RO5353

Protein target name:

MDM2

Mechanism of action:

Antagonist

Probe Information

Target and Probe Information

Target Details

Target class:

ACTFS, HDMX, hdm5

Probe Details

Chemical Probes Portal ID:

CPP1596

PubChem CID:

56649312

ChEMBL ID:

CHEMBL3109040

SMILES string:

CC(C)(C)C[C@H]1[C@@]2([C@H]([C@@H](N1)C(=O)NC3=C(C=C(C=C3)C(=O)N)OC)C4=C(C(=CC=C4)Cl)F)C5=C(C=C(S5)Cl)NC2=O

InChi Key:

UOVSZRTTWLJPHE-HNXAPSNLSA-N

Orthogonal probe:

AMG232AM-6761MI-77301

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

IC50

7 nM

Potency assay:

HTRF assay

PDB ID for probe-target interaction (3D structure):

4LWV

Structure-activity relationship:

Yes, see ACS Med Chem Lett paper

In cell validation

On target activity in cells

Potency in cells:

IC50

0.007 uM

Potency assay (cells):

Antiproliferative activity (MTT assay) averaged across multiple p53 wild-type cells (SJSA-1, HCT-116, RKO). RO5353 is 1023-fold selective for p53 wild type compared with p53 mutant cells.

In vivo validation

Organism:

Male C57 mice, Female nude mice

Dose:

10 mg/kg (oral)

Route of delivery:

Oral

Cmax:

1.3 ug/mL (oral)

Organ of Interest

Organ (O):

SJSA-1 xenografts (nude mice, 3-20 mg/kg)

Primary Reference

Reference (doi):

10.1021/ml400359z

Reference (PMID):

24900784

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

RO5353 is an excellent probe to study the in vitro and in vivo biology of p53-MDM2 inhibition. It is potent, has excellent cellular selectivity and robust in-vivo efficacy at a dose (10 mg/kg) that is well-tolerated in mice. Although characterization of the activities of this compound against other proteins (e.g., kinase or other protein family selectivity panels) has not been described, the structural complexity of the probe and the fact that there is rich SAR around this compound reduces the likelihood that off-target activities will be significant, as compared to its potent 7 nM inhibitory activity against p53-MDM2. Moreover, the mode of action as an inhibitor of a highly specific protein-protein interaction also renders confounding off-target activity in intact biological systems fairly unlikely.

Jul 27 2017 - 11:37am

RO5353 is potent against MDM2 (7 nM), showing equipotency (7 nM) in antiproliferative assays with multiple WT p53 cells lines (i.e., the expected on-target pharmacology). The average antiproliferative activity in two mutant p53 cell lines was ~7 uM, suggesting a concentration at which off-target pharmacology can be expected. The individual IC50's for those two cell lines were not reported, so a top concentration of 7 uM should probably not be approached. It is unknown if RO5353 has any off-target activities <7 uM. RO5353 has moderate PK and achieved complete tumor regression of a human tumor xenograft in male mice at 10 mg/kg oral (with statistically signficant reduction in tumor burden at 3 mg/kg) with chronic dosing over 3-4 weeks. Ortholog potency was not reported, so it is unclear what dose is necessary to study mouse MDM2 pharmacology in vivo. Significant hematologic toxicities were observed in humans with a related molecule, RG7112, which should be considered if chronically dosing animals. Two related compounds, RO8994 and RO2468, show tumor stasis or regression at 3 mg/kg, likely owing to improved DNAUC, so if material supply is a concern for in vivo studies, those two molecules could be considered. The overall balance of reasonable PK, cell potency and the window over general cytotoxicity puts RO5353 as the likely best tool for MDM2.

Aug 3 2017 - 3:40pm

RO5353

Probe Information

Target Details

ACTFS, HDMX, hdm5

Probe Details

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

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RO5353

Probe Information

Target Details

ACTFS, HDMX, hdm5

Probe Details

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?