Belumosudil (KD025)

Catalog No.S7936 Batch:S793605

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Technical Data

Formula

C26H24N6O2

Molecular Weight 452.51 CAS No. 911417-87-3
Solubility (25°C)* In vitro DMSO 91 mg/mL (201.1 mM)
Ethanol 91 mg/mL (201.1 mM)
Water Insoluble
In vivo (Add solvents to the product individually and in order)
Clear solution
5%DMSO 35%PEG300 2%Tween80 58%ddH2O
7.0mg/ml Taking the 1 mL working solution as an example, add 50 μL of 140 mg/ml clarified DMSO stock solution to 350 μL of PEG300, mix evenly to clarify it; add 20 μL of Tween80 to the above system, mix evenly to make it clear; then continue to add 580 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results. 
* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
* Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

Preparing Stock Solutions

Biological Activity

Description Belumosudil (KD025, SLx-2119) is an orally available, and selective ROCK2 inhibitor with IC50 and Ki of 60 nM and 41 nM, respectively. Phase 2.
Targets
ROCK2 [1]
(Cell-free assay)
ROCK2 [1]
(Cell-free assay)
41 nM(Ki) 60 nM
In vitro

KD025 down-regulates the IL-17 and IL-21 secretion in human PBMCs, and leads to down-regulation of STAT3 phosphorylation, IRF4, and RORγt expression in CD4+ T cells. [2]

KD025 inhibits the secretion of IL-21, IL-17, and IFNγ along with decreasing phosphorylated STAT3 and reduced protein expression of IRF4 and BCL6 in human peripheral blood mononuclear cells. [3]

In vivo

In mice, KD025 (200 mg/kg, p.o.) inhibits ROCK activity in brain and heart, as measured by the degree of MYPT1 phosphorylation. KD025 (200 mg/kg, p.o.) significantly reduces the area of perfusion defect, and reduces tissue loss in the ipsilateral hemisphere. [1]

In a collagen-induced arthritis (CIA) mouse model, KD025 (200 mg/kg, i.p.) downregulates the progression of collagen-induced arthritis via targeting of the Th17-mediated pathway. [2]

KD025 ()150 mg/kg, i.p. or p.o.) effectively ameliorates cGVHD in a full MHC-mismatch model of multi-organ system cGVHD with bronchiolitis obliterans syndrome and a minor MHC-mismatch model of sclerodermatous GVHD. [3]

Protocol (from reference)

Kinase Assay:

[1]

  • Recombinant ROCK1 and ROCK2 assays

    Compound dilutions and reactions are performed in 96-well polystyrene low-binding plates. Filtration is done in 96-well filter plates containing hydrophilic phospho-cellulose cation exchanger membranes. Enzymatic activity of the recombinant ROCK1 and ROCK2 is measured radiometrically in 50 μL of reaction mixture containing assay buffer (50 mmol/L Tris, pH 7.5, 0.1 mmol/L ethyleneglycoltetraacetic acid, 10 mmol/L magnesium acetate and 1 mmol/L dithiothreitol). Long S6 peptide (KEAKEKRQEQIAKRRRLSSLRASTSKSGGSQK, 30 μmol/L), ROCK (4 mU per reaction) and ATP (10 μmol/L; 1 μCi [[gamma]-33P]ATP) and test compound are diluted to a final dimethylsulfoxide concentration of 1%. The reaction is incubated for 45 min at room temperature and stopped with 25 μL of 3% phosphoric acid. Phosphorylated long S6 peptide is separated from unreacted [[gamma]-33P]ATP by filtration of the quenched reaction contents through a P30 phosphocellulose filter plate. Each filter is washed three times with 75 μL of 75 mmol/L phosphoric acid and one time with 30 μL of 100% methanol. Filter plates are allowed to dry and 30 μL of OptiPhase ‘SuperMix’ scintillation fluid is added to each well. 33Phosphorous is quantified in an I450 MicroBeta scintillation counter and corrected by subtracting the radioactivity associated with the background samples. Data are analyzed and expressed as percent inhibition using the formula ((U − B)/(C − B)) × 100 where U is the unknown value, B is the average of staurosporine background wells, and C is the average of control wells. Curve fitting is performed by GraphPad Prism software using sigmoidal dose-response (variable slope) equation type analysis to generate IC50 values. Ki values are calculated from an equation of Ki = IC50/(1 + [S]/Km)), where [S] and Km are the concentration of ATP and the Km value of ATP, respectively.

Cell Assay:

[4]

  • Cell lines

    A7r5 cells

  • Concentrations

    10 µM

  • Incubation Time

    2 h

  • Method

    Cells were treated with KD025 for 2 hours.

Animal Study:

[1]

  • Animal Models

    Young adult, aged, or type 2 diabetic mice

  • Dosages

    300 mg/kg

  • Administration

    p.o.

Selleck's Belumosudil (KD025) has been cited by 9 publications

Selectivity matters: selective ROCK2 inhibitor ameliorates established liver fibrosis via targeting inflammation, fibrosis, and metabolism [ Commun Biol, 2023, 10.1038/s42003-023-05552-0] PubMed: 37980369
ROCK1 mechano-signaling dependency of human malignancies driven by TEAD/YAP activation [ Nat Commun, 2022, 13(1):703] PubMed: 35121738
Inhibition of ROCK ameliorates pulmonary fibrosis by suppressing M2 macrophage polarisation through phosphorylation of STAT3 [ Clin Transl Med, 2022, 12(10):e1036] PubMed: 36178087
Phenotypic Screening for Small Molecules that Protect β-Cells from Glucolipotoxicity [ ACS Chem Biol, 2022, 10.1021/acschembio.2c00052] PubMed: 35439415
Pressure and stiffness sensing together regulate vascular smooth muscle cell phenotype switching [ Sci Adv, 2022, 8(15):eabm3471] PubMed: 35427166
The Rho kinase 2 (ROCK2)-specific inhibitor KD025 ameliorates the development of pulmonary arterial hypertension [ Biochem Biophys Res Commun, 2021, 534:795-801] PubMed: 33160621
Selective dysregulation of ROCK2 activity promotes aberrant transcriptional networks in ABC diffuse large B-cell lymphoma [ Sci Rep, 2020, 10(1):13094] PubMed: 32753663
KD025, an anti-adipocyte differentiation drug, enhances the efficacy of conventional chemotherapeutic drugs in ABCG2-overexpressing leukemia cells [ Oncol Lett, 2020, 20(6):309] PubMed: 33093918
Integrated proximal proteomics reveals IRS2 as a determinant of cell survival in ALK-driven neuroblastoma. [ Sci Signal, 2018, 11(557)] PubMed: 30459283

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SHIPPING AND STORAGE
Selleck products are transported at room temperature. If you receive the product at room temperature, please rest assured, the Selleck Quality Inspection Department has conducted experiments to verify that the normal temperature placement of one month will not affect the biological activity of powder products. After collecting, please store the product according to the requirements described in the datasheet. Most Selleck products are stable under the recommended conditions.

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