Second‐Generation Inhibitors for the Metalloprotease Neprilysin Based on Bicyclic Heteroaromatic Scaffolds: Synthesis,Biological Activity,and X‐Ray Crystal‐Structure Analysis

A new class of nonpeptidic inhibitors of the Zn^II‐dependent metalloprotease neprilysin with IC₅₀ values in the nanomolar activity range (0.034–0.30 μM ) were developed based on structure‐based de novo design (Figs. 1 and 2). The inhibitors feature benzimidazole and imidazo[4,5‐c]pyridine moieties as central scaffolds to undergo H‐bonding to Asn542 and Arg717 and to engage in favorable π‐π stacking interactions with the imidazole ring of His711. The platform is decorated with a thiol vector to coordinate to the Zn^II ion and an aryl residue to occupy the hydrophobic S1′ pocket, but lack a substituent for binding in the S2′ pocket, which remains closed by the side chains of Phe106 and Arg110 when not occupied. The enantioselective syntheses of the active compounds (+)‐ 1 , (+)‐ 2 , (+)‐ 25 , and (+)‐ 26 were accomplished using Evans auxiliaries (Schemes 2, 4, and 5). The inhibitors (+)‐ 2 and (+)‐ 26 with an imidazo[4,5‐c]pyridine core are ca. 8 times more active than those with a benzimidazole core ((+)‐ 1 and (+)‐ 25 ) (Table 1). The predicted binding mode was established by X‐ray analysis of the complex of neprilysin with (+)‐ 2 at 2.25‐Å resolution (Fig. 4 and Table 2). The ligand coordinates with its sulfanyl residue to the Zn^II ion, and the benzyl residue occupies the S1′ pocket. The 1H‐imidazole moiety of the central scaffold forms the required H‐bonds to the side chains of Asn542 and Arg717. The heterobicyclic platform additionally undergoes π‐π stacking with the side chain of His711 as well as edge‐to‐face‐type interactions with the side chain of Trp693. According to the X‐ray analysis, the substantial advantage in biological activity of the imidazo‐pyridine inhibitors over the benzimidazole ligands arises from favorable interactions of the pyridine N‐atom in the former with the side chain of Arg102. Unexpectedly, replacement of the phenyl group pointing into the deep S1′ pocket by a biphenyl group does not enhance the binding affinity for this class of inhibitors.     

Loss of H2O from M + t-Butyl Complex Ions of Benzyl Alcohol in Isobutane Chemical Ionization Mass Spectrometry

In isobutane chemical ionization mass spectrometry benzyl alcohol exhibits ions at m/z 147 (‘M + 39’) that arise by a loss of H₂O from [M + C₄H₉]⁺, i.e. ‘M + 57’ complex ions. Electrophilic aromatic substitution of a proton at an ortho-position of neutral C₆H₅CH₂OH with [t-C₄H₉]⁺ and, alternatively, nucleophilic substitution of H₂O at the benzylic carbon in \documentclass{article}\pagestyle{empty}\begin{document}$ {\rm C_6 H_5 CH_2}\mathop {\rm O}\limits^+ {\rm H}_2 $\end{document} with CH₂?C (CH₃)₂ are discussed as possible pathways. Evidence in favor of the latter is derived from the analysis of C₆D₅CH₂OH and C₆H₅CD₂OH for the origin of the H-atoms lost in H₂O. The inferred ion structure of m/z 147 is verified by mass-analyzed ion kinetic energy (MIKE.) measurements of its collision-activated (CA.) decomposition. MIKE./CA. spectra of mass-selected m/z 147 ions, once generated by (CI(i-C₄H₁₀) from benzyl alcohol and, once, from 2-methyl-4-phenyl-2-butanol match closely and, thus, reflect identical ion structures. With reference to the simple genesis of this ion from the latter precursor, the structure in question can be concluded to be \documentclass{article}\pagestyle{empty}\begin{document}$ {\rm C_6 H_5 CH_2 CH_2}\mathop {\rm C}\limits^+ ({\rm CH}_3)_2 $\end{document} .     

Structural Comparison of Oligoribonucleotides and Their 2′-Deoxy-2′-fluoro Analogs by heteronuclear NMR spectroscopy

1-(2′-Deoxy-2′-fluororibofuranosyl)pyrimidines were synthesized and incorporated into an RNA oligonucleotide to give 5′-r[C_fGC_f(U_fU_fC_fG)GC_fG]-3′ (C_f: short form of C = 2′-deoxy-2′-fluorocytidine; U_f: short form of U = 2′-deoxy-2′-fluorouridine). The oligomer was investigated by means of UV, CD, and NMR spectroscopy to address the question of how F-labels can substitute ¹³C-labels in the ribose ring. Through-space (NOE) and through-bond (scalar couplings) experiments were performed that make use of the ameliorated chemical-shift dispersion induced by ¹⁹F as an alternative heteronucleus. A comparison of the structures of fluorinated vs. unmodified oligomer is given. It turns out that the fluorinated oligonucleotide exists in a 14:3 equilibrium between a hairpin and a duplex conformation, in contrast to the unmodified oligonucleotide which predominantly adopts the hairpin conformation. Furthermore, the fluorinated hairpin structure adopts two distinct conformations that differ in the sugar conformation of the U and C nucleoside units, as detected by the ¹⁹F-NMR chemical shifts. The role of the 2′-OH group as stabilizing element in RNA secondary structure is discussed.     

Simultaneous acquisition of X-ray spectra using a multi-wire, position-sensitive gas flow detector

A multi-wire, gas-filled position-sensitive detector has been developed for the simultaneous recording of wavelength-dispersed X-ray signals that enables X-ray fluorescence spectrometry with a limited multi-element capability in the low Z element range. Details of the modular construction of the detector are given. The detector performance was characterized using Al–K radiation and a variable slit system. The detector has been applied in a laboratory spectrometer equipped with an electron source and a double multilayer mirror device as the wavelength-dispersing element. Spectra from Al and Si obtained in the simultaneous acquisition mode show good agreement with calculations performed using a ray-tracing model.     

Anthropogenic and solar forcing in δ13C time pattern of coralline sponges

We present the results of a re-analysis of a previously published carbon isotope data-set related to coralline sponges in the Caribbean Sea. The original interpretation led to the discrimination between a pre-industrial period, with a signal controlled by solar-induced climatic variations, followed by the industrial era, characterized by a progressive δ¹³C negative shift due to the massive anthropogenic carbon emissions. Our re-analysis allowed to extract from the raw isotopic data evidence of a solar forcing still visible during the industrial era, with a particular reference to the 88-year Gleissberg periods. These signals are related to slope changes in both the δ¹³C versus time and the δ¹³C versus carbon emission curves.     

Shear thickening, temporal shear oscillations, and degradation of dilute equimolar CTAB/NaSal wormlike solutions

The rheological characterization of dilute and semi-dilute equimolar cetyltrimethylammonium bromide and sodium salicylate solutions is reported, including their linear and nonlinear responses. Start-up experiments and direct birefringence measurements suggest that even at a concentration of as low as 1.0 mM, temporal shear oscillations occur at low shear rates. At concentrations above 10 mM, those low-stress structures vanish and give way to shear-thickening and shear-banding behaviors as seen for other semi-dilute surfactant solutions. Also, the degradation of these solutions after exposure to rubber pump tubing is covered.