Snh-119014, a novel pyruvate kinase activator, enhances ATP production and reduces oxidative stress in erythroid cells from patients with β-thalassemia major

Abstract

Abstract Introduction: Oxidative stress contributes significantly to ineffective erythropoiesis and reduced red blood cell (RBC) survival in β-thalassemia major (β-TM). Enhancing adenosine triphosphate (ATP) production through pyruvate kinase (PK) activation may counteract metabolic stress. SNH-119014 is a novel allosteric PK activator with structural similarity to mitapivat (AG-348), a first-in-class agent known to ameliorate anemia in thalassemia. This study compared the pharmacodynamic effects of SNH-119014 and AG-348 in ex vivo models using erythroid cells from β-TM patients. Methods: PK activity was evaluated using recombinant human PKLR isoforms with graded concentrations of SNH-119014 or AG-348. RBCs were isolated from 30 β-TM patients and 30 healthy controls, treated ex vivo with 10 μM of either compound for 4 hours, and assessed for ATP levels. Additionally, CD34+ cells from β-TM bone marrow were cultured and differentiated into erythroid precursors. Treatments (SNH-119014 at 5 μM, AG-348 at 5 μM, or vehicle) were applied, followed by assessment of ATP levels, reactive oxygen species (ROS), reduced/oxidized glutathione, and metabolomics profiling. Results: SNH-119014 and AG-348 both increased PK activity, with AC 50 values of 10.42 nM and 8.08 nM, respectively. In RBCs, SNH-119014 increased ATP by a mean of 129% vs. 131% for AG-348 (p=0.0928). In erythroid precursors, ATP increased by 119% vs. 120% (p=0.5738), respectively. ROS levels decreased by 30.8% with SNH-119014 and 26.1% with AG-348. The glutathione /glutathione disulfide (GSH/GSSG) ratio improved with both agents. Metabolomics revealed 181 differential metabolites between β-TM and controls, with glycolysis/gluconeogenesis pathways significantly altered by both treatments compared to the vehicle control. Conclusion: SNH-119014 activates PK, increases ATP production, and alleviates oxidative stress in erythroid cells from β-TM patients. Its comparable efficacy to AG-348 in ex vivo systems supports SNH-119014 as a promising candidate for therapeutic development in thalassemia.