The role of active metabolites isolated from Jasminum multiflorum flowers against hepatitis C virus infection and related hepatocellular carcinoma.

Jasminum multiflorum Burm. f. (J. multiflorum) is an ornamental plant with traditional medicinal importance. This study aims to evaluate the activity of J. multiflorum isolated compounds against hepatocellular carcinoma cells infected with hepatitis C virus (HCV) in vitro. The in vitro anti-viral and anti-oncogenic-related activity were validated by anchorage-independent assay plus transwell migration/invasion and spreading assay. In addition to chromatographic isolation of the active metabolites. The flower extract demonstrated a significant antiviral potential through reducing active viral replication by more than 90%. Study results credit this to specific reduction of viral NS5A and cellular EphA2 protein levels. Molecular docking analysis proved the role of the isolated compounds especially multifloroside, jasfloroside A and jasfloroside B as possible anti HCV molecules.

Antioxidant, Anti-Inflammatory and Cytotoxic Activities of Jasminum multiflorum (Burm. F.) Andrews Leaves towards MCF-7 Breast Cancer and HCT 116 Colorectal Cell Lines and Identification of Bioactive Metabolites.

BACKGROUND: The plants of high phenolic contents are perfect antioxidant and anti-inflammatory agents and participate in biological studies as effective agents towards different cancer cell lines. OBJECTIVE: To investigate the antioxidant, anti-inflammatory, and cytotoxic activities of the hydromethanolic leaf extract of Jasminum multiflorum (Burm. f.) Andrews. (J. multiflorum), and phenolic profiling of the extract. METHODS: The antioxidant activity for the extract was estimated using ß-Carotene-linoleic and Ferric Reducing Antioxidant Power (FRAP) assays. The anti-inflammatory activity was evaluated by histamine release assay. Cytotoxicity of J. multiflorum was performed using a neutral red uptake assay towards breast cancer (MCF-7) and colorectal cancer (HCT 116) cell lines. Phenolic profiling of the leaves was characterized using high performance liquid chromatography coupled to photodiode array detector-mass spectroscopy-mass spectroscopy (HPLC-PDA-MS/MS), and chromatographic isolation and identification of the isolated compounds were performed using spectroscopic and NMR data, and virtual docking was performed to the isolated compounds against HSP90 (HEAT SHOCK PROTEIN 90). RESULTS: At a concentration of 75 µg mL-1, J. multiflorum extract showed high antioxidant power; 68.23±0.35 % inhibition and 60.30±0.60 a TEAC (µmol Trolox g-1) for ß-Carotene-linoleic assay and FRAP assay; respectively, and possessed anti-inflammatory activity with IC50 67.2 µg/ml. J. multiflorum showed high cytotoxic activity with IC50 of 24.81 µg/ml and 11.38 µg/ml for MCF-7 and HCT 116 cell lines, respectively. HPLC-PDA-MS/MS analysis tentatively identified 39 compounds; major compounds are secoiridoid glycosides, kaempferol, and quercetin glycosides, in addition to simple phenylethanoid compounds. Isolation of active metabolites was performed and led to the isolation and identification of four compounds. On the basis of docking study using HSP90 legend, kaempferol neohesperidoside showed a high cytotoxic potential supported by a high affinity score towards HSP90 legend protein. CONCLUSION: Jasminum multiflorum is a good candidate to isolate cytotoxic agents.

Phenolic profiling of different Jasminum species cultivated in Egypt and their antioxidant activity.

This study focused on the profiling of phenolic constituents in 80% methanolic extracts of the leaves of some Jasminum species cultivated in Egypt and their antioxidant activities. Phenolic profiling was performed by total phenolic contents, total flavonoid contents and HPLC-DAD for selected species in comparison to authentic standards. DPPH assay was used to estimate the antioxidant activities of Jasminum azoricum L., Jasminum humile L., Jasminum multiflorum (Burm.f.) Andrew, Jasminum officinale L., Jasminum sambac (Ait) L. "Arabian Nights cultivar" and Jasminum sambac (Ait) L. "Grand Duke of Tuscany cultivar". Jasminum multiflorum showed the highest antioxidant activity among selected species with IC50 of 34.8 µg/ml. J. multiflorum showed high concentrations of hydroxytyrosol, protocatechuic acid, hydroxybenzoic acid, kaempferol-3-O- neohesperidoside, and quercetin-3-O-glucoside with concentrations of 977.1 µg/g, 2224.7 µg/g, 714.8 µg/g, 1738.8 µg/g, and 4356.1 µg/g, respectively.

