99. Arsonic and stibonic acids derived from quinoline and acridine. Part I

Individual roles and overlapping functionalities of 12 human caspases during apoptosis and other cellular processes remain poorly resolved primarily due to a lack of chemical tools. Here we present a new selective caspase-3 inhibitor, termed Ac-ATS010-KE, with rapid and irreversible binding kinetics. Relative to previously designed caspase-3-selective molecules that have tremendously abated inhibitory rates and thus limited use in biological settings, the improved kinetics of Ac-ATS010-KE permits its use in a cell-based capacity. We demonstrate that Ac-ATS010-KE prevents apoptosis with comparable efficacy to the general caspase inhibitor Ac-DEVD-KE and surprisingly does so without side-chain methylation. This observation is in contrast to the well-established peptide modification strategy typically employed for improving cellular permeability. Ac-ATS010-KE protects against extrinsic apoptosis, which demonstrates the utility of a thiophene carboxylate leaving group in biological settings, challenges the requisite neutralization of free carboxylic acids to improve cell permeability, and provides a toollike compound to interrogate the role of caspase-3 in a variety of cellular processes.

show abstract

“…Rfree is Rcryst with 5.0% test set structure factors. 6) Cross-validated (CV) Luzzati coordinate errors.…”

Section: S8mentioning

confidence: 99%

“…The following supplemental protocols are based on previously reported procedures used to generate cysteine protease inhibitors. [4][5][6]…”

Section: General Synthetic Scheme For Fmoc-asp(otbu)-ke Semicarbazidmentioning

confidence: 99%

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

2019

View full text Add to dashboard Cite

show abstract

“…[9][10][11] The sensitivity of this mass spectrometric method is further enhanced when nanoscale electrospray ionization is used. 12 This technique turns solutions of analytes into a fine mist; further evaporation of solvent from these minute charged droplets reduces their size, thereby increasing their surface charge density.…”

Section: Resultsmentioning

confidence: 99%

“…2-AMAC was synthesised using a modified procedure of Bamet et al 9 Derivatisation of glycans with 2-AMAC was carried out using a procedure outlined by us previously. 8 The pure derivatives were isolated after passage through an Oasis cartridge (Waters, UK), followed by lyophilisation.…”

Section: Methodsmentioning

confidence: 99%

Profiling of 2-aminoacridone derivatised glycans by electrospray ionization mass spectrometry

Charlwood

Langridge

Tolson

et al. 1999

Rapid Commun. Mass Spectrom.

View full text Add to dashboard Cite

Three different types of N-glycans, high-mannose, complex and hybrid, have been derivatised with 2-aminoacridone and analysed by electrospray mass spectrometry and tandem mass spectrometry (MS/MS) using a hybrid quadrupole orthogonal acceleration time-of-flight mass spectrometer, fitted with a nanoflow electrospray ion source. Relatively simple MS/MS fragmentation patterns have been observed for each type of glycan, allowing rapid elucidation of the order in which the monosaccharide residues making up these glycans are linked to one another.

show abstract

“…Detailed protocols for the synthesis of the semicarbazide resin, AMAC-substrates and inhibitors are outlined in the Supporting Information available [insert hyperlink]. [23][24][25] Preparation of recombinant caspases. Casp-3, casp-6, casp-7, casp-8, casp-9 and casp-10 were expressed and purified as previously described.…”

Section: Methodsmentioning

confidence: 99%

A selective and rapid cell-permeable inhibitor of human caspase-3

Solania

González-Páez

Wolan

2019

Preprint

View full text Add to dashboard Cite

The individual roles and overlapping functionalities the twelve human caspases have during apoptosis and other cellular processes remain poorly resolved primarily due to a lack of chemical tools. Here we present a new selective caspase-3 inhibitor, termed Ac-ATS010-KE, with rapid and irreversible binding kinetics. Relative to previously designed caspase-3-selective molecules that have tremendously abated inhibitory rates and thus limited use in biological settings, the improved kinetics of Ac-ATS010-KE permit its use in a cell-based capacity. We demonstrate that Ac-ATS010-KE prevents apoptosis with comparable efficacy to the general caspase inhibitor Ac-DEVD-KE and surprisingly does so without side-chain methylation. This observation is in contrast to the well-established peptide modification strategy typically employed for improving cellular permeability. Ac-ATS010-KE protects against extrinsic apoptosis, which demonstrates the utility of a thiophene carboxylate leaving group in biological settings, challenges the requisite neutralization of free carboxylic acids to improve cell permeability, and provides a tool-like compound to interrogate the role of caspase-3 in a variety of cellular processes.Cell Culture. Jurkat A3 cells were cultured as described by ATCC using 10% FBS and pen/strep/glutamine at 37 °C with 5% CO2. For all experiments, Jurkat cells were grown to near confluence, harvested and resuspended in fresh media to the indicated density. Apoptosis was induced using 10 ng/mL MegaFasL (AdipoGen® Life Sciences).Apoptotic protection assays. Cells were plated into sterile Nunc Edge 96-well tissue culture-treated plates at a density of 10,000 cells/well and preincubated (or chased) with 20 µM of the indicated inhibitor or DMSO vehicle for the indicated time before (or after) apoptotic induction. 3 h after induction, cellular viability was measured using CellTiter-Glo® on a PerkinElmer EnVision plate reader.

show abstract

99. Arsonic and stibonic acids derived from quinoline and acridine. Part I

Cited by 11 publications

References 0 publications

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

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

Profiling of 2-aminoacridone derivatised glycans by electrospray ionization mass spectrometry

A selective and rapid cell-permeable inhibitor of human caspase-3

Contact Info

Product

Resources

About