¹H-NMR Metabolic Profiling, Antioxidant Activity, and Docking Study of Common Medicinal Plant-Derived Honey.

The purpose of this investigation was to determine ¹H-NMR profiling and antioxidant activity of the most common types of honey, namely, citrus honey (HC1) (Morcott tangerine L. and Jaffa orange L.), marjoram honey (HM1) (Origanum majorana L.), and clover honey (HT1) (Trifolium alexandrinum L.), compared to their secondary metabolites (HC2, HM2, HT2, respectively). By using a ¹H-NMR-based metabolomic technique, PCA, and PLS-DA multivariate analysis, we found that HC2, HM2, HC1, and HM1 were clustered together. However, HT1 and HT2 were quite far from these and each other. This indicated that HC1, HM1, HC2, and HM2 have similar chemical compositions, while HT1 and HT2 were unique in their chemical profiles. Antioxidation potentials were determined colorimetrically for scavenging activities against DPPH, ABTS, ORAC, 5-LOX, and metal chelating activity in all honey extract samples and their secondary metabolites. Our results revealed that HC2 and HM2 possessed more antioxidant activities than HT2 in vitro. HC2 demonstrated the highest antioxidant effect in all assays, followed by HM2 (DPPH assay: IC50 2.91, 10.7 µg/mL; ABTS assay: 431.2, 210.24 at 50 ug/mL Trolox equivalent; ORAC assay: 259.5, 234.8 at 50 ug/mL Trolox equivalent; 5-LOX screening assay/IC50: 2.293, 6.136 ug/mL; and metal chelating activity at 50 ug/mL: 73.34526%, 63.75881% inhibition). We suggest that the presence of some secondary metabolites in HC and HM, such as hesperetin, linalool, and caffeic acid, increased the antioxidant activity in citrus and marjoram compared to clover honey.

LC/MS Profiling and Gold Nanoparticle Formulation of Major Metabolites from Origanum majorana as Antibacterial and Antioxidant Potentialities.

Origanum majoranum L. is a Lamiaceae medicinal plant with culinary and ethnomedical applications. Its biological and phytochemical profiles have been extensively researched. Accordingly, this study aimed to investigate the chemical composition and the antibacterial and antioxidant properties of O. majoranum high features, as well as to search for techniques for activity optimization. A metabolomics study of the crude extract of O. majoranum using liquid chromatography-high-resolution electrospray ionization mass spectrometry (LC ± HR ± ESI ± MS) was conducted. Five fractions (petroleum ether, dichloromethane, ethyl acetate, n-butanol, and aqueous) were derived from the total extract of the aerial parts. Different chromatographic methods and NMR analysis were utilized to purify and identify the isolated phenolics (high features). Moreover, the antimicrobial, antibiofilm, and antioxidant activity of phenolics were performed. Results showed that metabolomic profiling of the crude extract of O. majoranum aerial parts revealed the presence of a variety of phytochemicals, predominantly phenolics, resulting in the isolation and identification of seven high-feature compounds comprising two phenolic acids, rosmarinic and caffeic acids, one phenolic diterpene, 7-methoxyepirosmanol, in addition to four flavonoids, quercetin, hesperitin, hesperidin, and luteolin. On the other hand, 7-methoxyepirosmanol (OM1) displayed the most antimicrobial and antioxidant potential. Such a phenolic principal activity improvement seems to be established after loading on gold nanoparticles.

In Silico-Based Discovery of Natural Anthraquinones with Potential against Multidrug-Resistant E. coli.

