Síntese e atividade anti-HCV, anti-candida e antibacteriana de nitrochalconas

Pós-graduação em Química - IBILCE

Synthesis and characterization of the atropisomeric relationships of a substituted N-phenyl-bipyrazole derivative with anti-inflammatory properties

This work describes the atropisomeric relationships of 3-methyl-5-(3-methyl-5-phenyl-1H-pyrazol-1-yl)-1-phenyl-1H-pyrazol-4-amine (2d), which belongs to series 4-aminobipyrazole derivatives designed as anti-inflammatory agents. The 1H nuclear magnetic resonance spectra obtained in the presence of a chiral lanthanide shift salt associated to chiral high-performance liquid chromatography analysis, X-ray diffraction, and molecular modeling tools confirmed that ortho bis-functionalized bipyrazole 2d exists as a mixture of aR,aS-atropisomers. These results provide useful information to understand the pharmacological profile of this derivative and of other 4-aminobipyrazole analogs. Chirality 24:463470, 2012. (c) 2012 Wiley Periodicals, Inc.

Self assembled monolayers for engineering of structured inorganic materials in the micrometer and submicrometer range

Deutsch:In der vorliegenden Arbeit konnten neue Methoden zur Synthese anorganischer Materialien mit neuartiger Architektur im Mikrometer und Nanometer Maßstab beschrieben werden. Die zentrale Rolle der Formgebung basiert dabei auf der templatinduzierten Abscheidung der anorganischen Materialien auf selbstorganisierten Monoschichten. Als Substrate eignen sich goldbedampfte Glasträger und Goldkolloide, die eine Mittelstellung in der Welt der Atome bzw. Moleküle und der makroskopischen Welt der ausgedehnten Festkörper einnehmen. Auf diesen Substraten lassen sich Thiole zu einer monomolekularen Schicht adsorbieren und damit die Oberflächeneigenschaften des Substrates ändern. Ein besonderer Schwerpunkt bei dieser Arbeit stellt die Synthese speziell auf die Bedürfnisse der jeweiligen Anwendung ausgerichteten Thiole dar.Im ersten Teil der Arbeit wurden goldbedampfte Glasoberflächen als Template verwendet. Die Abscheidung von Calciumcarbonat wurde in Abhängigkeit der Schichtdicke der adsorbierten Monolage untersucht. Aragonit, eine der drei Hauptphasen des Calciumcarbonat Systems, wurde auf polyaromatischen Amid - Oberflächen mit Schichtdicken von 5 - 400 nm Dicke unter milden Bedingung abgeschieden. Die einstellbaren Parameter waren dabei die Kettenlänge des Polymers, der w-Substituent, die Bindung an die Goldoberfläche über Verwendung verschiedener Aminothiole und die Kristallisationstemperatur. Die Schichtdickeneinstellung der Polymerfilme erfolgte hierbei über einen automatisierten Synthesezyklus.Titanoxid Filme konnten auf Oberflächen strukturiert werden. Dabei kam ein speziell synthetisiertes Thiol zum Einsatz, das die Funktionalität einer Styroleinheit an der Oberflächen Grenze als auch eine Möglichkeit zur späteren Entfernung von der Oberfläche in sich vereinte. Die PDMS Stempeltechnik erzeugte dabei Mikrostrukturen auf der Goldoberfläche im Bereich von 5 bis 10 µm, die ihrerseits über die Polymerisation und Abscheidung des Polymers in den Titanoxid Film überführt werden konnten. Drei dimensionale Strukturen wurden über Goldkolloid Template erhalten. Tetraethylenglykol konnte mit einer Thiolgruppe im Austausch zu einer Hydroxylgruppe monofunktionalisiert werden. Das erhaltene Molekül wurde auf kolloidalem Gold selbstorganisiert; es entstand dabei ein wasserlösliches Goldkolloid. Die Darstellung erfolgte dabei in einer Einphasenreaktion. Die so erhaltenen Goldkolloide wurden als Krstallisationstemplate für die drei dimensionale Abscheidung von Calciumcarbonat verwendet. Es zeigte sich, dass Glykol die Kristallisation bzw. den Habitus des krsitalls bei niedrigem pH Wert modifiziert. Bei erhöhtem pH Wert (pH = 12) jedoch agieren die Glykol belegten Goldkolloide als Template und führen zu sphärisch Aggregaten. Werden Goldkolloide langkettigen Dithiolen ausgesetzt, so führt dies zu einer Aggregation und Ausfällung der Kolloide aufgrund der Vernetzung mehrer Goldkolloide mit den Thiolgruppen der Alkyldithiole. Zur Vermeidung konnte in dieser Arbeit ein halbseitig geschütztes Dithiol synthetisiert werden, mit dessen Hilfe die Aggregation unterbunden werden konnte. Das nachfolgende Entschützten der Thiolfunktion führte zu Goldkolloiden, deren Oberfläche Thiol funktionalisiert werden konnte. Die thiolaktiven Goldkolloide fungierten als template für die Abscheidung von Bleisulfid aus organisch/wässriger Lösung. Die Funktionsweise der Schutzgruppe und die Entschützung konnte mittels Plasmonenresonanz Spektroskopie verdeutlicht werden. Titanoxid / Gold / Polystyrol Komposite in Röhrenform konnten synthetisiert werden. Dazu wurde ein menschliches Haar als biologisches Templat für die Formgebung gewählt.. Durch Bedampfung des Haares mit Gold, Assemblierung eines Stryrolmonomers, welches zusätzlich eine Thiolfunktionalität trug, Polymerisation auf der Oberfläche, Abscheidung des Titanoxid Films und anschließendem Auflösen des biologischen Templates konnte eine Röhrenstruktur im Mikrometer Bereich dargestellt werden. Goldkolloide fungierten in dieser Arbeit nicht nur als Kristallisationstemplate und Formgeber, auch sie selbst wurden dahingehend modifiziert, dass sie drahtförmige Agglormerate im Nanometerbereich ausbilden. Dazu wurden Template aus Siliziumdioxid benutzt. Zum einen konnten Nanoröhren aus amorphen SiO2 in einer Sol Gel Methode dargestellt werden, zum anderen bediente sich diese Arbeit biologischer Siliziumoxid Hohlnadeln aus marinen Schwämmen isoliert. Goldkolloide wurden in die Hohlstrukturen eingebettet und die Struktur durch Ausbildung von Kolloid - Thiol Netzwerken mittels Dithiol Zugabe gefestigt. Die Gold-Nanodrähte im Bereich von 100 bis 500 nm wurden durch Auflösen des SiO2 - Templates freigelegt.

