An Overview of the Evidence and Mechanisms of Herb–Drug Interactions

Despite the lack of sufficient information on the safety of herbal products, their use as alternative and/or complementary medicine is globally popular. There is also an increasing interest in medicinal herbs as precursor for pharmacological actives. Of serious concern is the concurrent consumption of herbal products and conventional drugs. Herb–drug interaction (HDI) is the single most important clinical consequence of this practice. Using a structured assessment procedure, the evidence of HDI presents with varying degree of clinical significance. While the potential for HDI for a number of herbal products is inferred from non-human studies, certain HDIs are well established through human studies and documented case reports. Various mechanisms of pharmacokinetic HDI have been identified and include the alteration in the gastrointestinal functions with consequent effects on drug absorption; induction and inhibition of metabolic enzymes and transport proteins; and alteration of renal excretion of drugs and their metabolites. Due to the intrinsic pharmacologic properties of phytochemicals, pharmacodynamic HDIs are also known to occur. The effects could be synergistic, additive, and/or antagonistic. Poor reporting on the part of patients and the inability to promptly identify HDI by health providers are identified as major factors limiting the extensive compilation of clinically relevant HDIs. A general overview and the significance of pharmacokinetic and pharmacodynamic HDI are provided, detailing basic mechanism, and nature of evidence available. An increased level of awareness of HDI is necessary among health professionals and drug discovery scientists. With the increasing number of plant-sourced pharmacological actives, the potential for HDI should always be assessed in the non-clinical safety assessment phase of drug development process. More clinically relevant research is also required in this area as current information on HDI is insufficient for clinical applications

HPLC-MS identification and expression of Candida drug-resistance proteins from African HIV-infected patients

The objective of this study was to elucidate the proteomic mechanisms of drug resistance in HIV-infected African patients. Cell membrane fractions from forty oral Candida isolates isolated from African HIV-positive patients were analysed using HPLC-MS with the aim of identifying proteins associated with their pathogenicity and drug resistance. Heat shock proteins that mediate the fungicidal activity of salivary peptides were found in all tested Candida fractions, with pH-responsive proteins associated with increased pathogenicity only being present in the three most commonly isolated species. ABC multidrug transporter efflux pumps and estrogen binding proteins were only found in C. albicans fractions, while ergosterol biosynthesis proteins were identified in four species. The combination of various adherence, invasion, upregulation and efflux pump mechanisms appear to be instrumental for the Candida host colonization and drug resistance emergence in HIV-infected individuals.This material is based upon work partially supported financially by the National Research Foundation of South Africa [Grant number TTK2008052700013]

The GenTree Platform: growth traits and tree-level environmental data in 12 European forest tree species

Background: Progress in the field of evolutionary forest ecology has been hampered by the huge challenge of phenotyping trees across their ranges in their natural environments, and the limitation in high-resolution environmental information. Findings: The GenTree Platform contains phenotypic and environmental data from 4,959 trees from 12 ecologically and economically important European forest tree species: Abies alba Mill. (silver fir), Betula pendula Roth. (silver birch), Fagus sylvatica L. (European beech), Picea abies (L.) H. Karst (Norway spruce), Pinus cembra L. (Swiss stone pine), Pinus halepensis Mill. (Aleppo pine), Pinus nigra Arnold (European black pine), Pinus pinaster Aiton (maritime pine), Pinus sylvestris L. (Scots pine), Populus nigra L. (European black poplar), Taxus baccata L. (English yew), and Quercus petraea (Matt.) Liebl. (sessile oak). Phenotypic (height, diameter at breast height, crown size, bark thickness, biomass, straightness, forking, branch angle, fructification), regeneration, environmental in situ measurements (soil depth, vegetation cover, competition indices), and environmental modeling data extracted by using bilinear interpolation accounting for surrounding conditions of each tree (precipitation, temperature, insolation, drought indices) were obtained from trees in 194 sites covering the species’ geographic ranges and reflecting local environmental gradients. Conclusion: The GenTree Platform is a new resource for investigating ecological and evolutionary processes in forest trees. The coherent phenotyping and environmental characterization across 12 species in their European ranges allow for a wide range of analyses from forest ecologists, conservationists, and macro-ecologists. Also, the data here presented can be linked to the GenTree Dendroecological collection, the GenTree Leaf Trait collection, and the GenTree Genomic collection presented elsewhere, which together build the largest evolutionary forest ecology data collection available

Between but not within species variation in the distribution of fitness effects

New mutations provide the raw material for evolution and adaptation. The distribution of fitness effects (DFE) describes the spectrum of effects of new mutations that can occur along a genome, and is therefore of vital interest in evolutionary biology. Recent work has uncovered striking similarities in the DFE between closely related species, prompting us to ask whether there is variation in the DFE among populations of the same species, or among species with different degrees of divergence, i.e., whether there is variation in the DFE at different levels of evolution. Using exome capture data from six tree species sampled across Europe we characterised the DFE for multiple species, and for each species, multiple populations, and investigated the factors potentially influencing the DFE, such as demography, population divergence and genetic background. We find statistical support for there being variation in the DFE at the species level, even among relatively closely related species. However, we find very little difference at the population level, suggesting that differences in the DFE are primarily driven by deep features of species biology, and that evolutionarily recent events, such as demographic changes and local adaptation, have little impact

