Characterization of the main phase transition in 1,2-dipalmitoyl-phosphatidylcholine luvs by 1H NMR

The main phase transition (Tm) of 100 nm large unilamellar vesicles (LUVs) of 1,2-dipalmitoylphosphatidylcholine (DPPC) was investigated using 1H NMR (proton magnetic resonance) in deuterium oxide, and both DSC (differential scanning calorimetry) and IR (infrared) spectroscopy in water and deuterium oxide. The ability of 1H NMR to determine Tm was demonstrated and the values obtained were in general agreement with those observed with DSC and IR. However, the temperature range of the transition observed by NMR was significantly broader than that observed with either DSC or IR. The effect of deuterium oxide on Tm was studied by comparing results obtained in water and deuterium oxide with DSC and IR. The results showed no significant difference in Tm or temperature range of transition determined in these solvents.     

SYNTHESIS AND STRUCTURE DETERMINATION OF SOME OXADIAZOLE-2-THIONE AND TRIAZOLE-3-THIONE GALACTOSIDES

The syntheses of 5-pyridyl-3((β-D-galactopyranosyl)-1,3,4-oxadiazole-2-thiones 3a–3c and 5-pyridyl-2((β-D-galactopyranosyl)-4-benzyl-1,2,4-triazole-3-thiones 6a–6c are reported. The existence of N-galactosides – not S-galactosides – was proven by IR and ¹⁵N NMR spectroscopy. The structures of the final products and the intermediates were elucidated by IR, ¹H, ¹³C and ¹⁵N NMR spectroscopy and mass spectrometry.

     

Synthesis and anticandidal evaluation of new benzothiazole derivatives with hydrazone moiety

In this study, we have performed the synthesis of new N′-(arylidene)-4-[(benzothiazol-2-yl)thio]butanoylhydrazide derivatives (3a–s) bearing azole moiety and hydrazone group in a lipophilic structural framework. The target compounds were prepared by a three step synthetic procedure starting from 2-mercaptobenzothiazole. The structures of the target compounds were elucidated by IR, ¹H NMR, ¹³C NMR spectra and elemental analysis. The antifungal activity of the obtained compounds has been determined against a number of clinic and fluconazole-resistant Candida strains by using microdilution method. Compounds (3a–3s) exhibited anticandidal activity in different ratios varying between the range of MIC: 50 and 200?µg/mL.     

Effect of substitution of hydrogen oxide by deuterium oxide on thermotropic transition between the interdigitated gel phase and the ripple phase of dihexadecylphosphatidylcholine.

Thermotropic transitions of dihexadecylphosphatidylcholine (DHPC) dispersions in hydrogen oxide (1H2O) and deuterium oxide (2H2O) were investigated by differential scanning calorimetry (DSC). In DHPC dispersions, transition temperature between interdigitated gel phase (L beta I) and ripple phase (P beta') is lower in 2H2O than in 1H2O, and transition between the ripple phase (P beta') and fluid phase (L alpha) in 2H2O occurs at a temperature slightly higher than in 1H2O. In dipalmitoylphosphatidylcholine (DPPC) dispersions, on the other hand, transition temperature between lamellar gel phase (L beta') and ripple phase is higher in 2H2O than in 1H2O. These results suggest that the interdigitated gel phase is more stable in 1H2O than in 2H2O. To account for the shift of transition temperature by the water substitution, difference of interfacial energies between these aqueous environments is discussed.     

Synthesis,characterization, antiamoebic activity and cytotoxicity of new pyrazolo[3, 4-d]pyrimidine-6-one derivatives

A new series of pyrazolo[3,4-d]pyrimidine-6-one derivatives (2a–2j) were prepared by using the Biginelli multicomponent cyclocondensation of 3-methyl-1-phenyl-1H-pyrazol-5(4H)-one (1a), different aromatic aldehydes, and urea with a catalytic amount of HCl at reflux temperature. These compounds were characterized by IR, ¹H NMR, ¹³C NMR, and Mass spectral data. In vitro antiamoebic activity was performed against HM1:IMSS strain of Entamoeba histolytica. The results showed that the compounds 2b, 2i, and 2j with IC₅₀ values of 0.37 µM, 0.04 µM, and 0.06 µM, respectively, exhibited better antiamoebic activity than the standard drug metronidazole (IC₅₀?=?1.33 µM). The toxicological studies of these compounds on human breast cancer MCF-7 cell line showed that the compounds 2b, 2i, and 2j exhibited >80% viability at the concentration range of 1.56–50 µM.     