E. coli is a Gram-negative bacterium that causes different human infections. Additionally, it resists common antibiotics due to its outer protective membrane. Natural products have been proven to be efficient antibiotics. However, plant natural products are far less explored in this regard. Accordingly, over 16,000 structures covering almost all African medicinal plants in AfroDb in a structural-based virtual screening were used to find efficient anti-E. coli candidates. These drug-like structures were docked into the active sites of two important molecular targets (i.e., E. coli's Ddl-B and Gyr-B). The top-scoring hits (i.e., got docking scores < -10 kcal/mol) produced in the initial virtual screening (0.15% of the database structures for Ddl-B and 0.17% of the database structures for Gyr-B in the database) were further refined using molecular dynamic simulation-based binding free energy (ΔG) calculation. Anthraquinones were found to prevail among the retrieved hits. Accordingly, readily available anthraquinone derivatives (10 hits) were selected, prepared, and tested in vitro against Ddl-B, Gyr-B, multidrug-resistant (MDR) E. coli, MRSA, and VRSA. A number of the tested derivatives demonstrated strong micromolar enzyme inhibition and antibacterial activity against E. coli, MRSA, and VRSA, with MIC values ranging from 2 to 64 µg/mL. Moreover, both E. coli's Ddl-B and Gyr-B were inhibited by emodin and chrysophanol with IC50 values comparable to the reference inhibitors (IC50 = 216 ± 5.6, 236 ± 8.9 and 0.81 ± 0.3, 1.5 ± 0.5 µM for Ddl-B and Gyr-B, respectively). All of the active antibacterial anthraquinone hits showed low to moderate cellular cytotoxicity (CC50 > 50 µM) against human normal fibroblasts (WI-38). Furthermore, molecular dynamic simulation (MDS) experiments were carried out to reveal the binding modes of these inhibitors inside the active site of each enzyme. The findings presented in this study are regarded as a significant step toward developing novel antibacterial agents against MDR strains.

A metabolomic approach to target antimalarial metabolites in the Artemisia annua fungal endophytes.

Fungal endophytes are a major source of anti-infective agents and other medically relevant compounds. However, their classical blinded-chemical investigation is a challenging process due to their highly complex chemical makeup. Thus, utilizing cheminformatics tools such as metabolomics and computer-aided modelling is of great help deal with such complexity and select the most probable bioactive candidates. In the present study, we have explored the fungal endophytes associated with the well-known antimalarial medicinal plant Artemisia annua for their production of further antimalarial agents. Based on the preliminary antimalarial screening of these endophytes and using LC-HRMS-based metabolomics and multivariate analyses, we suggested different potentially active metabolites (compounds 1-8). Further in silico investigation using the neural-network-based prediction software PASS led to the selection of a group of quinone derivatives (compounds 1-5) as the most possible active hits. Subsequent in vitro validation revealed emodin (1) and physcion (2) to be potent antimalarial candidates with IC50 values of 0.9 and 1.9 µM, respectively. Our approach in the present investigation therefore can be applied as a preliminary evaluation step in the natural products drug discovery, which in turn can facilitate the isolation of selected metabolites notably the biologically active ones.

Ecological HPLC method for analyzing an antidiabetic drug in real rat plasma samples and studying the effects of concurrently administered fenugreek extract on its pharmacokinetics.

Currently, the total number of diabetic people worldwide is constantly increasing. Metformin (MET) is known to be a first-line antidiabetic drug with varied, wide-reaching applications. Concurrent administration of phytomedicines such as fenugreek extract with synthetic drugs is very common. It is reported that concomitant administration of fenugreek extract with metformin maintains lower blood glucose levels than metformin alone. In this work, an ecofriendly RP-HPLC method was established to study and compare the pharmacokinetics of metformin with and without the contemporary administration of fenugreek extract using rat as an animal model. In the developed method, a solvent mixture of 0.5 mM KH2PO4 solution : methanol (65 : 35, v/v) was used as a mobile phase and guaiphenesin was used as an internal standard. The plasma concentration-time curve was plotted, and non-compartmental pharmacokinetic analysis was performed using PKSolver. The results of the pharmacokinetic study showed that concurrent administration of fenugreek significantly increased the bioavailability of metformin and doubled the time required to reach the peak plasma concentration (T max). Moreover, the volume of drug distribution decreased by about 70%, while its rate of clearance decreased by about 55.96%. Accordingly, the administration of fenugreek in combination with metformin significantly affected the pharmacokinetics of metformin, and this combination will be very useful in controlling blood glucose levels in diabetic patients. The greenness of the method was assessed using the Analytical Eco-Scale, Analytical Method Volume Intensity (AMVI), and National Environmental Method Index (NEMI), and all results affirmed that the method can be considered to be ecological.

Premna odorata extract as a protective agent on neurotoxic effect of aluminum: neurochemical, molecular, and histopathological alterations.

