Cell lines

ME-1 (human leukemia cell lines)

Preparation method

Limited solubility. General tips for obtaining a higher concentration: Please warm the tube at 37 ℃ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20℃ for several months.

Reaction Conditions

6 h

Applications

After 6 hours of treatment, AI-10-49 effectively dissociates RUNX1 from CBFb-SMMHC with 90% dissociation, by contrast, it has a mild effect on CBFb-RUNX1 association. The ChIP assays indicates that AI-10-49 treatment on ME-1 cells for 6 hours increases RUNX1 occupancy 8-, 2.2-, and 8-fold at the RUNX3, CSF1R, and CEBPA promoters.

Animal models

Transplanted mice with Cbfb+/MYH11:Ras+/G12D leukemic cells

Dosage form

200 mg/kg; 10 days

Applications

Mice treated with AI-10-49 survives markedly longer (median latency = 61 days) and transient AI-10-49 treatment also reduces leukemia spreading in vivo.

Other notes

Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal.

AI-10-49 is a selective inhibitor of CBFβ –SMMHC and RUNX1 interaction with a FRET IC50 value of 260nM.
AI-10-49 restores RUNX1 transcriptional activity, displays favorable pharmacokinetics, and delays leukemia progression in mice. Treatment of primary inv(16) AML patient with AI-10-49 triggers selective cell death. Direct inhibition of the oncogenic CBFβ-SMMHC fusion protein may be an effective therapeutic approach for inv(16) AML, and they provide support for transcription factor targeted therapy in other cancers. The stability of RUNX1, CBFb, and CBFb-SMMHC was not affected by AI-10-49 [1].
In 11 human leukemia cell lines, ME-1 cells were the only cell line highly sensitive to AI-10-49. In ME-1 cell, AI-10-49 has enhanced inhibitory activity on growth (IC50 = 0.6 mM) compared with the parent protonated bivalent compound AI-4-83 (IC50 of ~3 mM). In normal human bone marrow cells, AI-10-49 showed negligible activity (IC50 > 25 mM), which indicated a robust potential therapeutic window. In chromatin-immunoprecipitation (ChIP) assays, treatment of ME-1 cells for 6 hours with AI-10-49 increased RUNX1 occupancy 8-, 2.2-, and 8-fold at the RUNX3, CSF1R, and CEBPA promoters, respectively.
After treatment with AI-10-49 to leukemia mice significantly prolonged their lives, and after 7 days of administration of AI-10-49, we observe no evidence of toxicity [1].
Reference:
1.Illendula A, Pulikkan JA, Zong H et al. A small-molecule inhibitor of the aberrant transcription factor CBFβ-SMMHC delays leukemia in mice. Science. 2015 Feb 13;347(6223):779-784.