Enhancing intracellular accumulation and target engagement of PROTACs with reversible covalent chemistry

AbstractCurrent efforts in the proteolysis targeting chimera (PROTAC) field mostly focus on choosing an appropriate E3 ligase for the target protein, improving the binding affinities towards the target protein and the E3 ligase, and optimizing the PROTAC linker. However, due to the large molecular weights of PROTACs, their cellular uptake remains an issue. Through comparing how different warhead chemistry, reversible noncovalent (RNC), reversible covalent (RC), and irreversible covalent (IRC) binders, affects the degradation of Bruton’s Tyrosine Kinase (BTK), we serendipitously discover that cyano-acrylamide-based reversible covalent chemistry can significantly enhance the intracellular accumulation and target engagement of PROTACs and develop RC-1 as a reversible covalent BTK PROTAC with a high target occupancy as its corresponding kinase inhibitor and effectiveness as a dual functional inhibitor and degrader, a different mechanism-of-action for PROTACs. Importantly, this reversible covalent strategy is generalizable to improve other PROTACs, opening a path to enhance PROTAC efficacy.

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Lysine-Targeted Inhibitors and Chemoproteomic Probes

Annual Review of Biochemistry ◽

10.1146/annurev-biochem-061516-044805 ◽

2019 ◽

Vol 88 (1) ◽

pp. 365-381 ◽

Cited By ~ 19

Author(s):

Adolfo Cuesta ◽

Jack Taunton

Keyword(s):

Drug Target ◽

Chemical Biology ◽

Large Set ◽

Target Engagement ◽

Covalent Bonds ◽

Protein Targets ◽

Ligand Binding Sites ◽

Covalent Inhibitors ◽

Enzyme Cofactors ◽

Reversible Covalent

Covalent inhibitors are widely used in drug discovery and chemical biology. Although covalent inhibitors are frequently designed to react with noncatalytic cysteines, many ligand binding sites lack an accessible cysteine. Here, we review recent advances in the chemical biology of lysine-targeted covalent inhibitors and chemoproteomic probes. By analyzing crystal structures of proteins bound to common metabolites and enzyme cofactors, we identify a large set of mostly unexplored lysines that are potentially targetable with covalent inhibitors. In addition, we describe mass spectrometry–based approaches for determining proteome-wide lysine ligandability and lysine-reactive chemoproteomic probes for assessing drug–target engagement. Finally, we discuss the design of amine-reactive inhibitors that form reversible covalent bonds with their protein targets.

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Optimization of templated replication and macrocycle formation using tunable orthogonal reversible covalent bond chemistry

10.1021/scimeetings.0c06796 ◽

2020 ◽

Author(s):

Josef Maier ◽

Eric Anslyn ◽

Arjun Kumar Menta

Keyword(s):

Covalent Bond ◽

Reversible Covalent

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Decision letter for "Intracellular accumulation of advanced glycation end‐products induces osteoblast apoptosis via endoplasmic reticulum stress"

10.1002/jbmr.4053/v1/decision1 ◽

2020 ◽

Keyword(s):

Endoplasmic Reticulum ◽

Endoplasmic Reticulum Stress ◽

Advanced Glycation End Products ◽

Intracellular Accumulation ◽

Advanced Glycation ◽

Glycation End Products ◽

End Products ◽

Osteoblast Apoptosis

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SENSITIVITY OF THE RAT DIAPHRAGM TO GROWTH HORMONE.

Acta Endocrinologica ◽

10.1530/acta.0.0540645 ◽

1967 ◽

Vol 54 (4) ◽

pp. 645-662 ◽

Cited By ~ 8

Author(s):

Å. Hjalmarson ◽

K. Ahrén

Keyword(s):

Growth Hormone ◽

Inhibitory Effect ◽

Rat Diaphragm ◽

Intracellular Accumulation ◽

Slight Effect ◽

Kidney Capsule ◽

Muscle Tissues ◽

Hypophysectomized Rats ◽

Isolated Rat

ABSTRACT The effect of growth hormone (GH) in vitro on the rate of intracellular accumulation of the non-utilizable amino acid α-aminoisobutyric acid (AIB) was studied in the intact rat diaphragm preparation. Bovine or ovine GH (25 μg/ml incubation medium) markedly stimulated the accumulation of AIB-14C by diaphragms from hypophysectomized rats, while there was no or only a very slight effect on diaphragms from normal rats. In diaphragms from rats with the pituitary gland autotransplanted to the kidney capsule GH in vitro stimulated the accumulation of AIB-14C significantly more than in diaphragms from normal rats but significantly less than in diaphragms from hypophysectomized rats. Injections of GH intramuscularly for 4 days to hypophysectomized rats made the diaphragms from these rats less sensitive or completely insensitive to GH in vitro. These results indicate strongly that the relative insensitivity to GH in vitro of diaphragms from normal rats is due to the fact that the muscle tissues from these rats has been exposed to the endogenously secreted GH. The results show that GH can influence the accumulation of AIB-14C in the isolated rat diaphragm in two different ways giving an acute or »stimulatory« effect and a late or »inhibitory« effect, and that it seems to be a time-relationship between these two effects of the hormone.

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