TP-004

Protein target name:

METAP2

Mechanism of action:

Reversible Inhibitor

Recommended Concentration for use in cells:

200 nM to 1 uM

Probe Availability

MedChemExpress

Probe Information

Target and Probe Information

Target Details

Target class:

Other

Define other target class:

Enzyme

Subclass:

Methionine Aminopeptidase

HUGO ID:

HGNC:16672

Entrez gene ID:

10988

UNIPROT ID:

P50579

Target alias:

MAP2; MNPEP; p67eIF2

Probe Details

Chemical Probes Portal ID:

CPP1903

PubChem CID:

89794927

ChEMBL ID:

CHEMBL3799512

SMILES string:

Cc1nc(ncc1c2cc(cc3[nH]ncc23)C(F)(F)F)N4CC[C@@H](O)C4

InChi Key:

BNTAEJNPQLMGDJ-LLVKDONJSA-N

Control compound:

TPn-004

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

IC50

5nM with Mn; 31nM with Co

Potency assay:

Biochemical Activity

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

5JHU

Structure-activity relationship:

yes

In cell validation

On target activity in cells

Potency in cells:

EC50

15 nM

Potency assay (cells):

Accumulation of NMet14-3-3 in HUVEC cells

Off target activity in cells

Probe Selectivity in Cell:

> 100 fold. Evaluated in a Ricerca Comprehensive Pharmacological Profile panel of 100 biological targets (GPCRs, ion channels, transporters and enzymes) + panel of proteases and was found to have a clean selectivity profile at 10 µM (< 50% inhibition). hERG is 1 % at 10 µM.

Toxicity

Cytoxicity assay:

Yes

Notes on cytotoxicity:

Cytotoxicity readout (HepG2_72): EC50 >50µM Cell Viability : 100% at 3µM; 100% at 10µM; 98.4% at 30µM

In vivo validation

Organism:

RatMouse

Dose:

30 mg/Kg10 mg/Kg5 mg/Kg

Route of delivery:

Oral

Plasma half life:

6 h

Systemic clearance:

17.2 ml/min/Kg

Primary Reference

Reference (doi):

10.1016/j.bmcl.2016.04.072 10.1016/j.bmcl.2016.04.073

Reference (PMID):

27155900 27136719

External Resources

Available data:

SGC additional analysis

Under Review

This probe is in the process of SAB review. Please continue to check back for new reviews and commentary.

Part of the Donated Chemical Probes initiative from SGC.

SAB Rating

Rating for use in Cells

2 review(s)

Rating for use in Model Organisms

2 review(s)

SAB Members

SAB Comments

The probe was evaluated against a panel of 100 GPCR receptors, ion channels, transporters and enzymes, (referred to as the Ricerca Comprehensive Pharmacological Profile screen) and 26 proteases and displayed inhibition ≤ 50% at 10 µM suggesting a useful degree of selectivity; however, the specific molecular targets were not disclosed. The probe has been counter-screened against the other human methionine aminopeptidase MetAP1 (IC50 = 18 µM; doi.org/10.1124/jpet.119.259028) providing 900-fold isoform selectivity. Given the high potency of the probe against MetAP2 and the nearly 3-orders of magnitude selectivity against MetAP1 and a panel of 126 other targets, the probe is possesses a promising level of selectivity. Further details of the specific enzymes evaluated would be useful to affirm the selectivity. It would also be useful to screen against a large panel of metalloenzymes since the probe directly coordinates with both metal atoms in the active site of MetAP2.

The probe is reported to possess an EC50 of 15 nM (doi.org/10.1016/j.bmcl.2016.04.073) and 26 nM (doi.org/10.1124/jpet.119.259028) in a cellular assay (HUVEC cells) that measures accumulation of N-Met 14-3-3g, which is a client protein of MetAP. The high cellular potency is consistent with the strong enzyme inhibition in the cell-free biochemical assay.

The probe shows almost linear dose-response relationship in vivo with 3, 10, 30 and 100 mg/kg doses displaying 1%, 5%, 11% and 22% weight loss at the 2 week timepoints. Target engagement studies measuring accumulation of N-Met 14-3-3g in adipose tissue also showed clear dose-response relationships.

Jun 2 2020 - 4:21pm

My reasons for concern in recommending this probe are that much of the data reported for this compound is not publicly available. For example, a selectivity profile of >1000 is reported in this form for MetAP2 over MetAP1, but no mention of MetAP1 data is made in the Bioorg. Med. Chem. Lett. paper, nor in the associated patent. Western blots to show target engagement in HUVEC cells are not depicted anywhere, so we must simply “believe” the authors. While I have no reason not to believe them, I would feel more comfortable giving three (or more) stars to a probe where the data is available for all to see. Because I don’t give a 3 star recommendation for cells, I also don’t give it for in vivo work. Furthermore, no on-target activity was demonstrated in animals other than the expected phenotype (weight loss). One last note that goes along this line of thinking: It appears as if an error has been made in reporting the IC50 value when using Cobalt as a metal. In the patent, a pIC50 value of 7.8 for compound 413 (which is TP-004) is listed. A pIC50 value of 7.8 equates to an IC50 value of 16 nM, but 31 nM is reported in this form.

Jul 30 2020 - 1:26pm

TP-004

Probe Availability

Probe Information

Target Details

MAP2; MNPEP; p67eIF2

Probe Details

In vitro validation

In cell validation

Toxicity

In vivo validation

Primary Reference

External Resources

Supporting Organizations

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TP-004

Probe Availability

Probe Information

Target Details

MAP2; MNPEP; p67eIF2

Probe Details

In vitro validation

In cell validation

Toxicity

In vivo validation

Primary Reference

External Resources

Supporting Organizations

Interested in supporting the portal?