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Herein, a chemophenetic significance, based on the phenolic metabolite profiling of three Senecio (S. hercynicus, S. ovatus, and S. rupestris) and two Jacobaea species (J. pancicii and J. maritima), coupled to morphometric data, is presented. A set of twelve morphometric characters were recorded from each plant species and used as predictor variables in a linear discriminant analysis (LDA) model. From a total 75 observations (15 from each of the five species), the model correctly assumed their species’ membership, except for 2 observations. Among the studied species, S. hercynicus and S. ovatus presented the greatest morphological similarity. A phytochemical profiling of phenolic specialized metabolites by UHPLC-Orbitrap-MS revealed 46 hydroxybenzoic, hydroxycinnamic, and acylquinic acids and their derivatives, 1 coumarin and 21 flavonoids. Hierarchical and PCA clustering applied to the phytochemical data corroborated the similarity of S. hercynicus and S. ovatus, observed in the morphometric analysis. This study contributes to the phylogenetic relationships between the tribe Senecioneae taxa and highlights the chemophenetic similarity/dissimilarity of the studied species belonging to Senecio and Jacobaea genera.

Despite the early access procedures for marketing authorization (MA) valid throughout the European Union still in the most of the Member states patient access to innovative medicines depends on cost-effectiveness, budget impact assessment and negotiations for price discount with the public payers. Retrospective analysis on the availability and time to market access of medicines authorized under the European medicines agency’s specific procedures for early access shows that despite the shortening of the time to market access after 2013, for most medicines still exceeds 365 days. This is due to the fact that requirements for pricing and reimbursement across EU is fixed to some degree and medicines with MA for early access are subject to the same legal requirements as the medicines with standard centralized marketing authorization. Some specific national legal requirements for pricing and reimbursement decisions, population of interest and manufactures intentions to enter certain markets should also be considered.

Diclofenac is one of the most prescribed non-steroidal anti-inflammatory drugs (NSAIDs) for acute and chronic inflammatory conditions. Taking into consideration the extensive use of diclofenac, the crucial role of inflammation in tumorigenesis and data on the effects of NSAIDs on cancer cell lines, we investigated the cytotoxic potential of diclofenac on a panel of human cell lines originating from breast (MCF-7), cervical (HeLa) and colorectal cancer (HT-29). The cytotoxicity was assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) dye reduction assay and the half maximal inhibitory concentrations (IC50) were determined. Diclofenac had higher potency against MCF-7 and HT-29 than against HeLa. The cellular and nuclear changes were examined by fluorescent microscopy using 4′,6′-diamino-2-phenylindole (DAPI) stain and acridine orange/ethidium bromide (AO/EB). The wound-healing scratch assay showed significant reduction in the migration capacity of all tested cancer cell lines. The observed antineoplastic activity implies that the anticancer potential of NSAIDs and diclofenac, in particular, necessitates further investigation.

Oleraceins are a class of indoline amide glycosides found in Portulaca oleracea L. (Portulacaceae), or purslane. These compounds are characterized with 5,6-dihydroxyindoline-2-carboxylic acid N-acylated with cinnamic acid derivatives, and many are glucosylated. Herein, hydromethanolic extracts of the aerial parts of purslane were subjected to UHPLC-Orbitrap-HRMS analysis, conducted in negative ionization mode. Diagnostic ion filtering (DIF), followed by diagnostic difference filtering (DDF), were utilized to automatically filter out MS data and select plausible oleracein structures. After an in-depth MS2 analysis, a total of 51 oleracein compounds were tentatively identified. Of them, 26 had structure matching one of already known oleraceins and the other 25 are new, undescribed in the literature structures, belonging to the oleracein class. Moreover, diagnostic fragment ions were selected, based on which clustering algorithms and visualizations were employed. As we demonstrate, clustering methods can provide valuable insights into the mass fragmentation elucidation of natural compounds in complex mixtures.