Green Synthesis of Silver Nanoparticles Using Extract of Jasminum officinal L. Leaves and Evaluation of Cytotoxic Activity Towards Bladder (5637) and Breast Cancer (MCF-7) Cell Lines.

INTRODUCTION: Jasminum officinale L. is a very important medicinal and industrial flowering aromatic plant. METHODS: The present study deals with Jasminum officinale L. leaves extract (JOLE) as a reducing and capping agent for the synthesis of silver nanoparticles (AgNPs) by the green pathway. Phenolic profile of the extract was evaluated using HPLC-PDA/MS/MS technique. Jasminum officinale L. leaves extract silver nanoparticles (JOLE-AgNPs) were characterized by ultraviolet light (UV), Fourier transform infrared spectroscopy (FTIR), transmission electron microscopy (TEM), zeta potential and X-ray diffraction (XRD). JOLE-AgNPs were examined for their cytotoxic activities by neutral red uptake assay (NRU) against bladder (5637) and breast cancer (MCF-7) cell lines. RESULTS: HPLC-PDA/MS/MS tentatively identified 51 compounds of different chemical classes. UV spectra showed absorption peak at λmax = 363 nm. The biosynthesized AgNPs were predominantly spherical in shape with an average size of 9.22 nm by TEM. The face cubic center (fcc) nature of silver nanoparticles was proved by XRD diffractogram. JOLE-AgNPs exhibited high cytotoxic activity against 5637 and MCF-7 cell lines compared to the cytotoxic activities of JOLE with IC50 of 13.09 µg/mL and 9.3 µg/mL, respectively. DISCUSSION: The silver nanoparticles formed by Jasminum officinale L. showed high cytotoxic activities against MCF-7 and 5637 cell lines and can be introduced as a new alternative cytotoxic medication.

Aldose reductase inhibitor form Cassia glauca: A comparative study of cytotoxic activity with Ag nanoparticles (NPs) and molecular docking evaluation.

UPLC-MS/MS profiling of Cassia glauca leaves extract revealed the identification of 10 flavonoids. Kaempferol 3-O-ß-D-rutinoside was isolated and studied for its cytotoxic activity. It showed high cytotoxic effects against MCF-7 (IC50 of 4.6±0.038 µg/ml) and HepG-2 (IC50 of 8.2±0.024 µg/ml) cancer cell lines, compared to the leaves extracts, their Ag nanoparticles, and doxorubicin. Moreover, Kaempferol 3-O-ß-D-rutinoside exerted a synergistic cytotoxic effect with doxorubicin on MCF-7 cell lines. It was discovered as kinases and aldose reductase inhibitor while rationalizing its cytotoxic activity through molecular docking study. Thus, it is expected that the cardiotoxic effects of doxorubicin can be also decreased by using Kaempferol 3-O-ß-D-rutinoside due to its aldose reductase inhibitory effect. These findings suggested that Kaempferol 3-O-ß-D-rutinoside could be used in combination with chemotherapeutic drugs to increase the sensitivity to their cytotoxic activity and protect against their side effects.

A highly sensitive GC-MS method for simultaneous determination of anacardic acids in cashew (Anacardium occidentale) nut shell oil in the presence of other phenolic lipid derivatives.

The commercial value of cashew nut shell liquid (CNSL) has become a cornerstone of the agrowaste industry. It is the by-product of the cashew industry and has an 1/8 inch thickness of soft honeycomb structure. CNSL contains phenolic lipids with aliphatic chains such as anacardic acid, cardanol, cardol and methyl cardol, and their derivatives. The developed GC-MS method is rapid, accurate and selective using a selected derivatizing reagent, namely N-methyl-N-(trimethylsilyl)-trifluoroacetamide that was previously diluted 1:1% with anhydrous pyridine. The proposed GC-MS method was applied for the analysis of different CNSL samples. The results showed that all classes of CNSL compounds were detected. The four alkyl phenols were detected with their different alkyl sidechains without any interference. This method is also specified for the detection of fatty acids of saturated and unsaturated chains. Silylation did not cause any alteration in the chemical structure of CNSL compounds regardless of esterification action. Silylation is considered a safe derivatizing agent compatible with GC chromatography and specific for all volatile and nonvolatile polar and nonpolar CNSL compounds that could be detected in CNSL samples.

