AMG-510

Protein target name:

KRAS G12C

Mechanism of action:

Covalent InhibitorMutant G12C specific

Recommended Concentration for use in cells:

~~Up to 1 uM~~

Reviewer suggested Concentration for use in cells:

500 nM

Probe Availability

MedChemExpress

Probe Information

Target and Probe Information

Target Details

Target class:

Other

Define other target class:

GTPase

Subclass:

GTPase

HUGO ID:

HGNC:6407

Entrez gene ID:

3845

UNIPROT ID:

P01116

Target alias:

NS; NS3; OES; CFC2; RALD; K-Ras; KRAS1; KRAS2; RASK2; KI-RAS; C-K-RAS; K-RAS2A; K-RAS2B; K-RAS4A; K-RAS4B; K-Ras 2; 'C-K-RAS; c-Ki-ras; c-Ki-ras2

Probe Details

Chemical Probes Portal ID:

CPP1829

Probe alias:

AMG510, (R)-38, Sotorasib

PubChem CID:

137278711

SMILES string:

O=C(C=C)N1C[C@H](C)N(C2=NC(N(C3=C(C)C=CN=C3C(C)C)C4=C2C=C(F)C(C5=C(O)C=CC=C5F)=N4)=O)CC1

InChi Key:

NXQKSXLFSAEQCZ-SFHVURJKSA-N

Control compound:

Compound-8

Orthogonal probe:

ARS-1620MRTX849

Physico-chemical properties

NMR:

1H NMR (600 MHz, DMSO-d6) δ ppm 10.20 (s, 1H), 8.39 (d, J = 4.9 Hz, 1H), 8.30 (d, J = 8.9 Hz, 0.5H), 8.27 (d, J = 8.7 Hz, 0.5H), 7.27 (q, J = 8.4 Hz, 1H), 7.18 (d, J = 4.9 Hz, 1H), 6.87 (dd, J = 16.2, 10.8 Hz, 0.5H), 6.84 (dd, J = 16.2, 10.7 Hz, 0.5H), 6.74 (d, J = 8.4 Hz, 1H), 6.68 (t, J = 8.4 Hz, 1H), 6.21 (d, J = 16.2 Hz, 0.5H), 6.20 (d, J = 16.2 Hz, 0.5H), 5.76 (d, J = 10.8 Hz, 0.5H), 5.76 (d, J = 10.7 Hz, 0.5H), 4.91 (m, 1H), 4.41 (d, J = 12.2 Hz, 0.5H), 4.33 (d, J = 12.2 Hz, 1H), 4.28 (d, J = 12.2 Hz, 0.5H), 4.14 (d, J = 12.2 Hz, 0.5H), 4.02 (d, J = 13.6 Hz, 0.5H), 3.69 (m, 1H), 3.65 (d, J = 13.6 Hz, 0.5H), 3.52 (t, J = 12.2 Hz, 0.5H), 3.27 (d, J = 12.2 Hz, 0.5H), 3.15 (t, J = 12.2 Hz, 0.5H), 2.72 (m, 1H), 1.90 (s, 3H), 1.35 (d, J = 6.7 Hz, 3H), 1.08 (d, J = 6.7 Hz, 3H), 0.94 (d, J = 6.7 Hz, 3H).

19F NMR (376 MHz, DMSO-d6) δ −115.6 (d, J = 5.2 Hz, 1 F), −128.6 (br s, 1 F).

13C NMR (151 MHz, DMSO-d6) δ ppm 165.0 (1C), 163.4 (1C), 162.5 (1C), 160.1 (1C), 156.8 (1C), 153.7 (1C), 151.9 (1C), 149.5 (1C), 148.3 (1C), 145.2 (1C), 144.3 (1C), 131.6 (1C), 130.8 (1C), 127.9 (0.5C), 127.9 (0.5C), 127.8 (0.5C), 127.7 (0.5C), 123.2 (1C), 122.8 (1C), 111.7 (1C), 109.7 (1C), 105.7 (1C), 105.3 (1C), 51.4 (0.5C), 51.0 (0.5C), 48.9 (0.5C), 45.4 (0.5C), 44.6 (0.5C), 43.7 (0.5C), 43.5 (0.5C), 41.6 (0.5C), 29.8 (1C), 21.9 (1C), 21.7 (1C), 17.0 (1C), 15.5 (0.5C), 14.8 (0.5C).

