Synergistic Experimental and Computational Investigation of the Bioorthogonal Reactivity of Substituted Aryltetrazines

Tetrazines (Tz) have been applied as bioorthogonal agents for various biomedical applications including pretargeted imaging approaches. In radioimmunoimaging, pretargeting increases the target-to-background ratio while simultaneously reducing the radiation burden. We have recently reported a strategy to directly 18F-label highly reactive tetrazines based on a 3-(3-fluorophenyl)-Tz core structure. Herein, we report a kinetic study on this versatile scaffold. A library of 40 different tetrazines was prepared, fully characterized, and investigated with emphasis on second order rate constants for the reaction with trans-cyclooctene (TCO). Our results reveal the effects of various substitution patterns and moreover demonstrate the importance of measuring reactivities in the solvent of interest, as click rates in different solvents do not necessarily correlate well. In particular, we report that tetrazines modified in 2-position of the phenyl substituent show high intrinsic reactivity towards TCO, which is diminished in aqueous systems by unfavorable solvent effects. The obtained results enable the prediction of the bioorthogonal reactivity and thereby facilitate the development of the next-generation of substituted aryltetrazines for in vivo application.

Light-Driven CO2 Reduction by Co-Cytochrome b562

The current trend in atmospheric carbon dioxide concentrations is causing increasing concerns for its environmental impacts, and spurring the developments of sustainable methods to reduce CO2 to usable molecules. We report the light-driven CO2 reduction in water in mild conditions by artificial protein catalysts based on cytochrome b562 and incorporating cobalt protoporphyrin IX as cofactor. Incorporation into the protein scaffolds enhances the intrinsic reactivity of the cobalt porphyrin toward proton reduction and CO generation. Mutations around the binding site modulate the activity of the enzyme, pointing to the possibility of further improving catalytic activity through rational design or directed evolution.

Prioritization of antimicrobial targets by CRISPR-based oligo recombineering

SummaryNucleophilic amino acids are important in covalent drug development yet underutilized as antimicrobial targets. Over recent years, several chemoproteomic technologies have been developed to mine chemically-accessible residues via their intrinsic reactivity toward electrophilic probes. However, these approaches cannot discern which reactive sites contribute to protein function and should therefore be prioritized for drug discovery. To address this, we have developed a CRISPR-based Oligo Recombineering (CORe) platform to systematically prioritize reactive amino acids according to their contribution to protein function. Our approach directly couples protein sequence and function with biological fitness. Here, we profile the reactivity of >1,000 cysteines on ~700 proteins in the eukaryotic pathogen Toxoplasma gondii and prioritize functional sites using CORe. We competitively compared the fitness effect of 370 codon switches at 74 cysteines and identify functional sites in a diverse range of proteins. In our proof of concept, CORe performed >800 times faster than a standard genetic workflow. Reactive cysteines decorating the ribosome were found to be critical for parasite growth, with subsequent target-based screening validating the apicomplexan translation machinery as a target for covalent ligand development. CORe is system-agnostic, and supports expedient identification, functional prioritization, and rational targeting of reactive sites in a wide range of organisms and diseases.

Structural Characteristics of Lignite Char from Different Pyrolysis Reactors and the Influence on Their Gasification Reactivity

The gasification reactivity of coal char is affected by numerous experimental variables, and char structure is one of the dominant factors. In this work, Raman spectroscopy, powder X-ray diffraction (XRD), and N2 adsorption were used to investigate the physical and chemical structure of char prepared under different pyrolysis conditions. Three kinds of pyrolysis reactors, fluidizedbed reactor (FL), entrained-flow-bed reactor (EF), and fixed-bed reactor (PT), were designed and used to prepare the char samples. Lignite was pyrolyzed in a fixed-bed reactor with a heating rate of 10 ºC min-1, and the final temperature was 1000 ºC. The gasification reactivity of char was characterized in a quartz fixed-bed reactor under CO2, H2O, and their mixtures at 750 ºC. FB reactor produces chars with a smaller interlayer spacing of aromatic layers (d002), and FL reactor produces chars with a larger mean crystallite size along the c-axis (Lc) and aromaticity (ƒa) but inhibits the growth of mean crystallite size along the a-axis (La). The content of small aromatic rings, which is higher in the FL reactor, positively affects the initial intrinsic reactivity.

