Determination of copper by AAS in tear fluid of patients with keratoconus.

Keratoconus (KC) is the most common degenerative corneal disease and no single biomarker for KC has been discovered. Its causes have not yet been clarified and this work aims to be a contribution to the deepening of the knowledge of this disease and a preliminary data to the evaluation of the possibility of the use of copper (Cu) concentration in the tear fluid as a specific marker. A tear fluid sampling and Cu determination by spectrometric atomic absorption method was optimized to determine Cu levels in the tear fluid of patients with KC compared to that of healthy patients. Results demonstrate that in the KC subjects (n = 6) the concentration of Cu ions was 325.5 ± 110.7 ng/ml, while in the control group was 141.3 ± 71.1 ng/ml. A significant increase in Cu ion levels in the tear fluid was observed in the KC group compared to the control group (p value < 0.001).

Novel analgesic/anti-inflammatory agents: 1,5-diarylpyrrole nitrooxyalkyl ethers and related compounds as cyclooxygenase-2 inhibiting nitric oxide donors.

A series of 3-substituted 1,5-diarylpyrroles bearing a nitrooxyalkyl side chain linked to different spacers were designed. New classes of pyrrole-derived nitrooxyalkyl inverse esters, carbonates, and ethers (7-10) as COX-2 selective inhibitors and NO donors were synthesized and are herein reported. By taking into account the metabolic conversion of nitrooxyalkyl ethers (9, 10) into corresponding alcohols, derivatives 17 and 18 were also studied. Nitrooxy derivatives showed NO-dependent vasorelaxing properties, while most of the compounds proved to be very potent and selective COX-2 inhibitors in in vitro experimental models. Further in vivo studies on compounds 9a,c and 17a highlighted good anti-inflammatory and antinociceptive activities. Compound 9c was able to inhibit glycosaminoglycan (GAG) release induced by interleukin-1ß (IL-1ß), showing cartilage protective properties. Finally, molecular modeling and (1)H- and (13)C-NMR studies performed on compounds 6c,d, 9c, and 10b allowed the right conformation of nitrooxyalkyl ester and ether side chain of these molecules within the COX-2 active site to be assessed.

New platinum-oxicam complexes as anti-cancer drugs. Synthesis, characterization, release studies from smart hydrogels, evaluation of reactivity with selected proteins and cytotoxic activity in vitro.

The reaction of aqueous cis-[Pt(NH(3))(2)(H(2)O)(2)](NO(3))(2) with Na(+)HMEL(-) (H(2)MEL, meloxicam, 4-hydroxy-2-methyl-N-(5-methyl-1,3-thiazol-2-yl)-2H-1,2-benzothiazine-3-carboxamide-1,1-dioxide), and Na(+)HISO(-) (H(2)ISO, isoxicam, 4-hydroxy-2-methyl-N-(5-methylisoxazol-3-yl)-2H-1,2-benzothiazine-3-carboxamide-1,1-dioxide) at pH 7 produced micro-crystalline cis-[Pt(NH(3))(2)(N(1')-HMEL)(2)], 5 and cis-[Pt(NH(3))(2)(N(1')-HISO)(2)], 6. The X-ray diffraction structure of 5 shows two HMEL(-) anions donating through the thiazole nitrogen atoms and adopting a head-to-tail (HT) conformation. The (1)H NMR spectrum for 5 from DMSO-d(6) shows inertness of the complex up to at least 24h. Delivery studies for 5 and 6 from vinyl hydrogel based on L-phenylalanine (pH 6.5, 25 degrees C) show that concentrations of complexes ranging between 2.5 and 5 microM can be reached after a day. Compounds 5 and 6 show strong anti-proliferative effects on CH1 cells (ovarian carcinoma, human) in vitro, IC(50) values being 0.60 and 0.37 microM, respectively (0.16 microM for reference, cis-diamminodichloridoplatinum(II), cisplatin). ESI-MS measurements clearly documented that both 5 and 6 form adducts with the three model proteins ubiquitin (UBI), cytochrome c (CYT C) and superoxide dismutase (SOD), the HISO(-) complex being significantly more effective than the HMEL(-) one. Density functional methods help in finding rationale for the easiest dissociation of Pt-H(2)ISO/HISO bonds when compared to the Pt-N(1)(')-H(2)MEL/N(1)(')-HMEL linkages.

Properties of sesame oil by detailed 1H and 13C NMR assignments before and after ozonation and their correlation with iodine value, peroxide value, and viscosity measurements.

Gaseous ozone chemically reacts with unsaturated triglyceride substrates leading to ozonated derivatives with a wide potential applications, ranging from the petrochemical to the pharmaceutical industry. To date, an ultimate understanding of the ozone reactivity during sesame oil ozonation process as well as detailed (1)H and (13)C NMR assignments are lacking. A practical advantage of NMR is that a single NMR sample measurement can explain many issues, while similar analysis by traditional methods may require several independent and time-consuming measurements. Moreover, significant relationships among NMR spectra and both conventional chemical analysis and viscosity measurements have been found. Eventually, NMR could play an important role for quality attributes of ozonated oil derivatives.