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

kinact/KI

9900 ± 1800 M–1 s–1

Potency assay:

Mass Spectroscopy

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

6OIM6PGO6PGP

Structure-activity relationship:

yes

In cell validation

On target activity in cells

Potency in cells:

IC50

68 nM

Potency assay (cells):

ERK1/2 phosphorylation assay

Off target activity in cells

Potency assay, off target (cells):

Cysteine-proteome profiling

Probe Selectivity in Cell:

Cysteine-proteome profiling (MS) further revealed (R)-38 to be highly selective for covalent modification of KRASG12C: of 6451 cysteine-containing peptides profiled, only the KRASG12C C12 peptide was found to be significantly modified

In vivo validation

Organism:

MouseRatDog

Dose:

1 mg/Kg IV10 mg/Kg POReviewer suggest 30 and 100 mg/Kg PO

Route of delivery:

OralIntravenous

Plasma half life:

0.5 (M), 0.5 (R), 0.4 (D) h

Systemic clearance:

1.6 (M) 3.4 (R), 2.2 (D) L/h/kg

Fraction of probe bound to plasma protein:

22-40% (M)28% (R)34% (D)

Primary Reference

Reference (doi):

10.1021/acs.jmedchem.9b01180 10.1038/s41586-019-1694-1

Reference (PMID):

31820981 31666701

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

AMG-510 is the potent R-atropisomer [t1/2 (Racemization) >180 years at 25 °C, ΔG⧧rotation > 31 kcal/mol].

The non-specific reactivity of AMG 510 with GSH is relatively slow (t1/2 = 196 min/within the range of clinical acrylamides).

“Rat and dog pharmacokinetic parameters for (R)-38/AMG 510 are in line with those observed in mouse, with reasonable oral bioavailability observed in all species, relatively short half- in all species […] but durable covalent inhibition of KRASG12C could be achieved without continuous drug exposure” (extract from 10.1021/acs.jmedchem.9b01180).

AMG-510 was approved to start Phase 2 clinical trials in December 2019.

Jul 30 2020 - 1:41pm

AMG 510 (compound 38) is a potent and selective covalent inhibitor of KRASG12C. It was derived by SAR and pharmacological optimisation of a series of compounds built upon the principle of covalently targeting the reactive cysteine residue of KRASG12C. As such it is selective for KRASG12C over other RAS mutants. Utilising A549 (KRASG12S mutant) cells as a control comparator. AMG 510 was >7000x more potent at reducing the viability of KRASG12C PaCa-21 cells (IC50 36.5 vs 0.005 uM respectively). The IC50 for suppression of pERK in PaCa-21 cells is 68nM. In addition to detailed SAR and crystallography demonstrating an interaction with a cryptic pocket (H95, Y96, Q99) of KRASG12C, mass spectrometry excluded promiscuous cysteine reactivity. AMG 510 is orally available and shows in vivo efficacy that correlates with pERK suppression at the 30-100mg/kg dose range in nude mice bearing PaCa-2. Preliminary data from phase 1 human evaluation has demonstrated tolerability and antitumor activity when administered as monotherapy to patients with advanced KRASG12C solid organ malignancy.

Jul 30 2020 - 1:52pm

AMG-510 is a covalent inhibitor of KRASG12C, specifically targeting the mutant cysteine-12 residue. It is created by leveraging the ARS-1620 structure and the exploitation of the cryptic pocket with H95, Y96, and Q99 in KRASG12C. It is a highly potent, selective and orally bioavailable chemical probe suitable for studies with in vitro and in vivo model systems. Activity: AMG 510 inhibits pERK signalling in cells with IC50 of 68 nM, which is supported by the PD data showing its time-dependent inhibition of pERK in mouse models. It also showed potent in vivo activity with TGI of 86% at 10 mg/kg and significant tumor regression at 30 mg/kg in a MIA PaCa-2 mouse model. Target engagement: Target engagement was supported by the cocrystalization structure of AMG 510 in complex with GDP-KRASG12C where the quinazolinone core of AMG 510 occupies the KRAS switch II pocket and the acrylamide moiety making a covalent bond with C12. Ligand occupancy in vivo was demonstrated by efficient covalent modification of KRASG12C by AMG 510 to near completion within two hours. Selectivity: It is a highly selective compound for covalent modification of KRASG12C. In a cysteine proteome profiling studies, only the KRASG12C C12 peptide was covalently modified by AMG 510, of 6451 cysteine-containing peptides profiled. Cell viability assays showed selective killing of cancer cells with KRASG12C (MIA PaCa-2) over those with other KRAS mutations (KRAS p.G12S A549 cells).

Aug 13 2020 - 9:18am

AMG-510

Probe Availability

Probe Information

Target Details

NS; NS3; OES; CFC2; RALD; K-Ras; KRAS1; KRAS2; RASK2; KI-RAS; C-K-RAS; K-RAS2A; K-RAS2B; K-RAS4A; K-RAS4B; K-Ras 2; 'C-K-RAS; c-Ki-ras; c-Ki-ras2

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

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AMG-510

Probe Availability

Probe Information

Target Details

NS; NS3; OES; CFC2; RALD; K-Ras; KRAS1; KRAS2; RASK2; KI-RAS; C-K-RAS; K-RAS2A; K-RAS2B; K-RAS4A; K-RAS4B; K-Ras 2; 'C-K-RAS; c-Ki-ras; c-Ki-ras2

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?