Molecules

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Open AccessArticle

Virtual Screening against Phosphoglycerate Kinase 1 in Quest of Novel Apoptosis Inhibitors

by Jie Xia, Bo Feng, Qianhang Shao, Yuhe Yuan, Xiang Simon Wang, Naihong Chen and Song Wu

Molecules 2017, 22(6), 1029; https://doi.org/10.3390/molecules22061029 - 21 Jun 2017

Cited by 10 | Viewed by 5739

Inhibition of apoptosis is a potential therapy to treat human diseases such as neurodegenerative disorders (e.g., Parkinson’s disease), stroke, and sepsis. Due to the lack of druggable targets, it remains a major challenge to discover apoptosis inhibitors. The recent repositioning of a marketed [...] Read more.

Inhibition of apoptosis is a potential therapy to treat human diseases such as neurodegenerative disorders (e.g., Parkinson’s disease), stroke, and sepsis. Due to the lack of druggable targets, it remains a major challenge to discover apoptosis inhibitors. The recent repositioning of a marketed drug (i.e., terazosin) as an anti-apoptotic agent uncovered a novel target (i.e., human phosphoglycerate kinase 1 (hPgk1)). In this study, we developed a virtual screening (VS) pipeline based on the X-ray structure of Pgk1/terazosin complex and applied it to a screening campaign for potential anti-apoptotic agents. The hierarchical filters in the pipeline (i.e., similarity search, a pharmacophore model, a shape-based model, and molecular docking) rendered 13 potential hits from Specs chemical library. By using PC12 cells (exposed to rotenone) as a cell model for bioassay, we first identified that AK-918/42829299, AN-465/41520984, and AT-051/43421517 were able to protect PC12 cells from rotenone-induced cell death. Molecular docking suggested these hit compounds were likely to bind to hPgk1 in a similar mode to terazosin. In summary, we not only present a versatile VS pipeline for potential apoptosis inhibitors discovery, but also provide three novel-scaffold hit compounds that are worthy of further development and biological study. Full article

(This article belongs to the Section Medicinal Chemistry)

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Graphical abstract

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Full article ">Figure 1
The chemical structure of terazosin that binds to hPgk1. Full article ">Figure 2
The pharmacophore models (based on protein-ligand interactions) generated by DS2016/Catalyst: (a) the binding mode of terazosin to hPgk1 (left panel), as well as the model selected for pharmacophore filtering (right panel); and (b) the other four pharmacophore models. Color codes: light blue, hydrophobic group; gray, excluded volumes. Full article ">Figure 3
The shape-based model generated from terazosin by ROCS program. Color code: gray, shape-based model. Full article ">Figure 4
The chemical structures of 13 potential hits (note: the compounds labeled with “*” were experimentally validated as active). Full article ">Figure 5
The binding poses of 13 potential hits: (a) the binding pocket of hPgk1 by the surface representation; and, (b) protein-ligand interactions between the potential hits and hPgk1. Color codes: green, hPgk1; light blue, potential hits; yellow, terazosin; red, oxygen atom; dark blue, nitrogen atom. Full article ">Figure 6
The effects of 13 potential hits on PC12 cells in the absence of rotenone. Ctrl, PC12 cells incubated with 0.1% DMSO only; Ngf, treatment of 50 ng/mL nerve growth factor; terazosin, treatment of 10 μmol/L terazosin. Error bars represent SD. *** p <0.001 vs. control group; one-way analysis of variance was used (n = 9). Full article ">Figure 7
The predicted binding modes of three apoptosis inhibitors to hPgk1: (a) AK-918/42829299; (b) AN-465/41520984; and, (c) AT-051/43421517. The residues that interact with each hit compound are labeled. Color codes: green, hPgk1; light blue, apoptosis inhibitors; red, oxygen atom; dark blue, nitrogen atom; yellow, sulfur atom. Full article ">Scheme 1
The general workflow for the discovery of apoptosis inhibitors. Full article ">

498 KiB

Open AccessArticle

Synthesis and Bioactivity Characterization of Scutellarein Sulfonated Derivative

by Ting Gu, Yue Zhong, Yu-Ting Lu, Ying Sun, Ze-Xi Dong, Wen-Yu Wu, Zhi-Hao Shi, Nian-Guang Li, Xin Xue, Fang Fang, He-Min Li and Yu-Ping Tang

Molecules 2017, 22(6), 1028; https://doi.org/10.3390/molecules22061028 - 21 Jun 2017

Cited by 6 | Viewed by 4825

Abstract

Scutellarin (1) has been widely used to treat acute cerebral infarction in clinic, but poor aqueous solubility decreases its bioavailability. Interestingly, scutellarin (1) could be metabolized into scutellarein (2) in vivo. In this study, a sulfonic group [...] Read more.

Scutellarin (1) has been widely used to treat acute cerebral infarction in clinic, but poor aqueous solubility decreases its bioavailability. Interestingly, scutellarin (1) could be metabolized into scutellarein (2) in vivo. In this study, a sulfonic group was introduced at position C-8 of scutellarein (2) to enhance the aqueous solubility of the obtained derivative (3). DPPH (1,1-diphenyl-2-picrylhydrazyl)-radical scavenging ability and antithrombic activity were also conducted to determine its bioactivity. The result showed that scutellarein derivate (3) could be a better agent for ischemic cerebrovascular disease treatment. Full article

(This article belongs to the Special Issue Synthesis and Modification of Natural Product)

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9431 KiB

Open AccessArticle

Molecular Quantum Similarity, Chemical Reactivity and Database Screening of 3D Pharmacophores of the Protein Kinases A, B and G from Mycobacterium tuberculosis

by Alejandro Morales-Bayuelo

Molecules 2017, 22(6), 1027; https://doi.org/10.3390/molecules22061027 - 21 Jun 2017

Cited by 13 | Viewed by 4850

Abstract

Mycobacterium tuberculosis remains one of the world’s most devastating pathogens. For this reason, we developed a study involving 3D pharmacophore searching, selectivity analysis and database screening for a series of anti-tuberculosis compounds, associated with the protein kinases A, B, and G. This theoretical [...] Read more.

Mycobacterium tuberculosis remains one of the world’s most devastating pathogens. For this reason, we developed a study involving 3D pharmacophore searching, selectivity analysis and database screening for a series of anti-tuberculosis compounds, associated with the protein kinases A, B, and G. This theoretical study is expected to shed some light onto some molecular aspects that could contribute to the knowledge of the molecular mechanics behind interactions of these compounds, with anti-tuberculosis activity. Using the Molecular Quantum Similarity field and reactivity descriptors supported in the Density Functional Theory, it was possible to measure the quantification of the steric and electrostatic effects through the Overlap and Coulomb quantitative convergence (alpha and beta) scales. In addition, an analysis of reactivity indices using global and local descriptors was developed, identifying the binding sites and selectivity on these anti-tuberculosis compounds in the active sites. Finally, the reported pharmacophores to PKn A, B and G, were used to carry out database screening, using a database with anti-tuberculosis drugs from the Kelly Chibale research group (http://www.kellychibaleresearch.uct.ac.za/), to find the compounds with affinity for the specific protein targets associated with PKn A, B and G. In this regard, this hybrid methodology (Molecular Mechanic/Quantum Chemistry) shows new insights into drug design that may be useful in the tuberculosis treatment today. Full article

(This article belongs to the Section Molecular Diversity)

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2448 KiB

Open AccessArticle

Screening and Identification of the Metabolites in Rat Plasma and Urine after Oral Administration of Areca catechu L. Nut Extract by Ultra-High-Pressure Liquid Chromatography Coupled with Linear Ion Trap–Orbitrap Tandem Mass Spectrometry

by Lulu Li, Zhiqiang Luo, Yang Liu, Hao Wang, Aoxue Liu, Guohua Yu, Mengwei Li, Ruirui Yang, Xinjing Chen, Jialian Zhu and Baosheng Zhao

Molecules 2017, 22(6), 1026; https://doi.org/10.3390/molecules22061026 - 21 Jun 2017

Cited by 14 | Viewed by 5794

Abstract

Areca catechu L. nut, a well-known toxic traditional herbal medicine, has been widely used to treat various diseases in China and many other Asian countries for centuries. However, to date the in vivo absorption and metabolism of its multiple bioactive or toxic components [...] Read more.

Areca catechu L. nut, a well-known toxic traditional herbal medicine, has been widely used to treat various diseases in China and many other Asian countries for centuries. However, to date the in vivo absorption and metabolism of its multiple bioactive or toxic components still remain unclear. In this study, liquid chromatography coupled with tandem mass spectrometry was used to analyze the major components and their metabolites in rat plasma and urine after oral administration of Areca catechu L. nut extract (ACNE). A total of 12 compounds, including 6 alkaloids, 3 tannins and 3 amino acids, were confirmed or tentatively identified from ACNE. In vivo, 40 constituents, including 8 prototypes and 32 metabolites were identified in rat plasma and urine samples. In summary, this study showed an insight into the metabolism of ACNE in vivo, which may provide helpful chemical information for better understanding of the toxicological and pharmacological profiles of ACNE. Full article

(This article belongs to the Section Metabolites)

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2383 KiB

Open AccessReview

From Farm to Pharma: An Overview of Industrial Heparin Manufacturing Methods

by Jan-Ytzen Van der Meer, Edwin Kellenbach and Leendert J. Van den Bos

Molecules 2017, 22(6), 1025; https://doi.org/10.3390/molecules22061025 - 21 Jun 2017

Cited by 89 | Viewed by 13397

Abstract

The purification of heparin from offal is an old industrial process for which commercial recipes date back to 1922. Although chemical, chemoenzymatic, and biotechnological alternatives for this production method have been published in the academic literature, animal-tissue is still the sole source for [...] Read more.

The purification of heparin from offal is an old industrial process for which commercial recipes date back to 1922. Although chemical, chemoenzymatic, and biotechnological alternatives for this production method have been published in the academic literature, animal-tissue is still the sole source for commercial heparin production in industry. Heparin purification methods are closely guarded industrial secrets which are not available to the general (scientific) public. However by reviewing the academic and patent literature, we aim to provide a comprehensive overview of the general methods used in industry for the extraction of heparin from animal tissue. Full article

(This article belongs to the Special Issue Looking Forward to the Future of Heparin: New Sources, Developments and Applications)

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Figure 1
Major disaccharide found in heparin: (-4)-α-l-IdoA2S-(1-4)-α-d-GlcNS6S-(1-) [<a href="#B5-molecules-22-01025" class="html-bibr">5</a>]. Full article ">Figure 2
(a), Schematic representation of an industrial heparin purification process; (b) Discussed topics per section; (c) General process conditions and reagents; (d) Removed impurities per section. Full article ">Figure 3
Structures of quaternary ammonium salts used for heparin capture by precipitation. (a) General structure of an applicable salt; (b) Structure of benzethonium chloride (Hyamine® 1622). Full article ">Figure 4
Fractional precipitation of heparin, DS and CS at different concentrations of methanol. Data derived from Volpi et al. [<a href="#B36-molecules-22-01025" class="html-bibr">36</a>]. Full article ">Figure 5
Reducing end oxidized N-acetylglucosamine heparin modification as a result of potassium permanganate bleaching. Full article ">Figure 6
3-Acetyluronic acid heparin modification as a result of peracetic acid bleaching. Full article ">

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Open AccessReview

Deoxyelephantopin and Isodeoxyelephantopin as Potential Anticancer Agents with Effects on Multiple Signaling Pathways

by Tahir Mehmood, Amara Maryam, Hamed A. Ghramh, Muhammad Khan and Tonghui Ma

Molecules 2017, 22(6), 1013; https://doi.org/10.3390/molecules22061013 - 21 Jun 2017

Cited by 31 | Viewed by 7314

Abstract

Cancer is the 2nd leading cause of death worldwide. The development of drugs to target only one specific signaling pathway has limited therapeutic success. Developing chemotherapeutics to target multiple signaling pathways has emerged as a new prototype for cancer treatment. Deoxyelephantopin (DET) and [...] Read more.