In vitro assessment of the interaction potential of ocimum basilicum (L.) extracts on CYP2B6, 3A4, and rifampicin metabolism

CITATION: Kumar, S., Bouic, P. J. & Rosenkranz, B. 2020. In vitro assessment of the interaction potential of ocimum basilicum (L.) extracts on CYP2B6, 3A4, and rifampicin metabolism. Frontiers in Pharmacology, 11:517, doi:10.3389/fphar.2020.00517.The original publication is available at https://www.frontiersin.orgPublication of this article was funded by the Stellenbosch University Open Access Fund.ENGLISH ABSTRACT: Ocimum basilicum L. or basilicum is a common culinary herb, used as a traditional medicine for various medical conditions including HIV/AIDS and tuberculosis, in Africa. The objective of this study was to evaluate the effect of methanol, ethanol, aqueous and ethyl acetate extracts of the dried leaves and inflorescence of O. basilicum, on the activity of cytochrome P450 enzymes (CYPs) CYP2B6 and 3A4, as well as esterase-mediated metabolism of rifampicin to 25-O-desacetyl rifampicin (25ODESRIF). Human liver microsomes (HLM) were used to evaluate inhibition and CYP2B6/3A4 mRNA expression HepG2 assays were used to measure induction. Furthermore, the phytoconstituents likely involved in causing the observed effect were analyzed using biochemical tests and LC-MS. The aqueous and methanolic extracts showed reversible and time-dependent inhibition (TDI) of CYP2B6 with TDI-IC50s 33.35 μg/ml (IC50 shift-fold >1.5) and 4.93 μg/ml (IC50 shift-fold >7) respectively, while the methanolic and ethanolic extracts inhibited 25ODESRIF formation (IC50s 31 μg/ml, 8.94 μg/ml). In HepG2 assays, the methanolic and ethanolic extracts moderately induced CYP2B6, 3A4 mRNA with 38%-, 28%-fold shift, and 22%-, 44%-fold shift respectively. LC-MS full scans identified phenols rosmarinic acid [m/z 359 (M-H)-, approximately 2298 mg/L in aqueous extract] and caftaric acid along with flavones salvigenin [m/z 329 (M+H)+, approximately 1855 mg/L in ethanolic extract], eupatorin [m/z 345 (M+H)+, 668.772 mg/L in ethanolic extract], rutin [m/z 609 (M-H)-] and isoquercetin [m/z 463 (M-H)-] and other compounds—linalool [m/z 153 (M-H)-], hydroxyjasmonic acid [m/z 225 (M-H)-], eucommiol [m/z 187 (M-H)-] and trihydroxy octadecenoic acid [m/z 329 (M-H)-, 530 mg/L in ethanolic extract]. The putative gastrointestinal tract (GIT) concentration for all extracts was calculated as 2,400 μg/ml and hepatic circulation concentrations were estimated at 805.68 μg/ml for the aqueous extract, and 226.56 μg/ml for methanolic extract. Based on the putative GIT concentration, estimated hepatic circulation concentration [I] and inhibition constant Ki, the predicted percentile of inhibition in vivo was highest for the aqueous extract on CYP2B6 (96.7%). The observations indicated that O. basilicum extracts may have the potential to cause clinically relevant herb-drug interactions (HDI) with CYP2B6 and rifampicin metabolism in vivo, if sufficient hepatic concentrations are reached in humans.https://www.frontiersin.org/articles/10.3389/fphar.2020.00517/fullPublisher's versio

A Validated stable HPLC method for the simultaneous determination of rifampicin and 25-O-desacetyl rifampicin – evaluation of in vitro metabolism

CITATION: Kumar, S. Bouic, P. J. & Rosenkranz, B. AValidated stable HPLC method for the simultaneous determination of rifampicin and 25-O-desacetyl rifampicin – evaluation of in vitro metabolism. Acta Chromatographica: 1-7, doi:10.1556/1326.2018.00361.The original publication is available at https://akademiai.comPublication of this article was funded by the Stellenbosch University Open Access Fund.A simple, efficient, and stable high-performance liquid chromatography (HPLC) separation method for a combination of rifampicin (RIF), its major metabolite 25-O-desacetyl rifampicin (25ODESRIF), and neostigmine (NEO) was developed and validated. The drugs individually, and in combination, were analyzed using a Waters Alliance 2695 HPLC coupled with 2996 photodiode array detector (PDA). Successful separation of combined drugs was achieved by gradient elution on a reverse-phase C-18 Phenomenex Luna column, using a mobile phase consisting of water and methanol at detection wavelength of 254 nm. The HPLC retention times were consistent at ±7.70 min, ±8.25 min, and ±10.70 min for RIF, 25ODESRIF, and NEO, respectively. The regression data for the calibration plots exhibited linear relationship (R 2 = 0.995) in the range of 0–200 μM for both RIF and 25ODESRIF, and the lower limit of detection (LLOD) and lower limit of quantification (LLOQ) were calculated at 5.86 μM and 17.75 μM for RIF and 7.78 μM and 23.57 μM for 25ODESRIF, respectively. The method was evaluated using in vitro human liver microsomes (HLMs) assays, and linearity was established for the 15, 30, 45, and 60 min incubations (R2 = 0.99). The formation of 25ODESRIF was characterized by hyperbolic kinetics (Km 48.23 μM, Vmax 1.233 pmol/min/mg protein, and CLint 0.026 μl/min/mg protein). The method was applied in HLM assays to understand the herb–drug interaction (HDI) potential of Althaea officinalis, a popular African herb consumed by tuberculosis (TB) patients, with RIF. None of the extracts of A. officinalis inhibited the esterase-mediated metabolism pathway of RIF, compared to the positive control nelfinavir (IC50 = 9.59 μM). The method provides a tool for quantifying RIF and 25ODESRIF in in vitro drug metabolism assays as well as investigating herb– and drug–drug interactions (DDIs).https://akademiai.com/doi/abs/10.1556/1326.2018.00361Publisher's versio

