Filgotinib

Protein target name:

JAK1

Mechanism of action:

Inhibitor

Recommended Concentration for use in cells:

200 nM - 2 uM

Probe Availability

MedChem Express (1206161-97-8)

Probe Information

Target and Probe Information

Target Details

Target class:

Protein kinase

Subclass:

HUGO ID:

HGNC:6190

Entrez gene ID:

3716

UNIPROT ID:

P23458

Target alias:

Janus kinase 1, JAK1AB, JTK3, JAK1

Probe Details

Chemical Probes Portal ID:

CPP1235

Probe alias:

GLPG0634

PubChem CID:

49831257

SMILES string:

C1CC1C(=O)NC2=NN3C(=N2)C=CC=C3C4=CC=C(C=C4)CN5CCS(=O)(=O)CC5

InChi Key:

RIJLVEAXPNLDTC-UHFFFAOYSA-N

Orthogonal probe:

BaricitinibRuxolitinib

Physico-chemical properties

NMR:

H (400 MHz, DMSO-d6) δ 11.15 (1H, br s, NH), 8.11 (2H, m, ArH), 7.78 (2H, m, ArH), 7.76 (1H, dd, J = 8.8, 6.5 Hz, ArH), 7.73 (1H, dd, J = 8.8, 2.1 Hz, ArH), 7.36 (1H, dd, J = 6.5, 2.1 Hz, ArH), 4.44 (2H, s, CH2), 3.53 (8H, m, 4 × CH2), 2.03 2.03 (1H, br, CH), 0.82−0.81 (4H, m, 2 × CH2). LCMS m/z 426.0 (M + H+ ).

13C (400 MHz, DMSO-d6) δ 171.5, 157.5, 149.9, 138.8, 132.7, 132.1, 131.1, 130.6, 129.3, 114.1, 113.7, 57.7, 49.7, 47.8, 13.8, 7.7. LCMS m/z 426 (M + H+ ). CHN analysis C%, 48.64; H%, 5.81; N%, 13.41; Cl%, 6.88; S%, 6.37 (theoretical: C%, 48.88; H%, 8.86; N%, 13.57; Cl%, 6.87; S%, 6.21)

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

JAK1: 11 nM, JAK2: 32 nM, JAK3: 300 nM

Potency assay:

KDs were measured by displacement of a fluorescent-derivative of ATP. In enzyme activity assays using recombinant enzymes, IC50s were JAK1: 10 nM, JAK2 28 nM, TYK2 116 nM, JAK3 810 nM

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

4P7E

Structure-activity relationship:

Yes, see J Med Chem paper

Off target activity

Potency (off target):

IC50

Off target protein and potency:

FLT3: 338 nM, FLT4: 274 nM; CSF1R: 489 nM

Potency assay (off target):

In a panel of 170 kinases at 1 uM, filgotinib had notable activity against the kinases listed below.

Probe Selectivity in Vitro:

In a panel containing 99 proteins (GPCRs, ion channels, transporters or non-kinase enzymes), no notable activity was reported. Filgotinib had no detectable impact on the activity of CYP450 enzymes (IC50>70 uM).

In cell validation

On target activity in cells

Potency in cells:

IC50

JAK1: 629 nM; JAK2: 17.4 uM

Potency assay (cells):

In whole blood assays, filgotinib inhibited IL-6-induced phosphorylation of STAT1 (JAK1-dependent activity) ~27-fold better than GM-CSF-induced phosphorylation of STAT5 (JAK2-specific activity), consistent with filgotinib showing selectivity for JAK1 in cells.

Target engagement assay (cells):

Indirect, inhibition of substrate phosphorylation

Off target activity in cells

Potency assay, off target (cells):

In hERG patch clamp assay, IC50>100 uM

In vivo validation

Organism:

Rat

Dose:

1 mg/kg (IV); 5 mg/kg (oral)

Route of delivery:

OralIntravenous

Plasma half life:

1.6 h (IV), 3.9 h (oral)

Systemic clearance:

1.35 L/h/kg (IV)

Cmax:

1,407 ng/mL (IV), 310 ng/mL (oral)

Tmax:

2.2 h (oral)

Primary Reference

Reference (doi):

10.4049/jimmunol.1201348 10.1021/jm501262q

Reference (PMID):

24006460 25369270

Endorsed probe

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

SAB Rating

Rating for use in Cells

3 review(s)

Rating for use in Model Organisms

3 review(s)

SAB Members

SAB Comments

Filgotinib is one of a number of JAK inhibitors available with varying selectivity within the JAK family and across other kinases. I recommend using more than one JAK inhibitor as a tool in cellular assays. Filgotinib is reported to have geater functional selectivity for JAK1 over JAK2 in cells but biochemical selectivity is only about three-fold. Mouse pharmacokinetic data are not found in the literature.

Mar 30 2017 - 8:19pm

The probe appears to show a significant degree of selectivity of JAK1 in cellular assays. I would hence recommend this compound as a fairly selective probe for JAK1 but not for the other kinases listed as targets (JAK2, JAK3, TYK2). The compounds appears to be very selective but has "only" been screened against 170 kinases. Screening of a wider panel would build even more confidence that this compound does not inhibit other kinases. The pharmokineted (PK) data clearly show that the compound is suitable for rat and dog in vivo experiments. It may be suitable for mouse in vivo experiments as well, but I could not find any mouse PK data. I thus recommend assessing PK in mice before using it in mice.

Apr 12 2017 - 12:41pm

Filgotinib is an orally available, selective JAK1 inhibitor and is one of the best JAK inhibitors at this moment. However, the in vitro selectivity index to JAK2 is only 2.8, and the results should be interpreted with caution.

Jun 16 2017 - 2:07pm

Filgotinib

Probe Availability

Probe Information

Target Details

Janus kinase 1, JAK1AB, JTK3, JAK1

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

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Filgotinib

Probe Availability

Probe Information

Target Details

Janus kinase 1, JAK1AB, JTK3, JAK1

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?