Fluorescence polarization-based competition binding assay for c-Jun N-terminal kinases 1 and 2

Ansideri, Francesco; Dammann, Marcel; Boeckler, Frank M.; Koch, Pierre

doi:10.1016/j.ab.2017.05.022

Cited by 8 publications

(11 citation statements)

References 18 publications

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“…In 2016, we published compound 1a as a balanced dual JNK3/p38α MAPK inhibitor, which served as a precursor for the synthesis of a fluorescent probe used in fluorescence polarization-based binding assays. 19 , 20 As it is evident from the biological activity of 1a in comparison to the activity of previous inhibitors, modifying the substitution pattern around the pyridinylimidazole scaffold can contribute to a shift in selectivity toward the JNK3.…”

Section: Introductionmentioning

confidence: 99%

Structural Optimization of a Pyridinylimidazole Scaffold: Shifting the Selectivity from p38α Mitogen-Activated Protein Kinase to c-Jun N-Terminal Kinase 3

et al. 2018

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Starting from known p38α mitogen-activated protein kinase (MAPK) inhibitors, a series of inhibitors of the c-Jun N-terminal kinase (JNK) 3 was obtained. Altering the substitution pattern of the pyridinylimidazole scaffold proved to be effective in shifting the inhibitory activity from the original target p38α MAPK to the closely related JNK3. In particular, a significant improvement for JNK3 selectivity could be achieved by addressing the hydrophobic region I with a small methyl group. Furthermore, additional structural modifications permitted to explore structure–activity relationships. The most potent inhibitor 4-(4-methyl-2-(methylthio)-1H-imidazol-5-yl)-N-(4-morpholinophenyl)pyridin-2-amine showed an IC50 value for the JNK3 in the low triple digit nanomolar range and its binding mode was confirmed by X-ray crystallography.

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Section: Introductionmentioning

confidence: 99%

Structural Optimization of a Pyridinylimidazole Scaffold: Shifting the Selectivity from p38α Mitogen-Activated Protein Kinase to c-Jun N-Terminal Kinase 3

et al. 2018

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“…The phage-encoded bicyclic peptide library was subjected to three iterative rounds of affinity selection against c-Jun N-terminal kinase 3 (JNK3) using this modification protocol. JNK3 was pragmatically chosen as a model target based on a variety of active and successful drug discovery projects (Goettert et al, 2010 ; Lange et al, 2015 ; Muth et al, 2015 , 2017 ; Ansideri et al, 2018 ) and assay developments (Ansideri et al, 2016 , 2017 ) reported recently by the involved laboratories. After each selection, the captured phages were eluted by treatment with low pH buffer.…”

Section: Resultsmentioning

confidence: 99%

“…For the assessment of target binding, fluorescence polarization assays with direct and competitive titration regimes were applied (Ansideri et al, 2016 ). For the competitive fluorescence polarizations assays, the previously established, fluorescein-labeled fluorescence polarization (FP) probe PIT0105006 was applied, binding with a K D -value of 3.0 ± 0.2 nM to JNK3 (Ansideri et al, 2017 ). All of the selected, TPSMB modified sequences exhibited affinities to JNK3 in the one-digit micromolar range.…”

Section: Resultsmentioning

confidence: 99%

Switching Between Bicyclic and Linear Peptides — The Sulfhydryl-Specific Linker TPSMB Enables Reversible Cyclization of Peptides

et al. 2018

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Phage display-selected bicyclic peptides have already shown their great potential for the development as bioactive modulators of therapeutic targets. They can provide enhanced proteolytic stability and improved membrane permeability. Molecular design of new linker molecules has led to a variety of new synthetic approaches for the generation of chemically constrained cyclic peptides. This diversity can be useful for the development of novel peptide-based therapeutic, diagnostic, and scientific tools. Herein, we introduce 1,3,5-tris((pyridin-2-yldisulfanyl)methyl)benzene (TPSMB) as a planar, trivalent, sulfhydryl-specific linker that facilitates reversible cyclization and linearization via disulfide bond formation and cleavage of bicyclic peptides of the format CXnCXnC, where X is any proteinogenic amino acid except cysteine. The rapid and highly sulfhydryl-specific reaction of TPSMB under physiological conditions is demonstrated by selecting bicyclic peptide binders against c-Jun N-terminal kinase 3 (JNK3) as a model target. While model peptides remain stably cyclized for several hours in presence of typical blood levels of glutathione in vitro, high cytosolic concentrations of glutathione linearize these peptides completely within 1 h. We propose that reversible linkers can be useful tools for several technical applications where target affinity depends on the bicyclic structure of the peptide.

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“…) . The latter serves as a reporter molecule in fluorescence polarization‐based competition binding assays to determine the affinity of potential inhibitors for p38α MAP kinase as well as for all JNK isoforms .…”

Section: Synthesis Of 245‐tri‐substituted Imidazolesmentioning

confidence: 99%

2‐Alkylsulfanyl‐4(5)‐aryl‐5(4)‐heteroarylimidazoles: An Overview on Synthetic Strategies and Biological Activity

Koch

Ansideri

2017

Archiv der Pharmazie

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2-Alkylsulfanyl-4(5)-aryl-5(4)-heteroarylimidazoles represent an important class of ATP-competitive protein kinase inhibitors, offering the possibility of multiple interactions with different regions of the target enzyme. The necessity of exploring the effects of diverse chemical decorations around the imidazole core prompted the design of several synthetic routes aimed at achieving both efficiency and flexibility. Additionally, the optimization of established protocols and the extensive use of transition metal-catalyzed cross-coupling reactions have been broadening the spectrum of preparative methodologies within the last decade. This review summarizes the progress in the development of synthetic strategies leading to 2-alkylsulfanyl-4(5)-aryl-5(4)-heteroarylimidazoles and 1-alkyl-2-alkylsulfanyl-4(5)-aryl-5(4)-heteroarylimidazoles and offers a glance at the biological activities of this class of compounds.

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Fluorescence polarization-based competition binding assay for c-Jun N-terminal kinases 1 and 2

Cited by 8 publications

References 18 publications

Structural Optimization of a Pyridinylimidazole Scaffold: Shifting the Selectivity from p38α Mitogen-Activated Protein Kinase to c-Jun N-Terminal Kinase 3

Structural Optimization of a Pyridinylimidazole Scaffold: Shifting the Selectivity from p38α Mitogen-Activated Protein Kinase to c-Jun N-Terminal Kinase 3

Switching Between Bicyclic and Linear Peptides — The Sulfhydryl-Specific Linker TPSMB Enables Reversible Cyclization of Peptides

2‐Alkylsulfanyl‐4(5)‐aryl‐5(4)‐heteroarylimidazoles: An Overview on Synthetic Strategies and Biological Activity

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