Methanol-aqueous extracts from the aerial parts of Gypsophila glomerata (GGE), G. trichotoma (GTE) and G. perfoliata (GPE) were investigated for antioxidant potential using different in vitro models, as well as for phenolic and flavonoid contents. The possible anti-cholinesterase, anti-tyrosinase, anti-amylase and anti-glucosidase activities were also tested. The flavonoid variability was analyzed using ultra high-performance liquid chromatography (UHPLC) coupled with hybrid quadrupole-Orbitrap high resolution mass spectrometry (HRMS). Eleven C-glycosyl flavones and 4 O-glycosyl flavonoids, including 2”-O-pentosyl-6-C-hexosyl-apigenin/methylluteolin, as well as their mono(di)-acetyl derivatives were found in GGE. Both GGE and GTE shared 2”-pentosyl-6-C-hexosyl-luteolin together with the common saponarin, homoorientin, orientin, isovitexin and vitexin, while di C-glycosyl flavones were evidenced only in GPE. The highest radical scavenging in both ABTS and DPPH assays was noted in GPE, as well as ferric and cupric reducing abilities. However, GTE had the strongest metal chelating activity (17.44 ± 0.51 mg EDTAE/g extract). GPE and GGE were more potent as acetylcholinesterases inhibitors witnessed by 2.09 ± 0.02 mg GALAE/g extract and 1.59 ± 0.09 mgGALAE/g extract, respectively. All flavonoids were found in G. glomerata for the first time. Therefore, further isolation and structural elucidation of newly described acetylated flavonoids are needed in order to determine their relevance in the beneficial properties of the plant.

In-depth characterization of specialized metabolites in the endemic Gypsophila perfoliata L. “tekirae” (G. tekirae Stef.) by liquid chromatography – quadrupol-Orbitrap mass spectrometry allowed dereplication/annotation of 22 flavonoids including 11 2″-O-pentosyl/deoxyhexosyl/hexosyl-C-hexosyl-flavones in the aerial parts and 23 gypsogenin- and quillaic acid-bidesmosides in the roots. Saponins were mainly mono-and diacetyl, and methoxycinnamoyl derivatives of 16 core structures forming isobaric isomers. Three acetylated derivatives of 2″-O-deoxyhexosyl-6-C-hexosyl-flavones are annotated for the first time in the genus Gypsophila together with five quillaic acid-based saponins. Aerial parts extract revealed prominent antioxidant and tyrosinase inhibitory activity, while roots demonstrated higher capacity against acetylcholinesterase, butyrylcholinesterase and α-glucosidase. The chemophenetic significance of acetylated 2″-O-glycosyl-6-C-hexosyl-flavones and GOTCAB saponins with methoxycinnamoyl-substituted α-chains was discussed.

A new convenient method for preparation of 2-aroyl-[1]benzopyrano[4,3-c]pyrazol-4(1H)-one derivatives 5b-g and coumarin containing hydrazide-hydrazone analogues 4a-e was presented. The antimycobacterial activity against reference strain Mycobacterium tuberculosis H37Rv and cytotoxicity against the human embryonic kidney cell line HEK-293 were tested in vitro. All compounds demonstrated significant minimum inhibitory concentrations (MIC) ranging 0.28-1.69μM, which were comparable to those of isoniazid. The cytotoxicity (IC50>200µM) to the "normal cell" model HEK-293T exhibited by 2-aroyl-[1]benzopyrano[4,3-c]pyrazol-4(1H)-one derivatives 5b-e, was noticeably milder compared to that of their hydrazone analogues 4a-e (IC50 33-403µM). Molecular docking studies on compounds 4a-e and 5b-g were also carried out to investigate their binding to the 2-trans-enoyl-ACP reductase (InhA) enzyme involved in M. tuberculosis cell wall biogenesis. The binding model suggested one or more hydrogen bonding and/or arene-H or arene-arene interactions between hydrazones or pyrazole-fused coumarin derivatives and InhA enzyme for all synthesized compounds.

Series of aroyl hydrazones of 2H-chromene and coumarin carbaldehydes were synthesized and evaluated for their anticonvulsant activity and neurotoxicity. Further docking study on gamma-aminobutyric acid receptor was performed to elucidate their mechanisms of action. The highest protection was demonstrated by 2-furyl substituted 2H-chromene 8b in the maximal electroshock test (ED50 = 12.51 mg kg−1, PI MES > 23.98) and the subcutaneous pentylenetetrazole tests (ED50 = 127.10 mg kg−1). Furyl-substituted derivative 4b (ED50 = 68.66 mg kg−1) was the most active in the maximal electroshock test while methoxyphenyl-substituted derivate 4c was the most active in the 6-Hz test (ED50 = 94.34 mg kg−1). None of the compounds displayed neurotoxicity in the rota-rod test. In silico assessment of their blood-brain barrier permeability indicated them as central nervous system active agents. The results suggest that coumarin/2H-chromene aroyl hydrazones scaffold deserve further evaluation in models of epilepsy and derivatization.

