Development of the novel ACLY inhibitor 326E as a promising treatment for hypercholesterolemia

Area of Science:

• Biochemistry
• Pharmacology
• Cardiovascular Science

Background:

• Hepatic cholesterol accumulation contributes to hypercholesterolemia, atherosclerosis, and cardiovascular disease (CVD).
• ATP-citrate lyase (ACLY) is a critical enzyme linking mitochondrial oxidative phosphorylation to cytosolic *de novo* lipogenesis.
• ACLY inhibition is a potential therapeutic strategy for managing cholesterol levels.

Purpose of the Study:

• To develop and characterize a novel small molecule inhibitor of ACLY.
• To evaluate the efficacy of the novel inhibitor, 326E, in reducing *de novo* lipogenesis and improving lipid profiles.
• To assess the therapeutic potential of 326E in preclinical models of hypercholesterolemia and atherosclerosis.

Main Methods:

• Development of 326E, an enedioic acid-based ACLY inhibitor.
• In vitro assessment of 326E-CoA's inhibitory activity against ACLY (IC50 determination).
• In vitro and in vivo studies to evaluate the effects of 326E on lipogenesis and cholesterol efflux.
• Pharmacokinetic analysis of 326E after oral administration.
• Chronic treatment studies in hyperlipidemic animal models (hamsters, rhesus monkeys) and atherosclerosis models (ApoE-/- mice), comparing 326E with bempedoic acid (BA).

Main Results:

• 326E-CoA demonstrated potent in vitro inhibition of ACLY (IC50 = 5.31 ± 1.2 μmol/L).
• 326E treatment reduced *de novo* lipogenesis and enhanced cholesterol efflux in vitro and in vivo.
• 326E exhibited favorable pharmacokinetics with higher blood exposure compared to bempedoic acid.
• Chronic oral administration of 326E significantly improved hyperlipidemia in hamsters and rhesus monkeys.
• 326E demonstrated superior efficacy in preventing atherosclerosis in ApoE-/- mice compared to bempedoic acid.

Conclusions:

• 326E is a novel, orally bioavailable ACLY inhibitor with potent lipid-lowering effects.
• 326E effectively reduces *de novo* cholesterol synthesis and promotes cholesterol efflux.
• 326E represents a promising therapeutic strategy for managing hypercholesterolemia and preventing atherosclerosis, outperforming the approved drug bempedoic acid in preclinical models.