AG-270

Protein target name:

MAT2A

Mechanism of action:

Allosteric Inhibitor

Recommended Concentration for use in cells:

up to 1 uM

Probe Availability

MedChemExpress

Probe Information

Target and Probe Information

Target Details

Target class:

MATA2; MATII; SAMS2

Probe Details

Chemical Probes Portal ID:

CPP2359

Probe alias:

WBG

PubChem CID:

134307820

ChEMBL ID:

CHEMBL4573938

SMILES string:

O=C1C(c2ccc(OC)cc2)=C(Nc3ccccn3)Nc4n1nc(c5ccccc5)c4C6=CCCCC6

InChi Key:

HSVRUECLQUGTDS-UHFFFAOYSA-N

Control compound:

AGI-38053

Orthogonal probe:

PF-9366

Physico-chemical properties

Solubility - concentration:

<0.5/<1.0 uM @ 2/7.4 pH

NMR:

1H NMR (400 MHz, DMSO-d6): δ 9.09 (br s, 1H), 8.20 (d, J = 4.3 Hz, 1H), 7.82 (t, J = 7.3 Hz, 1H), 7.73 (d, J = 7.3 Hz, 2H), 7.38–7.51 (m, 3H), 7.28–7.38 (m, 3H), 7.10–7.17 (m, 1H), 7.03 (d, J = 8.5 Hz, 2H), 6.06 (br s, 1H), 3.82 (s, 3H), 2.34 (br s, 2H), 2.06 (br s, 2H), 1.65–1.79 (m, 4H).

Caco-2 permeability: A > B/efflux ratio 0.13/0.76 10E–6cm/s

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

IC50, % inh

14 nM, 91 %

Potency assay:

Enzymatic MAT2A inhibition assay

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

7KCC

Structure-activity relationship:

yes

Off target activity

Probe Selectivity in Vitro:

Outside target family: assessment of AG-270 specificity in in vitro binding assays at 10 μM using Eurofins' panel of 90 unrelated targets did not reveal any effects exceeding 50% inhibition

In cell validation

On target activity in cells

Potency in cells:

IC50

20 nM (S), 257 nM (C)

Potency assay (cells):

72 h Cell MTAP-null SAM assay (S), MTAP-null cell proliferation (C)

In vivo validation

Organism:

MouseRatDogCyno

Dose:

10-200 mg/Kg

Route of delivery:

Oral

Plasma half life:

5.9 (M), 4.2 (R), 4.8 (C) 21.3 (D)

Systemic clearance:

0.0804 (M), 0.47 (R), 0.06 (C), 0.04 (D) L/h/Kg

Vss:

0.527 (M) 2.26 (R) 0.34 (C) 0.88 (D) L/Kg

AUC:

0.527 (M), 2.26 (R), 0.34 (C), 0.88 (D) h*ng/mL

Bioavailability:

31 (M), 48 (D), 92 (R)

Organ of Interest

Organ (O):

Xenograft

Target engagement assay:

Led to dose-dependent reductions in tumor SAM levels and tumor growth of KP4 MTAP-null xenografts (TGI = 36% (10 mg/kg), 48% (30 mg/kg), 66% (100 mg/kg), 67% (200 mg/kg).

Primary Reference

Reference (doi):

10.1021/acs.jmedchem.0c01895 10.1016/j.ccell.2020.12.010

Reference (PMID):

33829783 33450196

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

AG-270 is a well-characterised tool compound for the study of allosteric MAT2A inhibition both in cells and in vivo mouse models at the recommended concentration and dose ranges. However, AG-270 has low aqueous solubility (< 1.0 uM at pH 7.4) and required a spray-dried dispersion formulation for oral pharmacokinetic studies in higher species. Oral dosing in mouse models bearing human tumour xenografts utilised a bespoke formulation (6.7 % w/w HPMCAS-MF, 1% w/w PVP K30, 2% w/w TPGS, 0.1% Simethicone) – see J. Med Chem. 2021, 64, 4430-4449 Supplementary Information. Thus, care is required in the formulation of AG-270 to ensure adequate exposure in both cell-based and especially in vivo studies.

Jun 21 2021 - 9:30am

The compound is poorly soluble thus special care must be taken when preparing formulations for in vivo work. Specifically, the compound must be prepared as a spray-dried dispersion.

Jun 25 2021 - 4:53pm

AG-270 was developed from a relatively weak binder and abundant SAR data and co-crystal structures with its intended target MAT2A, demonstrate the inhibition mechanism. Cellular and in vivo effects make sense with the expected phenotypes and it appears there are no obvious off-target effects in cellular assays. Negative control compounds for AG-270 exist. Proteome-wide selectivity has not yet been demonstrated by using MAT2A resistance alleles, however, this is a minor concern since collectively all the available data suggests that AG-270 is a potent and selective inhibitor of MAT2A.

Jul 7 2021 - 12:32pm

AG-270

Probe Availability

Probe Information

Target Details

MATA2; MATII; SAMS2

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

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AG-270

Probe Availability

Probe Information

Target Details

MATA2; MATII; SAMS2

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?