Identification of Novel Antileishmanial Chemotypes By High-Throughput Virtual and In Vitro Screening.

BACKGROUND: Leishmaniasis is a deadly protozoan parasitic disease and a significant health problem in underdeveloped and developing countries. The global spread of the parasite, coupled with the emergence of drug resistance and severe side effects associated with existing treatments, has necessitated the identification of new and potential drugs. OBJECTIVE: This study aimed to identify promising compounds for the treatment of leishmaniasis by targeting two essential enzymes of Leishmania donovani: trypanothione reductase (Try-R) and trypanothione synthetase (Try-S). METHODS: High-throughput virtual and in vitro screening of in-house and commercial databases was conducted. A pharmacophore model with seven features was developed and validated using the Guner-Henery method. The pharmacophore-based virtual screening yielded 690 hits, which were further filtered through Lipinski's rule, ADMET analysis, and molecular docking against Try-R and Try-S. Molecular dynamics studies were performed on selected compounds, and in vitro experiments were conducted to evaluate their activity against the promastigote and amastigote forms of L. donovani. RESULTS: The virtual screening and subsequent analysis identified 33 promising compounds. Molecular dynamics studies of two compounds (comp-1 and comp-2) demonstrated stable binding interactions with the target enzymes and high affinity. In vitro experiments revealed that 13 compounds exhibited moderate activity against both the promastigote (IC50, 41 µM-76 µM) and the amastigote (IC50, 44 µM-72 µM) forms of L. donovani. Compounds 1 and 2 showed the highest percent inhibition and the lowest IC50 values. CONCLUSION: The identified compounds demonstrated significant inhibitory activity against Leishmania donovani and stable interactions with target enzymes. These findings suggest that the compounds could serve as promising leads for developing new treatments for leishmaniasis.

Discovery and determination of misuse and chemotypes of Pogostemon cablin by liquid chromatography-quadrupole time-of-flight mass spectrometry and liquid chromatography with diode-array detector.

Traditional Chinese medicine (TCM) has garnered significant scientific interest in healthcare but faces increased regulatory scrutiny due to concerns about uncontrolled usage. This study focuses on characterizing Pogostemon cablin (PC) to mitigate potential misuse and identify chemotype differences. Leveraging untargeted metabolomics, we identified 222 distinctive features effectively differentiating PC from Agastache rugosa (AR), reducing misidentification risks. Pogostone and tilianin emerged as potential markers, leading to a high-performance liquid chromatography-diode array detection (HPLC-DAD) method development for PC and AR discrimination. Evaluation of PC chromatograms revealed notable profile and pogostone level differences among samples, suggesting chemotype associations. Untargeted metabolic profiling identified 78 features with significant differences, highlighting 7,3',4'-tri-O-methyleriodictyol as a potential discriminatory marker between PC chemotypes. The developed HPLC-DAD method quantified pogostone and 7,3',4'-tri-O-methyleriodictyol, effectively discriminating PC chemotypes. This platform differentiates PC and AR and distinguishes chemical types within PC, like pogostone-type and patchoulol-type. Applied to local TCM stores, it ensures PC authenticity. This approach addresses TCM control concerns, enhancing understanding and application of herbal medicine by providing insights into PC chemical composition and discrimination.

Biochemical characterisation of a cassava (Manihot esculenta crantz) diversity panel for post-harvest physiological deterioration; metabolite involvement and environmental influence.