Geigeria alata Benth. & Hook.f. ex Oliv. & Hiern (Asteraceae) is used in Sudanese folk medicine for treatment of diabetes. The study aimed to estimate the acute oral toxicity of trans-3,5-dicaffeoylquinic acid (3,5-diCQA) from G. alata roots and to assess its antihypeglycemic, antioxidant and antihypertensive effects on chemically-induced diabetic spontaneously hypertensive rats (SHRs). The structure of 3,5-diCQA was established by NMR and HRMS spectra. Type 2 diabetes was induced by intraperitoneal injection of streptozotocin. 3,5-diCQA was slightly toxic with LD50 = 2154 mg/kg. At 5 mg/kg 3,5-diCQA reduced significantly (p

The purpose of this study was to reveal different subgroups of patients with at least moderate risk of developing diabetes in the next 10 years, based on clustering of cardiovascular risk factors.We performed a one-center cross-sectional study of adult patients (n = 109, median age 45 years) with Findrisc score of above 11 out of 26 maximum. We included in the cluster analysis anthropometrics, lipid and carbohydrate parameters obtained in oral glucose tolerance test (OGTT), insulin, C-peptide, creatinine, C-reactive protein, liver enzymes, beta-cell function, insulin sensitivity and insulin resistance (HOMA calculations). We also evaluated the atherogenic index of plasma (AIP).We identified three metabolic phenotypes of patients with at least moderate Findrisc score-one 'male' (cluster AM, n = 24), and two 'female' phenotypes (cluster AW, n = 9 and cluster BW, n = 76). Men were almost homogenous for their metabolic phenotype, with lower fat percentage than women (p .05). Most of the women (cluster BW, n = 76) presented with better metabolic pattern i.e. lower insulin resistance, lower C-reactive protein, lower degree of obesity and visceral fat rating (p .05), despite the higher fat percentage (p .05). Some of the women, however, (cluster AW, n = 9) presented with parameters very similar to that of men (cluster AM) and significantly higher than in cluster BW. Despite the lack of significant differences in lipid parameters among clusters, AIP was significantly lower in cluster BW (p .05).Most of the women presented with clearly less unfavorable atherogenic risk than men. Two different phenotypes of obese women with at least moderate Findrisc score were revealed, and the level of inflammation seems to be the main discriminant factor. Larger prospective studies are required to elucidate whether those are really two different pathogenically phenotypes or if they belong to the same phenotype's continuum.

A newly discovered Ne-feruloyl lysine (1) together with 16 known hydroxycinnamic acid amides (HCAAs) were identified in the roots of Sudanese plant Solanum schimperianum Hochst (syn. Solanum carense Dunal, Solanaceae). The structure of 1 was established on the basis of 1D (1H, 13C NMR) and 2D (1H-1H COSY, 1H-13C HSQC) NMR analysis, together with UV⿿vis, HR-ESI⿿MS and ESI⿿MS2 fragmentation analyses. HCAAs were analyzed using liquid chromatography coupled to an Orbitrap mass spectrometer (ThermoScientific) using parallel-reaction monitoring (PRM) mode. For the first time, eight HCAAs including agmatine and cadaverine amides, as well as Ne-feruloyl lysine and sinapoyl putrescine, were reported in genus Solanum. Ne-feruloyl lysine and the hydromethanolic extract of S. schimperianum roots were evaluated for their antioxidant activity using free radical DPPH and ABTS scavenging, and ferric reducing power (FRAP) methods. The antimicrobial activity was estimated on a panel of pathogenic bacteria and fungi by the broth microdilution method (BMD). Ne-feruloyl lysine demonstrated stronger DPPH (IC50 85.83 μg/ml), ABTS (IC50 255.54 μg/ml) and FRAP (0.44 ± 0.02 mM TE/mg dw) activity compared to the root extract. Ne-feruloyl lysine exhibited significantly higher antimicrobial activity against both Staphylococcus aureus strains NBIMCC 3359 (MIC/MBC 112.5 μg/ml) and penicillin resistant ATCC 6538P (MIC 112.5 μg/ml) than the total S. schimperianum extract (MIC 1.25 and 2.5 mg/ml, respectively). The extract showed antimicrobial activity towards Streptococcus pyogenes, Listeria monocytogenes and Candida albicans (MIC 1.25 mg/ml). Ne-feruloyl lysine had function for the observed protection against oxidative stress and antimicrobial activity.

