PT2385

Protein target name:

HIF-2α

Mechanism of action:

Antagonist

Recommended Concentration for use in cells:

up to 1 uM

Probe Availability

MedChem Exopress

SelleckChem

Probe Information

Target and Probe Information

Target Details

Target class:

Transcription factor

Subclass:

Hypoxia-Inducible Factor

HUGO ID:

HGNC:3374

Entrez gene ID:

2034

UNIPROT ID:

Q99814

Target alias:

EPAS1; HLF; MOP2; ECYT4; HIF2A; PASD2; bHLHe73

Probe Details

Chemical Probes Portal ID:

CPP1882

Probe alias:

79A, PT-2385, cpd 10i

PubChem CID:

91754484

ChEMBL ID:

CHEMBL4173075

SMILES string:

N#CC1=CC(F)=CC(OC2=CC=C(S(=O)(C)=O)C3=C2CC(F)(F)[C@H]3O)=C1

InChi Key:

ONBSHRSJOPSEGS-INIZCTEOSA-N

Control compound:

Orthogonal probe:

M1001T1001PT2399

Physico-chemical properties

Potential reactivities:

not known

NMR:

1H NMR (400 MHz, CDCl3): δ 7.93 (d, 1 H), 7.27–7.24 (m, 1 H), 7.15–7.14 (m, 1 H), 7.07–7.03 (m, 1 H), 7.00 (d, 1 H), 5.63–5.58 (m, 1 H), 3.56–3.35 (m, 3 H), 3.24 (s, 3 H).

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

Kd, IC50

< 50 nM, 12±5 nM

Potency assay:

ITC Binding Affinity, KD 167 nM as measured by MST, Scintillation proximity assay (IC50) with PAS-B* domain

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

6E3S,5TBM

Structure-activity relationship:

Yes

Off target activity

Potency (off target):

HIF-1alpha

Off target protein and potency:

not detected

Potency assay (off target):

ITC

In cell validation

On target activity in cells

Potency in cells:

IC50

42-44 nM (VGEFa Elisa)27±6 nM (Luciferase)

Potency assay (cells):

Luciferase assay, VEGFa ELISA, Expression of HIF-2 target genes in 786-O cells (constitutively active HIF-2α and nonfunctional HIF-1α) after treatment with PT2385 at various concentrations; Co-IP of ARNT

Off target activity in cells

Probe Selectivity in Cell:

0/40 kinase selectivity (<70% activity remaining at 10 μM)

In vivo validation

Organism:

male CD1 miceRatDog

Dose:

3 mg/Kg (IV)10 mg/Kg (PO)

Route of delivery:

OralIntravenous

Plasma half life:

3.3 h (M, R), 11 h (Dog)

Systemic clearance:

19.9 (M), 34.6 (R), 21.5. (D) mL/min/kg

Fraction of probe bound to plasma protein:

110 (M) %40 (R) %87(D) %

Primary Reference

Reference (doi):

10.1021/acs.jmedchem.8b01196 10.1158/0008-5472.CAN-16-0473

Reference (PMID):

30289716 27635045

SAB Rating

Rating for use in Cells

3 review(s)

Rating for use in Model Organisms

3 review(s)

SAB Members

SAB Comments

There are multiple lines of convincing evidence to support meaningful HIF2α modulation by PT-2385 in cell-free, cellular, and in vivo assays. This includes nanomolar cell-free target inhibition, x-ray crystal structures, ITC, convincing SAR, and activity consistent with HIF2α engagement in multiple cellular and in vivo assays. I recommend including an inactive control analog in cellular assays, and in vivo assays if possible.

May 20 2021 - 12:13pm

This is a promising HiF-2a antagonist with a high affinity to target, high cell potency, expected cellular effects (in vitro and in vivo), good in vivo PK and toxicology profiles, and very good selectivity properties.

Jun 4 2021 - 11:52am

PT2385

Probe Availability

Probe Information

Target Details

EPAS1; HLF; MOP2; ECYT4; HIF2A; PASD2; bHLHe73

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

PT2385

Probe Availability

Probe Information

Target Details

EPAS1; HLF; MOP2; ECYT4; HIF2A; PASD2; bHLHe73

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?