Cassava (Manihot esculenta Crantz) produces edible roots, a major carbohydrate source feeding more than 800 million people in Africa, Latin America, Oceania and Asia. Post-harvest physiological deterioration (PPD) renders harvested cassava roots unpalatable and unmarketable. Decades of research on PPD have elucidated several genetic, enzymatic and metabolic processes involved. Breeding populations were established to enable verification of robust biomarkers for PPD resistance. For comparison, these PPD populations have been cultivated concurrently with diversity population for carotenoid (ß-carotene) content. Results highlighted a significant variation of the chemotypes due to environmental factors. Less than 3% of the detected molecular features showed consistent trends between the two harvest years and were putatively identified as phenylpropanoid derived compounds (e.g. caffeoyl rutinoside). The data corroborated that ∼20 µg ß-carotene/g DW can reduced the PPD response of the cassava roots to a score of ∼1. Correlation analysis showed a significant correlation of ß-carotene content at harvest to PPD response (R2 -0.55). However, the decrease of ß-carotene over storage was not significantly correlated to initial content or PPD response. Volatile analysis observed changes of apocarotenoids derived from ß-carotene, lipid oxidation products (alkanes, alcohols and carbonyls and esters) and terpenes. The majority of these volatiles (>90%) showed no significant correlation to ß-carotene or PPD. Observed data indicated an increase (∼2-fold) of alkanes in varieties with ß-carotene >10 µg/g DW and a decrease (∼60%) in varieties with less ß-carotene. Fatty acid methyl esters with a chain length > C9 were detected solely after storage and show lower levels in varieties with higher ß-carotene content. In combination with correlation values to PPD (R2 ∼0.3; P-value >0.05), the data indicated a more efficient ROS quenching mechanism in PPD resistant varieties.

Potential Anti-Infectious Activity of Essential Oil Chemotypes of Lippia origanoides Kunth on Antibiotic-Resistant Staphylococcus aureus Strains.

Staphylococcus aureus infections are prevalent in healthcare and community environments. Methicillin-resistant S. aureus is catalogued as a superbug of high priority among the pathogens. This Gram-positive coccus can form biofilms and produce toxins, leading to persistent infection and antibiotic resistance. Limited effective antibiotics have encouraged the development of innovative strategies, with a particular emphasis on resistance mechanisms and/or virulence factors. Medicinal aromatic plants have emerged as promising alternative sources. This study investigated the antimicrobial, antibiofilm, and antihemolysis properties of three different chemotypes of Lippia origanoides essential oil (EO) against susceptible and drug-resistant S. aureus strains. The chemical composition of the EO was analyzed using GC-MS, revealing high monoterpene concentrations, with carvacrol and thymol as the major components in two of the chemotypes. The third chemotype consisted mainly of the sesquiterpene ß-caryophyllene. The MIC values for the two monoterpene chemotypes ranged from 62.5 to 500 µg/mL for all strains, whereas the sesquiterpene chemotype showed activity against seven strains at concentrations of 125-500 µg/mL, which is the first report of its anti-S. aureus activity. The phenolic chemotypes inhibited biofilm formation in seven S. aureus strains, whereas the sesquiterpene chemotype only inhibited biofilm formation in four strains. In addition, phenolic chemotypes displayed antihemolysis activity, with IC50 values ranging from 58.9 ± 3.8 to 128.3 ± 9.2 µg/mL. Our study highlights the importance of L. origanoides EO from the Yucatan Peninsula, which has the potential for the development of anti-S. aureus agents.

Unpredictable Chemical Diversity of Essential Oils in Cinnamomum burmanni (Lauraceae) Living Collections: Beyond Maternally Inherited Phylogenetic Relationships.

The genus Cinnamomum encompasses diverse species with various applications, particularly in traditional medicine and spice production. This study focuses on Cinnamomum burmanni, specifically on a high-D-borneol-content chemotype, known as the Meipian Tree, in Guangdong Province, South China. This research explores essential oil diversity, chemotypes, and chloroplast genomic diversity among 28 C. burmanni samples collected from botanical gardens. Essential oils were analyzed, and chemotypes classified using GC-MS and statistical methods. Plastome assembly and phylogenetic analysis were conducted to reveal genetic relationships. Results showed distinct chemotypes, including eucalyptol and borneol types, with notable variations in essential oil composition. The chloroplast genome exhibited conserved features, with phylogenetic analysis revealing three major clades. Borneol-rich individuals in clade II suggested a potential maternal inheritance pattern. However, phylogenetic signals revealed that the composition of essential oils is weakly correlated with plastome phylogeny. The study underscores the importance of botanical gardens in preserving genetic and chemical diversity, offering insights for sustainable resource utilization and selective breeding of high-yield mother plants of C. burmanni.

Chemical Composition of Essential Oils from Natural Populations of Artemisia scoparia Collected at Different Altitudes: Antibacterial, Mosquito Repellent, and Larvicidal Effects.

