AC-4-130

Protein target name:

STAT5B

Mechanism of action:

SH2 Domain Specific Inhibitor

Recommended Concentration for use in cells:

up to 2 uM

Probe Information

Target and Probe Information

Target Details

Target class:

Transcription factor

Subclass:

Signal transducer and transcription activator

HUGO ID:

HGNC:11367

Entrez gene ID:

6777

UNIPROT ID:

P51692

Target alias:

STAT5

Probe Details

Chemical Probes Portal ID:

CPP1876

Probe alias:

AC4130

PubChem CID:

154701370

SMILES string:

FC1=C(F)C(F)=C(S(=O)(N(CC(N(CC2=CC(C(C)(C)C)=CC(C(C)(C)C)=C2)C3=CC=C(C(O)=O)C=C3)=O)CC4=CC=C(Cl)C=C4)=O)C(F)=C1F

InChi Key:

NMQUSJQZQWKQBL-UHFFFAOYSA-N

Orthogonal probe:

PimozidePiceatannol

Physico-chemical properties

Solubility - preferred solvent:

DMSO

NMR:

1H NMR (400 MHz, Chloroform-d) δ 1.24 (d, J=1.1 Hz, 10H), 4.62 (s, 1H), 4.75 (s, 1H), 6.77 (d, J=8.0 Hz, 1H), 6.85 (s, 1H), 7.15 (d, J=8.1 Hz, 1H), 7.18 – 7.28 (m, 2H), 7.28 – 7.34 (m, 1H), 7.96 (d, J=8.0 Hz, 1H).

13C NMR (101 MHz, Chloroform-d) δ 31.32, 34.72, 47.87, 50.42, 53.54, 77.21, 121.73, 123.33, 128.41, 129.09, 130.15, 131.74, 132.69, 134.51, 146.24, 151.19, 165.39, 169.84

Storage Recommendations

Storage (solution):

-20 C

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency assay:

Thermal Shift Assay, 19F NMR

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

1Y1U

In cell validation

On target activity in cells

Target engagement assay (cells):

Dual-Luciferase Reporter Assay

Toxicity

Cytoxicity assay:

Yes

Notes on cytotoxicity:

3-fold less cytotoxic in MDA-MB-231 (IC50 = 10 μM) than in the high pSTAT5 leukemic cell line.

In vivo validation

Organism:

Mouse

Dose:

25 mg/Kg

Route of delivery:

Intraperitoneal

Primary Reference

Reference (doi):

10.1038/s41375-017-0005-9 10.3390/cancers12041021 10.1021/ml500165r

Reference (PMID):

29472718 32326377 25419444

SAB Rating

Rating for use in Cells

3 review(s)

Rating for use in Model Organisms

3 review(s)

SAB Members

SAB Comments

This compound is potent and has nice selectivity for STAT5B over STAT3, but there is no analysis in the literature of other targets of this molecule, so it should be used with some caution. As for in vivo testing, the compound appears to be well tolerated by mice, but I have not seen a pharmacokinetic analysis to determine, for example, exposure and how well correlated that is to the cellular IC50 values.

Jul 26 2020 - 10:17am

In the “Leukemia” key publication (doi: 10.1038/s41375-017-0005-9) the authors present the STAT5 inhibitor AC-4-130 as a new therapeutic agent for haematological malignancies. They showed an effect in mono therapy in different cellular models as well as in vivo and also demonstrated a potential of synergistic effects in combination with other therapeutic agents. Despite these impressive pre-clinical results, in my opinion, the small molecule AC-4-130 misses some critical requirements for the suitability as chemical probe. An in vitro activity was only shown with a non-quantitative thermal shift assay, instead of a more common Kd-value. Moreover, the fluorine NMR experiment suggesting the non-covalent binding to STAT5 also shows an irreversible interaction with STAT3 demonstrating a certain promiscuity of the molecule. Also, the binding mode was only proposed by docking models. A final validation in form of a co-crystal structure would be more applicable (but is also not mandatory).

In summary, with the current available data I do not recommend AC-4-130 as a chemical probe for STAT5, unless more sophisticated data on on-target activity (e.g. Kd) and selectivity are available.

Aug 28 2020 - 7:28am

The probe represents one of the first STAT5 inhibitors targeting the SH2 domain. However, while cellular data are convincing, current literature does not characterise the probe well in vitro using binding and selectivity assays that would be expected for a high quality probe molecule. Direct interaction with the STAT5 Sh2 domain is only demonstrated by an NMR STD experiment at high concentration, no direct binding data (eg ITC) are available and “on target” activity has not been demonstrated in a cellular context. Structural information of the binding mode of this probe is only provided by docking but not by experimental data.

Oct 12 2020 - 10:23am

AC-4-130

Probe Information

Target Details

STAT5

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

Toxicity

In vivo validation

Primary Reference

Supporting Organizations

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AC-4-130

Probe Information

Target Details

STAT5

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

Toxicity

In vivo validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?