannabichromenic acid. The K(m) values for both substrates were in the same order of magnitude although the Vmax value for the former was higher than that for the latter. These results suggested that cannabichromenic acid is predominantly formed from cannabigerolic acid rather than cannabinerolic acid. The enzyme required neither molecular oxygen nor hydrogen peroxide, indicating that the cannabichromenic acid synthase reaction proceeds through direct dehydrogenation without hydroxylation.
Several prenylphenols from basidiocarps of European and Chinese Albatrellus spp., namely grifolin (1), neogrifolin (2), confluentin (3), scutigeral (4), and albaconol (5) were investigated concerning their activities in test models for vanilloid receptor modulation. The isolation of these compounds from A. confluens and structure elucidation of the novel natural product confluentin (3) are described. The effects of scutigeral and neogrifolin on vanilloid receptors were studied by means of electrophysiological methodology on rat dorsal root ganglion neurons as well as on recombinant cell lines expressing the rat VR1 receptor. Concurrently, the effects of compounds 1-5 on a reporter cell line expressing the human vanilloid receptor VR1 were measured. In contrast to previous studies reported in the literature, the results of these investigations suggest that fungal prenylphenols act as weak antagonists (activity in the microM range), rather than exhibiting agonistic activities.
Four new prenylated orcinol derivatives, daurichromenes A-D (1-4), along with three known compounds, confluentin (5), grifolin (6), and orcinol (7), have been isolated from the Chinese medicinal plant Rhododendron dauricum. Their structures were established as 2R-(7'-hydroxy-4',8'-dimethyl-3'E,8'-nonadienyl)-5-hydroxy-2,7-dimethyl-2H-chromene (1), 2R-(3'-hydroxy-8'-methyl-4'-methyliden-7'-nonaenyl)-5-hydroxy-2,7-dimethyl-2H-chromene (2), 2R-(8'-hydroxy-4',8'-dimethyl-3'E,6'Z-nonadienyl)-5-hydroxy-2,7-dimethyl-2H-chromene (3), and 2R-(9'-hydroxy-4',8'-dimethyl-3'E,7'E-nonadienyl)-5-hydroxy-2,7-dimethyl-2H-chromene (4) by analysis of spectral data. The absolute configuration of the asymmetric carbons at the chromene ring in 1-5 was determined as R from their circular dichroism spectra. Compounds 1-6 significantly inhibited compound 48/80-induced histamine release from rat peritoneal mast cells.
Chemical classification of plants. XXXI. Hashish. 10. Cannabichromene, a new hashish component
- U Claussen
- F Spulak F V, Korte
Claussen U, Spulak F v, Korte F. (1966) Chemical classification of plants. XXXI. Hashish. 10. Cannabichromene, a new hashish component.
Tetrahedron, 22, 1477-1479.
Recent Development in the Chemistry of Natural Substances
- W D Ollis
- I O Sutherland
Ollis WD, Sutherland IO. (1961) Recent Development in the Chemistry of Natural Substances, p 84, Pergamon Press Oxford 1961.
Examination of a 140 year old ethanolic extract of Cannabis: Identification of new cannabitriol homologues and ethyl homologue of cannabinol
- D J Harvey
Harvey DJ. (1985) Examination of a 140 year old ethanolic extract of Cannabis: Identification of new cannabitriol homologues and ethyl
homologue of cannabinol. Proceeding of the Oxford Symposium on Cannabis, 23-30.