The current study aimed to evaluate the presence of chemical variations in essential oils (EOs) extracted from Artemisia scoparia growing at different altitudes and to reveal their antibacterial, mosquito larvicidal, and repellent activity. The gas chromatographic-mass spectrometric analysis of A. scoparia EOs revealed that the major compounds were capillene (9.6-31.8%), methyleugenol (0.2-26.6%), ß-myrcene (1.9-21.4%), γ-terpinene (1.5-19.4%), trans-ß-caryophyllene (0.8-12.4%), and eugenol (0.1-9.1%). The EO of A. scoparia collected from the city of Attock at low elevation was the most active against Staphylococcus aureus, Bacillus subtilis, Escherichia coli, and Pseudomonas aeruginosa bacteria (minimum inhibitory concentration of 156-1250 µg/mL) and showed the best mosquito larvicidal activity (LC50, 55.3 mg/L). The EOs of A. scoparia collected from the high-altitude areas of Abbottabad and Swat were the most repellent for females of Ae. aegypti and exhibited repellency for 120 min and 165 min, respectively. The results of the study reveal that different climatic conditions and altitudes have significant effects on the chemical compositions and the biological activity of essential oils extracted from the same species.

New withanolides from leaves along with alkyl glucoside from roots and stem as well as variations in withanolide contents in leaves of accessions of Withania somnifera, Ashwagandha- the Indian ginseng$.

Based on the major components in the leaves, the ashwagandha has been found to exist in several chemotypic forms in India. From the leaves of various accessions of Withania somnifera, which were maintained in our institute, three new steroids namely, 4-acetoxy-20ß-hydroxy-1-oxo-witha-2,5,24-trienolide (7), 24,25-dihydro-14α-hydroxy withanolide D (9), 5α,6ß,17α,27-tetrahydroxy-1-oxo-witha-2,24-dienolide (12) together with thirteen known withanolides were identified by spectroscopic methods. From the roots and stem of one accession and leaves of another, a new alkyl ester glucoside (4) has also been isolated. The new withanolides 7, 9 and 12 have been tentatively named as withanolide 135 A, withanolide 135B and withanolide 108, respectively.

Datasets Construction and Development of QSAR Models for Predicting Micronucleus In Vitro and In Vivo Assay Outcomes.

In silico (quantitative) structure-activity relationship modeling is an approach that provides a fast and cost-effective alternative to assess the genotoxic potential of chemicals. However, one of the limiting factors for model development is the availability of consolidated experimental datasets. In the present study, we collected experimental data on micronuclei in vitro and in vivo, utilizing databases and conducting a PubMed search, aided by text mining using the BioBERT large language model. Chemotype enrichment analysis on the updated datasets was performed to identify enriched substructures. Additionally, chemotypes common for both endpoints were found. Five machine learning models in combination with molecular descriptors, twelve fingerprints and two data balancing techniques were applied to construct individual models. The best-performing individual models were selected for the ensemble construction. The curated final dataset consists of 981 chemicals for micronuclei in vitro and 1309 for mouse micronuclei in vivo, respectively. Out of 18 chemotypes enriched in micronuclei in vitro, only 7 were found to be relevant for in vivo prediction. The ensemble model exhibited high accuracy and sensitivity when applied to an external test set of in vitro data. A good balanced predictive performance was also achieved for the micronucleus in vivo endpoint.

Composition and Anti-Helicobacter pylori Properties of Essential Oils Obtained from Selected Mentha Cultivars.