Asteraceae species Tanacetum macrophyllum (Walds. & Kit.) Sch.Bip. (Tm) and Telekia speciosa (Schreb.) Baumg. (Ts) are renowned for their ethnomedicinal use. This work aims to provide an in-depth characterization of the methanol-aqueous extracts from the flower heads, aerial parts and roots of the species. Annotation and dereplication of 113 secondary metabolites was achieved in the studied samples; 63 compounds are reported in the species for the first time. Herein, 38 acylquinic acids analogues, 31 flavonols and flavones, 17 sesquiterpene lactones together with phenylethanoid glycosides and sugar esters were evidenced in the assayed extracts. The principal component and heat-map analysis identified 20 marker metabolites that revealed herbal drug-specific chemical patterns: phenylethanoid glycosides, quinic, chlorogenic and 3-feruloyl-4-caffeoylquinic acid, patuletin, eupatorin, quercetagetin-trimethyl ether, artecalin and artemisiifolin had the highest contribution in variance. The esters of caffeic/ferulic acid-hexoside were exclusively produced by T. speciosa while methoxylated flavonoids were evidenced in T. macrophyllum. According to the multivariate analysis of biological potential, radical scavenging activity and reducing power accounted mostly for the stronger antioxidant capacity of Tm in comparison with Ts. Telekia specosa aerial parts and roots had marked impact on acetyl- and butyrylcholinesterase (up to 4.82 ± 0.22 mg galantamine equivalent (GALAE)/g and 3.20 ± 0.30 mg GALAE/g, respectively) and tyrosinase (up to 119.00 ± 0.97 mg kojic acid equivalent/g) inhibition. Tanacetum macrophyllum aerial parts and flower heads were beneficial for attenuating carbohydrate metabolism disorders witnessed by α-glucosidase (1.45 ± 0.27 mmol acarbose equivalent (ACAE)/g) and α-amylase (0.65 ± 0.04 mmol ACAE/g) inhibitory activity. Tanacetum macrophyllum flower heads and T. speciosa roots can be recommended for production of raw material with antioxidant and enzyme inhibitory potential.

Background: The 1,8-Naphthalimides constitute an important class of biologically active, DNAbinding compounds. There are no available data on the synthesis of 1,8-naphthalimide derivatives with nonprotein amino acids and their biological activity. The aim of this paper was to determine the synthesis, structural characterization and cytotoxic activity of new 1-(1,3-dioxo-1H-benzo[de]isoquinolin-2(3H)-yl)cycloalkane-1- carboxylic acids with 5-, 6-, 7-, 8- and 12-membered rings as well as 2-(1,3-dioxo-1H-benzo[de]isoquinolin-2(3H)- yl)adamantane-2-carboxylic acid and 1-(1,3-dioxo-1H-benzo[de]isoquinolin-2(3H)-yl)-1,2,3,4-tetrahydronaphthalene- 1-carboxylic acid. Methods: The target compounds were obtained by an interaction of 1,8-naphthalic anhydride with a series of non-protein amino acids. The optimized geometry and harmonic vibrational frequencies have been calculated by DFT employing B3LYP functional using 6-31G(d,p) basis set. An ab initio (MP2 and Hartee-Fock) and DFT (different functionals) using several basis sets have been applied for NMR calculations. The cytotoxic effects of the synthesized compounds are assessed against two human tumor cell lines, namely K-562 (chronic myeloid leukemia) and HUT-78 (cutaneous T-cell lymphoma) after 72 h exposure, using the MTT-dye reduction assay. The apoptogenic effects and the ability to modulate the NFκB-signaling pathways were determined using commercially available ELISA kits. Results: All compounds inhibited the growth of malignant cells at micromolar concentrations whereby compound 4b (1-(1,3-dioxo-1H-benzo[de]isoquinolin-2(3H)-yl)cyclohexane-1-carboxylic acid) demonstrated superior activity in both cell lines with IC50 values comparable to those of the reference anticancer drug melphalan. Conclusion: New 1,8-naphthalimide derivatives with non-protein amino acids were successfully synthesized. Quantum-chemical calculations were performed to elucidate the structure of the newly synthesized compounds. There is a proper alignment between theoretical and experimental results. The cytotoxicity of the synthesized products against two human tumor cell lines, namely K-562 and HUT-78 was evaluated. All compounds inhibited the growth of malignant cells at micromolar concentrations. The pharmacodynamics evaluation of compound 4b showed that its cytotoxicity is mediated by induction of apoptosis and inhibition of NFκB-signaling.

