AI-10-49: CBFβ-SMMHC Inhibitor Workflow

Setup and principle overview

AI-10-49 is a small-molecule CBFβ-SMMHC inhibitor designed to disrupt the interaction between the leukemia-associated fusion protein CBFβ-SMMHC and the RUNX1 Runt domain. That distinction matters experimentally: rather than treating growth inhibition as an isolated endpoint, investigators can ask whether compound exposure first releases RUNX1, restores its chromatin occupancy, and then changes the transcriptional program that sustains leukemia cells.

The most relevant disease context is inv(16) acute myeloid leukemia, in which CBFB is fused to MYH11. CBFβ-SMMHC binds RUNX1 with abnormal avidity and can interfere with normal hematopoietic transcription. AI-10-49 therefore provides a useful perturbation for testing fusion-protein dependency in ME-1 cells, primary inv(16) samples, engineered systems, and carefully selected in vivo leukemia models.

According to the AI-10-49, a selective leukemia oncoprotein CBFβ-SMMHC inhibitor product information, the compound has an IC50 of 0.26 μM for the targeted interaction. In ME-1 human leukemia cells, treatment produced approximately 90% dissociation of RUNX1 from CBFβ-SMMHC after 6 hours. The same product documentation reports increased RUNX1 occupancy at the RUNX3, CSF1R, and CEBPA promoters after treatment, supporting a target-engagement-to-transcription workflow rather than a viability-only screen.

AI-10-49 is supplied as a research reagent only. It should not be interpreted as a diagnostic or medical product, and all cell, animal, and primary-sample work requires appropriate institutional approvals.

Protocol Parameters

• Stock preparation: Prepare a 10 mM DMSO stock at 20–25°C; this corresponds to approximately 6.61 mg/mL for a molecular weight of 660.52, below the reported solubility of at least 16.53 mg/mL. Warm briefly and sonicate for 1–5 minutes if visible material remains.
• Cell-based dose response: Seed 1 × 10⁴ to 3 × 10⁴ cells in 100 μL per well, apply 0.01–3 μM AI-10-49 across a logarithmic concentration series, and incubate for 48–72 hours with a matched DMSO control maintained at 0.1% or less.
• Early target engagement: Collect ME-1 cells after 2, 6, and ’heure? No.