Substrate Activity Screening with Kinases: Discovery of Small‐Molecule Substrate‐Competitive c‐Src Inhibitors

Breen, Meghan E.; Steffey, Michael E.; Lachacz, Eric J.; Kwarcinski, Frank E.; Fox, Christel C.; Soellner, Matthew B.

doi:10.1002/anie.201311096

Cited by 19 publications

(29 citation statements)

References 28 publications

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“…Fluorinating the selected substrate afforded the initial tetrafluorophenol inhibitor. Its further optimization resulted in a substrate‐competitive inhibitor of c‐Src (compound 7 , K i =16 μ m ) …”

Section: Sas Studiesmentioning

confidence: 96%

“…Although this, as mentioned, is interesting from a time/cost efficiency perspective, it can also be expected that related or homologous enzymes may return identical substrate “hits” . Partially addressing these issues, more recent studies have used hierarchical, 2D extended connectivity analysis to guide library design, and this has been shown to have potential for the domain . On the other hand, systematically implementing structure‐based design during the substrate or inhibitor optimization process has not only the potential to facilitate the optimization process, but also to resolve selectivity issues …”

Section: Substrate Library Designmentioning

confidence: 99%

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Substrate Activity Screening (SAS) and Related Approaches in Medicinal Chemistry

et al. 2016

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Substrate activity screening (SAS) was presented a decade ago by Ellman and co-workers as a straightforward methodology for the identification of fragment-sized building blocks for enzyme inhibitors. Ever since, SAS and variations derived from it have been successfully applied to the discovery of inhibitors of various families of enzymatically active drug targets. This review covers key achievements and challenges of SAS and related methodologies, including the modified substrate activity screening (MSAS) approach. Special attention is given to the kinetic and thermodynamic aspects of these methodologies, as a thorough understanding thereof is crucial for successfully transforming the identified fragment-sized hits into potent inhibitors.

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Section: Sas Studiesmentioning

confidence: 96%

Section: Substrate Library Designmentioning

confidence: 99%

Substrate Activity Screening (SAS) and Related Approaches in Medicinal Chemistry

et al. 2016

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“…We have successfully used this approach for the identification of selective low molecular weight inhibitors of therapeutically relevant proteases 27–32 and phosphatases, 33–35 and other labs have implemented related strategies to target kinases. 36–37 …”

Section: Introductionmentioning

confidence: 99%

Identification of Multiple Structurally Distinct, Nonpeptidic Small Molecule Inhibitors of Protein Arginine Deiminase 3 Using a Substrate-Based Fragment Method

et al. 2015

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The protein arginine deiminases (PADs) are a family of enzymes that catalyze the post-translational hydrolytic deimination of arginine residues. Four different enzymologically active PAD subtypes have been characterized and exhibit tissue-specific expression and association with a number of different diseases. In this Article we describe the development of an approach for the reliable discovery of low-molecular weight, nonpeptidic fragment substrates of the PADs that then can be optimized and converted to mechanism-based irreversible PAD inhibitors. The approach is demonstrated by the development of the first potent and selective inhibitors of PAD3, a PAD subtype implicated in the neurodegenerative response to spinal cord injury. Multiple structurally distinct inhibitors were identified with the most potent inhibitors having >10,000 min−1 M−1 kinact/KI values and ≥10-fold selectivity for PAD3 over PADs 1, 2, and 4.

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“…4 Non-ATP-competitive inhibitors possess higher degrees of selectivity, however, they generally suffer from a lack of potency. 7–9 …”

mentioning

confidence: 99%

“…8,9 To validate that our bivalent probe can identify non-ATP-competitive inhibitors, we examined MEB-SCI, a substrate-competitive c-Src kinase inhibitor recently reported by our laboratory, and obtained a K d = 15.6 µM, which is identical to the reported K i value obtained using an activity-based assay. 7 We envisioned that we could screen a library of putative non-ATP-competitive kinase inhibitors against our bisubstrate probe in a TR-FRET assay. A second screen against an ATP-competitive TR-FRET tracer removes any ATP-competitive compounds from the hits.…”

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confidence: 99%

Exquisitely Specific Bisubstrate Inhibitors of c-Src Kinase

et al. 2015

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We have developed a modular approach to bisubstrate inhibition of protein kinases. We apply our methodology to c-Src and and identify a highly selective bisubstrate inhibitor for this target. Our approach has yielded the most selective c-Src inhibitor to date, and methodology to render the bisubstrate inhibitor cell-permeable provides a highly valuable tool for the study of c-Src signaling. In addition, we have applied our bisubstrate inhibitor to develop a novel screening methodology to identify non-ATP-competitive inhibitors of c-Src. Using this methodology, we have discovered the most potent non-ATP-competitive inhibitor reported to date. Our methodology is designed to be general and could be applicable to additional kinases inhibited by the promiscuous ATP-competitive fragment used in our studies.

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Substrate Activity Screening with Kinases: Discovery of Small‐Molecule Substrate‐Competitive c‐Src Inhibitors

Cited by 19 publications

References 28 publications

Substrate Activity Screening (SAS) and Related Approaches in Medicinal Chemistry

Substrate Activity Screening (SAS) and Related Approaches in Medicinal Chemistry

Identification of Multiple Structurally Distinct, Nonpeptidic Small Molecule Inhibitors of Protein Arginine Deiminase 3 Using a Substrate-Based Fragment Method

Exquisitely Specific Bisubstrate Inhibitors of c-Src Kinase

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