The potential of Hypoxis hemerocallidea for herb-drug interaction

CITATION: Fasinu, P. S. et al. 2013. The potential of Hypoxis hemerocallidea for herb-drug interaction. Pharmaceutical Biology, 51(12):1499-1507, doi:10.3109/13880209.2013.796393.The original publication is available at https://www.tandfonline.comContext: Aqueous decoction of Hypoxis hemerocallidea Fisch. & C.A. Mey. (Hypoxidaceae) (Hypoxis) is widely consumed in Southern Africa by people living with HIV/AIDS, some of whom are on ARV and other medications. Objective: The aim of this study was to investigate the potential of the crude aqueous extracts of Hypoxis to inhibit major forms of CYP450 and transport proteins. Materials and methods: Corms of Hypoxis were water-extracted and incubated (in graded concentrations: 1–100 mg/mL) with human liver microsomes (20 min) to monitor the effects on phenacetin O-deethylation, coumarin 7-hydroxylation, bupropion hydroxylation, paclitaxel 6a-hydroxylation, diclofenac 40 -hydroxylation, S-mephenytoin 40 -hydroxylation, bufuralol 10 -hydroxylation, chlorzoxazone 6-hydroxylation, midazolam 10 -hydroxylation and testosterone 6b-hydroxylation as markers for the metabolic activities of CYP1A2, 2A6, 2B6, 2C8, 2C9, 2C19, 2D6, 2E1 and 3A4/5, respectively. The generation of metabolites were monitored and quantified with the aid of LC-MS/MS. The potential of the extracts to inhibit human ATPbinding cassette transporter activity was assessed using recombinant MDCKII and LLC-PK1 cells over-expressing human breast cancer resistant protein and human P-glycoprotein , respectively (with Ko143 and cyclosporin A as positive controls). Similar assessment was performed with human organic anion transporting polypeptide (OATP1B1 and OATP1B3) using recombinant HEK293 cells over-expressing OATP1B1 and OATP1B3, respectively (with rifamycin and 10 mM atorvastatin as positive controls). Results: Extracts of Hypoxis inhibited the production of the metabolites of the substrates of the following enzymes (as compared to controls) with the indicated IC50 values (mg/mL): CYP1A2 (120.6), CYP2A6 (210.8), CYP2B6 (98.5), CYP2C8 (195.2), CYP2C9 (156) and CYP3A4/5 (185.4). The inhibition of the uptake activity of OATP1B1 and OATP1B3 were also observed with IC50 values of 93.4 and 244.8 mg/mL, respectively. Discussion: Extract concentrations higher than the estimated IC50 values are achievable in the gastrointestinal tract when traditional doses of Hypoxis are considered. This may have profound effects on presystemic metabolism of the drug substrates. If absorbed, systemic inhibition of metabolic enzymes/transporters by Hypoxis may be expected. Conclusion: The result suggests that there is the potential for HDI between Hypoxis and the substrates of the affected enzymes/transporters, if sufficient in vivo concentration of Hypoxis extracts is attained.https://www.tandfonline.com/doi/full/10.3109/13880209.2013.79639

Major histocompatibility complex class II invariant chain expression in non-antigen-presenting cells

In contrast to the generally accepted belief, the major histocompatibility complex (MHC) class II invariant chain (Ii) is commonly expressed intracellularly in cells that do not present exogenous antigens. Such cells include resting peripheral blood T cells and natural killer (NK) cells. In T cells, the Ii is associated with a 77 000 molecular-weight molecule (p77) that has yet to be identified. This molecule is co-precipitated with the anti-Ii monoclonal antibody (mAb) VCD-1, but not with mAb BU-45. This suggests that in the p77–Ii complex, the extracellular epitope of Ii recognized by BU-45 is hidden, whereas the Ii epitope for VCD-1 remains exposed. In antigen-presenting cells (APCs), p77 association with the Ii was minimal, if detectable. The p77–Ii association in non-professional APCs suggests that the Ii may have another, more general, function other than the one accepted in antigen presentation