MD-224

Protein target name:

MDM2

Mechanism of action:

DegraderPROTAC

Recommended Concentration for use in cells:

up to 30 nM

Probe Availability

MedChemExpress

Probe Information

Target and Probe Information

Target Details

Target class:

HDMX; LSKB; hdm2; ACTFS

Probe Details

Chemical Probes Portal ID:

CPP2130

Probe alias:

cpd14, PROTAC

PubChem CID:

131986956

SMILES string:

FC1=C(Cl)C=CC=C1[C@@H]2[C@@]3(C(C=CC(Cl)=C4)=C4NC3=O)C5(CCCCC5)N[C@H]2C(NC6=CC=C(C(NCCCC#CC7=C(CN(C8C(NC(CC8)=O)=O)C9=O)C9=CC=C7)=O)C=C6)=O

InChi Key:

ZLGNYFOIDAVMHY-MPKOGUQCSA-N

Orthogonal probe:

MG-277

Physico-chemical properties

NMR:

1H NMR (400 MHz, MeOH-d4): δ 7.83–7.62 (m, 4H), 7.56 (dd, J = 8.3, 2.5 Hz, 1H), 7.54–7.47 (m, 2H), 7.47–7.32 (m, 3H), 7.19 (ddd, J = 8.9, 7.6, 3.3 Hz, 1H), 7.12 (dd, J = 8.2, 2.0 Hz, 1H), 6.80 (d, J = 1.9 Hz, 1H), 5.30 (d, J = 10.8 Hz, 1H), 5.13 (ddd, J = 13.0, 7.4, 5.2 Hz, 1H), 4.98 (d, J = 10.9 Hz, 1H), 4.49–4.25 (m, 2H), 3.57 (t, J = 6.6 Hz, 2H), 2.97–2.83 (m, 2H), 2.83–2.73 (m, 1H), 2.68–2.42 (m, 3H), 2.28–2.13 (m, 2H), 2.13–1.84 (m, 5H), 1.84–1.68 (m, 2H), 1.57 (ddd, J = 16.3, 6.2, 3.0 Hz, 1H), 1.34–1.19 (m, 2H)

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

Ki, IC50

0.16 nM (Ki), 4.4 (IC50) Mi-1061 binding MDM2

Potency assay:

Fluorescence Polarization-Based (FP-Based) Protein Binding Assay

Structure-activity relationship:

partial, limited to linker

In cell validation

On target activity in cells

Potency in cells:

IC50

1.5 nM

Potency assay (cells):

cell growth inhibition assay, western blots assessing MDM2 degradation and p53 activation; Cell Growth Inhibition, Activity of MD-224 in a Panel of Human AML or ALL Cell Lines MOLM-13 7.3 nM, OL-14, 10.5 nM, SIG-M5 19.8 nM, ML-2 4.4 nM, OCL-AML-5 33.1 nM, mono-Man-6 >10 uM, KG-1 >10 uM. MD-224 is highly selective over leukemia cell lines with mutated p53.

Phenotypic Control Experiment:

Lenalidomide effectively blocks MDM2 degradation and accumulation of p53 protein induced by MD-224 as shown in western blotting analysis.

Off target activity in cells

Potency assay, off target (cells):

Cell Growth Inhibition Activity

Probe Selectivity in Cell:

Cell Growth Inhibition Activity of MD-224 in a Panel of Human AML or ALL Cell Lines MOLM-13 7.3 nM, OL-14, 10.5 nM, SIG-M5 19.8 nM, ML-2 4.4 nM, OCL-AML-5 33.1 nM, mono-Man-6 >10 uM, KG-1 >10 uM. MD-224 are highly selective over leukemia cell lines with mutated p53.

In vivo validation

Organism:

Mice bearing RS4;11 xenograft tumor

Dose:

25 mg/Kg IV100 mg/Kg PO

Route of delivery:

OralIntravenous

Primary Reference

Reference (doi):

10.1021/acs.jmedchem.8b00909

Reference (PMID):

30525597

Under Review

This probe is in the process of SAB review. Please continue to check back for new reviews and commentary.

SAB Rating

Rating for use in Cells

2 review(s)

Rating for use in Model Organisms

2 review(s)

SAB Members

SAB Comments

For degrader molecules, selectivity is often assessed through proteomic studies that explore the relative stability of the presumed target (here MDM2) compared to other possible targets. Although that standard has not, at present, been met for MD-224, other ways of rigorously addressing target engagement and selectivity have been reported.

Feb 9 2021 - 8:42pm

MG-277 has also been described as an optimized MDM2 degrader modified from MD-222. Ref: https://doi.org/10.1016/j.apsb.2020.01.003

Feb 18 2021 - 8:49pm

MD-224

Probe Availability

Probe Information

Target Details

HDMX; LSKB; hdm2; ACTFS

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?

Search form

MD-224

Probe Availability

Probe Information

Target Details

HDMX; LSKB; hdm2; ACTFS

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?