Organization of chlorophyll a in bilayer membranes. The chlorophyll a/dimyristoylphosphatidylcholine system

In order to investigate the relative importance of the hydrophobic and headgroup interactions of chlorophyll a in phospholipid bilayers, we have carried out differential scanning calorimetry (DSC) and deuterium (²H) and phosphorus (³¹P) nuclear magnetic resonance (NMR) experiments on the multilamellar system of chlorophyll a in dimyristoylphosphatidylcholine (DMPC). Compared to the phytol chain of chlorophyll and the previously reported distearoylphosphatidylcholine (DSPC), the acyl chains of DMPC are shorter in length by three and four carbons, respectively. A lowering in the phase-transition temperature was observed for the DMPC multilayers in the presence of chlorophyll a in the DSC thermograms and in the ³¹P chemical shift anisotropy measurements. These results, together with data on hydrophobic interactions as measured by ²H-NMR and on headgroup interactions as evidenced from ³¹P-NMR, suggest a phase diagram for the chlorophyll a/DMPC system in which phase separation readily occurs between a chlorophyll-rich compound phase and a chlorophyll-poor phospholipid phase. Compound formation appears to be important in the stabilization of chlorophyll a in bilayers with shorter chains.     

Estimation of Differently Bound Water Molecules for the Gel Phase of Dimyristoylphosphatidylethanolamine-Water System as Studied by DSC and 2H-NMR Spectroscopy

A measurement of ²H spin-lattice relaxation time, T ₁, forD₂O was performed with a high resolution liquid NMR apparatus fortwo samples of dimyristoylphosphatidylethanolamine (DMPE)-D₂Osystem in a full hydration at varying temperatures of –20, –10, and 5 ^°C, and both components and compositions of differently boundfreezable water molecules were estimated from a best-fitted curve toexperimental inversion recovery data. A choice of the best-fitted curve wasbased on a distribution of weighted residuals for the experimental data. Asingle component was found for a temperature of –20 ^°C. At 5 ^°C, where all the freezable water exists in the liquid state, threecomponents were observed to be characterized by T ₁ values ofapproximately 20, 100, and 200 ms, respectively. By comparingcompositions of these individual components with those obtained in ourprevious DSC study, it was revealed that the first and secondarycomponents are members of freezable interlamellar water and the last oneis comparable to bulk water.     

The Nature and Origin of Chemical Shift for Intracellular Water Nuclei in Artemia Cysts

We investigated the possible existence of chemical shift of water nuclei in Artemia cysts using high resolution nuclear magnetic resonance (NMR) methods. The results conducted at 60, 200, and 500 MHz revealed an unusually large chemical shift for intracellular water protons. After correcting for bulk susceptibility effects, a residual downfield chemical shift of 0.11 ppm was observed in fully hydrated cysts. Similar results have been observed for the deuterium and ¹⁷O nuclei.

We have ruled out unusual intracellular pH, diamagnetic susceptibility of intracellular water, or interaction of water molecules with lipids, glycerol, and/or trehalose as possible origins of the residual chemical shift. We conclude that the residual chemical shift observed for water nuclei (¹H, ²H, and ¹⁷O) is due to significant water-macromolecular interactions.

     

