Spiro-indans from Cannabis sativa

Three new spiro-indans were isolated from hashish (Cannabis sativa). Their chemical structures were established by spectroscopic means, and by chemical correlation with cannabispiran and iso-cannabispiran. They were found to be 7-hydroxy-5-methoxyindan-1-spiro-cyclohexane, its isomer 5-hydroxy-7-methoxyindan-1-spirocyclo-hexane and 5,7-dihydroxyindan-1-spiro-cyclohexane.     

In silicio expression analysis of PKS genes isolated from Cannabis sativa L

Cannabinoids, flavonoids, and stilbenoids have been identified in the annual dioecious plant Cannabis sativa L. Of these, the cannabinoids are the best known group of this plant's natural products. Polyketide synthases (PKSs) are responsible for the biosynthesis of diverse secondary metabolites, including flavonoids and stilbenoids. Biosynthetically, the cannabinoids are polyketide substituted with terpenoid moiety. Using an RT-PCR homology search, PKS cDNAs were isolated from cannabis plants. The deduced amino acid sequences showed 51%-73% identity to other CHS/STS type sequences of the PKS family. Further, phylogenetic analysis revealed that these PKS cDNAs grouped with other non-chalcone-producing PKSs. Homology modeling analysis of these cannabis PKSs predicts a 3D overall fold, similar to alfalfa CHS2, with small steric differences on the residues that shape the active site of the cannabis PKSs.     

NMR assignments of the major cannabinoids and cannabiflavonoids isolated from flowers of Cannabis sativa

The complete 1H- and 13C-NMR assignments of the major Cannabis constituents, delta9-tetrahydrocannabinol, tetrahydrocannabinolic acid, delta8-tetrahydrocannabinol, cannabigerol, cannabinol, cannabidiol, cannabidiolic acid, cannflavin A and cannflavin B have been determined on the basis of one- and two-dimensional NMR spectra including 1H- and 13C-NMR, 1H-1H-COSY, HMQC and HMBC. The substitution of carboxylic acid on the cannabinoid nucleus (as in tetrahydrocannabinolic acid and cannabidiolic acid) has a large effect on the chemical shift of H-1" of the C5 side chain and 2'-OH. It was also observed that carboxylic acid substitution reduces intermolecular hydrogen bonding resulting in a sharpening of the H-5' signal in cannabinolic acid in deuterated chloroform. The additional aromaticity of cannabinol causes the two angular methyl groups (H-8 and H-9) to show identical 1H-NMR shifts, which indicates that the two aromatic rings are in one plane in contrast to the other cannabinoids. For the cannabiflavonoids, the unambiguous assignments of C-3' and C-4' of cannflavin A and B were determined by HMBC spectra.     

Water-soluble glycoproteins from Cannabis sativa (Thailand)

Glycoproteins were extracted with water from leaves of Cannabis sativa grown from seeds of Thailand origin. By ion exchange chromatography the material was separated into a neutral and an acidic fraction. Both glycoprotein fractions contained arabinose, galactose, glucose, mannose and xylose, and in addition rhamnose and galacturonic acid were present in the acidic fraction. The carbohydrate moieties were investigated by methylation analysis and Smith-degradation, whereas the glycopeptide linkage was studied by alkaline hydrolysis in the presence of NaBH₄ and Na₂SO₃, respectively. This linkage was shown to be of the serine-O-galactoside type. The carbohydrate structure is highly branched, the majority of branches terminating in arabinofuranose end groups. Arabinose is also present in the chain, predominantly (1 → 4)- and/or (1 → 5)-linked. Galactose makes up most of the main chain as (1 → 3)-linked residues but also constitutes end groups and branch points, as do mannose and/or glucose. Xylose and rhamnose are present as (1 → 4)- and (1 → 2)-linked units, respectively. Galacturonic acid is assumed to be (1 → 4)- linked with some branching at 3 position. The amino acid hydroxyproline, present in the glycoprotein of South African Cannabis leaves, was absent in the corresponding Thailand material.     

Cannabicitran: A new naturally occurring tetracyclic diether from lebanese Cannabis sativa

A new cannabinoid, cannabicitran, was isolated from Lebanese hashish. This component was found to be identical to synthetic citrylidene-cannabis.     

