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Structure-Based Design of AG-946, a Pyruvate Kinase Activator.

Tao Liu¹, Anil K Padyana², Evan T Judd³

¹Ensem Therapeutics, 880 Winter St, Waltham, MA 02451, USA.

|December 18, 2023

Summary

Researchers designed AG-946, a potent activator for pyruvate kinase (PK) and its red blood cell form (PKR). This small molecule advanced to clinical trials for its potential in treating PK-related conditions.

Keywords:

AG-946 allosterism medicinal chemistry pharmacokinetics pyruvate kinase activator

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Area of Science:

Biochemistry
Enzymology
Drug Discovery

Background:

Pyruvate kinase (PK) is a key glycolytic enzyme regulating cell metabolism.
PKR is the red blood cell-specific isoform of PK.
Dysregulation of PK activity is linked to various metabolic disorders.

Purpose of the Study:

To describe the structure-based design of AG-946, a novel allosteric activator of PK isoforms.
To enhance potency and selectivity towards PKR compared to previous scaffolds.
To evaluate the efficacy of AG-946 in activating wild-type and mutant PK enzymes.

Main Methods:

Structure-based drug design utilizing pseudo-C2-symmetry principles.
Biochemical assays to determine activation potency (AC50) and kinetics.
Enzyme stability assays under denaturing conditions.

Main Results:

AG-946 demonstrated potent activation of wild-type PK (AC50 = 0.005 μM) and mutants K410E (AC50 = 0.0043 μM) and R510Q (AC50 = 0.0069 μM).
AG-946 exhibited a significantly longer activation half-time (>150-fold) compared to a reference compound.
AG-946 stabilized the catalytic activity of the unstable PKR R510Q mutant under denaturing conditions.

Conclusions:

AG-946 is a potent, orally available, small-molecule allosteric activator of wild-type and mutant PKR.
The drug's design improved potency and reduced off-target activity.
AG-946 has been advanced to human clinical trials for potential therapeutic applications.