Cancer is the 2nd leading cause of death worldwide. The development of drugs to target only one specific signaling pathway has limited therapeutic success. Developing chemotherapeutics to target multiple signaling pathways has emerged as a new prototype for cancer treatment. Deoxyelephantopin (DET) and isodeoxyelephantopin (IDET) are sesquiterpene lactone components of “Elephantopus scaber and Elephantopus carolinianus”, traditional Chinese medicinal herbs that have long been used as folk medicines to treat liver diseases, diabetes, diuresis, bronchitis, fever, diarrhea, dysentery, cancer, and inflammation. Recently, the anticancer activity of DET and IDET has been widely investigated. Here, our aim is to review the current status of DET and IDET, and discuss their anticancer activity with specific emphasis on molecular targets and mechanisms used by these compounds to trigger apoptosis pathways which may help to further design and conduct research to develop them as lead therapeutic drugs for cancer treatments. The literature has shown that DET and IDET induce apoptosis through multiple signaling pathways which are deregulated in cancer cells and suggested that by targeting multiple pathways simultaneously, these compounds could selectively kill cancer cells. This review suggests that DET and IDET hold promising anticancer activity but additional studies and clinical trials are needed to validate and understand their therapeutic effect to develop them into potent therapeutics for the treatment of cancer. Full article

(This article belongs to the Collection Bioactive Compounds)

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Figure 1
Chemical structure and natural sources of deoxyelephantopin (DET) and isodeoxyelephantopin (IDET). Full article ">Figure 2
A schematic representation of DET- and IDET-induced cell cycle arrest at multiple phases in different cancer cell lines. DET and IDET inhibit S phase and/or G2/M phase transition by down-regulating the expression of cyclin-dependent kinase 1 (CDK1), CDK2, CDK4, CDK6, cyclin A2, cyclin D1, cyclin D3, cyclin E2, cyclin B1, and cdc2 while up-regulating the expression of p21 (CDK inhibitor) and p53, a tumor suppresser gene which regulates cell cycle arrest at different phases by inducing p21;├ Inhibition, ↑ Up-regulation. Full article ">Figure 3
A schematic representation of DET- and IDET-induced apoptosis in different cancer cell lines. DET and IDET trigger the activation of extrinsic apoptosis by activating caspase-8 which in turn either initiates the type 1 extrinsic apoptotic pathway by activating downstream effector caspase-3 or the type 2 extrinsic apoptotic pathway by truncation of Bid. DET and IDET induce intrinsic apoptosis by dissipating mitochondrial membrane potential and modulating the expression of Bcl-2 family proteins which results in the activation of caspase-3. Subsequently, activated caspase-3 leads to apoptosis by substrate cleavage. DET activates c-Jun N-terminal kinase (JNK) and p38 and inhibits the activation of Phosphatidylinositol-3-Kinase (PI3K)/AKT/ mammalian target of rapamycin (mTOR). DET inhibits the activation of extracellular signal-regulated kinase (ERK)-mitogen-activated protein kinase (MAPK) and activates caspase-9 and induces apoptosis; ├ Inhibition, ↑ Up-regulation, ↓ Down-regulation. Full article ">Figure 4
Proposed mechanism by which DET and IDET inhibit nuclear factor kappa B (NF-κB) and signal transducers and activators of transcription 3 (STAT3) activation and NF-κB and STAT3-regulated gene expression involved in cell proliferation and invasion. DET and IDET inhibited the activation of NF-κB induced by various inflammatory stimuli such as (tumor necrosis factor (TNF), lipopolysaccharide (LPS), interleukin-1 beta (IL-1β) which activates NF-κB through different pathways. DET and IDET inhibited NF-κB activation at the same step of all these stimuli. Inhibition of NF-κB activation was associated with decreased phosphorylation and the degradation of IκB-α and upstream activation of IκB kinase-alpha/-beta (IKK-α/-β) and IKK kinase activities. DET and IDET down-regulated the expression of NF-κB-regulated gene products involved in invasion such as matrix metalloproteinase (MMP-9) and intercellular adhesion molecule 1 (ICAM-1); cell proliferation such as cyclooxygenase-2 (COX-2), cyclin D1, and cellular-Myc (c-Myc); and anti-apoptosis such as inhibitor of apoptosis protein-1/-2 (IAP-1/-2), B-cell lymphoma 2 (Bcl-2), B-cell lymphoma-extra-large (Bcl-xL), Bcl-2-related protein A1 also known as Bfl-1/A1, TNF receptor-associated factor (TRAF1), FADD-like IL-1beta-converting enzyme (FLICE)/caspase-8-inhibitory protein (FLIP), and survivin). DET inhibits the activation of STAT3 by reducing phosphorylation at tyrosine-705 and up-regulates the suppressor of cytokine signaling 3 (SOCS3) which is a major component for the negative regulation of the IL-6 signaling cascade; ├ Inhibition, ↑ Up-regulation. Full article ">

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Open AccessReview

Insights into Penicillium brasilianum Secondary Metabolism and Its Biotechnological Potential

by Jaqueline Moraes Bazioli, Luciana Da Silva Amaral, Taícia Pacheco Fill and Edson Rodrigues-Filho

Molecules 2017, 22(6), 858; https://doi.org/10.3390/molecules22060858 - 20 Jun 2017

Cited by 30 | Viewed by 9318

Abstract

Over the past few years Penicillium brasilianum has been isolated from many different environmental sources as soil isolates, plant endophytes and onion pathogen. All investigated strains share a great ability to produce bioactive secondary metabolites. Different authors have investigated this great capability and [...] Read more.

Over the past few years Penicillium brasilianum has been isolated from many different environmental sources as soil isolates, plant endophytes and onion pathogen. All investigated strains share a great ability to produce bioactive secondary metabolites. Different authors have investigated this great capability and here we summarize the metabolic potential and the biological activities related to P. brasilianum’s metabolites with diverse structures. They include secondary metabolites of an alkaloid nature, i.e., 2,5-diketopiperazines, cyclodepsipeptides, meroterpenoids and polyketides. Penicillium brasilianum is also described as a great source of enzymes with biotechnological application potential, which is also highlighted in this review. Additionally, this review will focus on several aspects of Penicillium brasilianum and interesting genomic insights. Full article

(This article belongs to the Special Issue Expanding the Potential of Metabolomics for Under Investigated Organisms and in Drug Discovery)

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3692 KiB

Open AccessArticle

The Antiproliferative Effect of Cyclodipeptides from Pseudomonas aeruginosa PAO1 on HeLa Cells Involves Inhibition of Phosphorylation of Akt and S6k Kinases

by Laura Hernández-Padilla, Dolores Vázquez-Rivera, Luis A. Sánchez-Briones, Alma L. Díaz-Pérez, José Moreno-Rodríguez, Mario A. Moreno-Eutimio, Victor Meza-Carmen, Homero Reyes-De la Cruz and Jesús Campos-García

Molecules 2017, 22(6), 1024; https://doi.org/10.3390/molecules22061024 - 20 Jun 2017

Cited by 22 | Viewed by 6429

Abstract

Pseudomonas aeruginosa PAO1, a potential pathogen of plants and animals, produces the cyclodipeptides cyclo(l-Pro-l-Tyr), cyclo(l-Pro-l-Phe), and cyclo(l-Pro-l-Val) (PAO1-CDPs), whose effects have been implicated in inhibition of human tumor cell line proliferation. Our purpose was to investigate in depth in the mechanisms of HeLa [...] Read more.

Pseudomonas aeruginosa PAO1, a potential pathogen of plants and animals, produces the cyclodipeptides cyclo(l-Pro-l-Tyr), cyclo(l-Pro-l-Phe), and cyclo(l-Pro-l-Val) (PAO1-CDPs), whose effects have been implicated in inhibition of human tumor cell line proliferation. Our purpose was to investigate in depth in the mechanisms of HeLa cell proliferation inhibition by the PAO1-CDPs. The results indicate that PAO1-CDPs, both purified individually and in mixtures, inhibited HeLa cell proliferation by arresting the cell cycle at the G0–G1 transition. The crude PAO1-CDPs mixture promoted cell death in HeLa cells in a dose-dependent manner, showing efficacy similar to that of isolated PAO1-CDPs (LD₅₀ of 60–250 µM) and inducing apoptosis with EC₅₀ between 0.6 and 3.0 µM. Moreover, PAO1-CDPs showed a higher proapoptotic activity (~10³–10⁵ fold) than their synthetic analogs did. Subsequently, the PAO1-CDPs affected mitochondrial membrane potential and induced apoptosis by caspase-9-dependent pathway. The mechanism of inhibition of cells proliferation in HeLa cells involves inhibition of phosphorylation of both Akt-S473 and S6k-T389 protein kinases, showing a cyclic behavior of their expression and phosphorylation in a time and concentration-dependent fashion. Taken together our findings indicate that PI3K–Akt–mTOR–S6k signaling pathway blockage is involved in the antiproliferative effect of the PAO1-CDPs. Full article

(This article belongs to the Special Issue Peptide-Based Drugs and Drug Delivery Systems)