Helicobacter pylori infections are highly common amongst the global population. Such infections have been shown to be the cause of gastric ulcers and stomach carcinoma and, unfortunately, most cases are asymptomatic. Standard treatment requires antibiotics such as metronidazole or azithromycin to which many strains are now resistant. Mentha species have been used as a natural treatment for gastrointestinal diseases throughout history and essential oils (EOs) derived from these plants show promising results as potential antimicrobial agents. In this study, EOs obtained from the leaves and flowers of five cultivars of Mentha × piperita and M. spicata were examined by GC-MS. The investigated mints are representatives of four chemotypes: the menthol chemotype (M. × piperita 'Multimentha' and M. × piperita 'Swiss'), the piperitenone oxide chemotype (M. × piperita 'Almira'), the linalool chemotype (M. × piperita 'Granada'), and the carvone chemotype (M. spicata 'Moroccan'). The chemical composition of EOs from mint flowers and leaves was comparable with the exception of the Swiss cultivar. Menthol was the most abundant component in the leaves while menthone was highest in flowers. The H. pylori ATCC 43504 reference strain and 10 other H. pylori clinical strains were examined for their sensitivity to the EOs in addition to their major monoterpenoid components (menthol, menthone, carvone, dihydrocarvone, linalool, 1,8-cineole, and limonene). All tested mint EOs showed inhibitory activity against both the reference H. pylori ATCC 43504 strain (MIC 15.6-31.3 mg/L) and clinical H. pylori strains (MIC50/90 31.3-250 mg/L/62.5-500 mg/L). Among the reference monoterpenes, menthol (MIC50/90 7.8/31.3 mg/L) and carvone (MIC50/90 31.3/62.5 mg/L) had the highest anti-H. pylori activity, which also correlated with a higher activity of EOs containing these compounds (M. × piperita 'Swiss' and M. spicata 'Moroccan'). A synergistic and additive interaction between the most active EOs/compounds and antibiotics possibly points to a new plant-based anti-H. pylori treatment.

Comparative developmental toxicities of zebrafish towards structurally diverse per- and polyfluoroalkyl substances.

Structurally diverse per- and polyfluoroalkyl substances (PFASs) are increasingly detected in ecosystems and humans. Therefore, the clarification of their ecological and health risks is urgently required. In the present study, the toxicity of a series of PFASs, including PFOS, PFBS, Nafion BP1, Nafion BP2, F53B, OBS, PFOA, PFUnDA, PFO5DoDA, HFPO-TA was investigated. Similarities and differences in the developmental toxicity potentials were revealed. Our results demonstrated that PFUnDA exhibited the highest toxicity with the lowest EC50 value of 4.36 mg/L (for morphological abnormality); this was followed by F53B (5.58 mg/L), PFOS (6.15 mg/L), and OBS (10.65 mg/L). Positive correlations with volatility/solubility and chemotypes related to specific biological activity, including the bioconcentration factor (LogBCF), and negative correlations with lipid solubility and carbon chain component-related chemotypes, including the number of carbon and fluorine atoms, provided a reasonable explanation in the view of molecular structures. Furthermore, comparative transcriptome analysis provided molecular evidence for the relationship between PFASs exposure and malformations. Common differentially expressed genes (DEGs) involved in spine curve development, pericardial edema, and cell/organism growth-related pathways presented common targets, leading to toxic effects. Therefore, the present results provide novel insights into the potential environmental risks of structurally diverse PFASs and contribute to the selection of safer PFAS replacements.

Bioprospecting of Five Ocimum sp. Cultivars from Croatia: New Potential for Dietary and Dermatological Application with Embryotoxicity Tests.

Ocimum basilicum L. is the most common Ocimum species, and it is used as an ornamental plant and in food condiments. This unique study examined the chemical composition and biological activities of six extracts from five basil cultivars, including their antimicrobial, antidiabetic, antilipidemic, neuroprotective, and anticollagenase activity. Moreover, their toxicological effects were studied using the zebrafish Danio rerio. Volatile components were determined using HS-SPME and GC-MS, while total polyphenols were detected using HPLC and the spectrophotometric Folin-Ciocalteu method. Spectrophotometric assays (DPPH, ABTS, ORAC, FRAP) were performed to determine antioxidant activity, collagenase inhibition, acetylcholinesterase inhibition, and pancreatic lipase inhibition. Antimicrobial activity was determined using the broth microdilution test. The study found that the biological activities of different basil cultivars varied depending on the proportion of active compounds, as determined by chemical analyses. All six basil extracts significantly inhibited α-amylase, while Purple basil extract most significantly inhibited the activity of collagenase, acetylcholinesterase, and pancreatic lipase. Purple basil and Dark Opal basil I extracts exhibited the highest antimicrobial activity, while the Dark Opal basil II extract had the most significant antioxidant potential. The findings in this study suggest that ethanolic basil extracts have the potential to be used as dietary drugs and implemented in antiaging products. This study is unique in its aims to compare the chemical composition and biological activities of basil cultivars from Croatia and to evaluate potential toxicological effects through embryotoxicity tests on zebrafish Danio rerio embryos, and it reports the first evidence of anticollagenase, antidiabetic, and antilipidemic activities for these cultivars.