First novel synthesis of triazole thioglycosides as ribavirin analogues

This study reports a novel and efficient method for the synthesis of the first reported novel class of triazole thioglycosides. These series of compounds were designed through the reaction of potassium cyanocarbonimidodithioate 2 with hydrazine derivatives 3a-d in EtOH at room temperature to give the corresponding potassium 5-amino-1H-1,2,4-triazole-3-thiolates 4a-d. The latter compounds were treated with tetra-O-acetyl-α-D-glucopyranosyl bromide 6a and tetra-O-acetyl-α-D-galactopyranosyl bromide 6b in DMF at room temperature to give in high yields the corresponding triazole thioglycosides 7a-h. Treatment of triazole salts 4a–d with hydrochloric acid afforded the corresponding 3-mercaptotriazoles 5a-d. Compounds 5a-d were then reacted with bromoperacetylated sugars 6a,b in sodium hydride-DMF at ambient temperature to afford the thioglycosyl compounds 7a-h. Ammonolysis of the triazole thioglycosides 7a-h afforded the corresponding free thioglycosides 8a-h. The scope and limitation of the method is demonstrated. The structure of the reaction products was confirmed on the basis of their elemental analysis and spectral data (IR, ¹H NMR, MS and ¹³C NMR).     

Synthesis,Structural Characterization,and Chiroptical Studies of Bidentate Salen‐Type Lanthanide (III) Complexes

The salen‐type ligand prepared with (R,R) diphenylethan‐1,2‐diamine and salicylaldehyde provides stable and inert complexes KLnL₂ upon simple reaction with lanthanide halides or pseudohalides LnX₃ (Ln = Tb³⁺‐Lu³⁺; X = Cl^? or TfO^?) of its potassium salt. All the complexes were completely characterized through nuclear magnetic resonance (NMR), electronic circular dichroism (ECD) in the UV and some (Er³⁺, Tm³⁺, Yb³⁺) also with Near‐IR ECD (NIR‐ECD) and luminescence (Tb³⁺, Tm³⁺). Careful analysis of the NMR shifts demonstrated that the complexes are isostructural in solution and afforded an accurate geometry. This was further confirmed by means of Density Functional Theory (DFT) optimization of the Lu³⁺ complex, and by comparing the ligand‐centered experimental and time‐dependent TD‐DFT computed UV‐ECD spectra. As final validation, we used the NIR‐ECD spectrum of the Yb³⁺ derivative calculated by means of Richardson's equations. The excellent match between calculated and experimental ECD spectra confirm the quality of the NMR structure.  Chirality 27:857–863, 2015. © 2015 Wiley Periodicals, Inc.     

New pentadienone oxime ester derivatives: synthesis and anti-inflammatory activity

To develop novel anti-inflammatory agents, a series of new pentadienone oxime ester compounds were designed and synthesized. The structures were determined by IR, ¹H NMR, ^13?C NMR, and HRMS. All compounds have been screened for their anti-inflammatory activity by evaluating their inhibition against LPS-induced nitric oxide (NO) release in RAW 264.7 cell. Among them, compound 5j was found to be one of the most potent compounds in inhibiting NO and IL-6 (IC₅₀ values were 6.66?µM and 5.07?µM, respectively). Preliminary mechanism studies show that title compound 5j could significantly suppress expressions of nitric oxide synthase, COX-2, and NO, IL-6 through Toll-like receptor 4/mitogen-activated protein kinases/NF-κB signalling pathway. These data support further studies to assess rational design of more efficient pentadienone oxime ester derivatives with anti-inflammatory activity in the future.     

Microwave accelerated solvent-free synthesis and antifungal evaluations of flavanones

Microwave irradiation of 2-hydroxy chalcones under solvent-free conditions resulted in a “green-chemistry” procedure for the preparation of flavanones in good yields, using an unmodified household microwave oven and silica as solid support. By irradiation of 2-hydroxy chalcones with trifluoroacetic acid over silica gel, 11 known flavanones were prepared in high yields. The synthesised compounds were characterised using spectroscopic techniques, namely, ¹H NMR, ¹³C NMR and IR, and screened for their antifungal activity in vitro against Sclerotium rolfsii and Rhizoctonia solani by poisoned food technique. The compounds tested were found to be more active against R. solani, whereas against S. rolfsii, moderate activity was observed, as evident from LC₅₀ values. The most potent compound 2-(4-fluorophenyl)-2,3-dihydrochromen-4-one (4a) had LC₅₀ value of 12.0 mg L^?1 followed by 11, 11a, 3a, 9a, 8a, 10a and 10 having LC₅₀ values 18.21, 18.3, 32.9, 50.7, 88.8, 118.8 and 119.7 mg L^?1, respectively.     

