Reactive deep eutectic solvents for EDC-mediated amide synthesis.

The sustainability of amide bond formation is an ever-present topic in the pharmaceutical industry, as it represents the common motif in many clinically approved drugs. Despite many procedures for accomplishing eco-friendly amide synthesis having been developed, this transformation still remains a contemporary challenge. Herein, we report a greener approach for amide synthesis by using Reactive Deep Eutectic Solvents (RDESs) acting as both the reaction medium and reactants. The procedure not only avoids the use of hazardous solvents but also provides operationally simple product recovery with high purity and efficiency, without chromatographic purification. This approach was efficiently applied to the synthesis of a key intermediate in the production of an active pharmaceutical ingredient like atenolol. The green metrics of the gram-scale procedure were compared to the conventional industrial strategy showing an advancement in the greening of amide synthesis.

A Picrocrocin-Enriched Fraction from a Saffron Extract Affects Lipid Homeostasis in HepG2 Cells through a Non-Statin-like Mode.

Dyslipidemia is a lipid metabolism disorder associated with the loss of the physiological homeostasis that ensures safe levels of lipids in the organism. This metabolic disorder can trigger pathological conditions such as atherosclerosis and cardiovascular diseases. In this regard, statins currently represent the main pharmacological therapy, but their contraindications and side effects limit their use. This is stimulating the search for new therapeutic strategies. In this work, we investigated in HepG2 cells the hypolipidemic potential of a picrocrocin-enriched fraction, analyzed by high-resolution 1H NMR and obtained from a saffron extract, the stigmas of Crocus sativus L., a precious spice that has already displayed interesting biological properties. Spectrophotometric assays, as well as expression level of the main enzymes involved in lipid metabolism, have highlighted the interesting hypolipidemic effects of this natural compound; they seem to be exerted through a non-statin-like mechanism. Overall, this work provides new insights into the metabolic effects of picrocrocin, thus confirming the biological potential of saffron and paving the way for in vivo studies that could validate this spice or its phytocomplexes as useful adjuvants in balancing blood lipid homeostasis.

Synthesis and Antiproliferative Activity of Novel Dehydroabietic Acid-Chalcone Hybrids.

Dehydroabietic Acid (DHA, 1) derivatives are known for their antiproliferative properties, among others. In the context of this work, DHA was initially modified to two key intermediates bearing a C18 methyl ester, a phenol moiety at C12, and an acetyl or formyl group at C13 position. These derivatives allowed us to synthesize a series of DHA-chalcone hybrids, suitable for structure-activity relationship studies (SARS), following their condensation with a variety of aryl-aldehydes and methyl ketones. The antiproliferative evaluation of the synthesized DHA-chalcone hybrids against three breast cancer cell lines (the estrogen-dependent MCF-7 and the estrogen-independent MDA-MB-231 and Hs578T) showed that eight derivatives (33, 35, 37, 38, 39, 41, 43, 44) exhibit low micromolar activity levels (IC50 2.21-11.5 µΜ/MCF-7). For instance, some of them showed better activity compared to the commercial anticancer drug 5-FU against MCF-7 cells (33, 41, 43, 44) and against MDA-MB231 (33 and 41). Hybrid 38 is a promising lead compound for the treatment of MCF-7 breast cancer, exhibiting comparable activity to 5-FU and being 12.9 times less toxic (SI = 22.7). Thus, our findings suggest that DHA-chalcone hybrids are drug candidates worth pursuing for further development in the search for novel breast cancer therapies.

Recovery of Biophenols from Olive Vegetation Waters by Carbon Nanotubes.

In this work, the possibility of using carbon nanotubes for the treatment of olive vegetation waters (OVWs) was investigated. In general, the disposal of OVWs represents an important environmental problem. The possibility of considering these waters no longer just as a problem but as a source of noble substances, thanks to the recovery of biophenols from them, was tested. In particular, predetermined quantities of olive vegetation waters were treated with carbon nanotubes. The quantities of adsorbed biophenols were studied as a function of the quantities of carbon nanotubes used and the contact time. The experimental conditions for obtaining both the highest possible quantities of biophenol and a purer adsorbate with the highest percentage of biophenols were studied. After the adsorption tests, the vegetation waters were analyzed by UV spectrophotometry to determine, in particular, the variation in the concentration of biophenols. The carbon nanotubes were weighed before and after each adsorption test. In addition, kinetic studies of the adsorption processes were considered. Carbon nanotubes proved their effectiveness in recovering biophenols.