Chemical Variability of the Essential Oils from Two Portuguese Apiaceae: Coriandrum sativum L. and Foeniculum vulgare Mill.

Coriandrum sativum L. and Foeniculum vulgare Mill. are two aromatic and medicinal Apiaceae species commonly grown in Portugal, whose essential oils (EOs) are used in the food, pharmaceutical, and cosmetics industries. The present study evaluated EOs isolated from the fruits and vegetative aerial parts (VAPs) of 11 samples of Coriandrum sativum L. and from the fruits of 19 samples of Foeniculum vulgare Mill. The plant material was grown in experimental fields, after collection from several regions of mainland Portugal. The EOs were isolated by hydrodistillation and analyzed by gas chromatography and gas chromatography-mass spectrometry. The coriander EOs analysis evidenced two main clusters, with the first containing the fruits' EOs dominated by linalool (60-73%), γ-terpinene, and α-pinene and the second with the VAPs' EOs, which showed 2-trans-decenal (37-63%) and n-decanal (13-30%) as the main compounds. The fennel EOs analysis revealed two well correlated clusters, the first dominated by estragole (34-76%) and fenchone (16-30%) and the other dominated by trans-anethole (37-56%) and fenchone (14-34%). The present data suggest coriander EOs' chemical descriptors as linalool for the fruits' EOs and 2-trans-decenal with n-decanal for the VAPs' EOs. For the fennel fruit EOs, the putative descriptors were trans-anethole and estragole, with variable fenchone content. The gathered data reinforce the relevance of clarifying variability of these species' EOs, particularly when considering aromatic and medicinal plants with such a wide range of applications.

Chemical Composition, Antioxidant Properties, and Antibacterial Activity of Essential Oils of Satureja macrostema (Moc. and Sessé ex Benth.) Briq.

Satureja macrostema is a plant that is located in various regions of Mexico and is used in a traditional way against illness. Essential oils (EOs) were obtained from leaves Satureja macrostema and the chemical composition was evaluated by gas chromatography-mass spectrometry (GC-MS). The antioxidant effect of the oil was assayed by 2,2-diphenyl-1-picrylhydrazyl (DPPH) and by Trolox Equivalent Antioxidant Capacity (TEAC). In vitro antibacterial activity against Escherichia coli and Staphylococcus aureus was determined using a broth microdilution assay and thin layer chromatography-direct bioautography (TLC-DB) to identify active antibacterial compounds. The EOs analysis showed 21 compounds, 99% terpenes, and 96% oxygenated monoterpenes, with trans-piperitone epoxide (46%), cis-piperitone epoxide (22%), and piperitenone oxide (11%) as more abundant compounds. Likewise, S. macrostema EOs showed an antioxidant activity of DPPH = 82%, with 50% free radical scavenging (IC50) = 7 mg/mL and TEAC = 0.005, an antibacterial effect against E. coli of 73% inhibition, and 81% over S. aureus at dose of 100 µL of undiluted crude oil. The TLC-DB assay showed that the most active compounds were derived from piperitone. The comparison with other studies on S. macrostema shows variability in the compounds and their abundances, which can be attributed to climatic factors and the maturity of plants with similar antioxidant and antibacterial activities.

Chemical and Olfactory Analysis of the Volatile Fraction of Ocimum gratissimum Concrete from Madagascar.

The chemical composition of the volatile fraction of Ocimum gratissimum concrete (romba) from Madagascar has been determined for the first time by GC/MS and GC-FID. A methyl cinnamate chemotype has been determined for this material, along with a set of compounds typical in essential oils and extracts from plants of the Ocimum genus. Variability was mostly observed on terpenes and terpenoids components. GC-O-MS was also used for a sensory evaluation of this material performed by a master perfumer. The chemical composition of this O. gratissimum extract was then compared with literature data to assess subtle differences between chemotypes of the same species and other species of the same genus within natural variability. A mapping illustrates the occurrence of the cinnamate chemotype in Eastern Africa, India and now Madagascar, while other origins generally present eugenol, thymol, camphor, or linalool chemotypes.

