A High Energy X-ray Diffraction Study of Amorphous Indomethacin.

Amorphous pharmaceuticals often possess a wide range of molecular conformations and bonding arrangements. The x-ray pair distribution function (PDF) method is a powerful technique for the characterization of variations in both intra-molecular and inter-molecular packing arrangements. Here, the x-ray PDF of amorphous Indomethacin is shown to be particularly sensitive to the preferred orientations of the chlorobenzyl ring found in isomers in the crystalline state. In some cases, the chlorobenzyl ring has no preferred torsional angle in the amorphous form, while in others evidence of distinct isomer orientations are observed. Amorphous samples with no preferred torsion angles of the chlorobenzyl ring are found to favor enhanced inter-molecular hydrogen bonding, and this is reflected in the intensity of the first sharp diffraction peak. These significant variations in structure rule out amorphous Indomethacin as a possible standard for x-ray PDF measurements. At high humidity, time resolved PDF's for >40 h reveal water molecules forming hydrogen bonds with Indomethacin molecules. A simple linear hydrogen bond model indicates that water molecules in the wet amorphous form have similar hydrogen bond strengths to those found between Indomethacin dimers or chains in the dry amorphous form.

Hard x-ray methods for studying the structure of amorphous thin films and bulk glassy oxides.

High-energy photon diffraction minimizes many of the corrections associated with laboratory x-ray diffractometers, and enables structure factor measurements to be made over a wide range of momentum transfers. The method edges us closer toward an ideal experiment, in which coordination numbers can be extracted without knowledge of the sample density. Three case studies are presented that demonstrate new hard x-ray methods for studying the structure of glassy and amorphous materials. First, the methodology and analysis of high-energy grazing incidence on thin films is discussed for the case of amorphous In2O3. The connectivity of irregular InO6polyhedra are shown to exist in face-, edge- and corner-shared configurations in the approximate ratio of 1:2:3. Secondly, the technique of high-energy small and wide angle scattering has been carried out on laser heated and aerodynamically levitated samples of silica-rich barium silicate (20BaO:80SiO2), from the single phase melt at 1500oC to the phase separated glass at room temperature. Based on Ba-O coordination numbers of 6 to 7, it is argued that the although the potential of Ba is ionic, it is weak enough to cause the liquid-liquid immiscibility to become metastable. Lastly, high-energy small and wide angle scattering has also been applied to high water content (up to 12 wt.%) samples of hydrous SiO2glass quenched from 1500oC at 4 GPa. An increase of Si1-O2correlations at 4.3 Å is found to be consistent with an increase in the population of three-membered SiO4rings at the expense of larger rings.

Reorientation Times for Solid-State Electrolyte Solvents and Electrolytes from NMR Spin-Lattice Relaxation Studies.

Ionic and molecular plastic crystals have been studied recently as solid electrolytes or solvents, but the specific role of molecular reorientation has not been clarified. We use NMR spin-lattice relaxation times (T1 minima) to compare the time scale for magnetic fluctuations in a plastic crystal solvent to the molecular reorientation times, as established by dielectric spectroscopy. We focus on a mixture of succinonitrile and glutaronitrile, in which the rotationally disordered phase is stabilized against crystallization. Reorientation times can then be studied over 13 orders of magnitude, down to the glass transition temperature at 144 K. For each nucleus, 1H and 13C, the most probable magnetic fluctuation time is found to be slightly shorter than the reorientation time, but with practically indistinguishable temperature dependence. This facilitates investigation of the relation of solvent reorientation to ion conductivity relaxation times in ionic conducting systems in which the conductivity swamps the dielectric signature of solvent reorientation.

Using containerless methods to develop amorphous pharmaceuticals.

BACKGROUND: Many pipeline drugs have low solubility in their crystalline state and require compounding in special dosage forms to increase bioavailability for oral administration. The use of amorphous formulations increases solubility and uptake of active pharmaceutical ingredients. These forms are rapidly gaining commercial importance for both pre-clinical and clinical use. METHODS: Synthesis of amorphous drugs was performed using an acoustic levitation containerless processing method and spray drying. The structure of the products was investigated using in-situ high energy X-ray diffraction. Selected solvents for processing drugs were investigated using acoustic levitation. The stability of amorphous samples was measured using X-ray diffraction. Samples processed using both spray drying and containerless synthesis were compared. RESULTS: We review methods for making amorphous pharmaceuticals and present data on materials made by containerless processing and spray drying. It was shown that containerless processing using acoustic levitation can be used to make phase-pure forms of drugs that are known to be difficult to amorphize. The stability and structure of the materials was investigated in the context of developing and making clinically useful formulations. CONCLUSIONS: Amorphous compounds are emerging as an important component of drug development and for the oral delivery of drugs with low solubility. Containerless techniques can be used to efficiently synthesize small quantities of pure amorphous forms that are potentially useful in pre-clinical trials and for use in the optimization of clinical products. GENERAL SIGNIFICANCE: Developing new pharmaceutical products is an essential enterprise to improve patient outcomes. The development and application of amorphous pharmaceuticals to increase absorption is rapidly gaining importance and it provides opportunities for breakthrough research on new drugs. There is an urgent need to solve problems associated with making formulations that are both stable and that provide high bioavailability. This article is part of a Special Issue entitled "Science for Life" Guest Editor: Dr. Austen Angell, Dr. Salvatore Magazù and Dr. Federica Migliardo.

