TH1760

Protein target name:

NUDT15

Mechanism of action:

Inhibitor

Recommended Concentration for use in cells:

up to 10 uM

Probe Information

Target and Probe Information

Target Details

Target class:

Epigenetics

Subclass:

Nudix Hydrolase

HUGO ID:

HGNC:23063

Entrez gene ID:

55270

UNIPROT ID:

Q9NV35

Target alias:

MTH2; NUDT15D

Probe Details

Chemical Probes Portal ID:

CPP2037

Probe alias:

MKB

SMILES string:

O=C1Nc2ccc(cc2O1)[S](=O)(=O)N3CCN(CC3)C(=O)c4ccc5[nH]ccc5c4

InChi Key:

HNCVDRFFJZJQRW-UHFFFAOYSA-N

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

IC50

25 nM dGDP, 57 nM 6-thio-dGTP

Potency assay:

MG (Malachite Green) assay

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

6T5J

Structure-activity relationship:

yes

Off target activity

Potency assay (off target):

@ 100 uM, TH1760 showed selectivity over a panel of related proteins with sequential or functional resemblance to NUDT15, including other human NUDIX proteins (MTH1, NUDT2, NUDT5, NUDT9, NUDT12, NUDT14, NUDT18 and NUDT22) and nucleotide pyrophosphatases (dCTPase, dUTPase and ITPase)

Probe Selectivity in Vitro:

The selectivity of TH1760 was further scrutinized and confirmed with the Eurofins Cerep SafetyScreen44 panel and a curated library of 44 kinases, where TH1760 at 10 or 12 µM, respectively, did not demonstrate significant interaction and/or inhibition of the tested targets

In cell validation

On target activity in cells

Potency in cells:

T (agg)

Potency assay (cells):

CETSA/Western Blot, DARTS assay

Target engagement assay (cells):

Two orthogonal cellular target engagement assays using both endogenous and HA (human influenza hemagglutinin protein, aa 98-106)-tagged NUDT15

Primary Reference

Reference (doi):

10.1038/s41589-020-0592-z

Reference (PMID):

32690945

Endorsed probe

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

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 top-quality chemical probe for in vitro and cellular studies of NUDT15. Nanomolar affinity to NUDT15 is shown, and direct engagement with the protein has clearly been demonstrated (TSA, X-ray). Also, cellular target engagement has been shown (CETSA, DARTS). The compound is selective to NUDT15 over closely related protein homologues (TH1760 here tested at 100 µM), and is only little active against two broad panels of counter-targets (TH1760 tested at 10 µM in a safety screen and a range of kinases). TH1760 has been shown to induce the expected biochemical outcome in various cell lines (10 µM). A close analogue (TH7285) has been identified, which can serve an excellent negative control compound. This is the first bona fide small-molecule NUDT15 inhibitor. Currently, there is no information about in vivo activity of TH1760 available in literature.

Dec 2 2020 - 9:51am

TH1760 chemical probe for NUDT15 was extensively characterized in vitro and in cells. It is highly potent in vitro and shows a remarkable selectivity within the Nudix enzyme family. The cellular characterization of this probe includes CETSA and DARTS assays. The biological sensitization to thiopurines via accumulation of 6-thio-dGTP was shown as mechanism for the compound and NUDT15 knockdown sensitization; thus, providing additional evidence of compound activity in cells.

Dec 2 2020 - 9:53am

Sufficient efforts have been made to characterize the lead compound and compare it with its negative control, which is structurally similar to the parental by the difference of a methyl group at a key interacting site. The chemical probe has demonstrated target engagement and modes of interaction with multiple biochemical assays and in multiple cellular contexts and the on-target selectivity/minimal off-target effects. The biological outcomes of the chemical probe are consistent with the underlying mechanism and modes of action. Therefore, this chemical probe, paired with its negative control, is well justified for its use for cellular assays. The reviewer recommends this chemical probe to suppress the enzymatic activity of NUDT15 for biochemical and cellular assays. Given that the recommended dose of 10 uM is modest, the reviewer suggests pairing this chemical probe with its negative control to further avoid unexpected off-target effects. It is worth noting that this chemical probe is not ready for use in model organisms because its on/off-target effect has only been evaluated against human NUDT15 with human cell lines.

Dec 8 2020 - 11:47am

TH1760

Probe Information

Target Details

MTH2; NUDT15D

Probe Details

In vitro validation

In cell validation

Primary Reference

Supporting Organizations

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TH1760

Probe Information

Target Details

MTH2; NUDT15D

Probe Details

In vitro validation

In cell validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?