Structural Analysis of the Complex of Human Transthyretin with 3′,5′-Dichlorophenylanthranilic Acid at 1.5 Å Resolution

Human transthyretin (hTTR) can form amyloid deposits that accumulate in nerves and organs, disrupting cellular function. Molecules such as tafamidis that bind to and stabilize the TTR tetramer can reduce such amyloid formation. Here, we studied the interaction of VCP-6 (2-((3,5-dichlorophenyl)amino)benzoic acid) with hTTR. VCP-6 binds to hTTR with 5 times the affinity of the cognate ligand, thyroxine (T₄). The structure of the hTTR:VCP-6 complex was determined by X-ray crystallography at 1.52 Å resolution. VCP-6 binds deeper in the binding channel than T₄ with the 3′,5′-dichlorophenyl ring binding in the ‘forward’ mode towards the channel centre. The dichlorophenyl ring lies along the 2-fold axis coincident with the channel centre, while the 2-carboxylatephenylamine ring of VCP-6 is symmetrically displaced from the 2-fold axis, allowing the 2-carboxylate group to form a tight intermolecular hydrogen bond with Nζ of Lys15 and an intramolecular hydrogen bond with the amine of VCP-6, stabilizing its conformation and explaining the greater affinity of VCP-6 compared to T₄. This arrangement maintains optimal halogen bonding interactions in the binding sites, via chlorine atoms rather than iodine of the thyroid hormone, thereby explaining why the dichloro substitution pattern is a stronger binder than either the diiodo or dibromo analogues.     

Characterization and Authentication of “Ricotta” Whey Cheeses through GC-FID Analysis of Fatty Acid Profile and Chemometrics

The fatty acid (FA) profiles of 240 samples of ricotta whey cheese made from sheep, goat, cow, or water buffalo milk were analyzed by gas-chromatography (GC). Then, sequential preprocessing through orthogonalization (SPORT) was used in order to classify samples according to the nature of the milk they were made from. This strategy achieved excellent results, correctly classifying 77 (out of 80) validation samples. Eventually, since 36 (over 114) sheep ricotta whey cheeses were PDO products, a second classification problem, finalizing the discrimination of PDO and Non-PDO dairies, was faced. In this case, two classifiers were used, SPORT and soft independent modelling by class analogy (SIMCA). Both approaches provided more than satisfying results; in fact, SPORT properly assigned 63 (of 65) test samples, whereas the SIMCA model accepted 14 PDO individuals over 15 (93.3% sensitivity) and correctly rejected all the other samples (100.0% specificity). In conclusion, all the tested approaches resulted as suitable for the two fixed purposes. Eventually, variable importance in projection (VIP) analysis was used to understand which FAs characterize the different categories of ricotta. Among the 22 analyzed compounds, about 10 are considered the most relevant for the solution of the investigated problems.     

Chemically resolved structure of the surface

The structure and chemical states of the Sn/Ge(111) surface are characterized by x-ray standing waves combined with photoemission. For the room temperature square root 3xsquare root 3 phase two chemical components, approximately 0.4 eV apart, are observed for both Sn 3d and 4d core levels. Our model-independent, x-ray standing wave analysis shows unambiguously that the two components originate from Sn adatoms located at two different heights separated vertically by 0.23 A, in favor of a model composed of a fluctuating Sn layer. Contrary to the most accepted scenario, the stronger Sn 3d and 4d components, which appear at the lower binding-energy sides and account for 2/3 of the Sn adatoms, are identified to be associated with the higher Sn position, manifesting their filled valence state character.     

Structural Elucidation of Silver(I) Amides and Their Application as Catalysts in the Hydrosilylation and Hydroboration of Carbonyls

This study details the isolation and characterisation of three novel silver(I) amides in solution and solid-state, [Ag(Cy₃P)(HMDS)] 2 , [Ag(Cy₃P){N(TMS)(Dipp)}] 3 and [Ag(Cy₃P)₂(NPh₂)] 4 . Their catalytic abilities have proved successful in hydroboration and hydrosilylation reactions with a full investigation performed with complex 2 . Both protocols proceed under mild conditions, displaying exceptional functional-group tolerance and chemoselectivity, in excellent conversions at competitive reaction times. This work reveals the first catalytic hydroboration of aldehydes and ketones performed by a silver(I) catalyst.     

Cycloisomerization of Olefins in Water

Preparative reactions that occur efficiently under dilute, buffered, aqueous conditions in the presence of biomolecules find application in ligation, peptide synthesis, and polynucleotide synthesis and sequencing. However, the identification of functional groups or reagents that are mutually reactive with one another, but unreactive with biopolymers and water, is challenging. Shown here are cobalt catalysts that react with alkenes under dilute, aqueous, buffered conditions and promote efficient cycloisomerization and formal Friedel–Crafts reactions. The constraining conditions of bioorthogonal chemistry are beneficial for reaction efficiency as superior conversion at low catalyst concentration is obtained and competent rates in dilute conditions are maintained. Efficiency at high dilution in the presence of buffer and nucleobases suggests that these reaction conditions may find broad application.     