Isolation of Myricitrin and 3,5-di-O-Methyl Gossypetin from Syzygium samarangense and Evaluation of their Involvement in Protecting Keratinocytes against Oxidative Stress via Activation of the Nrf-2 Pathway.

The wax apple (Syzygium samarangense) is traditionally employed as an antibacterial and immunostimulant drug in traditional medicine. This plant is rich in different flavonoids and tannins. In this study, we isolated two compounds from S. samarangense leaves: myricitrin and 3,5-di-O-methyl gossypetin. Then, we investigated the mechanisms of action of the two compounds against oxidative stress (induced by sodium arsenite) and inflammation (induced by UV light) on human keratinocytes. We could clearly demonstrate that the pre-treatment of cells with both compounds was able to mitigate the negative effects induced by oxidative stress, as no alteration in reactive oxygen species (ROS) production, glutathione (GSH) level, or protein oxidation was observed. Additionally, both compounds were able to modulate mitogen-activated protein kinase (MAPK) signaling pathways to counteract oxidative stress activation. Finally, we showed that 3,5-di-O-methyl gossypetin exerted its antioxidant activity through the nuclear transcription factor-2 (Nrf-2) pathway, stimulating the expression of antioxidant proteins, such as HO-1 and Mn-SOD-3.

Chemical profiling of secondary metabolites of Eugenia uniflora and their antioxidant, anti-inflammatory, pain killing and anti-diabetic activities: A comprehensive approach.

ETHNOPHARMACOLOGICAL RELEVANCE: The red Brazilian cherry, Eugenia uniflora, is widely used in traditional medicine. The aim of this study was to investigate the phytochemical composition of a methanol extract from leaves of E. uniflora and characterization of the isolated compounds. In addition, we aimed to determine the antioxidant activities in vitro and in a cell-based (HaCaT cell) model. We also studied the anti-inflammatory, analgesic, antipyretic and antidiabetic activities in relevant rat models. The molecular mode of action of the antidiabetic activities was also investigated. MATERIALS AND METHODS: UV, MS, and NMR (1H, 13C, DEPT, COSY, HMQC, and HMBC) were used to identify the secondary metabolites. Antioxidant effects were determined in vitro and in HaCaT cells. The ani-inflammatory and antidibetic activities were studied in experimental animals. RESULTS: In this work, a new compound, gallic acid 3-O-[6'-O-acetyl-ß-D-glucoside], along with 16 known plant secondary metabolites (PSM) were isolated, characterized using UV, MS, and NMR (1H, 13C, DEPT, COSY, HMQC, and HMBC). Noticeable antioxidant effects were determined in HaCaT cells: The extract reduced the elevated levels of ROS and p38 phosphorylation and increased the reduced glutathione (GSH) content induced by UVA. The extract showed substantial anti-inflammatory activities in vivo: It diminished the edema thickness in carrageenan-induced hind-paw edema rat model and lowered the leukocyte migration into the peritoneal cavity. In rats, central and peripheral anti-nociceptive properties were also observed: The extract reduced the number of writhing in acid induced writhing and increased the latency time in hot plate test. Furthermore, adequate antipyretic effects were observed: The extract reduced the elevated rectal temperature in rats after intraperitoneal injection of Brewer's yeast. Moreover, the extract possessed robust anti-diabetic activity in streptozotocin (STZ) -diabetic rats: It markedly reduced the elevated serum glucose and lipid peroxidation levels and increased the insulin concentration in serum with higher potency than the positive control, glibenclamide. These effects might be associated with the interaction of PSM with the conserved amino acid residues of human pancreatic α-amylase (HPA), maltase glucoamylase (MGAM-C) and aldose reductase (ALR2) revealing considerable binding affinities. CONCLUSION: A plethora of substantial pharmacological properties indicates that Eugenia uniflora is a good antioxidant and a sustainable by-product with solid therapeutic potential for treating diabetes, inflammation, pain and related oxidative stress diseases.