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Full article ">Figure 1
Effects of PAO1 and synthetic CDPs on HeLa cell viability and apoptosis induction. HeLa cells were incubated in CM medium and treated with CDPs for 24 h as described in Materials and Methods. Determination of viability of HeLa cells by an MTT assay treated with CDPs from P. aeruginosa PAO1 (a) and synthetic analogous CDPs (b). Induction of apoptosis in human HeLa cells by CDPs was analyzed in cultures grown in the CM medium and after treatment with CDPs for 24 h. Human cell lines were stained with annexin V and propidium iodide and analyzed by flow cytometry. The percentage of viability was determined by fluorescent cell quantitation in the dot plots, values were graphed as bars. Apoptosis induction in HeLa cells treated with PAO1-CDPs (c) and with the mixture of synthetic analogs of CDPs (d); (e) Apoptosis induction in normal human lung fibroblasts by the PAO1-CDP mixture at different concentrations determined as in (c); (f) IL-8 induction in HEK-293 cells by ELISA assay as described in Materials and Methods. Bars represent mean ± standard error (SE) of three independent experiments, n = 6. One-way analysis of variance (ANOVA) was carried out, with Tukey’s post hoc test; statistical significance (p < 0.01) of differences between treatments is indicated with different lowercase letters. Full article ">Figure 2
Morphological changes in human cell cultures stimulated with the CDP mixture from P. aeruginosa PAO1. (a–n) Images of human cells taken by means of phase contrast and confocal microscopy after treatment; (a–c) HeLa cell line; (d–f) Human lung fibroblast cell line. (a,d) DMSO (0.05%; negative control); (b,e) Actinomycin D (50 mg/mL; positive apoptosis control); (c,f) PAO1-CDP mixture (10 mg/mL) treatment for 2 h; (g) HeLa cells without treatment; (h) Determination of HeLa cells’ membrane potential (ΔΨm) using Rhodamine 123 without treatment. (i) HeLa cell membrane potential (ΔΨm) and superoxide (O2•−) quantification (using DHE probe) without treatment; (j) Same as in (i); but examination by dark field microscopy; (k–n) Conditions as in (g–j) but with treatment with the PAO1-CDP mixture. Images of the cells were taken at 40× magnification, using inverted phase-contrast microscope (HB0-50, Carl-Zeiss, San Diego, CA, USA) or confocal microscope (FV1000, Olympus, Center Valley, PA, USA). Full article ">Figure 3
Apoptosis induction in HeLa cells by CDPs from P. aeruginosa PAO1. HeLa cells were incubated in the CM medium after treatment with PAO1-CDPs (various doses) as a function of time. Cells were stained with annexin V, 7-AAD, or propidium iodide and analyzed by flow cytometry. (a–d) The percentage of fluorescent cells determined in the dot plots is shown, corresponding to HeLa cells treated for 6 h with (a) DMSO; (b) actinomycin D (50 mg/mL), (c) PAO1-CDP mixture (1 μg/mL); (d) PAO1-CDP mixture (10 μg/mL). Q1, early apoptosis; Q2, late apoptosis; Q3, necrotic cells; Q4, viable cells; (e,f) Kinetic of induction early and late apoptosis stages in HeLa cells treated with the PAO1-CDP mixture; (g,h) Kinetics of induction of early and late apoptosis stages in blood mononuclear human cells treated with the PAO1-CDP mixture. Points of the plots represent mean ± standard error (SE) of three independent experiments; (i) Effects of inhibitors of apoptosis on HeLa cells in the presence of the PAO1-CDP mixture. The cells were incubated in the CM medium after treatment with 10 μg/mL PAO1-CDP mixture with the addition of caspases inhibitors. Cells were revealed with annexin V and analyzed by flow cytometry. The percentage of fluorescent cells (apoptotic cells) after each treatment was determined by means of the dot plots and is shown as bars in the graph. The inhibitors tested are indicated on the X-axis. Bars of the plots represent mean ± SE of three independent experiments, n = 3. One-way ANOVA was carried out, with Tukey’s post hoc test; statistical significance (p < 0.01) of differences between treatments is showed with different lowercase letters; (j) Effects of the PAO1-CDP mixture on the cell cycle in HeLa cells. HeLa cells were incubated in serum-free medium (SS) and serum-enriched medium (CM) after treatment with various doses of the PAO1-CDPs for 24 h. Cells were fixed with paraformaldehyde (4%) for 10 min on ice. Then, the cells were incubated with DAPI (1:1000) for 10 min at room temperature and analyzed by flow cytometry for DNA quantitation. Full article ">Figure 4
Membrane potential and superoxide quantification in HeLa cells treated with the CDP mixture from P. aeruginosa PAO1. Cell cultures were grown in the CM medium, harvested, and incubated in the CM or SS medium for 2 h. After that, the cell suspensions were incubated with or without PAO1-CDPs for 1 h at 37 °C. At the indicated time points, samples (100 μL) were resuspended in PBS and mixed with the Rhodamine 123 and DHE probes for quantitation of mitochondrial membrane potential and superoxide, respectively. The samples were incubated for 30 min and washed, and fluorescence was measured in real-time by flow cytometry. (a) Mitochondrial membrane potential and (b) superoxide quantification as percentage of fluorescent cells. Values are the mean of three independent experiments with 20,000 cells counted by flow cytometry per data point. SEM values are indicated as bars (n = 3), one-way ANOVA with Tukey’s post hoc test, significant differences (p < 0.01) are indicated by different lowercase letters. Full article ">Figure 5
Effects of the CDP mixture from P. aeruginosa PAO1 on Akt and S6k phosphorylation and expression in HeLa cells. HeLa cells were incubated in SS and CM media and treated with 0.01 or 0.1 μg/mL PAO1-CDP mixture. At the indicated time points, cells were harvested and disrupted by sonication, and the solubilized proteins were separated by denaturing polyacrylamide gel electrophoresis (SDS-PAGE). Gels were electroblotted to PVDF membranes, and protein bands immunodetected using the indicated antibodies [anti-Akt (C-20-R), anti-Akt-phosphoryled 1/2/3 (Ser 473-R), anti-p70 S6 kinase α (H-160), anti-phosphoryled-p70 S6 kinase α (Thr 389)-R, and anti-β-actin] as the first antibody and a horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG antibody as the second antibody. Images correspond to representative gels from at least three independent treatments (left). Data correspond to the mean of three independent assays; the band intensity was determined by densitometry using the Image J software (right). (a) Immunodetection using the anti-phosphorylated Akt-S473 and anti-Akt antibodies after 5 and 15 min of treatment with the PAO1-CDP mixture; (b) HeLa cells extracts were obtained of cultures grown in CM media and treated with 0.01 or 0.1 mg/mL PAO1-CDP mixture. The same membranes reveled with anti-phosphorylated Akt-S473 were after immunodetected with the next antibodies: anti-Akt, anti-S6k, anti-phosphorylated-S6k-T389 and anti-β-actin. The assay was repeated at least three times using cell extracts from different cultures and treatments. A representative immunodetection assay is shown, and their plots of band intensity quantitation are shown to the right of the images. Bars represent mean of three densitometry determinations. Two-ways ANOVA was carried out, with Tukey’s post hoc test; statistical significance (p < 0.05) of differences between treatments is indicated with different lowercase letters. Full article ">Figure 5 Cont.
Effects of the CDP mixture from P. aeruginosa PAO1 on Akt and S6k phosphorylation and expression in HeLa cells. HeLa cells were incubated in SS and CM media and treated with 0.01 or 0.1 μg/mL PAO1-CDP mixture. At the indicated time points, cells were harvested and disrupted by sonication, and the solubilized proteins were separated by denaturing polyacrylamide gel electrophoresis (SDS-PAGE). Gels were electroblotted to PVDF membranes, and protein bands immunodetected using the indicated antibodies [anti-Akt (C-20-R), anti-Akt-phosphoryled 1/2/3 (Ser 473-R), anti-p70 S6 kinase α (H-160), anti-phosphoryled-p70 S6 kinase α (Thr 389)-R, and anti-β-actin] as the first antibody and a horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG antibody as the second antibody. Images correspond to representative gels from at least three independent treatments (left). Data correspond to the mean of three independent assays; the band intensity was determined by densitometry using the Image J software (right). (a) Immunodetection using the anti-phosphorylated Akt-S473 and anti-Akt antibodies after 5 and 15 min of treatment with the PAO1-CDP mixture; (b) HeLa cells extracts were obtained of cultures grown in CM media and treated with 0.01 or 0.1 mg/mL PAO1-CDP mixture. The same membranes reveled with anti-phosphorylated Akt-S473 were after immunodetected with the next antibodies: anti-Akt, anti-S6k, anti-phosphorylated-S6k-T389 and anti-β-actin. The assay was repeated at least three times using cell extracts from different cultures and treatments. A representative immunodetection assay is shown, and their plots of band intensity quantitation are shown to the right of the images. Bars represent mean of three densitometry determinations. Two-ways ANOVA was carried out, with Tukey’s post hoc test; statistical significance (p < 0.05) of differences between treatments is indicated with different lowercase letters. Full article ">

596 KiB

Open AccessArticle

Phenolic Glycosides from Capsella bursa-pastoris (L.) Medik and Their Anti-Inflammatory Activity

by Joon Min Cha, Won Se Suh, Tae Hyun Lee, Lalita Subedi, Sun Yeou Kim and Kang Ro Lee

Molecules 2017, 22(6), 1023; https://doi.org/10.3390/molecules22061023 - 20 Jun 2017

Cited by 31 | Viewed by 9404

Abstract

A new sesquilignan glycoside 1, together with seven known phenolic glycosides 2–8 were isolated from the aerial parts of Capsella bursa-pastoris. The chemical structure of the new compound 1 was elucidated by extensive nuclear magnetic resonance (NMR) data (¹ [...] Read more.

A new sesquilignan glycoside 1, together with seven known phenolic glycosides 2–8 were isolated from the aerial parts of Capsella bursa-pastoris. The chemical structure of the new compound 1 was elucidated by extensive nuclear magnetic resonance (NMR) data (¹H- and ¹³C-NMR, ¹H-¹H correlation spectroscopy (¹H-¹H COSY), heteronuclear single-quantum correlation (HSQC), heteronuclear multiple bond correlation (HMBC), and nuclear overhauser effect spectroscopy (NOESY)) and HR-FABMS analysis. The anti-inflammatory effects of 1–8 were evaluated in lipopolysaccharide (LPS)-stimulated murine microglia BV-2 cells. Compounds 4 and 7 exhibited moderate inhibitory effects on nitric oxide production in LPS-activated BV-2 cells, with IC₅₀ values of 17.80 and 27.91 µM, respectively. Full article

(This article belongs to the Collection Bioactive Compounds)

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6331 KiB

Open AccessArticle

Complex Coacervation of Soy Proteins, Isoflavones and Chitosan

by Yu-Hsuan Hsiao, Sheng-Yang Hsia, Yin-Ching Chan and Jung-Feng Hsieh

Molecules 2017, 22(6), 1022; https://doi.org/10.3390/molecules22061022 - 20 Jun 2017

Cited by 9 | Viewed by 7874

Abstract

In this study, the chitosan-induced coacervation of soy protein-isoflavone complexes in soymilk was investigated. Most of the soymilk proteins, including ?-conglycinin (7S), glycinin (11S), and isoflavones, were found to coacervate into the soymilk pellet fraction (SPF) following the addition of 0.5% chitosan. The [...] Read more.

In this study, the chitosan-induced coacervation of soy protein-isoflavone complexes in soymilk was investigated. Most of the soymilk proteins, including ?-conglycinin (7S), glycinin (11S), and isoflavones, were found to coacervate into the soymilk pellet fraction (SPF) following the addition of 0.5% chitosan. The total protein in the soymilk supernatant fraction (SSF) decreased from 18.1 ± 0.3 mg/mL to 1.6 ± 0.1 mg/mL, and the pH values decreased slightly, from 6.6 ± 0.0 to 6.0 ± 0.0. The results of SDS-PAGE revealed that the 7S ?’, 7S ?, 7S ?, 11S A3, and 11S acidic subunits, as well as the 11S basic proteins in the SSF, decreased to 0.7 ± 0.5%, 0.2 ± 0.1%, 0.1 ± 0.0%, 0.2 ± 0.2%, 0.2 ± 0.2% and 0.3 ± 0.2%, respectively. We also found that isoflavones in the SSF, including daidzein, glycitein, and genistein, decreased to 9.6 ± 2.3%, 5.7 ± 0.9% and 5.9 ± 1.5%, respectively. HPLC analysis indicated that isoflavones mixed with soy proteins formed soy protein-isoflavone complexes and were precipitated into the SPF by 0.5% chitosan. Full article

(This article belongs to the Special Issue Protein-Carbohydrate Interactions)

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Figure 1
Changes in the total protein content of soymilk resulting following the addition of various quantities of chitosan (0.0%, 0.1%, 0.2%, 0.3%, 0.4%, or 0.5%). Samples were incubated at 30 °C for 1 h. ○: soymilk supernatant fraction (SSF); ●: soymilk pellet fraction (SPF). - - -: pH value (SSF). Vertical bars represent standard deviations. Full article ">Figure 2
Changes in the SDS-PAGE profiles of soymilk following the addition of different amounts of chitosan (0.0%, 0.1%, 0.2%, 0.3%, 0.4%, or 0.5%). Samples were incubated at 30 °C for 1 h. (A) soymilk supernatant fraction (SSF); (B) soymilk pellet fraction (SPF); M: protein marker. Full article ">Figure 2 Cont.
Changes in the SDS-PAGE profiles of soymilk following the addition of different amounts of chitosan (0.0%, 0.1%, 0.2%, 0.3%, 0.4%, or 0.5%). Samples were incubated at 30 °C for 1 h. (A) soymilk supernatant fraction (SSF); (B) soymilk pellet fraction (SPF); M: protein marker. Full article ">Figure 3
Densitograms corresponding to the SDS-PAGE analysis of soymilk proteins containing various concentrations of chitosan (0.0%, 0.1%, 0.2%, 0.3%, 0.4% or 0.5%). Samples were incubated at 30 °C for 1 h. (A) soymilk supernatant fraction (SSF); (B) soymilk pellet fraction (SPF). Full article ">Figure 4
HPLC chromatograms: (A) isoflavone standards; (B) isoflavones extracted from soymilk. Full article ">Figure 5
Changes in the percentage of isoflavone content (%) in soymilk samples with various quantities of chitosan (0.0%, 0.1%, 0.2%, 0.3%, 0.4% or 0.5%). Samples were incubated at 30 °C for 1 h. (A) soymilk supernatant fraction (SSF); (B) soymilk pellet fraction (SPF). Vertical bars represent standard deviations. Full article ">Figure 6
Changes in the isoflavone contents with different amounts of chitosan (0.0%, 0.1%, 0.2%, 0.3%, 0.4% or 0.5%) at 30 °C for 1 h. Vertical bars represent standard deviations. Full article ">Figure 7
Reaction scheme illustrating the effects of chitosan on the coacervation of β-conglycinin, glycinin, and isoflavones in soymilk. Full article ">

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Open AccessArticle

The Effects of Selected Sesquiterpenes from Myrica rubra Essential Oil on the Efficacy of Doxorubicin in Sensitive and Resistant Cancer Cell Lines

by Martin Ambrož, Petra Matoušková, Adam Skarka, Martina Zajdlová, Kateřina Žáková and Lenka Skálová

Molecules 2017, 22(6), 1021; https://doi.org/10.3390/molecules22061021 - 20 Jun 2017

Cited by 32 | Viewed by 6044

Abstract

?-caryophyllene oxide (CAO), ?-humulene (HUM), trans-nerolidol (NER) and valencene (VAL) are constituents of the essential oil of Myrica rubra (MEO), which has significant antiproliferative effect in various cancer cell lines. In the present study, we compared the antiproliferative effect of these sesquiterpenes alone [...] Read more.