Premna odorata Blanco (Lamiaceae) is an ethnomedicinal plant, where some reports claimed their anti-inflammatory, cytotoxic, and antituberculosis effects, without investigating its role on the brain. Therefore, forty mature male rats were equally divided into 4 groups; the 1st was kept as control. Rats in groups 2 and 4 were orally given P. odorata extract daily at a dose of 500 mg/kg B.W., while those in groups 3 and 4 were daily administrated aluminum chloride "AlCl3" (70 mg/kg B.W.). The treatments extended for 30 successive days. At the end of the experimental period, brain samples were collected for biochemical assay of glutathione reductase (GSH), catalase, malondialdehyde (MDA), and acetylcholinesterase activity (AChE). Besides, monoamines (norepinephrine, dopamine, serotonin), amino acids (glutamine, serine, arginine, taurine and gamma-aminobutyric acid (GABA)), neurotransmitters, DNA damage, cyclooxygenase-2 (COX-2), and tumor necrosis factor (TNF)-α genes were estimated. Moreover, brain samples were obtained for histopathological investigation. Aluminum toxicity resulted in a decline of GSH concentration, elevation of MDA, and AChE activity. Except for GABA which exhibited a significant decrease, there was a marked increase in the measured amino acid and monoamine neurotransmitters. Also, an increase in mRNA expressions of TNF-α and COX-2 was detected. It was noticed that Premna odorata extract reduced the oxidative stress and counteracted the augmentations in AChE caused by AlCl3. Marked improvements in most measured neurotransmitters with downregulation of pro-inflammatory gene expression were recorded in P. odorata + AlCl3 group. Premna odorata restores the altered histopathological feature induced by AlCl3. In conclusion, the present findings clarify that P. odorata extract could be important in improving and treatment of neurodegenerative disorders as it was able to reduce oxidative stress, DNA damage, biochemical alterations, and histopathological changes in rats exposed to AlCl3 toxicity.

Testicular Caspase-3 and ß-Catenin Regulators Predicted via Comparative Metabolomics and Docking Studies.

Many routes have been explored to search for effective, safe, and affordable alternatives to hazardous female contraceptives. Herbal extracts and their secondary metabolites are some of the interesting research areas to address this growing issue. This study aims to investigate the effects of ten different plant extracts on testicular spermatogenesis. The correlation between the chemical profile of these extracts and their in vivo effect on male reproductive system was evaluated using various techniques. Approximately 10% of LD50 of hydro-methanolic extracts were orally administrated to rats for 60 days. Semen parameters, sexual organ weights, and serum levels of male sex hormones in addition to testes histopathology, were evaluated. Moreover, metabolomic analysis using (LC-HRESIMS), multivariate analysis (PCA), immunohistochemistry (caspase-3 and ß-catenin), and a docking study were performed. Results indicated that three plant extracts significantly decreased epididymal sperm density and motility. Moreover, their effects on testicular cells were also assured by histopathological evaluations. Metabolomic profiling of the bioactive plant extracts showed the presence of diverse phytochemicals, mostly oleanane saponins, phenolic diterpenes, and lupane triterpenes. A docking study on caspase-3 enzyme showed that oleanane saponins possessed the highest binding affinity. An immunohistochemistry assay on ß-catenin and caspase-3 indicated that Albizzia lebbeck was the most active extract for decreasing immunoexpression of ß-catenin, while Rosmarinus officinalis showed the highest activity for increasing immunoexpression of caspase-3. The spermatogenesis decreasing the activity of A. lebbeck, Anagallis arvensis, and R. officinalis can be mediated via up-regulation of caspase-3 and down-regulation of ß-catenin existing in testis cells.

Pharmacological and natural products diversity of the brown algae genus Sargassum.

Sargassum (F. Sargassaceae) is an important seaweed excessively distributed in tropical and subtropical regions. Different species of Sargassum have folk applications in human nutrition and are considered a rich source of vitamins, carotenoids, proteins, and minerals. Many bioactive compounds chemically classified as terpenoids, sterols, sulfated polysaccharides, polyphenols, sargaquinoic acids, sargachromenol, and pheophytin were isolated from different Sargassum species. These isolated compounds and/or extracts exhibit diverse biological activities, including analgesic, anti-inflammatory, antioxidant, neuroprotective, anti-microbial, anti-tumor, fibrinolytic, immune-modulatory, anti-coagulant, hepatoprotective, and anti-viral activities. This review covers the literature from 1974 to 2020 on the genus Sargassum, and reveal the active components together with their biological activities according to their structure to create a base for additional studies on the clinical applications of Sargassum.