Metabolomics for the Quality Assessment of Lycium chinense Fruits

Aibellin is a 20-residue peptide antibiotic that has been isolated from the fungus Verticimonosporium ellipticum. Sequence-specific assignment of the ¹H- and ¹³C-NMR signals of aibellin in a methanol solution was achieved by using the two-dimensional NMR technique. Furthermore, its secondary structure was characterized by circular dichroism (CD) and NOESY spectra. The observed NOEs, ³J_NHCαH coupling constants and amide hydrogen–deuterium (H–D) exchange rates show that the peptide consisted of two α-helices and a bent structure around a Pro-14 residue.     

Insights into Sphingolipid Miscibility: Separate Observation of Sphingomyelin and Ceramide N-Acyl Chain Melting

Ceramide produced from sphingomyelin in the plasma membrane is purported to affect signaling through changes in the membrane’s physical properties. Thermal behavior of N-palmitoyl sphingomyelin (PSM) and N-palmitoyl ceramide (PCer) mixtures in excess water has been monitored by ²H NMR spectroscopy and compared to differential scanning calorimetry (DSC) data. The alternate use of either perdeuterated or proton-based N-acyl chain PSM and PCer in our ²H NMR studies has allowed the separate observation of gel-fluid transitions in each lipid in the presence of the other one, and this in turn has provided direct information on the lipids’ miscibility over a wide temperature range. The results provide further evidence of the stabilization of the PSM gel state by PCer. Moreover, overlapping NMR and DSC data reveal that the DSC-signals parallel the melting of the major component (PSM) except at intermediate (20 and 30 mol %) fractions of PCer. In such cases, the DSC endotherm reports on the presumably highly cooperative melting of PCer. Up to at least 50 mol % PCer, PSM and PCer mix ideally in the liquid crystalline phase; in the gel phase, PCer becomes incorporated into PSM:PCer membranes with no evidence of pure solid PCer.     

Ionic liquid catalyzed synthesis and characterization of heterocyclic and optically active poly (amide-imide)s incorporating <Emphasis Type="SmallCaps">l</Emphasis>-amino acids

N,N′-Pyromelliticdiimido-di-l-alanine (1), N,N′-Pyromelliticdiimido-di-l-phenylalanine (2), and N,N′-Pyromelliticdiimido-di-l-leucine (3) were prepared from the reaction of Pyromellitic dianhydride with corresponding l-amino acids in a mixture of glacial acetic acid and pyridine solution (3/2 ratio) under refluxing conditions. A series of poly (amide-imide)s containing l-amino acids were prepared from the synthesized dicarboxylic acids with two synthetic aromatic diamines in an ionic liquid (IL) as a green, safe and eco-friendly medium and also reactions catalysis agent. Evaluation of data shows that IL is the better polyamidation medium than the reported method and the catalysis stand on the higher inherent viscosities of the obtained PAIs and the rate of polymerizations beyond the greener reaction conditions and deletion of some essential reagents in conventional manners. Characterization were performs by means of IR, MS and ¹H NMR spectroscopy, elemental analysis, specific rotation, thermogravimetric analysis and differential scanning calorimetric techniques. Molecular weights of the obtained polymers were evaluated viscometrically, and the measured inherent viscosities were in the range 0.43–0.85 dL/g. These polymers were readily soluble in many organic solvents. These polymers still kept good thermal stability with glass transition temperatures in the range of 94–154°C, and the decomposition temperature under the nitrogen atmosphere for 10% weight-loss temperatures in excess of 308°C.     

Deuterium NMR study of the interaction of alpha-tocopherol with a phospholipid model membrane

The effects of 5, 10, and 20 mol % incorporation of alpha-tocopherol (vitamin E) on 50 wt % aqueous multilamellar dispersions of sn-2-substituted [2H31]palmitoylphosphatidylcholine (PC-d31), a saturated, deuterated phospholipid prepared from egg lysophosphatidylcholine, have been studied by deuterium nuclear magnetic resonance (2H NMR) and differential scanning calorimetry (DSC). Moment analysis of the 2H NMR spectra as a function of temperature and DSC heating curves demonstrate that the main gel to liquid-crystalline phase transition is progressively broadened and its onset temperature lowered by increasing concentrations of alpha-tocopherol. Below the transition temperature (40 degrees C) for PC-d31 bilayers, the 2H NMR spectra indicate that acyl chain motion is increased by addition of alpha-tocopherol and that this effect extends to lower temperatures with higher alpha-tocopherol content. Above the transition, average carbon-deuterium bond order parameters calculated from the first spectral moment establish that alpha-tocopherol increases acyl chain ordering within the PC-d31 bilayer by as much as 17% at 20 mol % incorporation. Profiles of order parameter vs. chain position, constructed from 2H NMR spectra following application of the depaking technique, show that despite higher order the general form of the profile is not significantly altered by alpha-tocopherol.     