?-caryophyllene oxide (CAO), ?-humulene (HUM), trans-nerolidol (NER) and valencene (VAL) are constituents of the essential oil of Myrica rubra (MEO), which has significant antiproliferative effect in various cancer cell lines. In the present study, we compared the antiproliferative effect of these sesquiterpenes alone and in combination with the cytostatic drug doxorubicin (DOX) in cancer cell lines with different sensitivity to DOX. Two ovarian cancer cell lines (sensitive A2780 and partly resistant SKOV3) and two lymphoblast cancer cell lines (sensitive CCRF/CEM and completely resistant CEM/ADR) were used. The observed effects varied among sesquiterpenes and also differed in individual cell lines, with only VAL being effective in all the cell lines. A strong synergism of DOX with NER was found in the A2780 cells, while DOX acted synergistically with HUM and CAO in the SKOV3 cells. In the CCRF/CEM cells, a synergism of DOX with CAO and NER was observed. In resistant CEM/ADR cells, sesquiterpenes did not increase DOX efficacy, although they significantly increased accumulation of DOX (up to 10-times) and rhodamine-123 (substrate of efflux transporter ABCB1) within cancer cells. In conclusion, the tested sesquiterpenes were able to improve DOX efficacy in the sensitive and partly resistant cancer cells, but not in cells completely resistant to DOX. Full article

(This article belongs to the Special Issue Essential Oils: Chemistry and Bioactivity)

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Figure 1
Effect of sesquiterpenes on the proliferation of ovarian cell lines A2780 (a) and SKOV3 (b). The number of viable cells was assayed using NRU. Data presented as a percentage share of controls (=100%) represent the mean ± S.D. calculated from 3 independent measurements (6 parallels in each). The asterisk indicates a significant difference from the control cells (p < 0.05). Full article ">Figure 2
Effect of sesquiterpenes on proliferation of lymphoblast cell lines CCRF/CEM (a) and CEM/ADR (b). The number of viable cells was assayed using AB. Data presented as a percentage share of controls (=100%) represent the mean ± S.D. calculated from 3 independent measurements (6 parallels in each). The asterisk indicates a significant difference from the control cells (p < 0.05). Full article ">Figure 3
Combination indexes (CI) of DOX and sesquiterpenes in dependence of the fraction of affected cells (Fa) in ovarian cancer cell lines A2780 (a) and SKOV3 (b) and lymphoblast cell line CCRF/CEM (c). Incubations lasted for 72 h. Data were calculated using CalcuSyn software. Full article ">Figure 3 Cont.
Combination indexes (CI) of DOX and sesquiterpenes in dependence of the fraction of affected cells (Fa) in ovarian cancer cell lines A2780 (a) and SKOV3 (b) and lymphoblast cell line CCRF/CEM (c). Incubations lasted for 72 h. Data were calculated using CalcuSyn software. Full article ">Figure 4
Immuno-quantification of ABCB1 level in the tested cancer cell lines. Protein β-actin was used as a reference housekeeping one. Full article ">Figure 5
Effect of sesquiterpenes (40 and 200 µM) on intracellular DOX concentration. Data presented DOX intracellular concentration (in percentages), with DOX alone serving as control (100%). The colored bars represent the mean ± S.D. calculated from 3 independent measurements. The asterisk indicates a significant difference from the control cells (p < 0.05). Full article ">Figure 6
Fluorescence of RHO 123 in the CEM/ADR cell line. Untreated control cells represent 100%. Verapamil 20 µM was used as a positive control. Inhibition of efflux transporters is indicated by the increase of fluorescence. Data represent the mean ± S.D. calculated from 3 independent measurements (6 parallels in each). The asterisk indicates a significant difference from the control cells (p < 0.05). Full article ">

3608 KiB

Open AccessArticle

Determination of Structural Requirements of N-Substituted Tetrahydro-?-Carboline Imidazolium Salt Derivatives Using in Silico Approaches for Designing MEK-1 Inhibitors

by Jingwei Liang, Mingyang Wang, Xinyang Li, Xin He, Chong Cao and Fanhao Meng

Molecules 2017, 22(6), 1020; https://doi.org/10.3390/molecules22061020 - 19 Jun 2017

Cited by 7 | Viewed by 5824

Abstract

Novel N-substituted tetrahydro-?-carboline imidazolium salt derivatives proved to have potent antitumor activity in past research. The Topomer CoMFA and CoMSIA function in Sybyl-X 2.0 software was applied for the identification of important features of N-substituted tetrahydro-?-carboline-imidazolium salt derivative moieties. In the [...] Read more.

Novel N-substituted tetrahydro-?-carboline imidazolium salt derivatives proved to have potent antitumor activity in past research. The Topomer CoMFA and CoMSIA function in Sybyl-X 2.0 software was applied for the identification of important features of N-substituted tetrahydro-?-carboline-imidazolium salt derivative moieties. In the case of Topomer CoMFA, all the compounds were split into two fragments which were used to generate a 3D invariant representation, the statistical results of the Topomer CoMFA model: q² value of 0.700; r² value of 0.954; with 5 optimum components. The database alignment was utilized for building the CoMSIA model, and the CoMSIA model had q² and r² values of 0.615 and 0.897, with 4 optimum components. Target fishing of the PharmMapper platform was utilised for finding potential targets, the human mitogen-activated protein kinase 1 (MEK-1) was found to be the primary potential target for the three compounds with the fit scores of 6.288, 5.741, and 6.721. The molecular docking technique of MOE 2015 was carried out to identify the interactions of amino acids surrounding the ligand, and correlating QASR contour maps were used to identify structural requirements of N-substituted tetrahydro-?-carboline imidazolium salt moieties. Molecular dynamics and simulation studies proved that the target protein was stable for 0.8–5 ns. The pivotal moieties of N-substituted tetrahydro-?-carboline imidazolium salt derivatives and its potential targets were verified by the QASR study, PharmMapper, and the molecular docking study which would be helpful to design novel MEK-1 inhibitors for anticancer drugs. Full article

(This article belongs to the Collection Efficient Pharmaceutical and Chemical Approaches for Anticancer Therapy: Design, Preliminary Evaluations, and Further Developments)

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1849 KiB

Open AccessReview

Leptadenia reticulata (Retz.) Wight & Arn. (Jivanti): Botanical, Agronomical, Phytochemical, Pharmacological, and Biotechnological Aspects

by Sudipta Kumar Mohanty, Mallappa Kumara Swamy, Uma Rani Sinniah and Maniyam Anuradha

Molecules 2017, 22(6), 1019; https://doi.org/10.3390/molecules22061019 - 19 Jun 2017

Cited by 87 | Viewed by 11965

Abstract

Leptadenia reticulata (Retz.) Wight & Arn. (Apocynaceae), is a traditional medicinal plant species widely used to treat various ailments such as tuberculosis, hematopoiesis, emaciation, cough, dyspnea, fever, burning sensation, night blindness, cancer, and dysentery. In Ayurveda, it is known for its revitalizing, rejuvenating, [...] Read more.

Leptadenia reticulata (Retz.) Wight & Arn. (Apocynaceae), is a traditional medicinal plant species widely used to treat various ailments such as tuberculosis, hematopoiesis, emaciation, cough, dyspnea, fever, burning sensation, night blindness, cancer, and dysentery. In Ayurveda, it is known for its revitalizing, rejuvenating, and lactogenic properties. This plant is one of the major ingredients in many commercial herbal formulations, including Speman, Envirocare, Calshakti, Antisept, and Chyawanprash. The therapeutic potential of this herb is because of the presence of diverse bioactive compounds such as ?-amyrin, ?-amyrin, ferulic acid, luteolin, diosmetin, rutin, ?-sitosterol, stigmasterol, hentricontanol, a triterpene alcohol simiarenol, apigenin, reticulin, deniculatin, and leptaculatin. However, most biological studies on L. reticulata are restricted to crude extracts, and many biologically active compounds are yet to be identified in order to base the traditional uses of L. reticulata on evidence-based data. At present, L. reticulata is a threatened endangered plant because of overexploitation, unscientific harvesting, and habitat loss. The increased demand from pharmaceutical, nutraceutical, and veterinary industries has prompted its large-scale propagation. However, its commercial cultivation is hampered because of the non-availability of genuine planting material and the lack of knowledge about its agronomical practices. In this regard, micropropagation techniques will be useful to obtain true-to-type L. reticulata planting materials from an elite germplasm to meet the current demand. Adopting other biotechnological approaches such as synthetic seed technology, cryopreservation, cell culture, and genetic transformation can help conservation as well as increased metabolite production from L. reticulata. The present review summarizes scientific information on the botanical, agronomical, phytochemical, pharmacological, and biotechnological aspects of L. reticulata. This comprehensive information will certainly allow better utilization of this industrially important herb towards the discovery of lead drug molecules. Full article

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2393 KiB

Open AccessArticle

Systemic Induction of the Defensin and Phytoalexin Pisatin Pathways in Pea (Pisum sativum) against Aphanomyces euteiches by Acetylated and Nonacetylated Oligogalacturonides

by Sameh Selim, Jean Sanssené, Stéphanie Rossard and Josiane Courtois

Molecules 2017, 22(6), 1017; https://doi.org/10.3390/molecules22061017 - 19 Jun 2017

Cited by 26 | Viewed by 7878

Abstract

Oligogalacturonides (OGs) are known for their powerful ability to stimulate the plant immune system but little is known about their mode of action in pea (Pisum sativum). In the present study, we investigated the elicitor activity of two fractions of OGs, [...] Read more.

