Evaluation of Pyrones in Bioorthogonal Reactions: Correlation between Structure, Reactivity, and Bioorthogonality

The Journal of Organic Chemistry, Год журнала: 2025, Номер unknown

Опубликована: Фев. 19, 2025

Alpha-pyrones have been used for applications ranging from total synthesis to antibiotics. However, their application as dienes in bioorthogonal reactions has not extensively explored. In previous work, we demonstrated the promising of ester-functionalized pyrones protein labeling. Here, constructed a library substituted evaluate potential by exploring relationships among structure, reactivity, and bioorthogonality. We found that most pyrone derivatives with electron-withdrawing groups exhibited reactivity toward endo-bicyclo[6.1.0]nonyne (BCN), producing tricyclic tetracyclic products good yields. As expected, more stronger substituents showed faster reaction kinetics BCN. Bicyclic substantially decreased likely resulting increased steric effects. Counterintuitively, substitutions at positions 4 5 affected than those 3 6. To provide insights into both expected counterintuitive reactivities members, performed quantum chemical analysis. Additionally, evaluated each pyrone's L-cysteine no correlation between BCN cysteine-based Finally, collection popular dienophiles reactions.

Язык: Английский

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Amber Codon Mutational Scanning and Bioorthogonal PEGylation for Epitope Mapping of Antibody Binding Sites on Human Arginase-1

ACS Chemical Biology, Год журнала: 2025, Номер unknown

Опубликована: Апрель 1, 2025

Epitope mapping is crucial for understanding immunological responses to protein therapeutics. Here, we combined genetic code expansion and bacterial surface display incorporate S-allylcysteine (SAC) into human arginase-1 (hArg1) via Methanococcoides burtonii pyrrolysyl-tRNA synthetase. Using an amber codon deep mutational scanning sequencing workflow, mapped SAC incorporation efficiency across the hArg1 sequence, providing insights structural sequence dependencies of noncanonical amino acid incorporation. We used mutually bioorthogonal allyl/tetrazine azide/DBCO chemistries achieve site-specific PEGylation fluorescent labeling hArg1, revealing side chain reactivity solvent accessibility residues in hArg1. This system was further applied determine binding epitope a monoclonal antibody on high-resolution data impact residue position binding. Our method produces high dimensional efficiency, functionalization enabled by chemistries, therapeutic proteins.

Язык: Английский

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Convergence of the Orthogonal: A Tribute to the Imaginative Chemistry of George Barany

International Journal of Peptide Research and Therapeutics, Год журнала: 2025, Номер 31(3)

Опубликована: Март 25, 2025

Язык: Английский

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Amber Codon Mutational Scanning and Bioorthogonal PEGylation for Mapping Antibody Binding Sites on Human Arginase-1

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Авг. 20, 2024

Abstract Epitope mapping is crucial for understanding immunological responses to protein therapeutics. Here, we combined genetic code expansion and bacterial surface display incorporate S-allylcysteine (SAC) into human arginase-1 (hArg1) via Methanococcoides burtonii pyrrolysyl-tRNA synthetase. Using an amber codon deep mutational scanning sequencing workflow, mapped SAC incorporation efficiency across the hArg1 sequence, providing insights structural sequence dependencies of non-canonical amino acid incorporation. We used mutually bioorthogonal allyl/tetrazine azide/DBCO chemistries achieve site-specific PEGylation fluorescent labeling hArg1, revealing side chain reactivity solvent accessibility residues in hArg1. This system was further applied determine binding epitope a monoclonal antibody on high-resolution data impact binding. Our method produces high dimensional efficiency, functionalization enabled by chemistries, therapeutic proteins.

Язык: Английский

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Synthesis of 3-Chloro-6-(4-chloro-3,5-dimethyl-1H-pyrazol-1-yl)pyridazine and Its Deuterated Analogue

Russian Journal of General Chemistry, Год журнала: 2024, Номер 94(11), С. 2833 - 2840

Опубликована: Ноя. 1, 2024

Язык: Английский

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