URMC-099

Protein target name:

MLK1, MLK2, MLK3

Mechanism of action:

Inhibitor

Recommended Concentration for use in cells:

N/A

Probe Availability

Cayman (1229582-33-5)

MedChem Express (1229582-33-5)

Probe Information

Target and Probe Information

Target Details

Target class:

Protein kinase

Subclass:

TKL

HUGO ID:

HGNC:6861 HGNC:6849 HGNC:6850

Entrez gene ID:

4293 4294 4296

UNIPROT ID:

P80192 Q02779 Q16584

Probe Details

Chemical Probes Portal ID:

CPP87

PubChem CID:

54764565

ChEMBL ID:

CHEMBL2436978

SMILES string:

CN1CCN(CC1)CC2=CC=C(C=C2)C3=CN=C4C(=C3)C(=CN4)C5=CC6=C(C=C5)NC=C6

InChi Key:

QKKIWEILHCXECO-UHFFFAOYSA-N

PAINs evaluation:

PAINs free

Physico-chemical properties

NMR:

1H NMR (DMSO-d6, 300 MHz): δ 11.82 (bs, 1H), 11.07 (bs, 1H), 8.54 (d, J = 1.2 Hz, 1H), 8.41 (d, J = 1.2 Hz, 1H), 7.89 (s, 1H), 7.76 (d, J = 1.5 Hz, 1H), 7.69 (d, J = 5.1 Hz, 2H), 7.48 (m, 2H), 7.39 (d, J = 4.8 Hz, 2H), 7.35 (m, 1H), 6.49 (dd, J = 2.7, 1.2 Hz, 1H), 3.48 (s, 2H), 2.50 (m, 3H), 2.38 (m, 5H). ESI (m/z) 422 (M + H), calcd 421

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

IC50

MLK1 19 nM, MLK2 42 nM, MLK3 14 nM

Structure-activity relationship:

In our SAR studies, we observed that the pyrrolo NH and the pyridine nitrogen of the scaffold were required for potency. In addition, for the side chains, having an aromatic group such as an indole, aniline, or phenol was associated with high in vitro enzyme inhibition potency as they provided a hydrogen bond donor from the para position of an aromatic ring.

Off target activity

Potency (off target):

IC50

Off target protein and potency:

LRRK2 11 nMDLK 150 nMABL T315I 3 nMFLT3 4 nM

Potency assay (off target):

In ScanMax Assay (DiscoveRx), URMC-099 inhibited 111 of 442 kinases >90% at 1 uM. In radiometric assay, URMC-099 inhibited 36 of 342 kinases >99% at 1 uM.

Probe Selectivity in Vitro:

Negligible activity against the CYP P450s 2D6, 3A4, 2C9 (IC50s > 10 uM).

In cell validation

On target activity in cells

Potency in cells:

IC50

0.51 uM

Potency assay (cells):

TNFalpha release from microglia

Off target activity in cells

Probe Selectivity in Cell:

Negligible activity against hERG in automated Cerep whole cell patch clamp assays, IC50 = 21 uM.

In vivo validation

Organism:

C57BL/6 mice

Dose:

10 mg/kg

Route of delivery:

Intravenous

Plasma half life:

2.72 hr

Systemic clearance:

2.9 L/h/kg

Ratio of probe in organ to plasma (O:P):

1.6

Fraction of probe bound to plasma protein:

0.98

Cmax:

1848 ug/L

Organ of Interest

Organ (O):

Brain

Target engagement assay:

Indirect: inhibition of substrate phosphorylation in the brain

Primary Reference

Reference (doi):

10.1021/jm401094t

Reference (PMID):

24044867

SAB Rating

Rating for use in Cells

3 review(s)

Rating for use in Model Organisms

3 review(s)

SAB Members

SAB Comments

This probe is not very selective for MLK3. A large number of other kinases are significantly inhibited by this probe.

Jun 30 2016 - 4:04am

This compound is an orally bioavailable, brain-penetrant inhibitor of MLK, but it show poor selectivity against many other protein kinases.

Jun 30 2016 - 4:05am

This is a promiscuous kinase inhibitor with significant off-target activity.

Jun 30 2016 - 6:47pm

This compound is rather promiscuous and targets several kinases other than the MLK family with equal or even greater potency.

Jul 3 2016 - 3:45pm

URMC-099

Probe Availability

Probe Information

Target Details

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

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URMC-099

Probe Availability

Probe Information

Target Details

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

In vivo validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?