Deep Eutectic Solvents for Improving the Solubilization and Delivery of Dapsone.

Owing to a growing awareness toward environmental impact, the use of safer and eco-friendly solvents like deep eutectic solvents (DESs), has recently undergone important growth in the pharmaceutical field, with regard to their application as non-aqueous liquid administration vehicles, since they do not carry the same risks of toxicity and handling as traditional organic solvents. Major attention has been given to the development of advantageous transdermal drug delivery systems, because of their ease of use and better acceptability. Here, we report the use of two different DESs, based on choline chloride, used as hydrogen bond acceptor (HBA), and ascorbic acid or propylene glycol, used as hydrogen bond donors (HBDs), able to enhance the solubility and the topical delivery of dapsone, representing a class IV drug. The interactions between the DESs' components and the drug were studied by performing DSC, FT-IR, and NMR analysis of the eutectic systems and the pure drug, confirming the establishment of H-bonds between the drug and the DESs' components. Diffusion and permeability studies, carried out in a Franz cell, showed an increase in permeability, highlighting the great potential of DESs as dissolution and permeation enhancers in the development of novel and more effective drug delivery systems in topical administration.

Green solvents for the formation of amide linkages.

The formation of the amide bond is among the most commonly performed transformations in the pharmaceutical industry and the wider chemical industry. The current methods for its installation in organic compounds frequently rely on the use of large amounts of organic solvents, mainly N,N-dimethylformamide (DMF), N-methylpyrrolidone (NMP), and dichloromethane (DCM), which have been associated with adverse environmental and health concerns over the last decades. This fact led academia and industry to make significant efforts toward the development of synthetic routes with the aim to avoid, reduce or replace the use of hazardous solvents. The present review fits into this framework and discusses the literature existing over the past ten years on strategies for reducing and replacing hazardous solvents, focusing on the use of biobased and neoteric solvents, such as ionic liquids and deep eutectic solvents (ILs and DESs, respectively), and on the reaction media that proved to be greener alternatives for amide bond formation.

The Role of Carbon Nanotube Pretreatments in the Adsorption of Benzoic Acid.

Four different types of multi-walled carbon nanotubes (MWCNTs) were used and compared for the treatment of benzoic acid contaminated water. The types of nanotubes used were: (1) non-purified (CNTsUP), as made; (2) purified (CNTsP), not containing the catalyst; (3) oxidized (CNTsOX), characterized by the presence of groups such as, -COOH; (4) calcined (CNTs900), with elimination of interactions between nanotubes. In addition, activated carbon was also used to allow for later comparison. The adsorption tests were conducted on an aqueous solution of benzoic acid at concentration of 20 mg/L, as a model of carboxylated aromatic compounds. After the adsorption tests, the residual benzoic acid concentrations were measured by UV-visible spectrometry, while the carbon nanotubes were characterized by TG and DTA thermal analyses and electron microscopy (SEM). The results show that the type of nanotubes thermally treated at 900 °C has the best performances in terms of adsorption rate and amounts of collected acid, even if compared with the performance of activated carbons.

Analytical Strategy for MS-Based Thanatochemistry to Estimate Postmortem Interval.

An analytical strategy for a matrix-assisted laser desorption mass spectrometry-based untargeted metabolomic study on vitreous humor (VH) was developed, looking for statistically significant parameters correlated to death time estimation. Five incubation stages of VH, 0, 24, 48, 72, and 96 h, at physiological pH and controlled temperature, were adopted to monitor time-dependent changes and correlate them with the postmortem interval (PMI). Using two multivariate statistical approaches, principal component regression (PCR) and partial least squares regression (PLSR), the PMI was assessed, considering the m/z values from mass spectra and the incubation time (ISt) as predictors. An independent validation set was used to evaluate the predictive capability of the models through the coefficient of determination (R2) and the root-mean-square error (RMSE). Different pre-treatments were applied to the raw mass spectra, and their performance in assessing PMI was evaluated. Based on the best outcomes in terms of both R2 and RMSE, multiplicative scatter correction combined with a logarithmic transformation was chosen. The results of PCR and PLSR based on the selected pre-treatment are encouraging because validation R2 is about 0.95 for both models. Moreover, the prediction error is 6 h for both models, when PMI is lower than 1 day. Although these results are obtained by the uncritical application of the models, they are comparable to or even better than those reported in the literature. Notwithstanding, we consider that many in situ influences, such as passive diffusion, functional loss of tissues, and advanced autolytic processes, could not get captured in vitro. However, the developed approach was optimized using VH samples and overcomes the limitations of the vast majority of methods that require validation for serum and/or urine samples.