Non-cannabinoid constituents from a high potency Cannabis sativa variety

Six new non-cannabinoid constituents were isolated from a high potency Cannabis sativa L. variety, namely 5-acetoxy-6-geranyl-3-n-pentyl-1,4-benzoquinone (1), 4,5-dihydroxy-2,3,6-trimethoxy-9,10-dihydrophenanthrene (2), 4-hydroxy-2,3,6,7-tetramethoxy-9,10-dihydrophenanthrene (3), 4,7-dimethoxy-1,2,5-trihydroxyphenanthrene (4), cannflavin C (5) and β-sitosteryl-3-O-β-d-glucopyranoside-2′-O-palmitate (6). In addition, five known compounds, α-cannabispiranol (7), chrysoeriol (8), 6-prenylapigenin (9), cannflavin A (10) and β-acetyl cannabispiranol (11) were identified, with 8 and 9 being reported for the first time from cannabis. Some isolates displayed weak to strong antimicrobial, antileishmanial, antimalarial and anti-oxidant activities. Compounds 2-4 were inactive as analgesics.     

三种产地火麻仁中CannabisinA的含量测定

为建立同时测定广西,甘肃,云南三个产地火麻仁中Cannabisin A的含量测定方法,本文采用高效液相色谱法对三种产地火麻仁中Cannabisin A 进行含量测定.用Waters C18(250 mm×4.6 mm,5 μm)色谱柱,流动相:乙腈-0.1%磷酸溶液(22∶ 78);流速1.0 mL/min;检测波长:257 nm,柱温25 ℃.结果表明:Cannabisin A在0.1～0.50 μg范围内线性关系良好,r=0.9996,加样回收率100.31%,RSD=0.63%(n＝6).三种产地的火麻仁中Cannabisin A含量分别为0.343%,0.250%,0.455%.不同产地火麻仁中Cannabisin A含量有一定的差异,该法操作简便,准确,重复性好,可用于火麻仁中Cannabisin A的含量测定.     

Flavonoid glycosides and cannabinoids from the pollen of Cannabis sativa L

Chemical investigation of the pollen grain collected from male plants of Cannabis sativa L. resulted in the isolation for the first time of two flavonol glycosides from the methanol extract, and the identification of 16 cannabinoids in the hexane extract. The two glycosides were identified as kaempferol 3-O-sophoroside and quercetin 3-O-sophoroside by spectroscopic methods including high-field two-dimensional NMR experiments. The characterisation of each cannabinoid was performed by GC-FID and GC-MS analyses and by comparison with both available reference cannabinoids and reported data. The identified cannabinoids were delta9-tetrahydrocannabiorcol, cannabidivarin, cannabicitran, delta9-tetrahydrocannabivarin, cannabicyclol, cannabidiol, cannabichromene, delta9-tetrahydrocannabinol, cannabigerol, cannabinol, dihydrocannabinol, cannabielsoin, 6a, 7, 10a-trihydroxytetrahydrocannabinol, 9, 10-epoxycannabitriol, 10-O-ethylcannabitriol, and 7, 8-dehydro-10-O-ethylcannabitriol.     

Isolation of endophytic fungi from Cannabis sativa and study their antifungal potential

A systemic study of fungal endophytes associated with different plant parts of Cannabis sativa and their antifungal activity was investigated in the present study. A total of 281 plant segments, including 91 leaves, 93 stem and 97 petioles samples, were screened for the isolation of endophytic fungi. Totally, 212 (77.65%) segments were found colonised by different fungi. Highest colonisation frequency were observed in stem parts (84.94%), then leaves (82.41%) and lowest 59.79% in petiole. Total eight fungal genera belonging to 12 species were isolated. Aspergillus is recorded as the most frequently occurring genera with three species Aspergillus niger, Aspergillus flavus and Aspergillus nidulans followed by Penicillium with two species Penicillium chrysogenum and Penicillium citrinum, while Phoma, Rhizopus, Colletotrichum, Cladosporium and Curvularia with single species. The antifungal potential of A. niger and A. flavus – two most frequently isolated endophytic fungi – was evaluated against two common plant pathogen, Colletotrichum gloeosporioides and Curvularia lunata. Different plant and fungal extracts individually and in combinations showed variations in antifungal activity against both the pathogens. The primary results obtained on antifungal activity of endophytes show their possible role in plant defence mechanism but it is a preliminary approach and more extensive research is still required.     

Phytochemical Characterization of Cannabis sativa L. Roots from Northeastern Brazil

This article describes the phytochemical study of Cannabis sativa roots from northeastern Brazil. The dried plant material was pulverized and subjected to exhaustive maceration with ethanol at room temperature, obtaining the crude ethanolic extract (Cs-EEBR). The volatile compounds were analyzed by gas chromatography coupled with mass spectrometry (GC/MS), which allowed to identify 22 compounds by comparing the linear retention index (LRI), the similarity index (SI) and the fragmentation pattern of the constituents with the literature. By this technique the major compounds identified were: friedelan-3-one and β-sitosterol. In addition, two fractions were obtained from Cs-EEBR by classical column chromatography and preparative thin layer chromatography. These fractions were analyzed by NMR and IR and together with the mass spectrometry data allowed to identify the compounds: epifriedelanol, friedelan-3-one, β-sitosterol and stigmasterol. The study contributed to the phytochemical knowledge of Cannabis sativa, specifically the roots, as there are few reports on the chemical constituents of this part of the plant.     