Variations in Essential Oils from the Leaves of Cinnamomum bodinieri in China.

Cinnamomum plants are rich in natural essential oils, which are widely used as materials in the fragrance, insecticidal, antibacterial agent, pharmaceutical, and food industries; however, few studies have investigated the essential oil components of Cinnamomum bodinieri. Therefore, this study investigated the diversity of essential oils from the leaves of 885 individual C. bodinieri plants across 32 populations in five provinces. Essential oils were extracted by hydrodistillation, and then qualitative and quantitative analyses of the compounds were performed by GC-MS and GC-FID. A total of 87 chemical constituents were identified in the essential oils, including 33 monoterpenes, 48 sesquiterpenes, and six other compounds. The average oil yield was 0.75%, and individual oil yields ranged from 0.01% to 4.28%. A total of 16 chemotypes were classified according to variations in the essential oil chemical constituents of C. bodinieri, among which the camphor-type, citral-type, and eucalyptol-type were dominant. Moreover, the borneol-type, cymol-type, elemol-type, methylisoeugenol-type, and selina-6-en-4-ol-type were reported in C. bodinieri for the first time. The yield and principal components of the essential oils were mainly affected by altitude, temperature, and sunshine duration, among which altitude had the most significant effect; thus, low-altitude areas are more suitable for the synthesis and accumulation of essential oils. Based on the different characteristics of the essential oils in the leaves of C. bodinieri, several excellent populations and individuals were identified in this study. Moreover, the findings provide a foundation for breeding superior varieties and studying essential oil biosynthesis mechanisms in the future.

Creation of New Oregano Genotypes with Different Terpene Chemotypes via Inter- and Intraspecific Hybridization.

Oregano is a medicinal and aromatic plant of value in the pharmaceutical, food, feed additive, and cosmetic industries. Oregano breeding is still in its infancy compared with traditional crops. In this study, we evaluated the phenotypes of 12 oregano genotypes and generated F1 progenies by hybridization. The density of leaf glandular secretory trichomes and the essential oil yield in the 12 oregano genotypes varied from 97-1017 per cm2 and 0.17-1.67%, respectively. These genotypes were divided into four terpene chemotypes: carvacrol-, thymol-, germacrene D/ß-caryophyllene-, and linalool/ß-ocimene-type. Based on phenotypic data and considering terpene chemotypes as the main breeding goal, six oregano hybrid combinations were performed. Simple sequence repeat (SSR) markers were developed based on unpublished whole-genome sequencing data of Origanum vulgare, and 64 codominant SSR primers were screened on the parents of the six oregano combinations. These codominant primers were used to determine the authenticity of 40 F1 lines, and 37 true hybrids were identified. These 37 F1 lines were divided into six terpene chemotypes: sabinene-, ß-ocimene-, γ-terpinene-, thymol-, carvacrol-, and p-cymene-type, four of which (sabinene-, ß-ocimene-, γ-terpinene-, and p-cymene-type) were novel (i.e., different from the chemotypes of parents). The terpene contents of 18 of the 37 F1 lines were higher than those of their parents. The above results lay a strong foundation for the creating of new germplasm resources, constructing of genetic linkage map, and mapping quantitative trait loci (QTLs) of key horticultural traits, and provide insights into the mechanism of terpenoid biosynthesis in oregano.

Chemical Diversity of Artemisia rutifolia Essential Oil, Antimicrobial and Antiradical Activity.

This paper presents the results of the study of the composition of the essential oil (EO) of Artemisia rutifolia by the GC/MS method as well as its antimicrobial and antiradical activities. According to the PCA-analysis, these EOs can be conditionally divided into "Tajik" and "Buryat-Mongol" chemotypes. The first chemotype is characterized by the prevalence of α- and ß-thujone, and the second chemotype by the prevalence of 4-phenyl-2-butanone, camphor. The greatest antimicrobial activity of A. rutifolia EO was observed against Gram-positive bacteria and fungi. The EO showed high antiradical activity with an IC50 value of 17.55 µL/mL. The presented first data on the composition and activity of the EO of A. rutifolia of the Russian flora indicate the prospects of the species as a raw material for the pharmaceutical and cosmetic industry.