Copper binding to the prion protein: Structural implications of four identical cooperative binding sites

Evidence is growing to support a functional role for the prion protein (PrP) in copper metabolism. Copper ions appear to bind to the protein in a highly conserved octapeptide repeat region (sequence PHGGGWGQ) near the N terminus. To delineate the site and mode of binding of Cu(II) to the PrP, the copper-binding properties of peptides of varying lengths corresponding to 2-, 3-, and 4-octarepeat sequences have been probed by using various spectroscopic techniques. A two-octarepeat peptide binds a single Cu(II) ion with Kd ≈ 6 μM whereas a four-octarepeat peptide cooperatively binds four Cu(II) ions. Circular dichroism spectra indicate a distinctive structuring of the octarepeat region on Cu(II) binding. Visible absorption, visible circular dichroism, and electron spin resonance spectra suggest that the coordination sphere of the copper is identical for 2, 3, or 4 octarepeats, consisting of a square-planar geometry with three nitrogen ligands and one oxygen ligand. Consistent with the pH dependence of Cu(II) binding, proton NMR spectroscopy indicates that the histidine residues in each octarepeat are coordinated to the Cu(II) ion. Our working model for the structure of the complex shows the histidine residues in successive octarepeats bridged between two copper ions, with both the Nɛ2 and Nδ1 imidazole nitrogen of each histidine residue coordinated and the remaining coordination sites occupied by a backbone amide nitrogen and a water molecule. This arrangement accounts for the cooperative nature of complex formation and for the apparent evolutionary requirement for four octarepeats in the PrP.

Site-directed spin labeling and chemical crosslinking demonstrate that helix V is close to helices VII and VIII in the lactose permease of Escherichia coli.