Measurement of conductivity and permittivity on samples sealed in nuclear magnetic resonance tubes.

We present a broadband impedance spectroscopy instrument designed to measure conductivity and∕or permittivity for samples that are sealed in glass tubes, such as the standard 5 mm tubes used for nuclear magnetic resonance experiments. The calibrations and corrections required to extract the dielectric properties of the sample itself are outlined. It is demonstrated that good estimates of the value of dc-conductivity can be obtained even without correcting for the effects of glass or air on the overall impedance. The approach is validated by comparing data obtained from samples sealed in nuclear magnetic resonance tubes with those from standard dielectric cells, using glycerol and butylmethylimidazolium-hexafluorophosphate as respective examples of a molecular and an ionic liquid. This instrument and approach may prove useful for other studies of permittivity and conductivity where contact to the metal electrodes or to the ambient atmosphere needs to be avoided.

Total x-ray scattering of spider dragline silk.

Total x-ray scattering measurements of spider dragline silk fibers from Nephila clavipes, Argiope aurantia, and Latrodectus hesperus all yield similar structure factors, with only small variations between the different species. Wide-angle x-ray scattering from fibers orientated perpendicular to the beam shows a high degree of anisotropy, and differential pair distribution functions obtained by integrating over wedges of the equatorial and meridian planes indicate that, on average, the majority (95%) of the atom-atom correlations do not extend beyond 1 nm. Futhermore, the atom-atom correlations between 1 and 3 nm are not associated with the most intense diffraction peaks at Q=1-2 Å(-1). Disordered molecular orientations along the fiber axis are consistent with proteins in similar structural arrangements to those in the equatorial plane, which may be associated with the silk's greater flexibility in this direction.

Vitrification of a monatomic metallic liquid.

Although the majority of glasses in use in technology are complex mixtures of oxides or chalcogenides, there are numerous examples of pure substances-'glassformers'-that also fail to crystallize during cooling. Most glassformers are organic molecular systems, but there are important inorganic examples too, such as silicon dioxide and elemental selenium (the latter being polymeric). Bulk metallic glasses can now be made; but, with the exception of Zr50Cu50 (ref. 4), they require multiple components to avoid crystallization during normal liquid cooling. Two-component 'metglasses' can often be achieved by hyperquenching, but this has not hitherto been achieved with a single-component system. Glasses form when crystal nucleation rates are slow, although the factors that create the slow nucleation conditions are not well understood. Here we apply the insights gained in a recent molecular dynamics simulation study to create conditions for successful vitrification of metallic liquid germanium. Our results also provide micrographic evidence for a rare polyamorphic transition preceding crystallization of the diamond cubic phase.

Comment on 'Microscopic structural evolution during the liquid-liquid transition in triphenyl phosphite' by R Kurita, Y Shinohara, Y Amemiya and H Tanaka J. Phys.: Condens. Matter 19 (2007) 152101.

In the communication by Kurita et al 2007 J. Phys.: Condens. Matter 19 152101, peaks in the liquid diffraction pattern of triphenyl phosphite have been attributed to intermolecular phosphor-phosphor distances. Based on our previous neutron and x-ray diffraction studies we argue that this assignment is incorrect and the peak contributions are likely to be much more complex.

Analysis of high-energy x-ray diffraction data at high pressure: the case of vitreous As2O3 at 32 GPa.

The x-ray structure factor of vitreous As2O3 has been measured at 32 GPa in a laser-perforated diamond anvil cell using a monochromatic, micro-focused high-energy x-ray beam. The experimental correction procedures are discussed in detail, and they yield reliable data over the range Q = 0.3-13.5 Å-1. The use of modified form factors to analyse the scattering data is presented to account for charge transfer. Analysis of the radial distribution function yields an increase in the coordination number from 3.1 ± 0.3 oxygen atoms surrounding an arsenic atom at normal pressure to 4.8 ± 0.5 at 32 GPa with only a slight change in the As-O bond length. Substantial structural changes are observed at higher distances, extending up to 18 Å in real space.

Orientational correlations in the glacial state of triphenyl phosphite.

