AMG 176

Protein target name:

MCL1

Mechanism of action:

Antagonist

Recommended Concentration for use in cells:

~~300 nM, 1 uM max~~

Reviewer suggested Concentration for use in cells:

1 nM

Probe Availability

MedChemExpress

Probe Information

Target and Probe Information

Target Details

Target class:

Other

Define other target class:

Apoptosis Regulator

Subclass:

BCL2 family

HUGO ID:

HGNC:6943

Entrez gene ID:

4170

UNIPROT ID:

Q07820

Target alias:

TM; EAT; MCL1L; MCL1S; Mcl-1; BCL2L3; MCL1-ES; bcl2-L-3; mcl1/EAT

Probe Details

Chemical Probes Portal ID:

CPP1960

Probe alias:

AMG176, AMG-176, 11

PubChem CID:

118910268

SMILES string:

CO[C@H]1\C=C\C[C@H](C)[C@@H](C)S(=O)(=O)NC(=O)C2=CC3=C(OC[C@]4(CCCC5=C4C=CC(Cl)=C5)CN3C[C@@H]3CC[C@@H]13)C=C2

InChi Key:

JQNINBDKGLWYMU-GEAQBIRJSA-N

Orthogonal probe:

A-1210477S63845

Physico-chemical properties

Solubility - preferred solvent:

DMSO

Potential reactivities:

Not Known

Storage Recommendations

Storage (dry powder):

-20 °C

Storage (solution):

-80 °C

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

60 pM

Potency assay:

Time-resolved fluorescence resonance energy transfer binding assays

Structure-activity relationship:

yes

Off target activity

Potency assay (off target):

TR-FRET

Probe Selectivity in Vitro:

Selective with BCL-xL 0.7 uM And BCL2 0.95 uM

In cell validation

On target activity in cells

Potency assay (cells):

Split Luciferase Complementation Assay, immunoblot (these experiments were performed with close analogue AM-8621

In vivo validation

Organism:

MouseRatDogMonkey

Dose:

30 and 60 mg/Kg 50 and 100 mg/Kg

Route of delivery:

OralIntravenous

Systemic clearance:

0.03 (m), 0.05 (R), 0.02 (D), 0.5 (M) L/h/Kg

Fraction of probe bound to plasma protein:

32 (m), 34(R), 27(D), 70(M) %

Tmax:

14 (m), 20 (R), 11 (D), 1.8 (M) h

Primary Reference

Reference (doi):

10.1158/2159-8290.CD-18-0387 10.1158/1078-0432.CCR-19-1397 10.1016/j.blre.2020.100672

Reference (PMID):

30254093 31937611 32204955

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

Targeting hMCL-1 by a small molecule is a challenge. The interaction of hMCL-1 and AMG 176 has been characterised (PDB: 6OVC) and this should be added to the review form.

No information regarding the selectivity of AMG 176 outside the primary target is provided and this is a large gap in its biological properties. AMG 176 is a direct antagonist of MCL-1 but classified as an “apoptosis regulator” and this process involves many targets. Therefore, the AMG 176 selectivity profile outside the target family (e.g. kinase) needs to be included in the review form. This is of relevance to compounds developed for oncology as they often exhibit poly-pharmacology.

Further details of the assays (TR-FRET binding assay and HEK293M cellular) should be provided. The cellular assay appears to make use of AM-8621 which is a close analogue of AMG 761 and these findings may not translate to AMG 176.

AMG 176 is a complex molecule with potentially reactive groups and will have a relatively high cost for synthesis/purchase. It is therefore not clear whether its bio-activity is due to the parent molecule or metabolite/s. The results of appropriate stability studies should be included in the review form.

The physico-chemical properties of AMG 176 are not reported. The solubility (solvent and concentration), potential reactivities and QC (NMR, mass spec, mp, etc), storage recommendations and stability (dry powder and solution) and sensitivities (e.g. freeze/thaw cycles) should be provided in the review form.

With a 60 pM Ki, recommending the use of 1µM AMG 176 in a cellular assay is excessive (currently ~17,000x Ki).

A pubchem search for AMG 176 (https://pubchem.ncbi.nlm.nih.gov/compound/118910268#section=Names-and-Identifiers) reveals that its molecular weight (613.2 g/mol) is relatively high and when taken together with a high tPSA (93.3 Å²), makes it undesirable for oral delivery, although this route is mentioned in the review form. AMG 176 would also not be CNS penetrant (Pajouhesh and Lenz, Medicinal Chemical Properties of Successful Central Nervous System Drugs, NeuroRx. 2005 Oct; 2(4): 541–553. doi: 10.1602/neurorx.2.4.541).

Oct 27 2020 - 2:33pm

A very close analogue (AM-8621) was tested in several cell lines, and demonstrate activity in AML cell lines, therefore I decided to classify this probe as ‘good to use in cells’. DOI: 10.1158/2159-8290.CD-18-0387

Nov 4 2020 - 11:54am

AMG 176

Probe Availability

Probe Information

Target Details

TM; EAT; MCL1L; MCL1S; Mcl-1; BCL2L3; MCL1-ES; bcl2-L-3; mcl1/EAT

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

AMG 176

Probe Availability

Probe Information

Target Details

TM; EAT; MCL1L; MCL1S; Mcl-1; BCL2L3; MCL1-ES; bcl2-L-3; mcl1/EAT

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?