Site-directed chemical cleavage of lactose permease indicates that helix V is in close proximity to helices VII and VIII. To test this conclusion further, permease containing a biotin-acceptor domain and paired Cys residues at positions 148 (helix V) and 228 (helix VII), 148 and 226 (helix VII), or 148 and 275 (helix VIII) was affinity purified and labeled with a sulfhydryl-specific nitroxide spin label. Spin-spin interactions are observed with the 148/228 and 148/275 pairs, indicating close proximity between appropriate faces of helix V and helices VII and VIII. Little or no interaction is evident with the 148/226 pair, in all likelihood because position 226 is on the opposite face of helix VII from position 228. Broadening of the electron paramagnetic resonance spectra in the frozen state was used to estimate distance between the 148/228 and the 148/275 pairs. The nitroxides at positions 148 and 228 or 148 and 275 are within approximately 13-15 A. Finally, Cys residues at positions 148 and 228 are crosslinked by dibromobimane, a bifunctional crosslinker that is approximately 5 A. long, while no crosslinking is detected between Cys residues at positions 148 and 275 or 148 and 226. The results provide strong support for a structure in which helix V is in close proximity to both helices VII and VIII and is oriented in such a fashion that Cys-148 is closer to helix VII.

Distance determination in proteins using designed metal ion binding sites and site-directed spin labeling: application to the lactose permease of Escherichia coli.

As shown in the accompanying paper, the magnetic dipolar interaction between site-directed metal-nitroxide pairs can be exploited to measure distances in T4 lysozyme, a protein of known structure. To evaluate this potentially powerful method for general use, particularly with membrane proteins that are difficult to crystallize, both a paramagnetic metal ion binding site and a nitroxide side chain were introduced at selected positions in the lactose permease of Escherichia coli, a paradigm for polytopic membrane proteins. Thus, three individual cysteine residues were introduced into putative helix IV of a lactose permease mutant devoid of native cysteine residues containing a high-affinity divalent metal ion binding site in the form of six contiguous histidine residues in the periplasmic loop between helices III and IV. In addition, the construct contained a biotin acceptor domain in the middle cytoplasmic loop to facilitate purification. After purification and spin labeling, electron paramagnetic resonance spectra were obtained with the purified proteins in the absence and presence of Cu(II). The results demonstrate that positions 103, 111, and 121 are 8, 14, and > 23 A from the metal binding site. These data are consistent with an alpha-helical conformation of transmembrane domain IV of the permease. Application of the technique to determine helix packing in lactose permease is discussed.

Fluidity of the lipid domain of cell wall from Mycobacterium chelonae.

The mycobacterial cell wall contains large amounts of unusual lipids, including mycolic acids that are covalently linked to the underlying arabinogalactan-peptidoglycan complex. Hydrocarbon chains of much of these lipids have been shown to be packed in a direction perpendicular to the plane of the cell surface. In this study, we examined the dynamic properties of the organized lipid domains in the cell wall isolated from Mycobacterium chelonae grown at 30 degrees C. Differential scanning calorimetry showed that much of the lipids underwent major thermal transitions between 30 degree C and 65 degrees C, that is at temperatures above the growth temperature, a result suggesting that a significant portion of the lipids existed in a structure of extremely low fluidity in the growing cells. Spin-labeled fatty acid probes were successfully inserted into the more fluid part of the cell wall. Our model of the cell wall suggests that this domain corresponds to the outermost leaflet, a conclusion reinforced by the observation that labeling of intact cells produced electron spin resonance spectra similar to those of the isolated cell wall. Use of stearate labeled at different positions showed that the fluidity within the outer leaflet increased only slightly as the nitroxide group was placed farther away from the surface. These results are consistent with the model of mycobacterial cell wall containing an asymmetric lipid bilayer, with an internal, less fluid mycolic acid leaflet and an external, more fluid leaflet composed of lipids containing shorter chain fatty acids. The presence of the low-fluidity layer will lower the permeability of the cell wall to lipophilic antibiotics and chemotherapeutic agents and may contribute to the well-known intrinsic resistance of mycobacteria to such compounds.

Chemical composition of glauconite and montmorillonite group minerals from sediments and rocks of the Norwegian Sea

Two quadrupole splitting doublets with delta E_q = 0.74 and 1.62 mm/s were found in the montmorillonite spectra. The more intense doublet corresponds to iron in a somewhat distorted tetrahedral coordination, while the less intense corresponds to quadri-coordinated iron. The EPR spectrum also exhibits two lines with a q-factor of 3 and 4.3, which corresponds to transformed minerals.