Spallation neutron and high-energy X-ray diffraction experiments have been performed to investigate the local structure of the glacial and supercooled liquid states in triphenyl phosphite. The observed diffraction patterns have been interpreted using a Reverse Monte Carlo modeling technique. The results show that the glacial state forms unusually weak intermolecular hydrogen bonds between an oxygen atom connected to a phenyl ring and an adjacent phenyl ring aligned in an approximately antiparallel configuration. The structure is very different from the hexagonal crystal which is characterized by two weaker hydrogen bonds between linear arrays of molecules which are offset from each other and packed in a hexamer arrangement.

Formation and structure of a dense octahedral glass.

We have performed in situ x-ray and neutron-diffraction measurements, and molecular dynamics simulations, of GeO2, an archetypal network-forming glass under pressure. Below 5 GPa, additional atoms encroaching on the first tetrahedral shell are seen to be a precursor of local coordination change. Between 6 and 10 GPa, we observe structures with a constant average coordination of approximately 5, indicating a new metastable, intermediate form of the glass. At 15 GPa, the structure of a fully octahedral glass has been measured. This structure is not retained upon decompression and, therefore, must be studied in situ.

Studies on TMPD:TCNB; a donor-acceptor with room temperature paramagnetism and n-pi interaction.

A donor-acceptor compound based on N,N,N',N'-tetramethyl-p-phenylene-diamine and 1,2,4,5-tetracyanobenzene (TMPD:TCNB) has been synthesized. The crystal structure of the black 1:1 complex formed between TMPD and TCNB has been determined by single crystal X-ray diffraction at room temperature. The compound crystallizes in the triclinic space group P-1 with cell dimensions: a = 7.4986(15) å, b =7.6772(11) å, c = 8.0764(15) å, alpha = 78.822(12) degrees, beta = 83.3779(19) degrees, gamma = 86.289(17) degrees .TMPD and TCNB molecules are stacked alternately in infinite columns along the a-axis. The structure does not seem to show the usual pi-pi interaction between the two aromatic rings, but indicates an n-pi interaction localized between the nitrogen atoms of the donor and the cyano groups of the acceptor.

Intermediate-range order in permanently densified GeO2 glass.

Information about the partial structure factors of densified GeO2 glass has been obtained from neutron and x-ray diffraction measurements. Densification causes a reduction in the length scale of the intermediate range order (IRO). The difference structure factors obtained by combining the x-ray and neutron data so as to eliminate one partial structure factor at a time shows the greatest effects when the Ge-Ge correlations are eliminated and least when O-O correlations are eliminated. This implies that the reduced length scale results from a decrease in the next-nearest neighbor Ge-O and O-O distance caused by a rotation about the Ge-O-Ge bonds and a distortion of the GeO4 tetrahedra.

Electron spin density distribution in the polymer phase of CsC60: assignment of the NMR spectrum.

We present high resolution 133Cs-13C double resonance NMR data and 13C-13C NMR correlation spectra of 13C enriched samples of the polymeric phase of CsC60. These data lead to a partial assignment of the lines in the 13C NMR spectrum of CsC60 to the carbon positions on the C60 molecule. A plausible completion of the assignment can be made on the basis of an ab initio calculation. The data support the view that the conduction electron density is concentrated at the C60 "equator," away from the interfullerene bonds.

Synthesis, Crystal Structure, NMR Studies, and Thermal Stability of Mixed Iron-Indium Phosphates with Quasi-One-Dimensional Frameworks.

The hydrothermal synthesis, single-crystal structure analysis, spectroscopic studies, and thermal stability of the compounds Ca(2)(In(1)(-)(x)()Fe(x)())(PO(4))(HPO(4))(2).H(2)O (0

Safety of 25-hydroxycholecalciferol as a source of cholecalciferol in poultry rations.

We conducted two safety studies of 25-hydroxycholecalciferol [25(OH)D3] in poultry broilers at levels ranging from 1 to 200 times those commonly used for cholecalciferol (vitamin D3) supplementation in the industry. In the first experiment, 1-d-old male and female broiler chickens were fed commercial diets containing either vitamin D3 or 25(OH)D3 at concentrations of 69, 207, and 690 micrograms of 25(OH)D3/kg of feed. The second experiment compared effects of 25(OH)D3 and vitamin D3 on performance and survival of broilers at levels ranging from 1 to 200 times the basal level of 69 micrograms/kg feed. When 25(OH)D3 was fed in equal amounts (wt/wt) to vitamin D3, there was an increase in body weight and a decrease (improvement) in adjusted feed efficiency in both experiments, but the changes were significant only in the first experiment. In the first experiment, serum 25(OH)D3 concentrations increased from 13.3 +/- 4.3 to 42.5 +/- 18 ng/mL in birds fed vitamin D3 or 25(OH)D3, respectively, and rose to 246 +/- 38 ng/mL in birds fed the highest level of 25(OH)D3. Tissue 25(OH)D3 concentrations were much lower than serum concentrations and were highly correlated to the latter, regardless of dietary treatment. In Experiment 2, there was some evidence of renal calcification in birds fed 25(OH)D3 at 10 times the basal level, whereas dietary levels of vitamin D3 of 50 times the basal level were required to show some evidence of renal calcification. On the basis of both renal calcification and body weight, the present studies would suggest that 25(OH)D3 is 5 to 10 times more toxic than vitamin D3.