Development of antitubercular compounds based on a 4-quinolylhydrazone scaffold. Further structure-activity relationship studies.

A series of 4-quinolylhydrazones was synthesized and tested in vitro against Mycobacterium tuberculosis. At a concentration of 6.25microg/mL, most of the newly synthesized compounds displayed 100% inhibitory activity against M. tuberculosis in cellular assays. Further screening allowed the identification of very potent antitubercular agents. Compound 4c was also tested in a time-course experiment and against mtb clinical isolates, displaying interesting results.

Physico-chemical characterization of sesame oil derivatives.

Ozone treatment of commercially available vegetable oils gives rise to the formation of chemical species that are responsible for the therapeutic properties of ozonated oil derivatives in dermatological diseases. In the last years, these products have been successfully used as a topical disinfectant in a number of serious skin affections. The medical application of empirically prepared ozonated oil has yielded striking improvements with unexpected and rapid healing, compelling us to begin a long-range study aiming first to define the main characteristics of the most common ozonated vegetable oils, about which there is usually no medical consensus because of the lack of standardization of their technological parameters. Sesame oil was selected because of its great amount of polyunsaturated acyl groups, as well as natural antioxidants. Moreover, we have determined the kinetics and optimal conditions of ozonation (e.g., ozone concentrations, time of exposure, temperature) for obtaining an ozonated oil characterized by well-established technological and physico-chemical properties, namely an accurate peroxide value determination. On the basis of the results, we have gained an understanding of the modifications of the vegetable oils during the ozonation process.

[RuII(NO+)]3+-core complexes with 4-methyl-pyrimidine and ethyl-isonicotinate: synthesis, X-ray structure, spectroscopy, and computational and NO-release studies upon UVA irradiation.

The reaction of RuCl(3)(NO).H(2)O with 4-methylpyrimidine (MePYM) and ethylisonicotinate (EINT), in absolute ethanol at 40-55 degrees C afforded crystalline trans-[RuCl(3)(NO)L(2)] complexes. Structural studies via X-ray diffraction, and spectroscopic methods (NMR, IR, UV-visible (UV-Vis)) revealed that the molecular structures have the two Ls in trans positions (axial) and the chloride anions and the NO(+) cation as equatorial ligands; pyrimidine...pyrimidine pairing pattern via two weak C-H...N interactions occur. The molecular structures for the EINT derivative was inferred from spectroscopy and computations. Under irradiation at 366 nm several solutions of the title compounds deliver NO via first order processes. Visible light (420-700 nm) does not produce significant NO release from CH(2)Cl(2) and CH(3)CN solutions within 24h.

Specific targeting of hepatitis C virus NS3 RNA helicase. Discovery of the potent and selective competitive nucleotide-mimicking inhibitor QU663.

Hepatitis C virus (HCV) infection is an emerging global epidemic, and no effective cure is yet available. Interferon-alpha (INFalpha) and pegylated INFs, in combination or otherwise with ribavirin, have proven to be effective in no more than 50% of chronically infected patients. New and better therapeutic strategies are therefore needed. HCV nonstructural protein 3 (NS3) RNA helicase (h) is a promising target for developing new therapeutics. QU663 was discovered as a potent new selective inhibitor of the helicase reaction of HCV NS3 (K(i) = 0.75 microM), competing with the nucleic acid substrate without affecting ATPase function, even at high concentrations. QU663 is one of a new generation of small-molecule nucleotide-mimicking inhibitors which are potential anti-HCV agents. A thorough molecular modeling study was carried out to explain the molecular basis of NS3h inhibition by QU663. The resulting three-dimensional interaction model is discussed.

Structural study of ribonucleotide reductase inhibitor hydrazones. Synthesis and X-ray diffraction analysis of a copper(II)-benzoylpyridine-2-quinolinyl hydrazone complex.