Phytochemical Comparison of Medicinal Cannabis Extracts and Study of Their CYP-Mediated Interactions with Coumarinic Oral Anticoagulants.

Introduction: Treatment with cannabis extracts for a variety of diseases has gained popularity. However, differences in herb-drug interaction potential of extracts from different plant sources are poorly understood. In this study, we provide a characterization of cannabis extracts prepared from four cannabis chemotypes and an in vitro assessment of their Cytochrome P450 (CYP)-mediated herb-drug interaction profiles. Methods: Plant extracts were either commercially obtained or prepared using ethanol as solvent, followed by overnight decarboxylation in a reflux condenser system. The extracts were characterized for their cannabinoid content using NMR and HPLC-PDA-ELSD-ESIMS. CYP inhibition studies with the cannabis extracts and pure cannabinoids (tetrahydrocannabinol [THC] and cannabidiol [CBD]) were performed using pooled, mixed gender human liver microsomes. Tolbutamide and testosterone were used as specific substrates to assess the inhibitory potential of the extracts on CYP2C9 and CYP3A4, and the coumarinic oral anticoagulants warfarin, phenprocoumon, and acenocoumarol were studied as model compounds since in vivo herb-drug interactions have previously been reported for this compound class. Results: In accordance with the plant chemotypes, two extracts were rich in THC and CBD (at different proportions); one extract contained mostly CBD and the other mostly cannabigerol (CBG). Residual amounts of the corresponding acids were found in all extracts. The extracts with a single major cannabinoid (CBD or CBG) inhibited CYP2C9- and CYP3A4-mediated metabolism stronger than the extracts containing both major cannabinoids (THC and CBD). The inhibition of CYP3A4 and CYP2C9 by the extract containing mostly CBD was comparable to their inhibition by pure CBD. In contrast, the inhibitory potency of extracts containing both THC and CBD did not correspond to the combined inhibitory potency of pure THC and CBD. Although being structural analogs, the three coumarin derivatives displayed major differences in their herb-drug interaction profiles with the cannabis extracts and the pure cannabinoids. Conclusion: Despite the fact that cannabinoids are the major components in ethanolic, decarboxylated cannabis extracts, it is difficult to foresee their herb-drug interaction profiles. Our in vitro data and the literature-based evidence on in vivo interactions indicate that cannabis extracts should be used cautiously when co-administered with drugs exhibiting a narrow therapeutic window, such as coumarinic anticoagulants, regardless of the cannabis chemotype used for extract preparation.

Essential Oils from Origanum vulgare subsp. virens (Hoffmanns. & Link) Ietsw. Grown in Portugal: Chemical Diversity and Relevance of Chemical Descriptors.

Origanum vulgare L. is a well-known aromatic and medicinal plant, whose essential oil (EO) has recognised flavouring and medicinal properties. In this study, Origanum vulgare subsp. virens (Hoffmanns. & Link) Ietsw. EOs, isolated from accessions grown in experimental fields, were evaluated. The plant material was grown from rooted cuttings or nutlets (fruits), originally collected in 20 regions in mainland Portugal and harvesting for EO isolation was performed in two years. EOs were isolated by hydrodistillation and analysed by gas chromatography and gas chromatography-mass spectrometry, for EO quantification and identification, respectively. EO yields ranged from <0.05-3.3% for rooted cuttings, with oregano samples obtained in Portalegre and Alandroal, respectively. Ninety-one compounds were identified, mainly grouped in oxygen-containing monoterpenes and monoterpene hydrocarbons. EO agglomerative cluster analysis evidenced two main clusters, with the first subdivided into four subclusters. From the obtained data, the putative O. vulgare subsp. virens chemotypes are carvacrol, thymol and linalool, with γ-terpinene, p-cymene, cis- and trans-ß-ocimene also contributing as these EOs chemical descriptors. The comparison between the present data and a survey of the existing literature on Portuguese O. vulgare reinforces the major variability of this species' EOs and emphasises the importance of avoiding wild collections to obtain a defined chemical type of crop production of market relevance.