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Tunable Methacrylamides for Covalent Ligand Directed Release Chemistry.

Journal of the American Chemical Society (2021-03-26)
Rambabu N Reddi, Efrat Resnick, Adi Rogel, Boddu Venkateswara Rao, Ronen Gabizon, Kim Goldenberg, Neta Gurwicz, Daniel Zaidman, Alexander Plotnikov, Haim Barr, Ziv Shulman, Nir London
ABSTRACT

Targeted covalent inhibitors are an important class of drugs and chemical probes. However, relatively few electrophiles meet the criteria for successful covalent inhibitor design. Here we describe α-substituted methacrylamides as a new class of electrophiles suitable for targeted covalent inhibitors. While typically α-substitutions inactivate acrylamides, we show that hetero α-substituted methacrylamides have higher thiol reactivity and undergo a conjugated addition-elimination reaction ultimately releasing the substituent. Their reactivity toward thiols is tunable and correlates with the pKa/pKb of the leaving group. In the context of the BTK inhibitor ibrutinib, these electrophiles showed lower intrinsic thiol reactivity than the unsubstituted ibrutinib acrylamide. This translated to comparable potency in protein labeling, in vitro kinase assays, and functional cellular assays, with improved selectivity. The conjugate addition-elimination reaction upon covalent binding to their target cysteine allows functionalizing α-substituted methacrylamides as turn-on probes. To demonstrate this, we prepared covalent ligand directed release (CoLDR) turn-on fluorescent probes for BTK, EGFR, and K-RasG12C. We further demonstrate a BTK CoLDR chemiluminescent probe that enabled a high-throughput screen for BTK inhibitors. Altogether we show that α-substituted methacrylamides represent a new and versatile addition to the toolbox of targeted covalent inhibitor design.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
N-(But-3-yn-1-yl)-2-(((2-oxo-2H-chromen-7-yl)oxy)methyl)acrylamide
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SKUVendor SKUPack SizeAvailabilityPrice
EN300-27159994-50MG50 mg
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$327.60
EN300-27159994-100MG100 mg
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$490.10
EN300-27159994-250MG250 mg
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$699.40
EN300-27159994-500MG500 mg
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$1,101.10
EN300-27159994-1G1 g
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$1,411.80
EN300-27159994-2.5G2.5 g
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$2,766.40
EN300-27159994-5G5 g
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$4,091.10
EN300-27159994-10G10 g
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$6,067.10

To order products, please contact your local dealer.

Sigma-Aldrich
RIPA Buffer
Sign Into View Organizational & Contract Pricing
SKUVendor SKUPack SizeAvailabilityPrice
EN300-27159994-50MG50 mg
Ships in 7 days - from Aldrich Partner.
$327.60
EN300-27159994-100MG100 mg
Ships in 7 days - from Aldrich Partner.
$490.10
EN300-27159994-250MG250 mg
Ships in 7 days - from Aldrich Partner.
$699.40
EN300-27159994-500MG500 mg
Ships in 7 days - from Aldrich Partner.
$1,101.10
EN300-27159994-1G1 g
Ships in 7 days - from Aldrich Partner.
$1,411.80
EN300-27159994-2.5G2.5 g
Ships in 7 days - from Aldrich Partner.
$2,766.40
EN300-27159994-5G5 g
Ships in 7 days - from Aldrich Partner.
$4,091.10
EN300-27159994-10G10 g
Ships in 7 days - from Aldrich Partner.
$6,067.10

To order products, please contact your local dealer.