The aggregation of sodium dehydrocholate in water

 We used a battery of different methods to study the association in aqueous sodium dehydrocholate (NaDHC) solutions. This salt associates by a stepwise mechanism. Below (9.6 ± 4.2) × 10⁻⁴ mol dm⁻³ there is a molecular solution with some strongly insoluble dehydrocholic acid produced by hydrolysis. Between (9.6 ± 4.2) × 10⁻⁴ and (5.2 ± 2.2) × 10⁻³ mol dm⁻³, an aggregate similar to acid soap (NaDHC.HDHC) appears and its amount and the aggregate's size increase with concentration. At =(2.20 ± 0.85) × 10⁻² mol dm⁻³ the aggregates formed have properties usually associated with true micelles, such as solubilisation of water-insoluble dyes. These aggregates increase in size with concentration and change their shape at 8 × 10⁻² mol dm⁻³, giving nonsymmetrical aggregates. The changes in the solution physicochemical properties at these concentrations may be misinterpreted and this explains the different values of the critical micelle concentration reported in the literature for substances with similar structure, such as bile salts. Received: 14 May 2001 Accepted: 10 August 2001     

Use of the tubulin bound paclitaxel conformation for structure-based rational drug design

A new computational docking protocol has been developed and used in combination with conformational information inferred from REDOR-NMR experiments on microtubule bound 2-(p-fluorobenzoyl)paclitaxel to delineate a unique tubulin binding structure of paclitaxel. A conformationally constrained macrocyclic taxoid bearing a linker between the C-14 and C-3'N positions has been designed and synthesized to enforce this "REDOR-taxol" conformation. The novel taxoid SB-T-2053 inhibits the growth of MCF-7 and LCC-6 human breast cancer cells (wild-type and drug resistant) on the same order of magnitude as paclitaxel. Moreover, SB-T-2053 induces in vitro tubulin polymerization at least as well as paclitaxel, which directly validates our drug design process. These results open a new avenue for drug design of next generation taxoids and other microtubule-stabilizing agents based on the refined structural information of drug-tubulin complexes, in accordance with typical enzyme-inhibitor medicinal chemistry precepts.     

Controlling cell attachment selectively onto biological polymer-colloid templates using polymer-on-polymer stamping

A new patterning approach using polymer-on-polymer stamping (POPS) has been developed to fabricate polymer-colloid templates for controlling selective cell attachment. In this paper, a polyamine surface patterned onto a poly(acrylic acid)/poly(allylamine hydrochloride) (PAA/PAH) cell resistant multilayer platform serves as a template for the deposition of close- or loose-packed colloidal particles. Peptides containing the RGD adhesion sequence were used to modify the PAH/colloid surface for specific cell attachment. Cell behavior was studied by varying colloidal packing array density, pattern geometry, and surface chemistry. It was found that loose-packed RGD-modified colloidal arrays enhance cell adhesion, as observed through the development of focal adhesion contacts and orientation of actin stress fibers, but close-packed colloidal arrays induce a rounded and nonadhesive cell morphology and yield a smaller number of attached cells. On loose-packed arrays, cells adjust their shapes to the pattern geometry when the stripe width is smaller than 50 microm and increase their extent of attachment when the concentration of surface RGD peptides is increased. This new biomaterials system allows the examination of cell behavior as a function of RGD surface distribution on the molecular to micrometer scale and reveals cellular response to different surface roughnesses.     

Nucleic acid related compounds. 116. Nonaqueous diazotization of aminopurine nucleosides. Mechanistic considerations and efficient procedures with tert-butyl nitrite or sodium nitrite

Nonaqueous diazotization-dediazoniation of two types of aminopurine nucleoside derivatives has been investigated. Treatment of 9-(2,3,5-tri-O-acetyl-beta-D-ribofuranosyl)-2-amino-6-chloropurine (1) with SbCl(3)/CH(2)Cl(2) was examined with benzyltriethylammonium (BTEA) chloride as a soluble halide source and tert-butyl nitrite (TBN) or sodium nitrite as the diazotization reagent. Optimized yields (>80%) of the 2,6-dichloropurine derivative were obtained with SbCl(3). Combinations with SbBr(3)/CH(2)Br(2) gave the 2-bromo-6-chloropurine product (>60%), and SbI(3)/CH(2)I(2)/THF gave the 2-iodo-6-chloropurine derivative (>45%). Antimony trihalide catalysis was highly beneficial. Mixed combinations (SbX(3)/CH(2)X'(2); X/X' = Br/Cl) gave mixtures of 2-(bromo, chloro, and hydro)-6-chloropurine derivatives that were dependent on reaction conditions. Addition of iodoacetic acid (IAA) resulted in diversion of purine radical species into a 2-iodo-6-chloropurine derivative with commensurate loss of other radical-derived products. This allowed evaluation of the efficiency of SbX(3)-promoted cation-derived dediazoniations relative to radical-derived reactions. Efficient conversions of adenosine, 2'-deoxyadenosine, and related adenine nucleosides into 6-halopurine derivatives of current interest were developed with analogous combinations.     