Chemical composition, antioxidant and hepatoprotective activities of methanol extracts from leaves of Terminalia bellirica and Terminalia sericea (Combretaceae).

BACKGROUND: Plants belonging to the genus Terminalia such as Terminalia bellirica and Terminalia sericea are used traditionally to treat several diseases and health disorders. Up to this date, the roots of Terminalia sericea and the fruits of Terminalia bellirica are the mostly studied plant parts. The phytochemical composition and the biological activities of the leaves of both species are not well identified so far. METHODS: The secondary metabolites of Terminalia bellirica and Terminalia sericea leaves were identified using HPLC-PDA-MS/MS. The antioxidant activities of the leaves extracts were determined by DPPH and FRAP assays. The hepatoprotective potential was evaluated in rats with D-galactosamine induced liver damage. The effect of the extracts on the expression of the anti-apoptotic marker Bcl-2 was measured in an immunohistochemical study. The most abundant compounds identified in the studied extracts were docked into Bcl-2: Bim (BH3) interaction surface using molecular operating environment software. RESULTS: A total of 85 secondary metabolites were identified in the leaf extracts of both species. Ellagitannins such as corilagin, chebulagic acid, galloylpunicalagin, and digalloyl-hexahydroxydiphenoyl-hexoside were found to be the major components in Terminalia bellirica whereas flavonoid glycosides including quercetin rutinoside and quercetin galloyl-glucoside were highly abundant in Terminalia sericea. The studied extracts exhibited pronounced antioxidant activities, moderate anti-apoptotic and hepatoprotective potential. In silico docking experiments revealed that the compounds abundant in the extracts were able to bind to Bcl-2: Bim (BH3) interaction surface with an appreciable binding free energy. DISCUSSION: The antioxidant and hepatoprotective activities exhibited by the studied extracts might be attributed to the high content of the polyphenols. The anti-apoptotic activity could be due to the interference with the apoptotic pathway mediated by Bcl-2: Bim interaction. These findings support the medicinal relevance of Terminalia bellirica and Terminalia sericea and provide a rational base for their utilization in folk medicine.

Cytotoxic constituents from Vicia monantha subsp. monantha seeds.

Chemical investigation of Vicia monantha subsp. monantha Retz. revealed isolation of one new hydroxy- fatty acid (6) identified as (6-Z, 10-E)-9-hydroxy henicosa-6,10-dienoic acid in addition to six known metabolites; hexadecanoic acid (1), ß-sitosterol (2), ß-amyrin (3), ß-sitosterol-glucoside (4), 2,3-dihydroxypropyl tetradecanoate (5) and (Z)-9-hydroxypentadec-6-enoic acid (7). The cytotoxic effect of the isolated compounds was assessed by MTT assay using lung cancer A-549, prostate cancer PC3, breast cancer MCF-7, colon cancer HCT-116 and liver cancer HepG2 cell lines. Only compounds 1, 2, and 4 showed cytotoxic effect on HCT-116 cells where compound 2 was the most active with IC50 value of 22.61 µg/mL. In addition, compounds 1, 2, 3, and 4 showed promising cytotoxic effect on MCF-7 cells with IC50 values of 21.03, 15.42, 10.089, and 11.34 µg/mL, respectively.

Rheum palmatum root extract inhibits hepatocellular carcinoma in rats treated with diethylnitrosamine.

OBJECTIVES: The aim of this study was to investigate the potential anticancer properties of a methanol extract of Rheum palmatum roots against diethylnitrosamine (DENA)-induced hepatocellular carcinoma (HCC) in rats and to characterize its phytoconstituents. METHODS: HPLC-PDA-MS/MS was used to profile the secondary metabolites in R. palmatum root extract. HCC was induced using diethylnitrosamine (DENA). The activity of serum alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP) and gamma-glutamyl transferase (GGT), alpha-fetoprotein (AFP), total proteins, serum albumin and serum globulin was determined. DNA fragmentation and histopathological examination and GST-P immunostaining were also studied. KEY FINDINGS: LC-MS/MS analysis identified 16 compounds belonging to anthraquinones, flavonoids and tannins. The root extract significantly reduced the elevated liver enzymes ALT and AST and increased total proteins, albumin and globulin in HCC-rats. Also, the tumour markers AFP and GGT levels were significantly reduced in HCC-rats treated with the extract. In addition, the extract significantly reduced elevated DNA fragmentation and decreased the numbers and areas of GST-P positive putative foci in HCC-rats. CONCLUSIONS: Rheum palmatum is a potential candidate to be explored for the treatment of hepatocellular carcinoma.

