MPS1-IN-1

Reagent Authentication Certificate:

Chemical Probes Portal MPS1-IN-1.pdf

Protein target name:

MPS1

Recommended Concentration for use in cells:

1-10 uM

Probe Availability

MedChem Express (1125593-20-5)

Tocris (1125593-20-5)

Cayman (1125593-20-5)

Probe Information

Target and Probe Information

Target Details

Target class:

Protein kinase

Subclass:

Other

HUGO ID:

HGNC:12401

Entrez gene ID:

7272

UNIPROT ID:

P33981

Target alias:

YDL028C, PAC8, RPK1, TTK

Probe Details

Chemical Probes Portal ID:

CPP114

PubChem CID:

25195352

ChEMBL ID:

CHEMBL3330411

SMILES string:

CC(C)S(=O)(=O)C1=CC=CC=C1NC2=CC(=NC3=C2C=CN3)NC4=C(C=C(C=C4)N5CCC(CC5)O)OC

InChi Key:

NMJMRSQTDLRCRQ-UHFFFAOYSA-N

PAINs evaluation:

PAINs free

Physico-chemical properties

NMR:

1H NMR 600 MHz (CDCl3): 8.51 (s, 1H), 7.83 (d, J = 1.8 Hz, 1H), 7.48 (m, 2H), 7.19 (m, 3H), 7.12 (m, 2H), 6.85 (d, J = 3.0 Hz, 1H), 6.49 (m, 1H), 6.19 (m, 2H), 3.84 (m, 1H), 3.78 (s, 3H), 3.46 (m, 2H), 3.19 (m, 1H), 2.90 (m, 2H), 1.99 (m, 2H), 1.69 (m, 2H), 1.19 (d, J = 6.6 Hz, 6H);

13C NMR 600 MHz (DMSO-d6): 153.8, 150.0, 148.1, 146.6, 141.3, 141.2, 135.2, 131.2, 123.0, 122.9, 121.2, 121.0, 120.2, 119.5, 107.4, 103.5, 100.9, 96.7, 89.1, 66.1, 55.4, 54.2, 47.6, 34.1, 14.8

Validation in Cells and Model Organisms

In vitro validation

On target activity

Potency:

27 nM

Potency assay:

Ambit KinomeScan In vitro kinase assay IC50 = 367 nM

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

3GFW

Off target activity

Potency (off target):

Off target protein and potency:

ALK 21 nMERK2 900 nMPYK2 280 nMFAK 440 nMIGF1R 750 nMINSR 470 nMLTK 29 nM

Potency assay (off target):

Ambit KinomeScan

In cell validation

On target activity in cells

Potency in cells:

IC50

0.65 uM

Potency assay (cells):

Phosphorylation of protein substrate

Target engagement assay (cells):

Indirect: inhibition of protein substrate phosphorylation, and replacement of MPS1 with MPS-IN-1-insenstive mutant (MPS1 M602Q, gate-keep mutant) raises IC50 to 3.1 uM.

Primary Reference

Reference (doi):

10.1038/nchembio.345

Reference (PMID):

20383151

SAB Rating

Rating for use in Cells

3 review(s)

Rating for use in Model Organisms

3 review(s)

SAB Members

SAB Comments

MPS1-IN-1 shows cellular activity in the range of 1-10 µM. Ideally the potency would be well below micromolar, but the compound is still useful for interrogating cellular biology. However, given the tight binding of ATP to MPS1 (Km < 1 µM), further potency optimization may be challenging. I suggest that 10 µM is not exceeded. If possible, a minimal dose response experiment would be useful (e.g., 0.3, 1.0, 3.0, and 10 µM). There is no available pharmacokinetic data for this compound. The utility of MPS1-IN-1 to probe MPS1 function is enhanced by the availability of the dual MPS1/PLK1 inhibitor MPS1-IN-2. Both compounds are known to inhibit other kinases, and their concurrent usage will help to increase confidence in MPS1 as the molecular target if they phenocopy.

Jun 12 2016 - 2:25pm

MPS1-IN-1 is only a moderately potent kinase inhibitor. In vitro IC50 = 350-650 nM for enzyme inhibition. Doses of 2-10 uM are required to inhibit MPS1-dependent activities in cells. The probe needs to be more potent and is only useful in cells at micromolar concentrations. Kinase inhibition profile and potency would need to determined on the kinases from the model organism for use in vivo. Additional controls are required to ensure that phenotypic activity is not due to off targets - examples include an inactive analog or another inhibitor from a different chemotype.

Jun 12 2016 - 2:25pm

While MPS1-IN-1 is a relatively selective kinase inhibitor, it displays only moderate biochemical (~0.5 uM) and cellular (3-10 uM) potency. The original study does provide evidence of the engagement of MPS1 in cells using a wild-type vs M602Q-mutant cell system. Nonetheless, when used at the high concentrations (10 uM) in cells, interaction with additional (nonkinase) targets may occur. Additionally, no ADME, PK or in vivo data are available from the primary reference. Accordingly, MPS1-IN-1 should be used with caution in cellular assays (preferably in conjunction with additional MPS1 inhibitors based on alternative chemical scaffolds), and its use is not recommended for in vivo experiments. Multiple recent publications describe the identification and characterization of more advanced MPS1 inhibitors, likely better suited for interrogation of MPS1 function in cells and in vivo. For example, compound 27b from PMID 25625617 displays a biochemical IC50 of 2.7 nM and a cellular IC50 for pMPS1 of 0.7 nM. Its utility in vivo may be limited by toxicity at higher doses. Similarly, NMS-P715 (PMID 21723120), MPI-0479605 (PMID 22632936) and 34h (PMID 27055065) all provide good MPS1 potency and kinome selectivity while having been used in vivo.

Jul 3 2016 - 3:51pm

MPS1-IN-1

Probe Availability

Probe Information

Target Details

YDL028C, PAC8, RPK1, TTK

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

Primary Reference

Supporting Organizations

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MPS1-IN-1

Probe Availability

Probe Information

Target Details

YDL028C, PAC8, RPK1, TTK

Probe Details

Physico-chemical properties

In vitro validation

In cell validation

Primary Reference

Supporting Organizations

Interested in supporting the portal?