Researches in physical optics

pt. I. Researches in physical optics with especial reference to the radiation of electrons. 1913.--pt. II. Resonance radiation and resonance spectra. 1919.

Embracing ligands. Synthesis, characterisation and the correlation between 59Co NMR and ligand field parameters of Co(III) complexes with a new class of nitrogen-thioether multidentate ligand

The syntheses of the hexadentate ligands 2,2,10,10-tetra(methyleneamine)-4,8-dithiaundecane (PrN(4)S(2)amp), 2,2,11,11-tetra(methyleneamine)-4,9-dithiadodecane (BuN(4)S(2)amp), and 1,2-bis(4,4-methyleneamine)-2-thiapentyl)benzene (XyN(4)S(2)amp) are reported and the complexes [Co(RN(4)S(2)amp)](3+) (R = Pr, Bu, Xy) characterised by single crystal X-ray study. The low-temperature (11 K) absorption spectra have been measured in Nafion films. From the observed positions of both spin-allowed (1)A(1g) --> T-1(1g) and (1)A(1g) --> T-1(2g) and spin forbidden (1)A(1g) --> T-3(1g) and (1)A(1g) --> T-3(2g) bands, octahedral ligand-field parameters (10D(q), B and C) have been determined. DFT calculations suggest that significant interaction between the d-d and CT excitations occurs for the complexes. The calculations offer an explanation for the observed deviations from linearity of the relationship between Co-59 magnetogyric ratio and beta(DeltaE)(-1) (beta = the nephelauxetic ratio; DeltaE the energy of the (1)A(1g) --> T-1(1g) transition) for a series of amine and mixed amine/thioether donor complexes.

Synthesis and characterization of cobalt(III) complexes with sinambic amplector ligands

The synthesis of the hexadentate ligand 5,6-dimethyl-2,2,9,9-tetra(methyleneamine)-4,7-dithiadecane (1,2-Me(2)EtN(4)S(2)amp) is reported. The diastereiosomers were separated as cobalt(III) complexes using cation exchange chromatography. The rac and mesa isomers were characterized by NMR (C-13, H-1, Co-59), ESI-MS, UV-Vis spectroscopy and cyclic voltammetry. Single crystals of [Co(rac-1,2-Me(2)EtN(4)S(2)amp)] Cl-2(ClO4) (.) 2H(2)O were characterized by X-ray diffraction. The low-temperature (11 K) absorption spectra of the complexes have been measured in Nafion films and from the observed positions of both spin-allowed (1)A(1g) --> T-1(1g) and (1)A(1g) --> T-1(2g) and spin forbidden (1)A(1g) --> T-3(2g) bands, octahedral ligand-field parameters (10Dq, B and C) were determined. These results, in conjunction with the Co-59 NMR data, are used to further explore the relationship between the Co-59 magnetogyric ratio (gamma(Co)) and the product of the nephelauxetic ratio and the wavelength of the (1)A(1g) --> T-1(1g) transition (beta(DeltaE)(-1)) for complexes of mixed donor nitrogen-thioether ligands. (C) 2004 Elsevier Ltd. All rights reserved.

A cyclopropabenzenylidenethenone (Propadienone) via a new route to alkylidenecycloproparenes

Reaction of 1,1-dichloro-2,5-diphenylcyclopropabenzene 6 with Meldrum's acid 8 in the presence of pyridine leads to coupling of the cycloproparenyl cation 7 with the stabilized diketo anion 9. Subsequent, spontaneous, base-induced dehydrochlorination gives the alkylidenecyclopropabenzene 11 in a one-pot reaction. Flash vacuum thermolysis of 11 at 650 degreesC ejects acetone and carbon dioxide, giving cyclopropabenzenylldenethenone 12 that is isolated in an Ar matrix at 20 K and characterized by a strong ketene band at 2107 cm(-1) in the IR spectrum.