Single crystals as yellow needles of N-(4,8-dimethyl-quinolin-2-yl)-N'-(1-pyridin-2-yl-ethylidene)-hydrazine, HL(1), 1, and N-(4-methyl-quinolin-2-yl)-N'-(phenyl-pyridin-2-yl-methylene)-hydrazine, HL(2), 2, were obtained from methanol (MeOH) and analyzed via X-ray diffraction (XRD). HL(2) reacted with copper(II) acetate to produce a dark red powder that gave single crystals of [Cu(L(2))(OOCCH(3))].0.9C(6)H(5)CH(3), 3.0.9C(6)H(5)CH(3) when recrystallized from toluene. The conformation of the N(quinolinyl,q)-C(q)-N(H)-N(imine,i)-C-C(pyridine,p)-N(p) grouping is trans,trans,trans,trans or tttt, and ttcc for 1 and 2, respectively, at the solid state, as revealed via single crystal X-ray diffraction. Thus, the structure of 1 has the methyl (hydrazone) group syn to the N-H bond and syn to the N(q) and N(p) atom. On the other side, the structure of 2 is stabilized by a strong intra-molecular N-H...N hydrogen bond which involves the pyridyl nitrogen atom. The molecule 1 is almost planar, the torsion angles do not deviate more than 4 degrees from the idealized values of 0 degrees and 180 degrees . In the structure of 2 the pyridyl ring is almost coplanar with the N(q)-C(q)-NH-N(i)-C system, whereas the phenyl (Ph) ring is twisted by ca. 55 degrees . The structure of 3 has the L(2) ligand as deprotonated at the N-N function and in a cttc conformation as opposite to the ttcc one found for pure 2. The metal center is coordinated through N(q), N(i), N(p) and through an oxygen atom from a carboxylate anion. The molecular modeling analysis of 1 and 2 (semi-empirical molecular orbital at Zerner's intermediate neglect of differential overlap (ZINDO/1) level and density functional theory (DFT) methods) gave good agreement with the X-ray structures. Semi-empirical quantum mechanics analysis of 3 allowed to assign the UV-Vis spectrum that is characterized by strong absorptions in the visible, UVA and UVB regions. Owing to the ribonucleotide reductase inhibitory activity of the ligand, to the ascertained anticancer activity shown previously by related copper(II)-hydrazone complexes, and to the oxygen radical scavenger activity of several copper(II)-complexes, 3 is potentially anticancer and anti-inflammatory.

New alpha-(N)-heterocyclichydrazones: evaluation of anticancer, anti-HIV and antimicrobial activity.

A series of 3- and 5-methylthiophene-2-carboxaldehyde alpha-(N)-heterocyclichydrazones were synthesized and submitted to an in vitro investigation of their anticancer, anti-HIV and antimicrobial activities. Some of the newly synthesized compounds were found to possess antiproliferative properties, whereas no anti-HIV activity was seen; the most active of the series was the derivative 2i, which exhibited tumour growth inhibition activity against all cell lines displaying GI50 values between 1.63 and 26.5 microM. The title compounds were generally ineffective against Gram-positive and Gram-negative bacteria, while showed a moderate antifungal activity against C. albicans and A. fumigatus.

High affinity central benzodiazepine receptor ligands. Part 3: insights into the pharmacophore and pattern recognition study of intrinsic activities of pyrazolo[4,3-c]quinolin-3-ones.

Novel 2-phenyl-2,5-dihydropyrazolo[4,3-c]quinolin-3-(3H)-ones (PQs) endowed with high affinity for central benzodiazepine receptor (BzR) were synthesized. In particular, 9-fluoro-2-(2-fluorophenyl)-2,5-dihydro-3H-pyrazolo[4,3-c]quinolin-3-one (2(2)) showed binding affinity in the subnanomolar concentration range and proved to be in vitro a potent antagonist. This finding allowed the nature of the hydrogen bonding receptor site H(2) to be established, as located between the N-1 nitrogen of the PQ nucleus and the ortho position of the N-2-aryl group. [35S]tert-Butylbicyclophosphorothionate ([35S]TBPS) binding assays and electrophysiological measurements of the effects on GABA-evoked Cl(-) currents at recombinant human alpha(1)beta(2)gamma(2)(L) GABA(A) receptors, expressed in Xenopus laevis oocytes, were used to assess the intrinsic activities of a large series of PQs. With the aim of extracting discriminant information and distinguishing BzR ligands with different profiles of efficacy, 51 PQ derivatives, including full and partial agonists, antagonists, and inverse agonists, were analyzed in a multidimensional chemical descriptor space, defined by the lipophilicity parameter CLOG P and 3-D molecular WHIM descriptors, by means of principal component analysis, k-nearest neighbors (k-NN) method, and linear discriminant analysis (LDA). The classification methods were applied to subsets of pairs of efficacy classes, and lipophilicity and 3-D size descriptors were detected as the discriminant variables by a stepwise linear discriminant analysis. LDA proved to be superior to k-NN, especially in classifying PQ ligands (60-84% of success in prediction ability) into categories of efficacies which were contiguous and quite overlapped in the hyperspace of variables.

Specific targeting of acetylcholinesterase and butyrylcholinesterase recognition sites. Rational design of novel, selective, and highly potent cholinesterase inhibitors.

Tacrine-based AChE and BuChE inhibitors were designed by investigating the topology of the active site gorge of the two enzymes. The homobivalent ligands characterized by a nitrogen-bridged atom at the tether level could be considered among the most potent and selective cholinesterase inhibitors described to date. The nitrogen-containing homobivalent ligands 3e,g and the sulfur-containing 3h validated the hypothesis of extra sites of interaction in the AChE and BuChE active site gorges.

Synthesis and anti-tubercular evaluation of 4-quinolylhydrazones.

A series of 4-quinolylhydrazones were synthesized and tested against Mycobacterium tuberculosis H37Rv. Preparation of the title compounds was achieved by reaction of 4-quinolylhydrazine and aryl- or heteroaryl-carboxaldehyde. For the most of derivatives interesting antitubercular properties were showed; two compounds (3(2) and 3(25)), identified as the most active, were tested also against Mycobacterium avium.