Oligogalacturonides (OGs) are known for their powerful ability to stimulate the plant immune system but little is known about their mode of action in pea (Pisum sativum). In the present study, we investigated the elicitor activity of two fractions of OGs, with polymerization degrees (DPs) of 2–25, in pea against Aphanomyces euteiches. One fraction was nonacetylated (OGs ? Ac) whereas the second one was 30% acetylated (OGs + Ac). OGs were applied by injecting the upper two rachises of the plants at three- and/or four-weeks-old. Five-week-old roots were inoculated with 10⁵ zoospores of A. euteiches. The root infection level was determined at 7, 10 and 14 days after inoculation using the quantitative real-time polymerase chain reaction (qPCR). Results showed significant root infection reductions namely 58, 45 and 48% in the plants treated with 80 µg OGs + Ac and 59, 56 and 65% with 200 µg of OGs ? Ac. Gene expression results showed the upregulation of genes involved in the antifungal defensins, lignans and the phytoalexin pisatin pathways and a priming effect in the basal defense, SA and ROS gene markers as a response to OGs. The reduction of the efficient dose in OGs + Ac is suggesting that acetylation is necessary for some specific responses. Our work provides the first evidence for the potential of OGs in the defense induction in pea against Aphanomyces root rot. Full article

(This article belongs to the Special Issue Structure, Chemical Analysis, Biosynthesis, Metabolism, Molecular Engineering and Biological Functions of Phytoalexins)

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3378 KiB

Open AccessPerspective

Photophysics and Photochemistry of Canonical Nucleobases’ Thioanalogs: From Quantum Mechanical Studies to Time Resolved Experiments

by Serra Arslancan, Lara Martínez-Fernández and Inés Corral

Molecules 2017, 22(6), 998; https://doi.org/10.3390/molecules22060998 - 18 Jun 2017

Cited by 59 | Viewed by 7521

Abstract

Interest in understanding the photophysics and photochemistry of thiated nucleobases has been awakened because of their possible involvement in primordial RNA or their potential use as photosensitizers in medicinal chemistry. The interpretation of the photodynamics of these systems, conditioned by their intricate potential [...] Read more.

Interest in understanding the photophysics and photochemistry of thiated nucleobases has been awakened because of their possible involvement in primordial RNA or their potential use as photosensitizers in medicinal chemistry. The interpretation of the photodynamics of these systems, conditioned by their intricate potential energy surfaces, requires the powerful interplay between experimental measurements and state of the art molecular simulations. In this review, we provide an overview on the photophysics of natural nucleobases’ thioanalogs, which covers the last 30 years and both experimental and computational contributions. For all the canonical nucleobase’s thioanalogs, we have compiled the main steady state absorption and emission features and their interpretation in terms of theoretical calculations. Then, we revise the main topographical features, including stationary points and interstate crossings, of their potential energy surfaces based on quantum mechanical calculations and we conclude, by combining the outcome of different spectroscopic techniques and molecular dynamics simulations, with the mechanism by which these nucleobase analogs populate their triplet excited states, which are at the origin of their photosensitizing properties. Full article

(This article belongs to the Special Issue Experimental and Computational Photochemistry of Bioorganic Molecules)

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1406 KiB

Open AccessArticle

Structural Characterization of a Rhamnogalacturonan I Domain from Ginseng and Its Inhibitory Effect on Galectin-3

by Huimin Shi, Li Yu, Yun Shi, Jiaojiao Lu, He Teng, Yifa Zhou and Lin Sun

Molecules 2017, 22(6), 1016; https://doi.org/10.3390/molecules22061016 - 18 Jun 2017

Cited by 32 | Viewed by 6879

Abstract

A rhamnogalacturonan I domain, named RG-I-3A, was prepared from ginseng pectin by pectinase digestion and chromatography separation. Monosaccharide composition analysis revealed that it was mainly composed of galacturonic acid, rhamnose, galactose, and arabinose in a molar ratio of 32.5:11.2:31.9:16.5, with a molecular weight [...] Read more.

A rhamnogalacturonan I domain, named RG-I-3A, was prepared from ginseng pectin by pectinase digestion and chromatography separation. Monosaccharide composition analysis revealed that it was mainly composed of galacturonic acid, rhamnose, galactose, and arabinose in a molar ratio of 32.5:11.2:31.9:16.5, with a molecular weight of 50 kDa. Partial acid hydrolysis, monoclonal antibody detection, and NMR spectra analysis suggested RG-I-3A was composed of ?4)-?-GalpA-(1?2)-?-Rhap-(1?disaccharide repeating units as backbone, with ?-1,4-galactan, ?-1,5-arabinan, AG-I, and AG-II side chains substituted via the O-4 of Rhap. Galectin-3-mediated hemagglutination and biolayer interferometry assay indicated that RG-I-3A had inhibitory activity on galectin-3. These findings suggest the potential use of this ginseng RG-I domain as a galectin-3 inhibitor in drug development applications. Full article

(This article belongs to the Special Issue Natural Polysaccharides)

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13733 KiB

Open AccessArticle

Repositioning FDA Drugs as Potential Cruzain Inhibitors from Trypanosoma cruzi: Virtual Screening, In Vitro and In Vivo Studies

by Isidro Palos, Edgar E. Lara-Ramirez, Julio Cesar Lopez-Cedillo, Carlos Garcia-Perez, Muhammad Kashif, Virgilio Bocanegra-Garcia, Benjamin Nogueda-Torres and Gildardo Rivera

Molecules 2017, 22(6), 1015; https://doi.org/10.3390/molecules22061015 - 18 Jun 2017

Cited by 38 | Viewed by 7370

Abstract

Chagas disease (CD) is a neglected disease caused by the parasite Trypanosoma cruzi, which affects underdeveloped countries. The current drugs of choice are nifurtimox and benznidazole, but both have severe adverse effects and less effectivity in chronic infections; therefore, the need to [...] Read more.

Chagas disease (CD) is a neglected disease caused by the parasite Trypanosoma cruzi, which affects underdeveloped countries. The current drugs of choice are nifurtimox and benznidazole, but both have severe adverse effects and less effectivity in chronic infections; therefore, the need to discover new drugs is essential. A computer-guided drug repositioning method was applied to identify potential FDA drugs (approved and withdrawn) as cruzain (Cz) inhibitors and trypanocidal effects were confirmed by in vitro and in vivo studies. 3180 FDA drugs were virtually screened using a structure-based approach. From a first molecular docking analysis, a set of 33 compounds with the best binding energies were selected. Subsequent consensus affinity binding, ligand amino acid contact clustering analysis, and ranked position were used to choose four known pharmacological compounds to be tested in vitro. Mouse blood samples infected with trypomastigotes from INC-5 and NINOA strains were used to test the trypanocidal effect of four selected compounds. Among these drugs, one fibrate antilipemic (etofyllin clofibrate) and three ?-lactam antibiotics (piperacillin, cefoperazone, and flucloxacillin) showed better trypanocidal effects (LC₅₀ range 15.8–26.1 ?g/mL) in comparison with benznidazole and nifurtimox (LC₅₀ range 33.1–46.7 ?g/mL). A short-term in vivo evaluation of these compounds showed a reduction of parasitemia in infected mice (range 90–60%) at 6 h, but this was low compared to benznidazole (50%). This work suggests that four known FDA drugs could be used to design and obtain new trypanocidal agents. Full article

(This article belongs to the Special Issue Emerging Drug Discovery Approaches against Infectious Diseases)

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Full article ">Figure 1
Structural comparison of the 4W5B protein with 1F2A protein. In green the 4W5B protein, in pale cyan the 1F2A protein that contain the known catalytic triad. The catalytic amino acids indicated by arrows are colored as follow: 1F2A in magenta and 4W5B in red. The compound N-(1H-benzimidazol-2-yl)-1,3-dimethyl-pyrazole-4-carboxamide is shown in blue. Full article ">Figure 2
Clustering pattern based on ligand contact amino acid showing the four major groups. In cyan, group 1, in blue, group 2, in red, group 3, and in green, group 4. Arrows indicate the compound ZINC03830554 and ZINC03831344 mentioned in the text. Full article ">Figure 3
The four selected compounds docked in the 4W5B Cz active sites. (A) Flucloxacillin sodium, (B) Cefoperazone sodium, (C) Piperacillin sodium, (D) Etofyllin clofibrate. In the four figures arcs with red lines represent amino acid hydrophobic contacts, green dashed lines represent hydrogen bonds, the red circles represent the shared amino acids. The image was produced with LigPlot software [<a href="#B17-molecules-22-01015" class="html-bibr">17</a>]. Full article ">Figure 4
Effects of the compounds in reducing parasites on mice infected with INC-5 (A) and NINOA (B) strains from T. cruzi over a period of 6 h. Full article ">

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Open AccessReview

Cancer Chemoprevention by Resveratrol: The p53 Tumor Suppressor Protein as a Promising Molecular Target

by Danielly C. Ferraz da Costa, Eliane Fialho and Jerson L. Silva

Molecules 2017, 22(6), 1014; https://doi.org/10.3390/molecules22061014 - 18 Jun 2017

Cited by 57 | Viewed by 18573

Abstract

Increasing epidemiological and experimental evidence has demonstrated an inverse relationship between the consumption of plant foods and the incidence of chronic diseases, including cancer. Microcomponents that are naturally present in such foods, especially polyphenols, are responsible for the benefits to human health. Resveratrol [...] Read more.

Increasing epidemiological and experimental evidence has demonstrated an inverse relationship between the consumption of plant foods and the incidence of chronic diseases, including cancer. Microcomponents that are naturally present in such foods, especially polyphenols, are responsible for the benefits to human health. Resveratrol is a diet-derived cancer chemopreventive agent with high therapeutic potential, as demonstrated by different authors. The aim of this review is to collect and present recent evidence from the literature regarding resveratrol and its effects on cancer prevention, molecular signaling (especially regarding the involvement of p53 protein), and therapeutic perspectives with an emphasis on clinical trial results to date. Full article

(This article belongs to the Special Issue Improvements for Resveratrol Efficacy)

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4018 KiB

Open AccessArticle

Mechanistic Explanation of the Weak Carbonic Anhydrase’s Esterase Activity

by Paolo Piazzetta, Tiziana Marino and Nino Russo

Molecules 2017, 22(6), 1009; https://doi.org/10.3390/molecules22061009 - 18 Jun 2017

Cited by 8 | Viewed by 6193

Abstract

In order to elucidate the elementary mechanism of the promiscuous esterase activity of human carbonic anhydrase (h-CA), we present an accurate theoretical investigation on the hydrolysis of fully-acetylated d-glucose functionalized as sulfamate. This h-CA’s inhibitor is of potential relevance in cancer therapy. [...] Read more.

In order to elucidate the elementary mechanism of the promiscuous esterase activity of human carbonic anhydrase (h-CA), we present an accurate theoretical investigation on the hydrolysis of fully-acetylated d-glucose functionalized as sulfamate. This h-CA’s inhibitor is of potential relevance in cancer therapy. The study has been performed within the framework of three-layer ONIOM (QM-high:QM’-medium:MM-low) hybrid approach. The computations revealed that the hydrolysis process is not energetically favored, in agreement with the observed weak carbonic anhydrase’s esterase activity. Full article

(This article belongs to the Special Issue Metallopeptides)

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2868 KiB

Open AccessArticle

Metabolomics Characterization of Two Apocynaceae Plants, Catharanthus roseus and Vinca minor, Using GC-MS and LC-MS Methods in Combination

by Qi Chen, Xueyan Lu, Xiaorui Guo, Qingxi Guo and Dewen Li

Molecules 2017, 22(6), 997; https://doi.org/10.3390/molecules22060997 - 17 Jun 2017

Cited by 44 | Viewed by 8915

Abstract

Catharanthus roseus (C. roseus) and Vinca minor (V. minor) are two common important medical plants belonging to the family Apocynaceae. In this study, we used non-targeted GC-MS and targeted LC-MS metabolomics to dissect the metabolic profile of two plants [...] Read more.