Design and construction of an ultrasonic probe for use in a Cryo-Magnet NMR Spectrometer

SINNMR (Sonically Induced Narrowing of the Nuclear Magnetic Resonance spectra of solids), is a novel technique that is being developed to enable the routine study of solids by nuclear magnetic resonance spectroscopy. SINNMR aims to narrow the broad resonances that are characteristic of solid state NMR by inducing rapid incoherent motion of solid particles suspended in a support medium, using high frequency ultrasound in the range 2-10 MHz. The width of the normal broad resonances from solids are due to incomplete averaging of several components of the total spin Hamiltonian caused by restrictions placed on molecular motion within a solid. At present Magic Angle Spinning (MAS) NMR is the classical solid state technique used to reduce line broadening, but: this has associated problems, not least of which is the appearance of many spinning side bands which confuse the spectra. It is hoped that SlNNMR will offer a simple alternative, particularly as it does not reveal spinning sidebands The fundamental question concerning whether the use of ultrasound within a cryo-magnet will cause quenching has been investigated with success, as even under the most extreme conditions of power, frequency and irradiator time, the magnet does not quench. The objective of this work is to design and construct a SINNMR probe for use in a super conducting cryo-magnet NMR spectrometer. A cell for such a probe has been constructed and incorporated into an adapted high resolution broadband probe. It has been proved that the cell is capable of causing cavitation, up to 10 MHz, by running a series of ultrasonic reactions within it and observing the reaction products. It was found that the ultrasound was causing the sample to be heated to unacceptable temperatures and this necessitated the incorporation of temperature stabilisation devices. Work has been performed on the investigation of the narrowing of the solid state 23Na spectrum of tri-sodium phosphate using high frequency ultrasound. Work has also been completed on the signal enhancement and T1 reduction of a liquid mixture and a pure compound using ultrasound. Some preliminary "bench" experiments have been completed on a novel ultrasonic device designed to help minimise sample heating. The concept involves passing the ultrasound through a temperature stabilised, liquid filled funnel that has a drum skin on the end that will enable the passage of ultrasound into the sample. Bench experiments have proved that acoustic attenuation is low and that cavitation in the liquid beyond the device is still possible.

Cerebellar morphology in developmental dyslexia

Recent evidence has suggested cerebellar anomalies in developmental dyslexia. Therefore, we investigated cerebellar morphology in subjects with documented reading disabilities. We obtained T1-weighted magnetic resonance images in the coronal and sagittal planes from 11 males with prior histories of developmental dyslexia, and nine similarly-aged male controls. Proton magnetic resonance spectra (TE=136 ms, TR=2.4 s) were obtained bilaterally in the cerebellum. Phonological decoding skill was measured using non-word reading. Handedness was assessed using both the Annett questionnaire of hand preference and Annett’s peg moving task. Cerebellar symmetry was observed in the dyslexics but there was significant asymmetry (right grey matter>left grey matter) in controls. The interpretation of these results depended whether a motor- or questionnaire-based method was used to determine handedness. The degree of cerebellar symmetry was correlated with the severity of dyslexics’ phonological decoding deficit. Those with more symmetric cerebella made more errors on a nonsense word reading measure of phonological decoding ability. Left cerebellar metabolite ratios were shown to correlate significantly with the degree of cerebellar asymmetry (P<0.05) in controls. This relationship was absent in developmental dyslexics. Cerebellar morphology reflects the higher degree of symmetry found previously in the temporal and parietal cortex of dyslexics. The relationship of cerebellar asymmetry to phonological decoding ability and handedness, together with our previous finding of altered metabolite ratios in the cerebellum of dyslexics, lead us to suggest that there are alterations in the neurological organisation of the cerebellum which relate to phonological decoding skills, in addition to motor skills and handedness.

Competing anisotropies in exchange biased nano-structured thin films

The magnetic anisotropies of a patterned, exchange biased Fe50Mn50/Ni80Fe20 system are studied using ferromagnetic resonance, supplemented by Brillouin light scattering experiments and Kerr magnetometry. The exchange biased bi-layer is partially etched into an antidot geometry so that the system approximates a Ni80 Fe20 layer in contact with antidot structured Fe50 Mn50 . Brillouin light scattering measurements of the spin wave frequency dependence on the wave vector reveal a magnonic band gap as expected for a periodic modulation of the magnetic properties. Analysis of the ferromagnetic resonance spectra reveals 8-fold and 4-fold contributions to the magnetic anisotropy. Additionally, the antidot patterning decreases the magnitude of the exchange bias and modifies strongly its angular dependence. Softening of all resonance modes is most pronounced for the applied magnetic field aligned within 10◦ of the antidot axis, in the direction of the bias. Given the degree to which one can tailor the ground state, the resulting asymmetry at low frequencies could make this an interesting candidate for applications such as selective/directional microwave filtering and multi-state magnetic logic.