Extraction of Pb(II), Cd(II), and Hg(II) from aqueous solution by nitrogen and thiol functionality grafted to silica gel measured by calorimetry

The reaction of ethylene sulfide with 3-aminopropyltrimethoxysilane gave a new silylating agent, which was anchored onto a silica surface via the sol–gel procedure. This surface displayed a chelating moiety containing nitrogen and two sulfur basic centers potentially capable of extracting cations from aqueous solutions. The process of metal extraction was followed by a batch method, and fitted to a modified Langmuir equation. The maximum adsorption capacities found were: 2.06 ± 0.01, 3.72 ± 0.02, and 5.14 ± 0.02 mmol g⁻¹ for Pb(II), Cd(II), and Hg(II), respectively. The enthalpies of bending are: −1.16 ± 0.04, −3.60 ± 0.10, and −8.94 ± 0.03 kJ mol⁻¹ for Cd(II), Pb(II), and Hg(II), respectively. The Gibbs free energies of binding agree with the spontaneity of the proposed reactions between cations and basic centers.     

Contrasting behavior in azide pyrolyses: an investigation of the thermal decompositions of methyl azidoformate, ethyl azidoformate and 2-azido-N, N-dimethylacetamide by ultraviolet photoelectron spectroscopy and matrix isolation infrared spectroscopy

The thermal decompositions of methyl azidoformate (N3COOMe), ethyl azidoformate (N3COOEt) and 2-azido-N,N-dimethylacetamide (N3CH2CONMe2) have been studied by matrix isolation infrared spectroscopy and real-time ultraviolet photoelectron spectroscopy. N2 appears as an initial pyrolysis product in all systems, and the principal interest lies in the fate of the accompanying organic fragment. For methyl azidoformate, four accompanying products were observed: HNCO, H2CO, CH2NH and CO2, and these are believed to arise as a result of two competing decomposition routes of a four-membered cyclic intermediate. Ethyl azidoformate pyrolysis yields four corresponding products: HNCO, MeCHO, MeCHNH and CO2, together with the five-membered-ring compound 2-oxazolidone. In contrast, the initial pyrolysis of 2-azido-N,N-dimethyl acetamide, yields the novel imine intermediate Me2NCOCH=NH, which subsequently decomposes into dimethyl formamide (HCONMe2), CO, Me2NH and HCN. This intermediate was detected by matrix isolation IR spectroscopy, and its identity confirmed both by a molecular orbital calculation of its IR spectrum, and by the temperature dependence and distribution of products in the PES and IR studies. Mechanisms are proposed for the formation and decomposition of all the products observed in these three systems, based on the experimental evidence and the results of supporting molecular orbital calculations.     

Spectroscopic investigation and thermal behavior of new carbonyl complexes [M(CO)4(N—N)(CuX)], (M = W,Mo; N—N = 2,2′-bipyridine, 1,10-phenanthroline; X = Cl,SCN)

[M(CO)₄(N—N)] reacts with CuCl to give new heterobimetallic metal carbonyls of the type [M(CO)₄(N—N)(CuCl)], M = W, Mo; N—N = 2,2-bipyridine (bipy), 1,10-phenanthroline (phen). Reactions of [M(CO)₄(N—N)(CuCl)] with NaSCN produced the series of complexes of general formula [M(CO)₄(N—N)(CuSCN)]. The i.r. spectral of all the bimetallic carbonyls exhibited the general four (CO) band patterns of the precursors. The u.v.–vis. spectral data for precursors and products showed bands associated with * (nitrogen ligands), dd (intrametal), as well as MLCT d* (nitrogen ligands) and MLCT d *(CO) transitions. The [M(CO)₄(N—N)(CuX)] (X = Cl, SCN) emission spectra showed only one band associated with the MLCT transition. The t.g. curves revealed a stepwise loss of CO groups. The initial decomposition temperatures of the [M(CO)₄(N—N)(CuX)] series suggest that the bimetallic compounds are indeed thermally less stable than their precursors, and the X-ray data showed the formation of MO₃, CuMO₄, Cu₂O and CuO as final decomposition products, M = W, Mo. The spectroscopic data suggests that the heterobimetallic compounds are polymeric.     

