RG7112

Protein target name:

MDM2

Mechanism of action:

Antagonist

Recommended Concentration for use in cells:

Up to 1 uM

Probe Availability

MedChem Express (939981-39-2)

Probe Information

Target and Probe Information

Target Details

Target class:

ACTFS, HDMX, hdm2

Probe Details

Chemical Probes Portal ID:

CPP1589

Probe alias:

UNII-Q8MI0X869M; RG7112; RO-5045337

PubChem CID:

57406853

ChEMBL ID:

CHEMBL2386346

SMILES string:

CCOC1=C(C=CC(=C1)C(C)(C)C)C2=N[C@@]([C@@](N2C(=O)N3CCN(CC3)CCCS(=O)(=O)C)(C)C4=CC=C(C=C4)Cl)(C)C5=CC=C(C=C5)Cl

InChi Key:

QBGKPEROWUKSBK-QPPIDDCLSA-N

Orthogonal probe:

RO5353AMG232AM-6761

Physico-chemical properties

NMR:

1H NMR (400 MHz, DMSO-d6) δ 1.36 (t, J = 6.90 Hz, 3H), 1.42 (s, 10H), 1.66 (s, 3H), 1.71 - 1.82 (m, 2H), 1.95 (s, 3H), 2.11 - 2.20 (m, 2H), 2.26 (d, J = 10.79 Hz, 1H), 2.49 (d, J = 10.29 Hz, 1H), 2.85 - 3.12 (m, 9H), 3.21 (d, J = 12.30 Hz, 1 H), 3.37 (s, 1H), 3.95 - 4.09 (m, 1H), 4.30 (br. s, 1H), 6.97 - 7.23 (m, 10H), 7.65 (d, J = 7.78 Hz, 1H).

13C NMR (100 MHz, DMSO-d6) δ 14.2, 19.3, 20.2, 23.8, 31.1, 34.9, 41.7, 44.7, 51.0, 51.2, 51.5, 55.4, 63.6, 75.0, 76.2, 108.4, 117.0, 117.1, 126.4, 127.0, 128.6, 130.4, 130.8, 131.4, 141.2, 141.9, 154.8, 155.7, 156.8, 157.3.

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

IC50

0.018 uM

Potency assay:

Homogenous time-resolved fluoresence (HTRF) assay with N-terminal domain from MDM2 and a peptide from the p53 binding site.

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

4IPF

Structure-activity relationship:

Yes, see ACS Med Chem paper

In cell validation

On target activity in cells

Potency in cells:

IC50

0.4 uM

Potency assay (cells):

MTT assay, also comparing selectivity of the compound for p53 wild-type (HCT-116, SJSA-1 and RKO cells) and p53 mutant (MDA-MB-435 and SW480) cells: 33.4-fold selective for the wild-type cells. In a broader panels of cells (7 mutant and 15 wild type lines), the compound was ~14-fold selective for wild-type cell lines.

Target engagement assay (cells):

In cultured MDM2-amplified GBM cells, RG7112 restored p53 expression prior to the induction of cell death.

In vivo validation

Organism:

Nude mouse

Dose:

50 mg/kg

Route of delivery:

Oral

Plasma half life:

8.8 h

Ratio of probe in organ to plasma (O:P):

1.9 (Brain:plasma)

Cmax:

15.5 ug/mL

Organ of Interest

Organ (O):

Subcutaneous xenografts (GBM and osteosarcoma)Brain engraftment (GBM)

Target engagement assay:

In subcutaneous GBM xenografts, RG7112 increase expression of MDM2, p53 and p21.

Primary Reference

Reference (doi):

10.1021/ml4000657 10.1158/1078-0432.CCR-15-1015 10.1158/0008-5472.CAN-12-2807

Reference (PMID):

24900694 26482041 23400593

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

Public notes can be found at http://www.citeulike.org/user/cdsouthan/article/14369611. Note it will also go live in the Guide to Pharmacology by August 2017 (Ligand 9699).

Configuration of the active enantiomer (4S,5R: CID 57406853) is critical since it is ~ 200-fold more potent compared with (4R,5S: CID 74889557). There are many vendors included in the PubChem entry, but users are advised to ensure (even by direct QC) that the bottle in fact contains the 4R,5S isomer. Note that Compound 2l with an IC50=2,163 nM could be used as a low activity (quasi negative) control, and Nutlin 3a would also be a useful internal control.

There are 27 matches in PubMed https://www.ncbi.nlm.nih.gov/pubmed/RG7112 so other publications can be consulted to get a full picture before using the probe. I could not find data demonstrating in vitro specificity against paralogues. While the sequence similarity with O15151 MDM4_HUMAN is low at 33% over 490 residues, there are reports of complexing between MDM2 and MDM4.

Jun 28 2017 - 12:25pm

RG7112

Probe Availability

Probe Information

Target Details

ACTFS, HDMX, hdm2

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

RG7112

Probe Availability

Probe Information

Target Details

ACTFS, HDMX, hdm2

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?