TH5427

Protein target name:

NUDT5

Mechanism of action:

Inhibitor

Recommended Concentration for use in cells:

up to 1.5 uM

Probe Availability

TOCRIS

Probe Information

Target and Probe Information

Target Details

Target class:

Epigenetics

Subclass:

Nudix Hydrolase

HUGO ID:

HGNC:8052

Entrez gene ID:

11164

UNIPROT ID:

Q9UKK9

Target alias:

YSA1; YSA1H; YSAH1; hNUDT5

Probe Details

Chemical Probes Portal ID:

CPP1853

Probe alias:

PubChem CID:

132472992

SMILES string:

CN1C2=C(C(=O)N(C1=O)C)N(C(=N2)N3CCNCC3)CC4=NN=C(O4)C5=CC(=C(C=C5)Cl)Cl

InChi Key:

QXCXMVYVUHVFLP-UHFFFAOYSA-N

Physico-chemical properties

NMR:

1 H NMR (400 MHz, DMSO-d6 ) δ ppm 2.70 - 2.79 (m, 4 H), 3.12 - 3.21 (m, 7 H), 3.40 (s, 3 H), 5.69 (s, 2 H), 7.85 - 7.96 (m, 2 H), 8.15 (dd, J=1.90, 0.32 Hz, 1 H).

13C NMR (101 MHz, DMSO-d6) δ ppm 27.6, 29.7, 40.6, 45.1, 50.9, 104.1, 123.7, 126.8, 128.3, 132.2, 132.7, 135.2, 147.6, 151.1, 154.0, 156.8, 162.8, 163.3.

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

IC50

29 nM

Potency assay:

MG assay

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

5NWH

Structure-activity relationship:

yes

Off target activity

Probe Selectivity in Vitro:

Screened in a panel of NUDIX enzymes and other nucleotide phosphohydrolases: outside of NUDT5, had the strongest activity against MTH1 (82 % inhibition) and gave 39, 66, and 38 % inhibition against dCTPase, NUDT12 and NUDT14, respectively, at 100 µM

In cell validation

On target activity in cells

Potency in cells:

EC50

2.1 uM

Potency assay (cells):

ITDRF CETSA

Off target activity in cells

Probe Selectivity in Cell:

SafetyScreen 44 and ExpresSDiversity kinase panels from Eurofins Cerep Panlabs.

Primary Reference

Reference (doi):

10.1038/s41467-017-02293-7 10.3390/cancers11091337

Reference (PMID):

29343827 31510016

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

no rating

SAB Members

SAB Comments

TH5427 is potent and selective chemical probe for NUDT5, a NUDIX hydrolase enzyme that plays important roles in the metabolism of adenosine-5’-diphisphoribose (ADPR) metabolism and regulation of hormone signaling. NUDT5 has been implicated in breast cancer, and TH5427 should provide a valuable tool for the continued exploration of the role of this target in breast cancer and other diseases models. TH5427 is at least 690-fold selective for NUDT5 over other NUDIX hydrolases in vitro, with only limited demonstrated interactions outside of the NUDIX hydrolyase family. I recommend the use of this probe in cells at concentrations up to 1.5 µM, with demonstrated effectiveness in HL-60 and T47D cells. TH5427 does show a large cellular potency offset comparing potency in vitro (malachite green assay, 29 nm) vs in cellular target engagement assays (CETSA and DARTS, 0.75-2.1 µM), though this potency offset has not been specifically explained (i.e. cellular availability vs metabolite competition or other factors).

Jul 11 2020 - 3:00pm

TH5427 showed 690-fold selectivity for NUDT5 over MTH1 in direct assay of ADP-ribose hydrolysis in vitro.

Jul 26 2020 - 10:06am

TH5427

Probe Availability

Probe Information

Target Details

YSA1; YSA1H; YSAH1; hNUDT5

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

Primary Reference

Supporting Organizations

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TH5427

Probe Availability

Probe Information

Target Details

YSA1; YSA1H; YSAH1; hNUDT5

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?