Interaction and lipid-induced conformation of two cecropin-melittin hybrid peptides depend on peptide and membrane composition

The interaction of two hybrid peptides of cecropin A and melittin [CA(1-8)M(1-18) and CA(1-7)M(2-9)] with liposomes was studied by differential scanning calorimetry (DSC), circular dichroism (CD), and quasi-elastic light scattering (QELS). The study was carried out with large unilamellar vesicles (LUVs) of three different lipid compositions: 1,2-dimyristoil-sn-glycero-3-phosphocholine (DMPC), 1,2-dimyristoyl-sn-glycero-3-phospho-rac-(1-glycerol) (DMPG) and a binary mixture of DMPC/DMPG, in a wide range of peptide-to-lipid (P:L) molar ratios (0 to 1:7). DSC results indicate that, for both peptides, the interaction depends on membrane composition, with very different behavior for zwitterionic and anionic membranes. CD data show that, although the two peptides have different secondary structures in buffer (random coil for CA(1-7)M(2-9) and predominantly beta-sheet for CA(1-8)M(1-18)), they both adopt an alpha-helical structure in the presence of the membranes. Overall, results are compatible with a model involving a strong electrostatic surface interaction between the peptides and the negatively charged liposomes, which gives place to aggregation in the gel phase and precipitation after a threshold peptide concentration. In the case of zwitterionic membranes, a progressive surface coverage with peptide molecules destabilizes the membrane, eventually leading to membrane disruption. Moreover, delicate modulations in behavior were observed depending on the peptide.     

A solvent-controlled switch of manganese complex assemblies with a beta-diketonate-based ligand

The coordination properties of the new polynucleating ligand H(3)L1 (1,3-bis(3-oxo-3-phenylpropionyl)-2-hydroxy-5-methylbenzene) with Mn(II/III) are described. Depending on the solvent used, the reaction of H(3)L1 with Mn(OAc)(2) yields either of the two new multinuclear assemblies [Mn(2)(HL1)(2)(py)(4)] (1) and [Mn(3)(HL1)(3)] (2), as revealed by X-ray crystallography. The structure of 2 is remarkable in that it shows a unique asymmetric triple-stranded helicate. Complexes 1 and 2 can be interconverted by controlling the solvent of the reaction system, and therefore, this ensemble constitutes an interesting externally addressable switch. In the presence of Mn(III)/pyridine, partial degradation of H(3)L1 occurs via oxidative cleavage, and the new complex [Mn(2)(L2)(2)(py)(4)] (3) is formed. The crystal structure of this complex has shown the fully deprotonated form of the new donor H(3)L2 (3-(3-oxo-3-phenylpropionyl)-5-methylsalicylic acid). From the same reaction, the Mn(II) complex 1 is also obtained. A rational synthesis of H(3)L2 is reported, and this has been used to prepare 3 in high yields, directly from its components. Variable-temperature magnetic susceptibility (chi(m)) measurements were performed on complexes 1-3 under a magnetic field of 1 kG. The data for each complex were fit to the appropriate chi(m) vs T theoretical equation, respectively. In 1, the Mn(II) ions are uncoupled, with g = 2.01. The data from 2 were fit by assuming the presence of an exchange coupled Mn(II)...Mn(II) pair next to a magnetically isolated Mn(II) center. The fit gave J = -2.75 cm(-1), g(12) = 1.97, and g(3) = 1.92, respectively. In 3, two models fit the experimental data. In the most satisfactory, the Mn(III) ions are coupled antiferromagnetically with J = -1.48 cm(-1) and g = 1.98 and a term for weak ferromagnetic intermolecular exchange is included with zJ' = 0.39 cm(-1). The other model contemplates the presence of two uncoupled zero field split Mn(III) ions.     

Inclusion Complex between Local Anesthetic/2-hydroxypropyl-β-cyclodextrin in Stealth Liposome

The drugs delivery system in the treatment of diseases has advantages such as reduced toxicity, increased availability of the drug, etc. Therefore, studies of the supramolecular interactions between local anesthetics (LAs) butamben (BTB) or ropivacaine (RVC) complexed with 2-hydroxypropyl-β-cyclodextrin (HP-βCD) and carried in Stealth liposomal (SL) are performed. ¹H-NMR nuclear magnetic resonance (DOSY and STD) were used as the main tools. The displacements observed in the ¹H-NMR presented the complexion between LAs and HP-βCD. The diffusion coefficients of free BTB and RVC were 7.70 × 10⁻¹⁰ m² s⁻¹ and 4.07 × 10⁻¹⁰ m² s⁻¹, and in the complex with HP-βCD were 1.90 × 10⁻¹⁰ m² s⁻¹ and 3.64 × 10⁻¹⁰ m² s⁻¹, respectively, which indicate a strong interaction between the BTB molecule and HP-βCD (98.3% molar fraction and Ka = 72.279 L/mol). With STD-NMR, the encapsulation of the BTB/HP-βCD and RVC/HP-βCD in SL vesicles was proven. Beyond the saturation transfer to the LAs, there is the magnetization transfer to the hydrogens of HP-βCD. BTB and RVC have already been studied in normal liposome systems; however, little is known of their behavior in SL.