Selective and Rapid Cell-Permeable Inhibitor of Human Caspase-3

Solania, Angelo; González-Páez, Gonzalo E.; Wolan, Dennis W.

doi:10.1021/acschembio.9b00564

Cited by 25 publications

(23 citation statements)

References 34 publications

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“…(a) Drug-likeness of cmpd 7. Solubility in PBS [31][32] , caspase activity 33 , microsomal stability and cellular toxicity were measured as described 34 , LogP was predicted with ChemDraw Ultra 17.1. (b) cmpd 7 prevents S. pyogenes WT GAS5448 from neutrophil killing by the inhibition of SpeB; however, no effect is observed on the SpeB strain.…”

Section: Figurementioning

confidence: 99%

Sulfur(VI) Fluoride Exchange (SuFEx)-Enabled High-Throughput Medicinal Chemistry

Kitamura¹,

Zheng²,

Woehl³

et al. 2020

J. Am. Chem. Soc.

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138

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Optimization of small-molecule probes or drugs is a lengthy, challenging and resource-intensive process. Lack of automation and reliance on skilled medicinal chemists is cumbersome in both academic and industrial settings. Here, we demonstrate a high-throughput hit-to-lead process based on the biocompatible SuFEx click chemistry. A modest high-throughput screening hit against a bacterial cysteine protease SpeB was modified with a SuFExable iminosulfur oxydifluoride [RN=S(O)F2] motif, rapidly diversified into 460 analogs in overnight reactions, and the products directly screened to yield drug-like inhibitors with 300-fold higher potency. We showed that the improved molecule is drug-like and biologically active in a bacteria-host coculture. Since these reactions can be performed on a picomole scale to conserve reagents, we anticipate our methodology can accelerate the development of robust biological probes and drug candidates. The introduction of high-throughput screening (HTS) robotics, liquid handler systems, and assay miniaturization have revolutionized screening of bioactive molecules. Relatively inexpensive HTS processes are now routinely used in cell-based and in vitro assays against biomedically relevant targets. Nevertheless, compound optimization is typically necessary to improve target specificity, potency, and stability. Lead optimization relies heavily on medicinal chemists, and extensive time and labor costs remain significant hurdles for probe and drug development. Click chemistry has found broad applications in materials chemistry, chemical biology, and drug development since the concept was first introduced in 1999 1-2. The sulfur(VI) fluoride exchange (SuFEx) represents the most recent set of ideal click chemistry transformations 3. Specifically, aryl fluorosulfates (ArOSO2F) and iminosulfur oxydifluorides (RN=S(O)F2) are readily synthesized using two connective oxyfluoride gases, sulfuryl fluoride (SO2F2) and thionyl tetrafluoride (O=SF4), respectively 4. These two S VI −F motifs have been successfully used as connective linkers in polymer synthesis and for construction of various functional molecules 5-7. Sulfonyl fluoride (RSO2F) and aryl fluorosulfate moieties have been successfully introduced into bioactive molecules in chemical biology and drug discovery 8-11 , especially as covalently binding warheads 12. However, the potential of SuFEx to unite diverse modules using an O=SF4 hub has not been explored in medicinal chemistry. While the copper(I)catalyzed azide-alkyne cycloaddition (CuAAC) reaction has been used in proof-of-concept studies on lead Statistical analysis was performed using one-way ANOVA with Dunnett's multiple comparisons test, *≤ 0.05. Supporting Information Additional texts, figures, and tables are provided. AUTHOR INFORMATION # Authors contributed equally.

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Section: Figurementioning

confidence: 99%

Sulfur(VI) Fluoride Exchange (SuFEx)-Enabled High-Throughput Medicinal Chemistry

Kitamura¹,

Zheng²,

Woehl³

et al. 2020

J. Am. Chem. Soc.

Self Cite

138

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“…Caspase-3 and -7 are highly homologous and share 54% structural identity and 77% active site identity, explaining in part why caspase-3 probes also often target caspase-7 [ 15 ]. Both caspase-3 and -7 preferentially recognize and cleave the DxxD motif, a tetrapeptide with two aspartic acid residues, a motif at the basis of many inhibitors and substrates [ 15 , 16 , 17 , 18 ]. Each caspase preferentially cleaves a certain substrate.…”