Adsorption of Reactive Blue 116 Dye and Reactive Yellow 81 Dye from Aqueous Solutions by Multi-Walled Carbon Nanotubes.

The multi-walled carbon nanotubes obtained by catalytic chemical vapour deposition synthesis are used as a solid matrix for the adsorption of the Reactive Blue 116 dye and the Reactive Yellow 81 dye from aqueous solutions at different pH values. The batch tests carried out allowed us to investigate the different effects of pH (2, 4, 7, 9 and 12) and of the contact time (2.5 ÷ 240 min) used. The liquid phase was analysed using ultraviolet-visible spectrophotometry in order to characterise the adsorption kinetics, the transport mechanisms and the adsorption isotherms. The adsorption of the optimal dye was observed at pH 2 and 12. The pseudo-first order kinetic model provided the best approximation of experimental data compared to the pseudo-second order kinetic model. The predominant transport mechanism investigated with the Weber and Morris method was molecular diffusion for both Reactive Yellow 81 and Reactive Blue 116, and the equilibrium data were better adapted to the Langmuir isothermal model. The maximum adsorption capacity for Reactive Yellow 81 and Reactive Blue 116 occurred with values of 33.859 mg g-1 and 32.968 mg g-1, respectively.

Protein Extraction, Enrichment and MALDI MS and MS/MS Analysis from Bitter Orange Leaves (Citrus aurantium).

Citrus aurantium is a widespread tree in the Mediterranean area, and it is mainly used as rootstock for other citrus. In the present study, a vacuum infiltration centrifugation procedure, followed by solid phase extraction matrix-assisted laser desorption ionization tandem mass spectrometry (SPE MALDI MS/MS) analysis, was adopted to isolate proteins from leaves. The results of mass spectrometry (MS) profiling, combined with the top-down proteomics approach, allowed the identification of 78 proteins. The bioinformatic databases TargetP, SignalP, ChloroP, WallProtDB, and mGOASVM-Loc were used to predict the subcellular localization of the identified proteins. Among 78 identified proteins, 20 were targeted as secretory pathway proteins and 36 were predicted to be in cellular compartments including cytoplasm, nucleus, and cell membrane. The largest subcellular fraction was the secretory pathway, accounting for 25% of total proteins. Gene Ontology (GO) of Citrus sinensis was used to simplify the functional annotation of the proteins that were identified in the leaves. The Kyoto Encyclopedia of Genes and Genomes (KEGG) showed the enrichment of metabolic pathways including glutathione metabolism and biosynthesis of secondary metabolites, suggesting that the response to a range of environmental factors is the key processes in citrus leaves. Finally, the Lipase GDSL domain-containing protein GDSL esterase/lipase, which is involved in plant development and defense response, was for the first time identified and characterized in Citrus aurantium.

2,3-Diaminopropanols Obtained from d-Serine as Intermediates in the Synthesis of Protected 2,3-l-Diaminopropanoic Acid (l-Dap) Methyl Esters.

A synthetic strategy for the preparation of two orthogonally protected methyl esters of the non-proteinogenic amino acid 2,3-l-diaminopropanoic acid (l-Dap) was developed. In these structures, the base-labile protecting group 9-fluorenylmethyloxycarbonyl (Fmoc) was paired to the p-toluensulfonyl (tosyl, Ts) or acid-labile tert-butyloxycarbonyl (Boc) moieties. The synthetic approach to protected l-Dap methyl esters uses appropriately masked 2,3-diaminopropanols, which are obtained via reductive amination of an aldehyde prepared from the commercial amino acid Nα-Fmoc-O-tert-butyl-d-serine, used as the starting material. Reductive amination is carried out with primary amines and sulfonamides, and the process is assisted by the Lewis acid Ti(OiPr)4. The required carboxyl group is installed by oxidizing the alcoholic function of 2,3-diaminopropanols bearing the tosyl or benzyl protecting group on the 3-NH2 site. The procedure can easily be applied using the crude product obtained after each step, minimizing the need for chromatographic purifications. Chirality of the carbon atom of the starting d-serine template is preserved throughout all synthetic steps.