Kanchan Arsenic Filters and the Future of Fe0-Based Filtration Systems for Single Household Drinking Water Supply

Biological and chemical contamination of natural water bodies is a global health risk for more than one billion people, mostly living in low-income countries. Innovative, affordable, and efficient decentralized solutions for safe drinking water supply are urgently needed. Metallic iron (Fe0)-based filtration systems have been described as such an appropriate solution. This communication focuses on the Kanchan arsenic filter (KAF), presented in the early 2000s and widely assessed during the past decade. The KAF contains iron nails as the Fe0 source and is primarily designed to remove As from polluted tube well waters. Recent independent works assessing their performance have all reported on a high degree of variability in efficiency depending mostly on (1) the current operating conditions, (2) the design, and (3) the groundwater chemistry. This communication shows that the major problems of the KAF are twofold: (1) a design mistake as the Fe0 units disturb the operation and functionality of the biosand filter, and (2) the use of poorly characterized iron nails of unknown reactivity. This assertion is supported by the evidence that the very successful community filter designed by the Indian Institute of Technology Bombay works with iron nails and has been efficient for many years. Replacing iron nails by more reactive Fe0 materials (e.g., iron fillings and steel wool) should be tested in a new generation KAF. It is concluded that a methodological or systematic approach in introducing and monitoring the efficiency of KAF should be used to test and disseminate the next generation KAF worldwide. Moreover, better characterization of the Fe0 materials including their intrinsic reactivity is required.

Concise asymmetric syntheses of streptazone A and abikoviromycin

Streptazone A and abikoviromycin are related alkaloids that both feature an unusual arrangement of reactive functionalities within an underlying compact tricyclic ring system. Here, we report a highly concise asymmetric synthesis of both natural products. The developed route first constructs another member of the streptazones, streptazone B1, using a rhodium-catalyzed distal selective allene-ynamide Pauson-Khand reaction as the key transformation. A regio- and enantioselective epoxidation under chiral phase-transfer catalytic conditions was then achieved to directly make streptazone A in 8 steps overall. A chemoselective, iridium-catalyzed reduction of the enaminone-system was employed to make abikoviromycin in one additional step. Studies of the intrinsic reactivity of streptazone A towards the cysteine mimic, <i>N</i>-acetylcysteamine, revealed unanticipated transformations, resulting in thiol conjugation to both the hindered tertiary carbon of the double allylic epoxide and in bis-thiol conjugation which may proceed via formation of a cyclopentadienone intermediate. With flexible access to these compounds, studies aimed to identify their direct biological targets are now possible.

Concise asymmetric syntheses of streptazone A and abikoviromycin

Streptazone A and abikoviromycin are related alkaloids that both feature an unusual arrangement of reactive functionalities within an underlying compact tricyclic ring system. Here, we report a highly concise asymmetric synthesis of both natural products. The developed route first constructs another member of the streptazones, streptazone B1, using a rhodium-catalyzed distal selective allene-ynamide Pauson-Khand reaction as the key transformation. A regio- and enantioselective epoxidation under chiral phase-transfer catalytic conditions was then achieved to directly make streptazone A in 8 steps overall. A chemoselective, iridium-catalyzed reduction of the enaminone-system was employed to make abikoviromycin in one additional step. Studies of the intrinsic reactivity of streptazone A towards the cysteine mimic, <i>N</i>-acetylcysteamine, revealed unanticipated transformations, resulting in thiol conjugation to both the hindered tertiary carbon of the double allylic epoxide and in bis-thiol conjugation which may proceed via formation of a cyclopentadienone intermediate. With flexible access to these compounds, studies aimed to identify their direct biological targets are now possible.