Glucuronide Oleanane-type Triterpenoid Carboxylic Acid 3, 28-Bidesmosides (GOTCAB) saponins are bioactive natural compounds spread in Caryophyllidae. The high complexity of GOTCAB occurring as closely related isobaric and positional isomers is a challenge in their separation and identification. A new liquid chromatography – high resolution Orbitrap mass spectrometry acquisition strategy would be important for the structural elucidation of GOTCAB in plant extracts. In this study, the fragmentation behaviors of GOTCAB from methanol-aqueous root extract of Gypsophila glomerata Pall ex M. B. (Caryophyllaceae) were investigated using ultra high-performance liquid chromatography (UHPLC) coupled with hybrid quadrupole-Orbitrap high resolution mass spectrometry (HRMS). A new saponin was isolated and its structure was established by 1D and 2D-NMR spectroscopic experiments as 3-O-β-D-galactopyranosyl-(1→2)-[α-L-arabinopyranosyl-(1→3)]-β-D-glucuronopyranosyl gypsogenin 28-O-α-L-arabinopyranosyl-(1→3)-[β-D-xylopyranosyl-(1→4)]-α-L-rhamnopyranosyl-(1→2)-β-D-fucopyranosyl ester. On the basis of the accurate mass measurements, fragmentation patterns in MS/MS analyses and comparison with previously isolated authentic references, a total of 41 GOTCAB saponins were identified or tentatively elucidated in G. glomerata roots, including 14 pairs of isobars. Possible fragmentation pathways for three groups of GOTCAB are suggested. The group I appeared to be GOTCAB of gypsogenin with two carbohydrate chains: a branched trisaccharide at C-3 and tri- to hexa-saccharide attached to C-28 of the aglycone through a deoxyhexose residue. Saponins with monoacetylated (group II) or sulphated (group III) C-28 chain were evidenced, as well as quillaic and oleanolic acid GOTCAB. Sixteen GOTCAB were previously not described. The content of Gypsophila prosaponins, gypsogenin 3-O-glucuronide (7.4079 ± 0.0723 mg/g dry weight, dw) and quillaic acid 3-O-glucuronide (4.4593 ± 0.1207 mg/g dw), was determined by solid phase extraction – high-performance liquid chromatography (SPE-HPLC). In this study is presented the first systematic investigation on the fragmentation patterns and diagnostic fingerprints of the fragment ions in the MS/MS spectra of the gypsogenin -, quillaic acid - and oleanolic acid – bidesmosides. A LC-HRMS Orbitrap acquisition strategy could give an insight in the GOTCAB containing taxa.

The compound in the title was prepared by reaction between tryptamine and ibuprofen using N,N′-dicyclohexylcarbodiimide as a “dehydrating„ reagent. The structure of the newly synthesized compound was determined by nuclear magnetic resonance (NMR) (1H-NMR and 13C-NMR), UV, IR, and mass spectral data.

A simple and rapid method of excised root cultures from six Gypsophila species was performed allowing continuous growth without phytohormones. Established on MH3 medium from solid-grown seedlings these roots were subcultured for 1 year on a solid medium before being transferred in a liquid medium to obtain substantial biomass for saponin content analysis. Morphologically, the different root lines presented different growth behaviors and different physical aspects: some have linear growth by the tip of the main axial root from the original seedling; others have additional lateral root initiations producing a hairy root-system more or less dense. A marked increase of biomass was observed in the light by comparison with dark conditions. Significant growth for Gypsophila glomerata was achieved within 3 weeks on liquid medium; biomass grew up to 50-fold in batch cultures reaching 10 g DW. The fingerprints of the saponin HPLC profiles of the six Gypsophila species were drastically different with at least up to 30 different saponins detected for some of them. The roots of Gypsophila elegans accumulated saponins up to 65 mg/g DW. These amounts were higher than in Gypsophila paniculata roots classically found as the best producing ones. On the contrary the root lines of G. glomerata showed a smaller quantitative amount of saponins (between 1.3 and 7.10 mg/g DW) than those of G. elegans but nearly the same HPLC profiles as for root extracts of G. paniculata plants grown directly in the fields.

The theophylline-7-acetic acid (7-TAA) scaffold is a promising novel lead compound for antimycobacterial activity. Here, we derive a model for antitubercular activity prediction based on 14 7-TAA derivatives with amino acid moieties and their methyl esters. The model is applied to a combinatorial library, consisting of 40 amino acid and methyl ester derivatives of 7-TAA. The best three predicted compounds are synthesized and tested against Mycobacterium tuberculosis H37Rv. All of them are stable, non-toxic against human cells and show antimycobacterial activity in the nanomolar range being 60 times more active than ethambutol.

A series of amides were synthesized by condensation of theophylline-7-acetic acid and eight commercially available amino acid methyl ester hydrochlorides. Consecutive hydrolysis of six of the amido-esters resulted in the formation of corresponding amido-acids. The newly synthesized compounds were evaluated for their in vitro activity against Mycobacterium tuberculosis H37Rv. The activity varied depending on the amino acid fragments and in seven cases exerted excellent values with MICs 0.46-0.26 μM. Assessment of the cytotoxicity revealed that the compounds were not cytotoxic against the human embryonal kidney cell line HEK-293T. The theophylline-7-acetamides containing amino acid moieties appear to be promising lead compounds for the development of antimycobacterial agents.