Catharanthus roseus (C. roseus) and Vinca minor (V. minor) are two common important medical plants belonging to the family Apocynaceae. In this study, we used non-targeted GC-MS and targeted LC-MS metabolomics to dissect the metabolic profile of two plants with comparable phenotypic and metabolic differences. A total of 58 significantly different metabolites were present in different quantities according to PCA and PLS-DA score plots of the GC-MS analysis. The 58 identified compounds comprised 16 sugars, eight amino acids, nine alcohols and 18 organic acids. We subjected these metabolites into KEGG pathway enrichment analysis and highlighted 27 metabolic pathways, concentrated on the TCA cycle, glycometabolism, oligosaccharides, and polyol and lipid transporter (RFOS). Among the primary metabolites, trehalose, raffinose, digalacturonic acid and gallic acid were revealed to be the most significant marker compounds between the two plants, presumably contributing to species-specific phenotypic and metabolic discrepancy. The profiling of nine typical alkaloids in both plants using LC-MS method highlighted higher levels of crucial terpenoid indole alkaloid (TIA) intermediates of loganin, serpentine, and tabersonine in V. minor than in C. roseus. The possible underlying process of the metabolic flux from primary metabolism pathways to TIA synthesis was discussed and proposed. Generally speaking, this work provides a full-scale comparison of primary and secondary metabolites between two medical plants and a metabolic explanation of their TIA accumulation and phenotype differences. Full article

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Leaf phenotypes of Catharanthus roseus and Vinca minor. (a) C. roseus; (b) V. minor. Full article ">Figure 2
The multivariate analysis of primary metabolites in Catharanthus roseus and Vinca minor. (a) PCA score plot; (b) PLS-DA score plot. Blue indicates C. roseus, red indicates V. minor, and yellow indicates Quality Control. Full article ">Figure 3
The histogram of different pathway −log (p-value, 10). The horizontal line of 1.3 indicates p < 0.05, the horizontal line of 1.5 indicates p < 0.01. Full article ">Figure 4
The cluster of metabolites and energy Q values between Catharanthus roseus and Vinca minor. (a) Heat map visualization of relative differences of metabolites in C. roseus and V. minor. The content value of each metabolite was normalized to complete linkage hierarchical clustering. The metabolites were glycolic acid, threonic acid, threitol, anhydroglucitol, digalacturonic acid, galactonic acid, gentiobiose, octanal, sorbitol, itaconic acid, cellobiose, maleamate, methyl- galactopyranoside, glucopyranoside, glycocyamine, aminooxyacetic acid, loganin, threonine, threonine, glyceric acid, dihydroxybenzoic acid, tartaric acid, quinic acid, glucoheptonic acid, trehalose, hydroxybutyrate, lactitol, levoglucosan, gallic acid, ethanolamine, lyxose, aminobutyric acid, succinic acid, salicin, maltotriose, raffinose, saccharic acid, isoleucine, lucose-1-phosphate, hydroxypropionic acid, methionine, aminoisobutyric acid, serine, ornithine, dodecanol, methyl phosphate, erythrose, methoxytryptamine, mono phthalate, fucose, caffeic acid, xylitol, nicotinoylglycine, malic acid, citramalic acid, glycolic acid, pyruvic acid, allothreonine, aspartic acid, oxoproline, galactinol, chlorogenic acid, ribose, myo-inositol, tagatose, fructose, talose, mannitol, glucose, anhydrogalactose, phosphate, fumaric acid, gluconic lactone, dimethylsuccinic acid, glucose-6-phosphate, valine, tyrosine, amino-1-phenylethanol, glutamic acid, maleic acid, glutamine, isocitric acid, alpha-ketoglutaric acid, acetyl-glutamic acid, galactose, phytol, norleucine, oxalic acid, mannosylglycerate, glutaconic acid, alanine, lysine, lactic acid, methyl-amino-1,2-propanediol, glycerol, proline, hydroxynorvaline, from top to bottom, in turn. Red indicates high abundance, whereas low relative metabolites are green. V1, V2, V3, V4, V5, V6 indicate V. minor, C1, C2, C3, C4, C5, C6 indicate C. roseus; (b) The Q value of energy between Catharanthus roseus and Vinca minor. Full article ">Figure 4 Cont.
The cluster of metabolites and energy Q values between Catharanthus roseus and Vinca minor. (a) Heat map visualization of relative differences of metabolites in C. roseus and V. minor. The content value of each metabolite was normalized to complete linkage hierarchical clustering. The metabolites were glycolic acid, threonic acid, threitol, anhydroglucitol, digalacturonic acid, galactonic acid, gentiobiose, octanal, sorbitol, itaconic acid, cellobiose, maleamate, methyl- galactopyranoside, glucopyranoside, glycocyamine, aminooxyacetic acid, loganin, threonine, threonine, glyceric acid, dihydroxybenzoic acid, tartaric acid, quinic acid, glucoheptonic acid, trehalose, hydroxybutyrate, lactitol, levoglucosan, gallic acid, ethanolamine, lyxose, aminobutyric acid, succinic acid, salicin, maltotriose, raffinose, saccharic acid, isoleucine, lucose-1-phosphate, hydroxypropionic acid, methionine, aminoisobutyric acid, serine, ornithine, dodecanol, methyl phosphate, erythrose, methoxytryptamine, mono phthalate, fucose, caffeic acid, xylitol, nicotinoylglycine, malic acid, citramalic acid, glycolic acid, pyruvic acid, allothreonine, aspartic acid, oxoproline, galactinol, chlorogenic acid, ribose, myo-inositol, tagatose, fructose, talose, mannitol, glucose, anhydrogalactose, phosphate, fumaric acid, gluconic lactone, dimethylsuccinic acid, glucose-6-phosphate, valine, tyrosine, amino-1-phenylethanol, glutamic acid, maleic acid, glutamine, isocitric acid, alpha-ketoglutaric acid, acetyl-glutamic acid, galactose, phytol, norleucine, oxalic acid, mannosylglycerate, glutaconic acid, alanine, lysine, lactic acid, methyl-amino-1,2-propanediol, glycerol, proline, hydroxynorvaline, from top to bottom, in turn. Red indicates high abundance, whereas low relative metabolites are green. V1, V2, V3, V4, V5, V6 indicate V. minor, C1, C2, C3, C4, C5, C6 indicate C. roseus; (b) The Q value of energy between Catharanthus roseus and Vinca minor. Full article ">Figure 5
The relative content of major different metabolites and biochemical pathway map. (a) Relative content of trehalose, cellobise, gallic acid, digalacturonic acid, raffinose and chlorogenic acid. (b) Visualization of secondary metabolites, photosynthesis and TCA cycle in a biochemical pathway map. Common metabolites are written black, V. minor ones are written in yellow, and red metabolites represent no content to display, C. roseus are written in blue, full lines represent one step reactions, and dashed lines represent multi-step reactions. Full article ">Figure 5 Cont.
The relative content of major different metabolites and biochemical pathway map. (a) Relative content of trehalose, cellobise, gallic acid, digalacturonic acid, raffinose and chlorogenic acid. (b) Visualization of secondary metabolites, photosynthesis and TCA cycle in a biochemical pathway map. Common metabolites are written black, V. minor ones are written in yellow, and red metabolites represent no content to display, C. roseus are written in blue, full lines represent one step reactions, and dashed lines represent multi-step reactions. Full article ">

884 KiB

Open AccessArticle

Sensory Characteristics and Volatile Components of Dry Dog Foods Manufactured with Sorghum Fractions

by Brizio Di Donfrancesco and Kadri Koppel

Molecules 2017, 22(6), 1012; https://doi.org/10.3390/molecules22061012 - 17 Jun 2017

Cited by 13 | Viewed by 6551

Abstract

Descriptive sensory analysis and gas chromatography-mass spectrometry (GC-MS) with a modified headspace solid-phase microextraction (SPME) method was performed on three extruded dry dog food diets manufactured with different fractions of red sorghum and a control diet containing corn, brewer’s rice, and wheat as [...] Read more.

Descriptive sensory analysis and gas chromatography-mass spectrometry (GC-MS) with a modified headspace solid-phase microextraction (SPME) method was performed on three extruded dry dog food diets manufactured with different fractions of red sorghum and a control diet containing corn, brewer’s rice, and wheat as a grain source in order to determine the effect of sorghum fractions on dry dog food sensory properties. The aroma compounds and flavor profiles of samples were similar with small differences, such as higher toasted aroma notes, and musty and dusty flavor in the mill-feed sample. A total of 37 compounds were tentatively identified and semi-quantified. Aldehydes were the major group present in the samples. The total volatile concentration was low, reflecting the mild aroma of the samples. Partial least squares regression was performed to identify correlations between sensory characteristics and detected aroma compounds. Possible relationships, such as hexanal and oxidized oil, and broth aromatics were identified. Volatile compounds were also associated with earthy, musty, and meaty aromas and flavor notes. This study showed that extruded dry dog foods manufactured with different red sorghum fractions had similar aroma, flavor, and volatile profiles. Full article

(This article belongs to the Collection Recent Advances in Flavors and Fragrances)

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1828 KiB

Open AccessArticle

Synthesis and Evaluation of Phenylxanthine Derivatives as Potential Dual A2AR Antagonists/MAO-B Inhibitors for Parkinson’s Disease

by Xuebao Wang, Chao Han, Yong Xu, Kaiqi Wu, Shuangya Chen, Mangsha Hu, Luyao Wang, Yun Ye and Faqing Ye

Molecules 2017, 22(6), 1010; https://doi.org/10.3390/molecules22061010 - 17 Jun 2017

Cited by 16 | Viewed by 6271

Abstract

The aim of this research was to prove the speculation that phenylxanthine (PX) derivatives possess adenosine A2A receptor (A2AR)-blocking properties and to screening and evaluate these PX derivatives as dual A2AR antagonists/MAO-B inhibitors for Parkinson?s disease. To explore this hypothesis, two series of [...] Read more.

The aim of this research was to prove the speculation that phenylxanthine (PX) derivatives possess adenosine A2A receptor (A2AR)-blocking properties and to screening and evaluate these PX derivatives as dual A2AR antagonists/MAO-B inhibitors for Parkinson?s disease. To explore this hypothesis, two series of PX derivatives were prepared and their antagonism against A2AR and inhibition against MAO-B were determined in vitro. In order to evaluate further the antiparkinsonian properties, pharmacokinetic and haloperidol-induced catalepsy experiments were carried out in vivo. The PX-D and PX-E analogues acted as potent A2AR antagonists with Ki values ranging from 0.27 to 10 ?M, and these analogues displayed relatively mild MAO-B inhibition potencies, with inhibitor dissociation constants (Ki values) ranging from 0.25 to 10 ?M. Further, the compounds PX-D-P6 and PX-E-P8 displayed efficacious antiparkinsonian properties in haloperidol-induced catalepsy experiments, verifying that these two compounds were potent A2AR antagonists and MAO-B inhibitors. We conclude that PX-D and PX-E analogues are a promising candidate class of dual-acting compounds for treating Parkinson?s disease. Full article

(This article belongs to the Special Issue Polypharmacology and Multitarget Drug Discovery)

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2888 KiB

Open AccessArticle

Cholinesterase Inhibitory Activities of Adamantyl-Based Derivatives and Their Molecular Docking Studies

by Huey Chong Kwong, Siau Hui Mah, Tze Shyang Chia, Ching Kheng Quah, Gin Keat Lim and C. S. Chidan Kumar

Molecules 2017, 22(6), 1005; https://doi.org/10.3390/molecules22061005 - 17 Jun 2017

Cited by 9 | Viewed by 5371

Abstract

Adamantyl-based compounds are clinically important for the treatments of type 2 diabetes and for their antiviral abilities, while many more are under development for other pharmaceutical uses. This study focused on the acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibitory activities of adamantyl-based ester derivatives [...] Read more.