大麻植物中大麻素成分研究进展

大麻（Cannabis sativa）是一种古老的栽培植物, 它既是一种毒品原植物, 又是一种极具开发利用价值的经济作物。大麻素是大麻植物中特有的含有烷基和单萜分子结构的一类次生代谢产物, 目前已分离出70多种, 其中包含使人致幻成瘾的四氢大麻酚（THC）。该文就大麻植物中几种主要的大麻素成分： 四氢大麻酚、大麻二酚（CBD）和大麻环萜酚（CBC）的存在特征、含量变化、生物合成途径、各关键酶及其基因、遗传方式等方面的研究进行概括和归纳, 并展望了当前大麻素的主要研究方向, 对加快我国大麻素的相关研究及大麻育种具有参考意义。     

Purification and some properties of an aminopeptidase from the seeds of Cannabis sativa

An aminopeptidase (HSA) with a molecular mass of 78 kDa was purified from hemp (Cannabis sativa) seeds. The activity was inhibited by monoiodeacetic acid, p-chloromercuri-phenylsulfonic acid, and Zn2+ ion. The specificity of HSA was similar to that of a leucyl aminopeptidase [EC 3.4.11.1] from mammalian cytosol. However, other enzyme properties were different from these of leucyl aminopeptidase.     

Chemical Characterization of Leaves,Male and Female Flowers from Spontaneous Cannabis (Cannabis sativa L.) Growing in Hungary

Spontaneous forms of hemp (Cannabis sativa L., often reported as Cannabis sativa var. spontanea Vavilov ) with a low content of psychoactive cannabinoids can be considered as a valuable source of other phytoconstituents to be used in nutraceuticals or for their health promoting properties. Chemical data on this hemp variety are rather scarce. In this article, we report a comprehensive phytochemical characterization of leaves, male and female inflorescences of C. sativa growing wild in Hungary. For the purpose, the essential oil along with polar extracts were analyzed using GC/MS, NMR and LC‐DAD‐MS techniques, respectively. The results indicated that female inflorescence essential oil contains high amounts of the CB2 agonists, (E)‐caryophyllene (28.3 %) and cannabidiol (CBD; 24.9 %), whereas leaves and male inflorescence essential oils contained lower amounts of both compounds. HPLC/MS allowed to quantify cannabidiol (CBD) and cannabidiolic acid (CBD?A) in the ethyl acetate extracts from leaves, male and female inflorescences; they were 0.3, 0.8 and 0.9 %, and 0.2, 0.3 and 0.4 %, respectively. Flavonoids of this spontaneous form of hemp were formed by C‐glycosides and glucuronic acids of kaempferol and apigenin with a total content of 3.8, 6.1 and 7.8 mg/g in methanolic extracts from leaves, male and female inflorescences, respectively. Based on these results, spontaneous C. sativa may represent an important source of CB₂ agonists and bioflavonoids to be used in nutraceuticals, cosmetics and pharmaceuticals.     

Comparative proteomics of Cannabis sativa plant tissues.

Comparative proteomics of leaves, flowers, and glands of Cannabis sativa have been used to identify specific tissue-expressed proteins. These tissues have significantly different levels of cannabinoids. Cannabinoids accumulate primarily in the glands but can also be found in flowers and leaves. Proteins extracted from glands, flowers, and leaves were separated using two-dimensional gel electrophoresis. Over 800 protein spots were reproducibly resolved in the two-dimensional gels from leaves and flowers. The patterns of the gels were different and little correlation among the proteins could be observed. Some proteins that were only expressed in flowers were chosen for identification by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry and peptide mass fingerprint database searching. Flower and gland proteomes were also compared, with the finding that less then half of the proteins expressed in flowers were also expressed in glands. Some selected gland protein spots were identified: F1D9.26-unknown prot. (Arabidopsis thaliana), phospholipase D beta 1 isoform 1a (Gossypium hirsutum), and PG1 (Hordeum vulgare). Western blotting was employed to identify a polyketide synthase, an enzyme believed to be involved in cannabinoid biosynthesis, resulting in detection of a single protein.