AM-6761

Reagent Authentication Certificate:

Chemical Probes Portal AM-6761.pdf

Protein target name:

MDM2

Mechanism of action:

Antagonist

Recommended Concentration for use in cells:

20 nM - 1 uM

Probe Information

Target and Probe Information

Target Details

Target class:

ACTFS, HDMX, hdm5

Probe Details

Chemical Probes Portal ID:

CPP1594

Probe alias:

cpd 4

PubChem CID:

73386675

ChEMBL ID:

CHEMBL3236357

SMILES string:

C[C@@]1(C[C@@H]([C@H](N(C1=O)[C@H](CS(=O)(=O)C(C)(C)C)C2CC2)C3=CC(=C(C=C3)Cl)F)C4=CC(=CC=C4)Cl)CC5=NC=C(S5)CC(=O)O

InChi Key:

HNLHOKSDTBAERS-CWIPYCOQSA-N

Orthogonal probe:

RO5353AMG232MI-77301

Physico-chemical properties

NMR:

1H NMR (500 MHz, CDCl3) δ 7.78 (br. s., 1H), 6.96–7.17 (m, 5H), 6.89 (d, J = 6.60 Hz, 1H), 4.96 (d, J = 10.51 Hz, 1H), 4.34 (t, J = 11.86 Hz, 2H), 3.96 (s, 2H), 3.67–3.82 (m, 2H), 3.61 (br. s., 1H), 2.91 (d, J = 12.96 Hz, 1H), 2.69 (br. s., 1H), 2.32 (t, J = 13.94 Hz, 1H), 1.80–1.97 (m, 2H), 1.44 (s, 9H), 1.26 (s, 3H), 0.37 (d, J = 4.16 Hz, 2H), −0.29 (br. s., 1H), −0.98 (br. s., 1H).

13C NMR (150 MHz, (CD3)2SO) δ 175.4, 171.9, 165.8, 158.9, 158.6, 144.2, 141.9, 141.6, 133.3, 131.9, 130.9, 127.7, 127.4, 126.6, 119.6, 119.5, 118.7, 116.7, 114.8, 69.9, 59.9, 58.7, 58.6, 44.4, 43.9, 43.6, 41.2, 38.2, 33.4, 26.5, 14.6, 7.3, 4.6.

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

IC50

0.1 nM 0% HS, 0.8 nM 15% HS

Potency assay:

HTRF-based neutralization assay with and without Human Serum (HS)

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

4ODE

Structure-activity relationship:

yes

In cell validation

On target activity in cells

Potency in cells:

IC50

16 nM

Potency assay (cells):

cell proliferation assays (EdU using SJSA-1 tumor cells)

Target engagement assay (cells):

Tested the ability of AM-6761 to inhibit the proliferation of HCT116 p53wt and p53–/– tumor cells in vitro. Displaying robust dose-dependent cell growth inhibitions of HCT116 wild-type p53 cells (IC50 = 11 nM) and no inhibition of p53 deficient cells at doses <10 μM. Similarly, AM-6761 also exhibited a dose-dependent increase of p21 mRNA, a direct transcriptional readout of p53 activity, in HCT116 p53wt cells (IC50 = 58 nM). No induction was observed when HTC116 p53–/– tumor cells were treated with AM-6761 at concentrations up to 10 μM.

Toxicity

Cytoxicity assay:

Yes

Notes on cytotoxicity:

Cytotoxicity observed with AM-6761 is attributable to activation of the p53 pathway.

In vivo validation

Organism:

Mouse, Rat

Dose:

0.5 mg/Kg (IV), 5 mg/Kg (PO)

Route of delivery:

OralIntravenous

Plasma half life:

3.2 (M), 4.4 (R) h (IV)

Systemic clearance:

0.15 (M), 0.23 (R) L/h/Kg (IV)

Vss:

0.35 (M), 0.27 (R) L/Kg (IV)

Bioavailability:

56 %

Organ of Interest

Organ (O):

SJSA-1 osteosarcoma xenograft model

Target engagement assay:

ED50 = 11 mg/kg

Primary Reference

Reference (doi):

10.1021/jm401911v

Reference (PMID):

24601644

Endorsed probe

The Chemical Probes Portal endorses this compound as chemical probe for use in cells and in model organisms.

SAB Rating

Rating for use in Cells

3 review(s)

Rating for use in Model Organisms

3 review(s)

SAB Members

SAB Comments

This is a good probe for cell and mouse studies, but it is probably less useful than related Amgen compounds that have entered human testing, such as AMG232. In a mouse xenograft SJSA-1 osteosarcoma tumor model the compound caused 6% tumor reduction at the highest dose, 50 mg/kg.

Jun 26 2017 - 12:21pm

AM-6761 is broadly comparable to the previously reported compound AMG-232 in terms of high potency and robust effects in cells. AM-6761 would be a confirmatory molecule from the same chemical scaffold. High selectivity has been shown for MDM2 over MDMX, and for effects on cells expressing WT P53 over P53-deficient cells. Broader off-target selectivity data has not been published and would be beneficial to aid interpretation of the effects of the compound in wider cell-based screening. Pharmacokinetic and pharmacodynamics data support the use of AM-6761 in mouse models in vivo. AM-6761 shows lower efficacy (fewer tumour regressions at high dose) in xenograft tumours than AMG-232, which might, therefore, be preferred for in vivo studies.

Jun 27 2017 - 1:50pm

This is a high quality probe derived from AMG232 and broadly comparable to that clinical agent, with similar potency and in vivo anti-tumor activity in an SJSA-1 mouse model. Publications of AMG232 and AM-6761 are highly similar in assays and format and can be compared for further details. This compound is very selective for MDM2 compared to MDMX (IC50>100 uM). Due to chemical similarity to AMG232, this compound could be viewed as a confirmatory molecule with the same scaffold.

Jul 7 2017 - 12:21pm

AM-6761

Probe Information

Target Details

ACTFS, HDMX, hdm5

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

Toxicity

In vivo validation

Primary Reference

Supporting Organizations

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AM-6761

Probe Information

Target Details

ACTFS, HDMX, hdm5

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

Toxicity

In vivo validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?