Chemoselective and metal-free reduction of α,ß-unsaturated ketones by in situ produced benzeneselenol from O-(tert-butyl) Se-phenyl selenocarbonate.

The carbon-carbon double bond of arylidene acetones and chalcones can be selectively reduced with benzeneselenol generated in situ by reacting O-(tert-butyl) Se-phenyl selenocarbonate with hydrochloric acid in ethanol. This mild, metal-free and experimentally simple reduction procedure displays considerable functional-group compatibility, products are obtained in good to excellent yields, and the use of toxic Se/CO mixture and NaSeH, or the smelly and air-sensitive benzeneselenol, is avoided.

A rapid MALDI MS/MS based method for assessing saffron (Crocus sativus L.) adulteration.

We report on a sensitive and fast quantitative MALDI-MS/MS method used to assess saffron authenticity by direct analysis through the determination of picrocrocin as the saffron authenticity marker, and using curcumin as the non-isotopic isobaric internal standard. The internal standard curcumin yielded good linearity (R2 = 0.994), and with confidence intervals at 95% for intercept. The detectable maximum adulteration percentage (99.0%) was estimated interpolating the limit of detection (LOD) for the isobaric internal standard in linear regression. The LOD was 47.63 ppm, and LOQ was 56.53 ppm. Good accuracy and precision were obtained for all concentrations. The capability of the MS approach to monitor analytes in a specific, selective fashion was used to obtain a semi-quantitative adulteration percentage and to establish the adulterant by additional experiments. The detection of gardecin and its derivatives in commercial samples indicated that Gardenia jasminoides Ellis was used as the adulterant.

Cloning, Purification, and Characterization of the Catalytic C-Terminal Domain of the Human 3-Hydroxy-3-methyl glutaryl-CoA Reductase: An Effective, Fast, and Easy Method for Testing Hypocholesterolemic Compounds.

3-hydroxy-3-methyl glutaryl-CoA reductase, also known as HMGR, plays a crucial role in regulating cholesterol biosynthesis and represents the main pharmacological target of statins. In mammals, this enzyme localizes to the endoplasmic reticulum membrane. HMGR includes different regions, an integral N-terminal domain connected by a linker-region to a cytosolic C-terminal domain, the latter being responsible for enzymatic activity. The aim of this work was to design a simple strategy for cloning, expression, and purification of the catalytic C-terminal domain of the human HMGR (cf-HMGR), in order to spectrophotometrically test its enzymatic activity. The recombinant cf-HMGR protein was heterologously expressed in Escherichia coli, purified by Ni+-agarose affinity chromatography and reconstituted in its active form. MALDI mass spectrometry was adopted to monitor purification procedure as a technique orthogonal to the classical Western blot analysis. Protein identity was validated by MS and MS/MS analysis, confirming about 82% of the recombinant sequence. The specific activity of the purified and dialyzed cf-HMGR preparation was enriched about 85-fold with respect to the supernatant obtained from cell lysate. The effective, cheap, and easy method here described could be useful for screening statin-like molecules, so simplifying the search for new drugs with hypocholesterolemic effects.

Preparation of ETS-10 Microporous Phase Pellets with Color Change Properties.