Balanites aegyptiaca ameliorates insulin secretion and decreases pancreatic apoptosis in diabetic rats: Role of SAPK/JNK pathway.

SAPK-JNK pathway performs a significant role in the pathogenesis of type 2 diabetes. Balanites aegyptiaca (BA) is used as an anti-diabetic agent in folk medicine however its hypoglycemic mechanism is not fully elucidated. The current study aimed to evaluate the effect of crude extract, butanol, and dichloromethane fractions from BA on the stress-activated protein kinase/c-Jun N-terminal kinase (SAPK-JNK) pathway in experimental diabetic rats. Six groups of male Wistar rats were included: normal control, diabetic, diabetic rats treated with crude, butanol or dichloromethane fraction from BA (50 mg/kg BW) and diabetic rats treated with gliclazide as a reference drug for one month. Our results suggested a protective role of treatment of diabetic rats with BA against oxidative stress-induced SAPK-JNK pathway. Moreover, BA treatment produced a reduction in plasma glucose, HbA1c, lactic acid, lipid profile, malondialdehyde levels and produced an increase in insulin, reduced glutathione levels, catalase and superoxide dismutase activities compared with untreated diabetic rats. Moreover, it decreased apoptosis signal-regulating kinase 1, c-Jun N-terminal kinase 1, protein 53 and increased insulin receptor substrate 1 in rat pancreas while it increased glucose transporter 4 in rat muscle. Analysis of BA extracts by LC-HRMS revealed the presence of different saponins with reported hypoglycemic effect. In conclusion, BA exerted hypoglycemic, hypolipidemic, insulinotropic and antioxidant effects. Additionally, it reduced apoptosis in pancreatic ß-cells and increased glucose uptake in muscle. These results suggest that the hypoglycemic effect of BA is due to the inhibition of the SAPK-JNK pathway.

Acylated Iridoids and Rhamnopyranoses from Premna odorata (Lamiaceae) as Novel Mesenchymal-Epithelial Transition Factor Receptor Inhibitors for the Control of Breast Cancer.

Phytochemical investigation of Premna odorata Blanco, Lamiaceae, leaves afforded three new acylated iridoid glycosides 1-3 and two new acylated rhamnopyranoses 9 and 10, in addition to ten known compounds. The structures of the new compounds were confirmed using extensive 1D and 2D NMR analysis. Molecular modeling study suggested the potential of the acylated rhamnopyranoses to bind at the c-Met kinase domain. Cell-free Z'-LYTE™ assay testing revealed the good c-Met phosphorylation inhibitory activity of 9, followed by 8, and 10, with IC50 values of 2.5, 6.9, and 12.7 µM, respectively. The (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) cell proliferation assay testing against the human c-Met expressing highly invasive MDA-MB-231 suggested compound 9 as the most active with IC50 value of 13.3 µM. Testing of compound 9 against multiple phenotypic breast cancer cell lines including MCF-7, BT-474 cells, and MDA-MB-468 proved enhanced activity against the highly c-Met expressing triple-negative breast cancer cell lines. Acylated rhamnopyranoses are potential novel c-Met inhibitors appropriate for future optimizations to control c-Met-dependent breast malignancies. Copyright © 2017 John Wiley & Sons, Ltd.

Decorosides A and B, cytotoxic flavonoid glycosides from the leaves of Rhododendron decorum.

Bioassay and NMR-guided fractionation of the methanolic extract of Rhododendron decorum leaves resulted in the isolation of two new flavonoid glycosides, 5,7-dihydroxy-6,8-dimethyldihydroflavanone-7-O-alpha-L-arabinopyranosyl(1-->6)-beta-D-glucopyranoside (decoroside A, 1) and its 3-hydroxy congener (decoroside B, 2), along with five known compounds myricitrin (3), afzelin (4), (-)-epicatechin (5), (+)-catechin (6), and ampeloptin (7). The structures of the isolated compounds were elucidated by extensive interpretation of their spectral data. Biological evaluation using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay revealed promising cytotoxic activities of these compounds against different cancer cell lines.