A New Polyoxygenated Flavonol Gossypetin-3-O-ß-d-Robinobioside from Caesalpinia gilliesii (Hook.) D. Dietr. and In Vivo Hepatoprotective, Anti-Inflammatory, and Anti-Ulcer Activities of the Leaf Methanol Extract.

A hitherto unknown polyoxygenated flavonol robinobioside (gossypetin-3-O-ß-d-robinobioside) was isolated from the leaves of Caesalpinia gilliesii along with thirteen known phenolic secondary metabolites. The isolated compounds were characterized using spectroscopic analysis, including 1D and 2D NMR and mass spectrometry (MS) analyses. The extract reduced the level of liver damage in CCl4-induced liver injury in rats. A decrease of the liver biomarkers-aspartate aminotransferase (AST) and alanine aminotransferase (ALT) and an increase of total antioxidant capacity (TAC) levels-were observed similar to the liver protecting drug silymarin. In addition, the extract showed promising activity against carrageenan-induced paw edema in rats and protected their stomachs against ethanol-induced gastric ulcers in a concentration dependent fashion. The observed activities could be attributed to the high content of antioxidant polyphenols. Our results suggest that the C. gilliesii has the capacity to scavenge free radicals and can protect against oxidative stress, and liver and stomach injury.

Phenolic metabolites, biological activities, and isolated compounds of Terminalia muelleri extract.

CONTEXT: Terminalia muelleri Benth. (Combretaceae), is rich with phenolics that have antioxidant and cytotoxic activities. No screening studies were published before on T. muelleri. OBJECTIVE: The study focused on isolation and identification of secondary metabolites from aqueous methanol leaf extract of T. muelleri and evaluation of its biological activities. MATERIALS AND METHODS: The n-butanol extract was chromatographed on polyamide 6, and eluted with H2O/MeOH mixtures of decreasing polarity, then separated by different chromatographic tools that yielded 10 phenolic compounds. The antioxidant activity of the extract was evaluated by investigating its total phenolic and flavonoid content and DPPH scavenging effectiveness. The extract and the two acylated flavones were evaluated for their anticancer activity towards MCF-7 and PC3 cancer cell lines. Molecular docking study of the acylated flavones was performed against topoisomerase enzyme. RESULTS AND DISCUSSION: Two acylated flavonoids, apigenin-8-C-(2″-O-galloyl) glucoside 1 and luteolin-8-C-(2″-O-galloyl) glucoside 2, were isolated and identified for the second time in nature, with eight tannins (3-10), from the leaves of T. muelleri. The extract and compound 10 showed the most significant antioxidant activity (IC50 = 3.55 and 6.34 µg/mL), respectively. The total extract and compound 2 demonstrated cytotoxic effect against MCF-7 with IC50 = 29.7 and 45.2 µg/mL respectively, while compound 1 showed cytotoxic effect against PC3 (IC50 = 40.8 µg/mL). The docking study of compounds 1 and 2 confirmed unique binding mode in the active site of human DNA topoisomerase enzyme. CONCLUSIONS: Terminalia muelleri is a promising medicinal plant as it possesses high antioxidant activity and moderate cytotoxic activity against MCF-7.

A New Octadecenoic Acid Derivative from Caesalpinia gilliesii Flowers with Potent Hepatoprotective Activity.