The main scope of the present work is to synthesize pH-responsive Engelhard titanium silicate (ETS)-10 phase crystalline pellets through the smart modification of a synthetic process which was previously applied to the preparation of other phases. The original preparative method, which envisages the use of the same initial synthesis as a binder for the preparation of pellets, was modified by adding an appropriate pH indicator to a number of systems subject to this investigation. It should be noted that the modified process was never before used to give access to pH-responsive ETS-10 phase pellets, and it is disclosed here for the first time. The study started from the definition of the best experimental conditions, which were optimized by analyzing the effects of temperature and system composition. The addition of the pH indicator did not alter the physicochemical characteristics and reactivity of the system. The pH-responsive ETS-10 phase crystalline pellets were characterized by an adequate mechanical strength and by a high capability to change color. The latter aspect can be particularly useful when this material is used in catalytic processes whose performance is strictly dependent on the pH value. The amount of gel used, as well as the working temperature, were the main critical parameters to be controlled during the preparation of pH-responsive ETS-10 phase crystalline pellets. The pellets were fully characterized by X-ray diffraction in order to investigate and identify the possible phases, and by using a hardness tester to measure the compressive strength. Finally, toning tests were performed.

Breakthroughs in Medicinal Chemistry: New Targets and Mechanisms, New Drugs, New Hopes-5.

Breakthroughs in Medicinal Chemistry: New Targets and Mechanisms, New Drugs, New Hopes is a series of Editorials which is published on a biannual basis by the Editorial Board of the Medicinal Chemistry section of the journal Molecules [...].

Removal of unleaded gasoline from water by multi-walled carbon nanotubes.

This article displays an efficient and cost effective technique for the removal of unleaded gasoline from water. Multi-walled carbon nanotubes (MWCNTs) were used as the sorbent material. Nanotubes were synthesized according to a well-known procedure and successfully used avoiding cumbersome purifications from traces of catalyst. A series of lab-scale experiments was performed on dispersions of commercial unleaded gasoline (20 mL) in water (30 mL), which were subjected to the action of variable amounts of MWCNTs at room temperature. Physicochemical characteristics and sorbent capacity of nanotubes were investigated by thermal analysis and FT-IR spectroscopy. The highest percentage of removed unleaded gasoline was obtained using small amounts (0.7 g) of MWCNTs, over very short stirring times (5 min). The composition of residual organic materials in water was investigated by 1H and 13C high-resolution NMR spectroscopy, which confirmed the almost complete removal of unleaded gasoline hydrocarbon components from polluted waters.

A rapid method for the assay of methylxanthines alkaloids: Theobromine, theophylline and caffeine, in cocoa products and drugs by paper spray tandem mass spectrometry.

A fast method for the determination of methylxanthines in cocoa products and drugs based on PS-MS/MS under MRM condition has been developed. Analyte ions were generated by applying a high voltage on a paper substrate drenched of sample extract using a small volume (∼15 µL) of spray solvent. The gas phase chemistry of the molecules under investigation has been elucidated. The accuracy values of the methodology ranged from 95 to 110%, while the analytical parameters LOQ, LOD, recovery and reproducibility, calculated analyzing spiked samples, confirmed the consistency of the proposed method. Furthermore, real samples have been analyzed both by the developed methodology and by using a classical HPLC-UV approach obtaining comparable values.

Breakthroughs in Medicinal Chemistry: New Targets and Mechanisms, New Drugs, New Hopes-6.

Breakthroughs in Medicinal Chemistry: New Targets and Mechanisms, New Drugs, New Hopes is a series of Editorials that is published on a biannual basis by the Editorial Board of the Medicinal Chemistry section of the journal Molecules [...].

Human coelomic fluid investigation: A MS-based analytical approach to prenatal screening.

Coelomic fluid (CF) is the earliest dynamic and complex fluid of the gestational sac. CF contains maternal cells and proteins produced by embryonic cells, tissues and excretions. The biochemical composition of CF is modified throughout the first trimester of pregnancy and its protein profile reflects both physiological/pathological changes affecting the embryo and mother. Identification of variations in the balance of proteins might indicate particular types of pathologies, or ascertain specific genetic disorders. A platform utilizing protein enrichment procedures coupled with shotgun identification and iTRAQ differentiation provided the identification and quantitation of 88 unique embryonic proteins. It is relevant to note that chromosome X protein CXorf23 was found suggesting the embryo sex. Foetal sex was determined by Quantitative Fluorescent Polymerase Chain Reaction (QF-PCR) on coelomic cells, foetal tissues and maternal white blood cells, with a 100% concordance rate between iTRAQ-MS/MS and QF-PCR data. The functional associations among the identified proteins were investigated using STRING database. Open Targets Platform showed as significant the following therapeutic areas: nervous, respiratory, eye and head system disease.