BAY-549

Protein target name:

ROCK1, ROCK2

Mechanism of action:

Inhibitor

Recommended Concentration for use in cells:

~~up to 100 nM~~

Reviewer suggested Concentration for use in cells:

0.5 to 1 uM

Probe Availability

MedChemExpress

TOCRIS

Probe Information

Target and Probe Information

Target Details

Target class:

Protein kinase

Subclass:

AGC

HUGO ID:

HGNC:10251 HGNC:10252

Entrez gene ID:

6093 9475

UNIPROT ID:

Q13464 O75116

Target alias:

ROCK-I; P160ROCK; ROCK-II

Probe Details

Chemical Probes Portal ID:

CPP1863

Probe alias:

BAY549, Azaindole 1

PubChem CID:

11524200

SMILES string:

C(=C12)(C(=CC=N1)OC(=C(C=C(C1)NC3=CC(=NC(=N3)N)[Cl])F)C=1F)C(=CN2)C

InChi Key:

NRSGWEVTVGZDFC-UHFFFAOYSA-N

Control compound:

BAY-4900

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

IC50

0.6 nM ROCK1, 1.2 nM ROCK2

Potency assay:

biochemical ROCK assay

Structure-activity relationship:

yes

Off target activity

Potency (off target):

IC50

Off target protein and potency:

Trk-A 252 nM, Flt3 303 nM

Potency assay (off target):

Kinase panel

Probe Selectivity in Vitro:

selectivity >250 fold vs all other kinases (Upstate panel @ 10 µM)

In cell validation

On target activity in cells

Potency in cells:

IC50

65 nM

Potency assay (cells):

mechanistic tissue assay (arteria saphena rabbit)

Off target activity in cells

Probe Selectivity in Cell:

LeadProfilingScreen closest off-targets @ 10 µM: Dopamine transporter (DAT, human) IC50 = 0.4 µM

In vivo validation

Organism:

Wistar RatNMRI MiceBeagle Dog

Dose:

1 mg/Kg3 mg/Kg10 mg/Kg0.1-0.3 mg/Kg IV

Route of delivery:

OralIntravenous

Plasma half life:

1.50 (m) 1.24 (r) 2.50 (d) h

Systemic clearance:

1.90 (m) 1.20 (r) 0.233 (d) L/h*Kg

Fraction of probe bound to plasma protein:

48% Rat73% Dog

Cmax:

0.0808 (r), 0.0430 (d) mg/L (oral)

Tmax:

2.00 h (r)0.794 h (d)

Primary Reference

Reference (doi):

10.1038/sj.bjp.0707484 10.1002/cmdc.200800211

Reference (PMID):

17934515 18973168

External Resources

Available data:

SGC additional analysis

Endorsed probe

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

Part of the Donated Chemical Probes initiative from SGC.

SAB Rating

Rating for use in Cells

3 review(s)

Rating for use in Model Organisms

3 review(s)

SAB Members

SAB Comments

The selectivity screening only included 112 kinases, data not supplied. It is possible that wider screening would identify additional inhibited kinases, albeit that the closest relatives to ROCK1 and ROCK2 (MRCKs) have only ~38% sequence identity over the kinase domain.

Jun 7 2020 - 10:34am

This compound is also called ROCK-IN-2, Azaindole 1, Azaindole 1, TC-5 7001 and the CAS No. is 867017-68-3

Schirok et al ChemMedChem 3 (2008) 1893 – 1904 (Cmpd 32)

SAR development? Yes, clear SAR with optimization.

ROCK-2 IC₅₀ is given as 3 nM.

Arteria saphena IC₅₀ 65 nM (functional, rabbit) SAR approximately parallels kinase SAR.

Biomarker pMBS not undertaken.

Medium clearance mice and dogs; t_1/2rats 1.2 h, F 48%.

Kast et al BJP 152 (2020) 1070-1080

IC₅₀ ROCK-1 is given as 0.6 nM.

Functional inhibition: IC₅₀ (pMBS, human) is given as 3.7 nM.

IC₅₀ ROCK-2 is given as 1.1 nM.

Functional inhibition: IC₅₀ (pMBS, human) is given as 4.8 nM.

The compound is ATP-competitive.

Inactive against 89 kinases (IC₅₀ > 10uM), another 21 weakly inhibited with IC₅₀ 1-10 uM, only 2 sub-uM IC₅₀ (TRK 252 nM, FLT3 303 nM).

Inactive against 60 relevant cardiovascular enzymes/receptors and weak activity just against L-type calcium channel (6.7 uM) and sodium channel (6.8 uM).

Highly bioavailable rats, dogs (approximately 50%-75%).

Jun 28 2020 - 9:08am

BAY-549, also known as compound 32 in Schirok H, et al ChemMedChem 2008, or Azaindole 1 in Kast R, et al Br J Pharmacol 2007, is a high quality tool compound for ROCK1/2. It is active in rabbit isolated artery assay and reduces blood pressure when dosed in rats and dogs. The only limitation is that kinome selectivity data are limited to a panel of 112 kinases (Upstate), so it remains possible that the compound may have off-target kinase activities outside the tested panel. That said, ROCK1/2 kinases have unique active site structure as noted in the paper, so this is an unlikely scenario. Regardless, it would be prudent to test multiple ROCK1/2 inhibitors from orthogonal chemical scaffolds, for example also GSK429286A (https://lincs.hms.harvard.edu/db/datasets/20030/main).

Mar 24 2021 - 10:18am

BAY-549

Probe Availability

Probe Information

Target Details

ROCK-I; P160ROCK; ROCK-II

Probe Details

In vitro validation

In cell validation

In vivo validation

Primary Reference

External Resources

Supporting Organizations

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BAY-549

Probe Availability

Probe Information

Target Details

ROCK-I; P160ROCK; ROCK-II

Probe Details

In vitro validation

In cell validation

In vivo validation

Primary Reference

External Resources

Supporting Organizations

Interested in supporting the portal?