Deuterium isotope effects in carbohydrates revisited. Cryoprobe studies of the anomerization and NH to ND deuterium isotope induced 13C NMR chemical shifts of acetamidodeoxy and aminodeoxy sugars

Complete ¹H and ¹³C NMR chemical shift assignments have been generated from a series of acetamidodeoxy and aminodeoxy sugar derivatives. For free sugars, the enhanced sensitivity of an NMR cryoprobe allowed simple 1D and 2D NMR spectra to be obtained from essentially single anomers, before significant mutarotation had occurred. The NMR assignments have been used to characterize deuterium isotope effects on ¹³C chemical shifts measured under conditions of slow NH to ND exchange in single solutions. Within a range of 0 to −0.138 ppm, β, γ, δ, and ζ deuterium isotope effects have been observed, thus providing additional reference data for assignment of the ¹³C NMR spectra of nitrogenous saccharides.     

Benzimidazolium-based novel silver N-heterocyclic carbene complexes: synthesis,characterisation and in vitro antimicrobial activity

This study reports the synthesis, characterisation and antimicrobial activity of five novel silver N-heterocyclic carbene (Ag–NHC) complexes obtained by N-propylphthalimide and N-methyldioxane substituted benzimidazolium salts with silver oxide. The reactions were performed at room temperature for 24?h in the absence of light. The obtained complexes were identified and characterised by ¹H and ¹³C NMR, FT-IR and elemental analysis techniques. The minimum inhibitory concentration (MIC) of the complexes was determined for E. coli, P. aeruginosa, E. faecalis, S. aureus, C. tropicalis and C. albicans in vitro through agar and broth dilution. The results indicated that these complexes exhibit antimicrobial activity. In particular, complex 3 presented the significant broad spectrum antimicrobial activity.     

Biotransformation of 3β-hydroxy-5-en-steroids by Mucor silvaticus

Abstract

The biotransformation of four 3β-hydroxy-5-en-steroids with varying substituents at C-16 or/and C-17 by Mucor silvaticus was investigated. The characterization of the metabolites was performed by IR, MS, ¹H NMR, ¹³C NMR, and 2-D NMR. All the examined substrates were transformed, mainly by 7α-hydroxylation. Studies carried out with M. silvaticus demonstrated the versatility of this organism in introducing hydroxyl groups at the 7α-, 9α-, 11α-, and 14α-positions in 3-ol-5-ene steroids. The relationships between the substrate structures and hydroxylated positions are also discussed.     

Comparison of the diastereoisomeric excess of uridine,inosine and adenosine cyanohydrins determined by HPLC-DAD and 1H NMR.

The separation of the diastereoisomers of the nucleoside derivatives of uridine, inosine and adenosine was performed by HPLC using chiral and no chiral columns, it was observed with the no chiral columns the resolution was good enough to determine diastereoisomeric excess. These methods were compared with ¹H NMR, and no significant differences were observed between the three techniques. Diastereoisomeric uridine (3a), inosine (3b) and adenosine (4c) cyanohydrins were resolved by ¹H nuclear magnetic resonance (¹H NMR), chiral normal phase-high-performance liquid chromatography-diode array detector (NP-HPLC-DAD) and reversed phase (RP-HPLC-DAD); these methods allowed the assesment of the percent diastereoisomeric excess (% de) of the nucleosidic cyanohydrins of 3a (4, 6 and 4), 3b (10, 8 and 6) and 4c (4, 4 and 4). To the best of our knowledge, there are no reports using analytical techniques for the separation of the epimers of 3a, 3b and 4c.