Adamantyl-based compounds are clinically important for the treatments of type 2 diabetes and for their antiviral abilities, while many more are under development for other pharmaceutical uses. This study focused on the acetylcholinesterase (AChE) and butyrylcholinesterase (BChE) inhibitory activities of adamantyl-based ester derivatives with various substituents on the phenyl ring using Ellman’s colorimetric method. Compound 2e with a 2,4-dichloro electron-withdrawing substituent on the phenyl ring exhibited the strongest inhibition effect against AChE, with an IC₅₀ value of 77.15 µM. Overall, the adamantyl-based ester with the mono-substituent at position 3 of the phenyl ring exhibited good AChE inhibition effects with an ascending order for the substituents: Cl < NO₂ < CH₃ < OCH₃. Furthermore, compounds with electron-withdrawing groups (Cl and NO₂) substituted at position 3 on their phenyl rings demonstrated stronger AChE inhibition effects, in comparison to their respective positional isomers. On the other hand, compound 2j with a 3-methoxyphenyl ring showed the highest inhibition effect against BChE, with an IC₅₀ value of 223.30 µM. Molecular docking analyses were conducted for potential AChE and BChE inhibitors, and the results demonstrated that the peripheral anionic sites of target proteins were predominant binding sites for these compounds through hydrogen bonds and halogen interactions instead of hydrophobic interactions in the catalytic active site. Full article

(This article belongs to the Special Issue The Cholinesterases—Structure, Mechanism, Function and Drug Design: The 25th Anniversary of the Solution of the Crystal Structure of Acetylcholinesterase by Joel L. Sussman and Israel Silman)

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Figure 1
The percentage of AChE inhibition as means ± SD (n = 3). Means with different alphabet letters are significantly different (p < 0.05). Full article ">Figure 2
Differential validation in the GOLD package by docking the native ligands of AChE into their binding sites: (a) The native co-crystallized tacrine represented by green sticks; docked ligands are shown in the form of balls and sticks, colored by element. Putative binding modes of: (b) compound 2c; (c) compound 2m; (d) compound 2g; (e) compound 2j; and (f) compound 2f. Full article ">Figure 3
Putative binding modes in AChE enzymes of: (a) compound 2b; (b) compound 2d; and (c) compound 2e. Full article ">Figure 4
Differential validation in the GOLD package by docking the native ligands of BChE into their binding sites: (a) The native co-crystallized tacrine is represented by green sticks, while the docked ligands are shown in the form of balls and sticks, colored by element. Putative binding modes of: (b) compound 2e in BChE, and (c) active compound 2j in BChE. Full article ">Figure 5
General reaction scheme for the preparation of 2-(adamantan-1-yl)-2-oxoethyl benzoates, 2a–q, and 2-(adamantan-1-yl)-2-oxoethyl-2-pyridinecarboxylate, 2r. Full article ">

4010 KiB

Open AccessArticle

In Vitro Comparative Study of the Inhibitory Effects of Mangiferin and Its Aglycone Norathyriol towards UDP-Glucuronosyl Transferase (UGT) Isoforms

by Dan Sun, Chun-Ze Zhang, Rui-Xue Ran, Yun-Feng Cao, Zuo Du, Zhi-Wei Fu, Chun-Ting Huang, Zhen-Ying Zhao, Wei-Hua Zhang and Zhong-Ze Fang

Molecules 2017, 22(6), 1008; https://doi.org/10.3390/molecules22061008 - 16 Jun 2017

Cited by 27 | Viewed by 5246

Abstract

Mangiferin (MGF), the predominant constituent of extracts of the mango plant Mangifera Indica L., has been investigated extensively because of its remarkable pharmacological effects. In vitro recombinant UGTs-catalyzed glucuronidation of 4-methylumbelliferone (4-MU) was used to investigate the inhibition of mangiferin and aglycone norathyriol [...] Read more.

Mangiferin (MGF), the predominant constituent of extracts of the mango plant Mangifera Indica L., has been investigated extensively because of its remarkable pharmacological effects. In vitro recombinant UGTs-catalyzed glucuronidation of 4-methylumbelliferone (4-MU) was used to investigate the inhibition of mangiferin and aglycone norathyriol towards various isoforms of UGTs in our study, which evaluated the inhibitory capacity of MGF and its aglycone norathyriol (NTR) towards UDP-glucuronosyltransferase (UGT) isoforms. Initial screening experiment showed that deglycosylation of MGF into NTR strongly increased the inhibitory effects towards almost all the tested UGT isoforms at a concentration of 100 ?M. Kinetic experiments were performed to further characterize the inhibition of UGT1A3, UGT1A7 and UGT1A9 by NTR. NTR competitively inhibited UGT1A3, UGT1A7 and UGT1A9, with an IC₅₀ value of 8.2, 4.4, and 12.3 ?M, and a Ki value of 1.6, 2.0, and 2.8 ?M, respectively. In silico docking showed that only NTR could dock into the activity cavity of UGT1A3, UGT1A7 and UGT1A9. The binding free energy of NTR to UGT1A3, 1A7, 1A9 were ?7.4, ?7.9 and ?4.0 kcal/mol, respectively. Based on the inhibition evaluation standard ([I]/Ki < 0.1, low possibility; 0.1 < [I]/Ki < 1, medium possibility; [I]/Ki > 1, high possibility), an in vivo herb–drug interaction between MGF/NTR and drugs mainly undergoing UGT1A3-, UGT1A7- or UGT1A9-catalyzed metabolism might occur when the plasma concentration of NTR is above 1.6, 2.0 and 2.8 ?M, respectively. Full article

(This article belongs to the Section Metabolites)

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4081 KiB

Open AccessArticle

Comprehensive Identification of Guan-Xin-Shu-Tong Capsule via a Mass Defect and Fragment Filtering Approach by High Resolution Mass Spectrometry: In Vitro and In Vivo Study

by Xun Gao, Jingqing Mu, Qing Li, Shaoyi Guan, Ran Liu, Yiyang Du, Huifen Zhang and Kaishun Bi

Molecules 2017, 22(6), 1007; https://doi.org/10.3390/molecules22061007 - 16 Jun 2017

Cited by 12 | Viewed by 5913

Abstract

The Guan-Xin-Shu-Tong capsule (GXSTC) is a well-known traditional Chinese medicine that is used for the treatment of coronary heart disease. Despite its common use in China, basic pharmacological research on its active components is limited. A comprehensive analytical method using quadrupole-time-of-flight mass spectrometry [...] Read more.

The Guan-Xin-Shu-Tong capsule (GXSTC) is a well-known traditional Chinese medicine that is used for the treatment of coronary heart disease. Despite its common use in China, basic pharmacological research on its active components is limited. A comprehensive analytical method using quadrupole-time-of-flight mass spectrometry (Q-TOF/MS), specifically with the Triple TOF 5600 platform, was developed to characterize the compounds in the GXSTC powder itself (in vitro) as well as the active components in healthy and heart disease model rats after its oral administration (in vivo). The 5600 platform was operated in both positive and negative ion modes, before the raw data were processed using the extracted ion chromatography (EIC), mass defect filtering (MDF) and fragment filtering (FF) techniques. With the aid of reference compounds for retention time and fragment ion comparisons, 18 compounds were unambiguously identified in vitro. An additional 56 other compounds were tentatively characterized using the accurate quasi-molecular ion mass and Tandem mass spectrometry (MS/MS) fragmentation pattern strategies. Among them, 30 compounds were characterized based on the MDF and FF approaches. Normal rats in addition to hyperlipidemic (HL) and acute blood stasis (ABS) model rats were given a single oral dose of GXSTC solution for subsequent blood analysis at 1 and 2 h after administration. A total of 24 prototypecomponents and 20 metabolites derived from GXSTC were differentially detected across the three animal groups, including the absence of four phase II phenolic acid metabolites in the ABS group and the presence of three diterpenoid-related metabolites exclusive to the HL group. The use of reference compounds as well as the mass defect and fragment-filtering strategies were critical to identify GXSTC compounds in vitro and in vivo. This can be used for further quality control and pharmacological studies aimed at characterizing the active and potential beneficial compounds of this ancient medicine. Full article

(This article belongs to the Section Natural Products Chemistry)

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1976 KiB

Open AccessArticle

Novel Tacrine-Scutellarin Hybrids as Multipotent Anti-Alzheimer’s Agents: Design, Synthesis and Biological Evaluation

by Katarina Spilovska, Jan Korabecny, Vendula Sepsova, Daniel Jun, Martina Hrabinova, Petr Jost, Lubica Muckova, Ondrej Soukup, Jana Janockova, Tomas Kucera, Rafael Dolezal, Eva Mezeiova, Daniel Kaping and Kamil Kuca

Molecules 2017, 22(6), 1006; https://doi.org/10.3390/molecules22061006 - 16 Jun 2017

Cited by 43 | Viewed by 8146

Abstract

A novel series of 6-chlorotacrine-scutellarin hybrids was designed, synthesized and the biological activity as potential anti-Alzheimer’s agents was assessed. Their inhibitory activity towards human acetylcholinesterase (hAChE) and human butyrylcholinesterase (hBChE), antioxidant activity, ability to cross the blood-brain barrier (BBB) [...] Read more.

A novel series of 6-chlorotacrine-scutellarin hybrids was designed, synthesized and the biological activity as potential anti-Alzheimer’s agents was assessed. Their inhibitory activity towards human acetylcholinesterase (hAChE) and human butyrylcholinesterase (hBChE), antioxidant activity, ability to cross the blood-brain barrier (BBB) and hepatotoxic profile were evaluated in vitro. Among these compounds, hybrid K1383, bearing two methylene tether between two basic scaffolds, was found to be very potent hAChE inhibitor (IC₅₀ = 1.63 nM). Unfortunately, none of the hybrids displayed any antioxidant activity (EC₅₀ ? 500 ?M). Preliminary data also suggests a comparable hepatotoxic profile with 6-Cl-THA (established on a HepG2 cell line). Kinetic studies performed on hAChE with the most active compound in the study, K1383, pointed out to a mixed, non-competitive enzyme inhibition. These findings were further corroborated by docking studies. Full article

(This article belongs to the Special Issue Polypharmacology and Multitarget Drug Discovery)

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Figure 1
Design strategy for the 6-Cl-THA-scutellarin hybrids K1383–K1389. Full article ">Figure 2
Steady-state inhibition of AChE hydrolysis of substrate acetylthiocholine (ATCh) by compound K1383 at different concentrations. Lineweaver-Burk plots of initial velocity at increasing substrate concentrations (0.156–1.250 mM) are presented. Lines were derived from a linear regression of the data points. Full article ">Figure 3
Superimposition of K1383 (A,B), K1384 (C,D) and K1385 (E,F) in hAChE active site. Important amino acid residues involved in enzyme-ligand interaction are depicted in light-blue carbon atoms, ligands in salmon carbons and catalytic triad in yellow (A,C,E). Three-dimensional figures (A,C,E) were created by PyMol viewer 1.3, 2D figures (B,D,F) were built by BIOVIA Discovery Studio software. Full article ">Figure 4
Superimposition of K1383 (A,B), K1384 (C,D) and K1385 (E,F) in hBChE active site. Important amino acid residues involved in enzyme-ligand interaction are depicted in light-blue carbon atoms, ligands in salmon carbons and catalytic triad in yellow (A,C,E). Three-dimensional Figures (A,C,E) were created by PyMol viewer 1.3., 2D Figures (B,D,F) were built by BIOVIA Discovery Studio software. Full article ">Scheme 1
Synthetic procedure for 6-chlorotacrine-scutellarin hybrids K1383–K1389. Reagents and conditions: (i) sodium acetate, acetic anhydride, 110 °C; (ii) BF3·Et2O, acetic acid, 70 °C; (iii) 4-bromo-benzaldehyde, EtOH, 30% KOH, r.t.; (iv) I2, DMSO, 170 °C; (v) POCl3, 130°C; (vi) phenol, appropriate 1,ω-diamines, 130 °C; (vii) Pd2(dba)3 + BINAP + NaOtBu, dry toluene, 110 °C. Full article ">

529 KiB

Open AccessArticle

Hydronopylformamides: Modification of the Naturally Occurring Compound (-)-?-Pinene to Produce Insect Repellent Candidates against Blattella germanica

by Shengliang Liao, Yan Liu, Hongyan Si, Zhuanquan Xiao, Guorong Fan, Shangxing Chen, Peng Wang and Zongde Wang

Molecules 2017, 22(6), 1004; https://doi.org/10.3390/molecules22061004 - 16 Jun 2017

Cited by 9 | Viewed by 5129

Abstract

The development of a novel repellent plays an important role in the integrated control of Blattella germanica. A series of novel hydronopylformamides derivatives were synthesized from a naturally occurring compound (-)-?-pinene. The structures of these hydronopylformamides derivatives were characterized by Fourier transform [...] Read more.