BACKGROUND: Caesalpinia gilliesii Hook is an ornamental shrub with showy yellow flowers. It was used in folk medicine due to its contents of different classes of secondary metabolites. In our previous study, dichloromethane extract of C. gilliesii flowers showed a good antioxidant activity. AIM OF THE STUDY: Isolation and identification of bioactive hepatoprotective compounds from C. gilliesii flowers dichloromethane fraction. MATERIALS AND METHODS: The hepatoprotective activity of dichloromethane fraction and isolated compounds were studied in CCl4-intoxicated rat liver slices by measuring liver injury markers (alanine aminotransferase, aspartate aminotransferase and glutathione [GSH]). All compounds were structurally elucidated on the basis of electron ionization-mass spectrometry, one- and two-dimensional nuclear magnetic resonance. RESULTS: A new 12,13,16-trihydroxy-14(Z)-octadecenoic acid was identified in addition to the known ß-sitosterol-3-O-butyl, daucosterol, isorhamnetin, isorhamnetin-3-O-rhamnoside, luteolin-7,4'-dimethyl ether, genistein-5-methyl ether, luteolin-7-O-rhamnoside, isovanillic acid, and p-methoxybenzoic acid. Dichloromethane fraction and isorhamnetin were able to significantly protect the liver against intoxication. Moreover, the dichloromethane fraction and the isolated phytosterols induced GSH above the normal level. CONCLUSION: The hepatoprotective activity of C. gilliesii may be attributed to its high content of phytosterols and phenolic compounds. SUMMARY: Bioactive Hepatoprotective phytosterols and phenolics from chloroform extract of Caesalpinia gilliesii Abbreviations used: ALT: Alanine Aminotransferase; AST: Aspartate aminotransferase; GSH: Glutathione; SC50: Scavenging Capacity 50 (SC 50); COSY: Correlation spectroscopy; NMR: Nuclear Magnetic Resonance; CC: Column chromatography; EI-MS: Electron-impact mass spectrometry; HSQC: Heteronuclear single-quantum correlation.

Isolation of major phenolics from Launaea spinosa and their protective effect on HepG2 cells damaged with t-BHP.

CONTEXT: Some Launaea species (Asteraceae) are used traditionally to treat liver oxidative stress. OBJECTIVE: The present study investigates the protective effects of isolated compounds from Launaea spinosa Sch. Bip. (Asteraceae) against oxidative stress on t-BHP-induced HepG2 cells. MATERIALS AND METHODS: Major phenolic content from flowering aerial parts of L. spinosa was isolated and identified. The protective effects of isolated compounds (10 and 20 µM) against oxidative stress induced by tert-butyl hydroperoxide (t-BHP) in HepG2 cells were investigated through the measurement of aspartate aminotransferase (AST), alanine transaminase (ALT), and superoxide dismutase (SOD) levels. RESULTS: A new phenolic compound identified as 2,3-diferulyl R,R-(+) methyl tartrate (6), in addition to five known metabolites, esculetin (1), esculetin-7-O-d-glucoside (cichoriin) (2), fertaric acid (3), acacetin-7-O-d-glucoside (4), and acacetin-7-O-d-glucuronic acid (5), were isolated. Oxidant-induced damage by 200 µM t-BHP in HepG2 cells was inhibited by compounds 1, 4, and 5 (10 and 20 µM), or quercetin (10 µM; positive control). The protective effects of compounds 1, 4, and 5 were associated with decreasing in AST, ALT, and SOD levels. Compound 4 (20 µM) decreased the AST level from 128.5 ± 13.9 to 7.9 ±1.8 U/mL. Meanwhile, compound 1 (20 µM) decreased ALT activity from 20.3 ± 7.0 to 7.6 ± 2.4 U/mL, while compound 5 decreased SOD levels from 41.6 ± 9.0 to 28.3 ± 3.4 mU/mg. CONCLUSION: The major phenolic compounds isolated from L. spinosa displayed a significant cytoprotective effect against oxidative stress, leading to maintenance of the normal redox status of the cell.

A new pregnane glycoside from Gomphocarpus fruticosus growing in Egypt.

Phytochemical investigation of Gomphocarpus fruticosus (L.) Ait. of Egyptian origin afforded the new pregnane glycoside lineolon-3-O-[ß-D-oleandropyranosyl-(1-4)-ß-D-cymaropyranosyl-(1-4)-ß-D-cymaropyranoside], along with six known compounds. The structures of the isolated compounds were elucidated on the basis of extensive spectroscopic evidences derived from 1D, 2D NMR experiments, mass spectrometry and by comparing their physical and spectroscopic data to literature. These included the triterpenoids 3ß-taraxerol, 3ß-taraxerol acetate and betulinic acid, which are identified for the first time in G. fruticosus and the cardenolides uzarigenin, gomphoside and calotropin.