Rapid Determination of Kinetic Constants for Slow-Binding Inhibitors and Inactivators of Human Histone Deacetylase 8

International Journal of Molecular Sciences, Journal Year: 2024, Volume and Issue: 25(11), P. 5593 - 5593

Published: May 21, 2024

The kinetics and mechanism of drug binding to its target are critical pharmacological efficacy. A high throughput (HTS) screen often results in hundreds hits, which usually only simple IC50 values determined during reconfirmation. However, kinetic parameters such as residence time for reversible inhibitors the kinact/KI ratio, is measure evaluating covalent inactivators, early predictive measures assess chances success hits clinic. Using promising cancer human histone deacetylase 8 an example, we present a robust method that calculates concentration-dependent apparent rate constants inhibition or inactivation HDAC8 from dose–response curves recorded after different pre-incubation times. With these data, hit compounds can be classified according their action, relevant calculated highly parallel fashion. with known modes action were correctly assigned mechanism, mechanisms some internal screening campaign newly determined. oxonitriles SVE04 SVE27 fast moderate time-constant 4.2 2.6 µM, respectively. compound TJ-19-24 SAH03 behave like slow two-step inactivators inhibitors, very low reverse isomerization rate.

Language: Английский

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Leveraging metabolism for better outcomes in heart failure

Cardiovascular Research, Journal Year: 2024, Volume and Issue: 120(15), P. 1835 - 1850

Published: Oct. 1, 2024

Whilst metabolic inflexibility and substrate constraint have been observed in heart failure for many years, their exact causal role remains controversial. In parallel, of our fundamental assumptions about cardiac fuel use are now being challenged like never before. For example, the emergence sodium-glucose cotransporter 2 inhibitor therapy as one four 'pillars' is causing a revisit metabolism key mechanism therapeutic target failure. Improvements field metabolomics will lead to far more granular understanding mechanisms underpinning normal abnormal human use, an appreciation drug action, novel strategies. Technological advances expanding biorepositories offer exciting opportunities elucidate aspects these mechanisms. Methodologic include comprehensive accurate quantitation such stable-isotope fluxomics, improved access arterio-venous blood samples across determine consumption energy conversion, high quality tissue biopsies, biochemical analytics, informatics. Pairing technologies with recent discoveries epigenetic regulation, mitochondrial dynamics, organ-microbiome crosstalk garner critical mechanistic insights this state-of-the-art review, we focus on new insights, eye emerging strategies Our synthesis be valuable diverse audience interest metabolism.

Language: Английский

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Two-hit mouse model of heart failure with preserved ejection fraction combining diet-induced obesity and renin-mediated hypertension

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: June 9, 2024

ABSTRACT Heart failure with preserved ejection fraction (HFpEF) is increasingly common but its pathogenesis poorly understood. The ability to assess genetic and pharmacologic interventions hampered by the lack of robust preclinical mouse models HFpEF. We have developed a novel “2-hit” model, which combines obesity insulin resistance chronic pressure overload recapitulate clinical features C57BL6/NJ mice fed high fat diet for >10 weeks were administered an AAV8-driven vector resulting in constitutive overexpression Renin1d . Control mice, HFD only, Renin only HFD-Renin (aka “HFpEF”) littermates underwent battery cardiac extracardiac phenotyping. demonstrated resistance, 2-3-fold increase circulating renin levels that resulted 30-40% left ventricular hypertrophy, systolic function, diastolic dysfunction indicated altered E/e’, IVRT, strain measurements; increased atrial mass; elevated natriuretic peptides; exercise intolerance. Transcriptomic metabolomic profiling myocardium upregulation pro-fibrotic pathways downregulation metabolic pathways, particular branched chain amino acid catabolism, similar findings human Treatment these sodium-glucose cotransporter 2 inhibitor empagliflozin, effective incompletely understood HFpEF therapy, improved tolerance, heart enlargement, homeostasis. model recapitulates key will enable studies dissecting contribution individual pathogenic drivers this complex syndrome. Addition platform allows orthogonal validity assessment interventions. NEW & NOTEWORTHY disease study due limited models. rigorously characterize new two-hit contributions synergy major drivers, hypertension diet-induced obesity. results are consistent reproducible two independent laboratories. This high-fidelity pre-clinical increases available, needed improve our understanding causes treatments

Language: Английский

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Ryanodine receptor stabilization therapy suppresses Ca2+- based arrhythmias in a novel model of metabolic HFpEF

Journal of Molecular and Cellular Cardiology, Journal Year: 2024, Volume and Issue: 195, P. 68 - 72

Published: July 23, 2024

Language: Английский

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Integrated systems biology identifies disruptions in mitochondrial function and metabolism as key contributors to heart failure with preserved ejection fraction (HFpEF)

bioRxiv (Cold Spring Harbor Laboratory), Journal Year: 2024, Volume and Issue: unknown

Published: Oct. 25, 2024

ABSTRACT Background Heart failure with preserved ejection fraction (HFpEF) accounts for ∼50% of HF cases, no effective treatments. The ZSF1-obese rat model recapitulates numerous clinical features HFpEF including hypertension, obesity, metabolic syndrome, exercise intolerance, and LV diastolic dysfunction. Here, we utilized a systems-biology approach to define the early transcriptional signatures gain mechanistic insight into pathways contributing development. Methods Male ZSF1-obese, ZSF1-lean hypertensive controls, WKY (wild-type) controls were compared at 14w age extensive physiological phenotyping tissue harvesting unbiased metabolomics, RNA-sequencing, assessment mitochondrial morphology function. Utilizing enabled distinction between hypertension-driven molecular changes pathology, versus hypertension + syndrome. Results rats displayed HFpEF. Comparison vs (i.e., hypertension-exclusive effects) revealed remodeling suggestive increased aerobic glycolysis, decreased β-oxidation, dysregulated purine pyrimidine metabolism few changes. worsened robust highlighted by upregulation inflammatory genes downregulation structure/function cellular processes. Integrated network analysis metabolomic RNAseq datasets nearly all catabolic energy production, manifesting in marked decrease energetic state reduced ATP/ADP, PCr/ATP). Cardiomyocyte ultrastructure area, size, cristae density, as well lipid droplet content hearts. Mitochondrial function was also impaired demonstrated substrate-mediated respiration calcium handling. Conclusions Collectively, integrated omics applied here provides framework uncover novel genes, metabolites, underlying HFpEF, an emphasis on potential target intervention.

Language: Английский

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Cardiometabolic effects of sacubitril/valsartan in a rat model of heart failure with preserved ejection fraction

Biochemical Pharmacology, Journal Year: 2024, Volume and Issue: unknown, P. 116571 - 116571

Published: Oct. 1, 2024

Language: Английский

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