A General Approach to O-Sulfation by a Sulfur(VI) Fluoride Exchange Reaction

O-sulfation is an important chemical code widely existing in bioactive molecules, but the scalable and facile synthesis of complex bioactive molecules carrying O-sulfates remains challenging. Reported here is a general approach to O-sulfation by the sulfur(VI) fluoride exchange (SuFEx) reaction between aryl fluorosulfates and silylated hydroxy groups. Efficient sulfate diester formation was achieved through systematic optimization of the electronic properties of aryl fluorosulfates. The versatility of this O-sulfation strategy was demonstrated in the scalable syntheses of a variety of complex molecules carrying sulfate diesters at various positions, including monosaccharides, disaccharides, an amino acid, and a steroid. Selective hydrolytic and hydrogenolytic removal of the aryl masking groups from sulfate diesters yielded the corresponding O-sulfate products in excellent yields. This strategy provides a powerful tool for the synthesis of O-sulfate bioactive compounds.     

Giant deuteron migration during the isosymmetric phase transition in deuterated 3,5-pyridinedicarboxylic acid

Deuterated 3,5‐pyridinedicarboxylic acid exhibits reversible temperature‐induced deuteron migration of a magnitude unprecedented in this class of compounds. We used a combination of variable‐temperature powder and single‐crystal neutron diffraction and density functional theory (DFT)‐based computational methods to elucidate the origin of this remarkable behaviour. Single‐crystal neutron diffraction shows that between 15 and 300 K, the deuteron moves by 0.32(1) Å and the structure changes from a low‐temperature N? D???O form to a high‐temperature N???D? O form. Variable‐temperature powder neutron‐diffraction data, which was fitted by using parametric Rietveld refinement, show that this deuteron migration is due to an isosymmetric, first‐order phase transition that occurs by growth of the daughter phase in the parent‐phase matrix. Similar phase transitions are observed in two selectively deuterated forms of the material. DFT calculations demonstrate the role of phonons and show that vibrational free‐energy stabilisation, which plays a key role in the observed structural phase transitions, is more pronounced in the fully deuterated material and proportional to the mass of the molecule, that is, the level of deuteration. This is consistent with our experimental work, for which distinct crystallographic phase transitions were clearly observed for the three deuterated systems, but not for the fully protonated material.     

Use of N-(4-aminophenyl)piperidine derivatization to improve organic acid detection with supercritical fluid chromatography-mass spectrometry

The analysis of organic acids in complex mixtures by LC-MS can often prove challenging, especially due to the poor sensitivity of negative ionization mode required for detection of these compounds in their native (i.e., underivatized or untagged) form. These compounds have also been difficult to measure using supercritical fluid chromatography (SFC)-MS, a technique of growing importance for metabolomic analysis, with similar limitations based on negative ionization. In this report, the use of a high proton affinity N-(4-aminophenyl)piperidine derivatization tag is explored for the improvement of organic acid detection by SFC-MS. Four organic acids (lactic, succinic, malic, and citric acids) with varying numbers of carboxylate groups were derivatized with N-(4-aminophenyl)piperidine to achieve detection limits down to 0.5 ppb, with overall improvements in detection limit ranging from 25-to-2100-fold. The effect of the derivatization group on sensitivity, which increased by at least 200-fold for compounds that were detectable in their native form, and mass spectrometric detection are also described. Preliminary investigations into the separation of these derivatized compounds identified multiple stationary phases that could be used for complete separation of all four compounds by SFC. This derivatization technique provides an improved approach for the analysis of organic acids by SFC-MS, especially for those that are undetectable in their native form.     

Chiral ligand-exchange separation and resolution of extremely rigid glutamate analogs: 1-aminospiro[2.2]pentyl-1,4-dicarboxylic acids

Owing to their chelation ability, a series of fully constrained l-Glu analogs formed by the spiro-union of two cyclopropane rings (1-aminospiro[2.2]pentyl-1,4-dicarboxylic acids, ASPED A–D), was submitted to chiral ligand-exchange chromatographic (CLEC) analysis. As the initial step, two methodologically different chiral devices were evaluated. A chiral stationary phase (CSP) obtained by dynamic coating of C₁₈ chains with the S-trityl-(R)-cysteine ((R)-STC) was used first with this objective. The lack of separation of the enantiomers of ASPED C and D prompted us to utilize the chiral mobile phase (CMP) prepared from O-benzyl-(S)-serine ((S)-OBS). The latter afforded complete separation of the four pairs of enantiomers. For all the pairs, quantum mechanical investigations shed light on the main features responsible for the different enantiomer recognition mechanism with (S)-OBS. The validated analytical method was then fruitfully adopted for semi-preparative-scale isolation of the enantiomers of ASPED C.