Section: Challenges Of Caspase-3 Probesmentioning

confidence: 99%

“…On the contrary, ABPs contain an electrophilic warhead for covalent binding to the target. The added steric hindrance and electronics of an electrophilic warhead moiety of ABPs can also confer additional selectivity to the probes towards the desired target by interacting with the prime side of the enzyme when combined with a non-prime side directed peptide, as described by Vickers et al [ 17 , 45 ]. Common electrophilic warheads used in caspase-3 ABPs are acyloxymethyl ketone (AOMK) and fluoromethyl ketone (FMK) groups, as they are specific for cysteine proteases [ 19 ].…”

Section: Peptide-based Caspase-3 Probesmentioning

confidence: 99%

“…This promising inhibitor turned out to have a major limitation: its binding kinetics proved to be too slow for in vivo use. Therefore, a second generation of fast-binding selective caspase-3 inhibitor was designed, termed Ac-ATS010-KE, with the potential to be used as a potent, highly selective, fast-binding radiotracer [ 17 ].…”

Section: Peptide-based Caspase-3 Probesmentioning

confidence: 99%

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Molecular Imaging of Apoptosis: The Case of Caspase-3 Radiotracers

Beroske

Wyngaert

Stroobants

et al. 2021

IJMS

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The molecular imaging of apoptosis remains an important method for the diagnosis and monitoring of the progression of certain diseases and the evaluation of the efficacy of anticancer apoptosis-inducing therapies. Among the multiple biomarkers involved in apoptosis, activated caspase-3 is an attractive target, as it is the most abundant of the executioner caspases. Nuclear imaging is a good candidate, as it combines a high depth of tissue penetration and high sensitivity, features necessary to detect small changes in levels of apoptosis. However, designing a caspase-3 radiotracer comes with challenges, such as selectivity, cell permeability and transient caspase-3 activation. In this review, we discuss the different caspase-3 radiotracers for the imaging of apoptosis together with the challenges of the translation of various apoptosis-imaging strategies in clinical trials.

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“…We therefore carried out insilico knockdown experiments of eCaspase, as its activation is essential for the final steps of apoptosis execution (Mehal, Inayat and Flavell, 2006). In addition, effector caspase inhibitors are readily available for laboratory use (Perry et al, 1997;Solania, González-Paéz and Wolan, 2019). We hypothesized that each execution mode would exhibit different execution mechanisms when eCaspase was knocked down by 50%.…”

Section: Signal Execution Modes Respond Differently To Ecaspase Pertumentioning

confidence: 99%

Emergent signal execution modes in biochemical reaction networks calibrated to experimental data

Ortega

Wilson²,

Pino³

et al. 2021

Preprint

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Mathematical models of biomolecular networks are commonly used to study mechanisms of cellular processes, but their usefulness is often questioned due to parameter uncertainty. Here, we employ Bayesian parameter inference and dynamic network analysis to study dominant reaction fluxes in models of extrinsic apoptosis. Although a simplified model yields thousands of parameter vectors with equally good fits to data, execution modes based on reaction fluxes clusters to three dominant execution modes. A larger model with increased parameter uncertainty shows that signal flow is constrained to eleven execution modes that use 53 out of 2067 possible signal subnetworks. Each execution mode exhibits different behaviors to in silico perturbations, due to different signal execution mechanisms. Machine learning identifies informative parameters to guide experimental validation. Our work introduces a probability-based paradigm of signaling mechanisms, highlights systems-level interactions that modulate signal flow, and provides a methodology to understand mechanistic model predictions with uncertain parameters.

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Selective and Rapid Cell-Permeable Inhibitor of Human Caspase-3

Cited by 25 publications

References 34 publications

Sulfur(VI) Fluoride Exchange (SuFEx)-Enabled High-Throughput Medicinal Chemistry

Sulfur(VI) Fluoride Exchange (SuFEx)-Enabled High-Throughput Medicinal Chemistry

Molecular Imaging of Apoptosis: The Case of Caspase-3 Radiotracers

Emergent signal execution modes in biochemical reaction networks calibrated to experimental data

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