The development of a novel repellent plays an important role in the integrated control of Blattella germanica. A series of novel hydronopylformamides derivatives were synthesized from a naturally occurring compound (-)-?-pinene. The structures of these hydronopylformamides derivatives were characterized by Fourier transform infrared spectroscopy (FTIR), nuclear magnetic resonance spectroscopy (¹H-NMR and ¹³C-NMR), and electron impact mass spectrometry (EI-MS). Repellency of these hydronopylformamides derivatives against Blattella germanica was evaluated by the using petri dish arena method. The results showed that four derivatives (compounds 8a, 8b, 8c and 8e) exhibited repellency against Blattella germanica at a concentration of 20 mg/mL. Compound 8a was the most active compound among these derivatives, where the repelling ratios of compound 8a against Blattella germanica were 66.10%, 50.46%, 48.26%, at concentrations of 20 mg/mL, 10 mg/mL, and 5 mg/mL, respectively. In addition, compound 8a showed better repellency than the traditional insect repellent N, N-diethyl-3-methylbenzamide (DEET), which indicated that compound 8a had a good application prospect in the prevention of Blattella germanica. This research hopes to promote the value-added utilization of (-)-?-pinene and the development of novel German cockroach repellents. Full article

(This article belongs to the Special Issue Natural Product Inspired Scaffolds Designs)

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1847 KiB

Open AccessArticle

Anti-Inflammatory and Anti-Oxidant Potential of the Root Extract and Constituents of Doronicum austriacum

by Stefania Marzocco, Simona Adesso, Mostafa Alilou, Hermann Stuppner and Stefan Schwaiger

Molecules 2017, 22(6), 1003; https://doi.org/10.3390/molecules22061003 - 16 Jun 2017

Cited by 25 | Viewed by 4703

Abstract

Background: Doronicum austriacum Jacq., Asteraceae, is a plant which is used in traditional alpine medicine. Historical sources describe the medical use of the root, but up until now only a few studies evaluated its pharmacological properties. The evaluation of the dichloromethane extract, and [...] Read more.

Background: Doronicum austriacum Jacq., Asteraceae, is a plant which is used in traditional alpine medicine. Historical sources describe the medical use of the root, but up until now only a few studies evaluated its pharmacological properties. The evaluation of the dichloromethane extract, and its major compounds for their anti-inflammatory and anti-oxidant potential was performed in macrophages J774A.1 and C6 astrocytes. Nitric oxide (NO) and reactive oxygen species (ROS) release, as well as nitrotyrosine formation, were evaluated. Moreover, in order to evaluate the potential anti-proliferative activity, under the same experimental conditions, 3-(4,5-dimethyltiazol-2yl)-2,5-phenyl-2H-tetrazolium bromide (MTT) assay was also performed. Our results indicate that Doronicum austriacum has a significant effect in inhibiting both pro-inflammatory and pro-oxidative mediators. All isolated compounds were able to significantly inhibit NO and ROS release both in macrophage and in astrocytes cells, even if the effect was more pronounced in macrophage. In particular, among the tested compounds, 6,12-dihydroxy-(?)-2S-tremetone exerted stronger activity. Both extract and single compounds did not affect cellular viability. This study provides evidence for the pharmacological anti-inflammatory and anti-oxidant potential of Doronicum austriacum extract. These effects could be due to the activity of its major constituents and subsequent identification of benzofurans as a promising compound class to combat inflammation and related diseases. Full article

(This article belongs to the Section Natural Products Chemistry)

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Full article ">Figure 1
Chromatogramm of the HPLC-DAD analysis of the investigated dichloromethane (DCM) extract of the roots of Doronicum austriacum (2.5 mg/mL MeOH) at 205 nm. Analytical conditions: stationary phase: Phenomenex Synergi Max-RP 80 Å, 4 µm (4.6 mm × 150 mm); temperature: 35 °C; mobile phase: A = water + 0.025% TFA, B = acetonitrile; flow rate: 1.00 mL/min; detection: 205 nm; injection volume: 10 µL; solvent composition during analysis: 0′: 75% A; 10′: 60% A; 30′: 30% A; 35′: 2% A; 45′: stop; post time: 10′. Full article ">Figure 2
Chemical structure of compounds 1 to 3. Full article ">Figure 3
Measured (acetonitrile) and calculated ECD-spectra of compound 1. Full article ">Figure 4
Effect of Doronicum austriacum DCM root extract on nitrite release, index of NO production (A,D), measured by Griess reaction, iNOS expression, evaluated by cytofluorimetry (B,E), and on ROS production (C,F), measured by DCHF assay, in J774A.1 macrophages (A–C) and C6 cells (D–F). Data are expressed as mean ± S.E.M.; *** denotes p < 0.001 vs. LPS (1 μg/mL) or LPS + IFN-γ (1 μg/mL; 100 U/mL). L-NAME (1 μM) was used as positive control for inhibition of nitrite release and iNOS expression and inhibited nitrite production with a percentage of 49.00 ± 2.08% and with a percentage of 59.00 ± 0.74% iNOS expression in J774A.1 macrophages. In C6 cells, L-NAME inhibited nitrite release with a percentage of 39.33 ± 4.37% and iNOS expression with a percentage of 46.21 ± 2.65%. N-acetyl cysteine (NAC; 10 mM) was used as positive control for ROS release inhibition, and gave a percentage of inhibition of ROS production of 34.50 ± 0.50% in J774A.1 and of 43.33 ± 1.45% in C6 cells, respectively. Full article ">Figure 5
Effect of Doronicum austriacum DCM root extract (25 and 50 µg/mL) on nitrotyrosine formation evaluated by confocal microscopy in J774A.1 macrophages (A) and C6 astrocytes (B) alone, or in combination with LPS (1 μg/mL) or LPS + IFN-γ (1 μg/mL; 100 U/mL) after 24 h. Full article ">Figure 6
Effect of compounds 1–3 (5–50 µM) on nitrite release (A,B), iNOS expression (C,D), and on ROS production (E,F), in J774A.1 macrophages (A,C,E) and C6 astrocytes (B,D,F) after 24 h. Data are expressed as mean ± S.E.M.; *** denotes p < 0.001 vs. LPS (1 μg/mL) or LPS + IFN-γ (1 μg/mL; 100 U/mL). °°°, °°, ° denote p < 0.001, p < 0.01, p < 0.05 vs. 2. òòò, òò, ò denote p < 0.001, p < 0.01, p < 0.05 vs. 3. L-NAME (1 μM), used as positive control for NO and iNOS inhibition gave a percentage of inhibition of 49.00 ± 2.08% for nitrite production of and a percentage of inhibition of 59.00 ± 0.74% of iNOS expression in J774A.1 macrophages. In C6 astrocytes, L-NAME gave a percentage of inhibition of 39.33 ± 4.37% and 46.21 ± 2.65% of nitrite production and iNOS expression, respectively. NAC (10 mM) was used as positive control for ROS release and gave a percentage of inhibition of ROS production of 34.50 ± 0.50% in J774A.1 macrophages and 43.33 ± 1.45% in C6 astrocytes. Full article ">

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Open AccessArticle

Identification of Onosma visianii Roots Extract and Purified Shikonin Derivatives as Potential Acaricidal Agents against Tetranychus urticae

by Stefania Sut, Roman Pavela, Vladislav Kolarčik, Loredana Cappellacci, Riccardo Petrelli, Filippo Maggi, Stefano Dall’Acqua and Giovanni Benelli

Molecules 2017, 22(6), 1002; https://doi.org/10.3390/molecules22061002 - 16 Jun 2017

Cited by 30 | Viewed by 6152

Abstract

There is an increasing need for the discovery of reliable and eco-friendly pesticides and natural plant-derived products may play a crucial role as source of new active compounds. In this research, a lipophilic extract of Onosma visianii roots extract containing 12% of shikonin [...] Read more.

There is an increasing need for the discovery of reliable and eco-friendly pesticides and natural plant-derived products may play a crucial role as source of new active compounds. In this research, a lipophilic extract of Onosma visianii roots extract containing 12% of shikonin derivatives demonstrated significant toxicity and inhibition of oviposition against Tetranychus urticae mites. Extensive chromatographic separation allowed the isolation of 11 naphthoquinone derivatives that were identified by spectral techniques and were tested against Tetranychus urticae. All the isolated compounds presented effects against the considered mite and isobutylshikonin (1) and isovalerylshikonin (2) were the most active, being valuable model compounds for the study of new anti-mite agents. Full article

(This article belongs to the Section Natural Products Chemistry)

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Open AccessArticle

Discovery of Indeno[1,2-c]quinoline Derivatives as Potent Dual Antituberculosis and Anti-Inflammatory Agents

by Chih-Hua Tseng, Chun-Wei Tung, Chen-Hsin Wu, Cherng-Chyi Tzeng, Yen-Hsu Chen, Tsong-Long Hwang and Yeh-Long Chen

Molecules 2017, 22(6), 1001; https://doi.org/10.3390/molecules22061001 - 16 Jun 2017

Cited by 34 | Viewed by 6104

Abstract

A series of indeno[1,2-c]quinoline derivatives were designed, synthesized and evaluated for their anti-tuberculosis (anti-TB) and anti-inflammatory activities. The minimum inhibitory concentration (MIC) of the newly synthesized compound was tested against Mycobacterium tuberculosis H₃₇R_V. Among the tested compounds, [...] Read more.

A series of indeno[1,2-c]quinoline derivatives were designed, synthesized and evaluated for their anti-tuberculosis (anti-TB) and anti-inflammatory activities. The minimum inhibitory concentration (MIC) of the newly synthesized compound was tested against Mycobacterium tuberculosis H₃₇R_V. Among the tested compounds, (E)-N?-[6-(4-hydroxypiperidin-1-yl)-11H-indeno[1,2-c]quinolin-11-ylidene]isonicotino-hydrazide (12), exhibited significant activities against the growth of M. tuberculosis (MIC values of 0.96 ?g/mL) with a potency approximately equal to that of isoniazid (INH), an anti-TB drug. Important structure features were analyzed by quantitative structure–activity relationship (QSAR) analysis to give better insights into the structure determinants for predicting the anti-TB activity. The anti-inflammatory activity was induced by superoxide anion generation and neutrophil elastase (NE) release using the formyl-l-methionyl-l-leucyl-l-phenylalanine (fMLF)-activated human neutrophils method. Results indicated that compound 12 demonstrated a potent dual inhibitory effect on NE release and superoxide anion generation with IC₅₀ values of 1.76 and 1.72 ?M, respectively. Our results indicated that compound 12 is a potential lead compound for the discovery of dual anti-TB and anti-inflammatory drug candidates. In addition, 6-[3-(hydroxymethyl)piperidin-1-yl]-9-methoxy-11H-indeno[1,2-c]quinolin-11-one (4g) showed a potent dual inhibitory effect on NE release and superoxide anion generation with IC₅₀ values of 0.46 and 0.68 ?M, respectively, and is a potential lead compound for the discovery of anti-inflammatory drug candidates. Full article

(This article belongs to the Special Issue Frontiers in Antimicrobial Drug Discovery and Design)

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Molecules, Volume 22, Issue 6 (June 2017) – 171 articles

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