Comparison of dietary 25-hydroxycholecalciferol and cholecalciferol in broiler chickens.

We conducted a series of 10 feeding trials involving over 36,000 broilers. The effects of various dietary levels of cholecalciferol (vitamin D3) or 25-hydroxycholecalciferol (25-OH-D3) were compared using a "basal" dosage level of 69 micrograms/kg feed, as well as levels ranging from .5 to 1.5 times the basal level. For all 10 studies, average body weight increased by an average of .042 +/- .03 kg (P < .001) and adjusted feed efficiency decreased (improved) by an average of .026 +/- .0046 kg/kg (P < .001) in birds fed 25-OH-D3 in comparison to those fed vitamin D3 at the basal level. Changes in mortality were not detected. Evaluation of different dietary levels of 25-OH-D3 revealed a significant dose-response relationship, with maximal effects on weight gain, feed efficiency, and breast meat yield being observed in the range of 50 to 70 micrograms/kg feed. Preliminary studies with different levels of vitamin D3 suggested no additional benefits on weight gain or feed efficiency with higher dietary levels of vitamin D3. Serum 25-OH-D3 concentrations increased more rapidly in birds fed 25-OH-D3 than in birds fed vitamin D3. There were significant correlations with body weight, feed conversion, and serum 25-OH-D3 concentrations, with no correlations observed between serum 1,25-(OH)2D3 concentrations and these variables.

Farnesyl diphosphate synthetase. Molecular cloning, sequence, and expression of an essential gene from Saccharomyces cerevisiae.

Farnesyl diphosphate (FPP) synthetase is a key enzyme in isoprenoid biosynthesis which supplies C15 precursors for several classes of essential metabolites including sterols, dolichols, and ubiquinones. The structural gene for FPP synthetase was isolated on a 4.5-kilobase EcoRI genomic restriction fragment from the yeast Saccharomyces cerevisiae. The clone encodes a 40,483-dalton polypeptide of 342 amino acids with a high degree of similarity to the protein encoded by a putative rat liver clone of FPP synthetase (Clarke, C. F., Tanaka, R. D., Svenson, K., Wamsley, M., Fogelman, A. M., and Edwards, P. A. (1987) Mol. Cell Biol. 7, 3138-3146) and to an active site protein fragment from avian liver FPP synthetase (Brems, D. N., Bruenger, E., and Rilling, H. C. (1981) Biochemistry 20, 3711-3718). When cloned into the yeast shuttle vector YRp17, the 4.5-kilobase EcoRI fragment directed a 2-3-fold over-expression of FPP synthetase activity in transformed yeast cells. The levels of expression were independent of culture growth phase and orientation of the insert, indicative of a functional promoter in the clone. Disruption of the FPP synthetase gene from a diploid yeast strain, followed by dissection and analysis of tetrads, demonstrates that the gene is an essential, single copy number gene in yeast. The gene for FPP synthetase resides on chromosome XI as judged from Southern blots of separated yeast chromosomes.

Transcription terminator-like element within a Saccharomyces cerevisiae promoter region.

We analyzed a cloned fragment of the yeast URA3 promoter region that contains a sequence of DNA capable of functioning as a highly efficient transcription terminator. BAL 31 deletions have shown the signal for the transcription termination activity is less than or equal to 110 base pairs and resides between bases 45 and 155 upstream of the URA3 primary ATG codon at base 227. In our in vivo assay system, the DNA fragment is able to terminate transcripts very efficiently in either orientation. The terminated transcripts bind to oligodeoxythymidylate cellulose columns and promote the synthesis of full-length cDNAs, suggesting that the transcripts are polyadenylated. The 110-base-pair region contains no sequence resembling terminator consensus sequences described by Zaret and Sherman (K.S. Zaret and F. Sherman, Cell, 28:563-573, 1982) or Henikoff and Cohen (S. Henikoff and E.H. Cohen, Mol. Cell. Biol., 4:1515-1520, 1984). We discuss the possible physiological relevance of this sequence to bona fide termination of transcription and to URA3 regulation in Saccharomyces cerevisiae.