SB505124

Catalog No.S2186 Batch:S218602

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Technical Data

Formula

C20H21N3O2
Molecular Weight 335.4 CAS No. 694433-59-5
Solubility (25°C)* In vitro DMSO 67 mg/mL (199.76 mM)
Ethanol 67 mg/mL (199.76 mM)
Water Insoluble
In vivo (Add solvents to the product individually and in order)
Clear solution
5%DMSO 40%PEG300 5%Tween80 50%ddH2O
7.5mg/ml Taking the 1 mL working solution as an example, add 50 μL of 150 mg/ml clarified DMSO stock solution to 400 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to make it clear; then continue to add 500 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 SB505124 is a selective inhibitor of TGFβR for ALK4, ALK5 with IC50 of 129 nM and 47 nM in cell-free assays, respectively, also inhibits ALK7, but does not inhibit ALK1, 2, 3, or 6.
Targets
ALK5 [1]
(Cell-free assay)		 ALK4 [1]
(Cell-free assay)
47 nM 129 nM
In vitro SB505124 is identified as a reversible ATP competitive and selective ALK inhibitor of ALK4 and ALK5. SB505124 shows no toxicity to renal epithelial A498 cells at concentrations up to 100 μM for 48 hours, and blocks TGF-β–induced apoptosis of FaO cells and NRP 154 cells in a concentration-dependent manner. [1] In human umbilical vein endothelial cells (HUVEC), SB505124 (500 nM) blocks the changes of TGF-β1 on F-actin assembly and prevents ROS production induced by TGF-β. [2] By inhibiting TGF-beta1 signaling, SB505124 leads to decreased deferoxamine (DFO)-induced neurogenesis. [3] A recent study shows that SB505124 suppresses the migration and invasion of breast cancer MCF-7-M5 cells. [4]
In vivo In a rabbit GFS model, SB505124 decreased the intraocular pressure (IOP) levels and reduces subconjunctival cell infiltration and scarring at the surgical site in the GFS. [5] In tacrolimus (TAC)-treated mice and FK12EC KO mice, SB505124 prevents the activation of endothelial TGF-β receptors and induction of renal arteriolar hyalinosis. [6]

Protocol (from reference)

Kinase Assay:[1]
  • In Vitro Protein Kinase Assay

    Kinase assays are performed as described by Laping et al., 2002 using the kinase domain of ALK5 and full-length N-terminal fused GST-Smad3. Kinase assays are performed with 65 nM GST-ALK5 and 184 nM GST-Smad3 in 50 mM HEPES, 5 mM MgCl2, 1 mM CaCl2, 1 mM dithiothreitol, and 3 μM ATP. Reactions are incubated with 0.5 μCi of [33P]γATP for 3 hours at 30 °C. Phosphorylated protein is captured on P-81 paper , washed with 0.5% phosphoric acid, and counted by liquid scintillation. Alternatively, Smad3 or Smad1 protein is also coated onto FlashPlate Sterile Basic Microplates. Kinase assays are then performed in FlashPlates with same assay conditions using either the kinase domain of ALK5 with Smad3 as substrate or the kinase domain of ALK6 (BMP receptor) with Smad1 as substrate. Plates are washed three times with phosphate buffer and counted by TopCount.

Cell Assay:[1]
  • Cell lines

    A498, FaO and NRP 154 cells

  • Concentrations

    0-10 μM

  • Incubation Time

    48 hours

  • Method

    Cell viability is measured as described by Laping et al., 2002 or by using the modified tetrazolium salt WST-1. XTT assay: The cells are serum-deprived for 24 hours and then treated with SB505124 for 48 hours to assess the cellular toxicity. Cell viability is determined by incubating cells for 4 hours with XTT labeling and electron coupling reagent according to the manufacturer's directions. Live cells with active mitochondria produce an orange-colored product, formazan, which is detected using a plate reader at between A450 nm and A500 nm with a reference wavelength greater than 600 nm. The absorbance values correlate with the number of viable cells. Modified tetrazolium salt WST-1: Approximately 2000 cells are seeded in 96-well dishes in 100 μL of 0.2% FBS phenol red-free media overnight. The cells are treated with 50 μL of SB505124 (to achieve the final concentrations indicated) for 30 minutes before being treated with or without TGF-β1 and TNF-α to a final volume of 200 μL. Cell growth is measured at the indicated time points by incubating each well with 10 μL of WST-1 for 3 hours at 37 °C. Metabolically active cells cleave WST-1 to water-soluble formazan, which is directly quantitated with an enzyme-linked immunosorbent assay plate reader. Each experiment is done at least twice, and treatment for each cell line is done in triplicate.
Animal Study:[5]
  • Animal Models

    New Zealand White (NZW) rabbits after glaucoma filtration surgery (GFS).

  • Dosages

    Tablets containing 5 mg of SB505124.

  • Administration

    Administered via p.o.

Customer Product Validation

Data from [Data independently produced by , , Stem Cells, 2016, 34:2635-2647.]

Data from [Data independently produced by , , Osteoarthritis Cartilage, 2017, 25(11):1868-1879]

Data from [Data independently produced by , , Oncotarget, 2016, 7(47):77495-77507]

Data from [Data independently produced by , , Osteoarthritis Cartilage, 2016, 24(2):345-53]

Selleck's SB505124 has been cited by 34 publications

Single-cell transcriptomics of human cholesteatoma identifies an activin A-producing osteoclastogenic fibroblast subset inducing bone destruction [ Nat Commun, 2023, 14(1):4417] PubMed: 37537159
In vivo induction of activin A-producing alveolar macrophages supports the progression of lung cell carcinoma [ Nat Commun, 2023, 14(1):143] PubMed: 36650150
Single-cell transcriptomics of human cholesteatoma identifies an activin A-producing osteoclastogenic fibroblast subset inducing bone destruction [ Nat Commun, 2023, 14(1):4417] PubMed: 37537159
Macrophages promote anti-androgen resistance in prostate cancer bone disease [ J Exp Med, 2023, 220(4)e20221007] PubMed: 36749798
Unravelling the Basic Calcium Phosphate crystal-dependent chondrocyte protein secretome; a role for TGF-β signaling [ Osteoarthritis Cartilage, 2023, S1063-4584(23)00755-0] PubMed: 37075856
secDrug: a pipeline to discover novel drug combinations to kill drug-resistant multiple myeloma cells using a greedy set cover algorithm and single-cell multi-omics [ Blood Cancer J, 2022, 12(3):39] PubMed: 35264575
TNFα activation and TGFβ blockage act synergistically for smooth muscle cell calcification in patients with venous thrombosis via TGFβ/ERK pathway [ J Cell Mol Med, 2022, 26(16):4479-4491] PubMed: 35808901
Establishment and characterization of immortalized sweat gland myoepithelial cells [ Sci Rep, 2022, 12(1):7] PubMed: 34997030
Dual mitigation of immunosuppression combined with photothermal inhibition for highly effective primary tumor and metastases therapy [ Biomaterials, 2021, 274:120856] PubMed: 33984634
Inhibition of TGFβ improves hematopoietic stem cell niche and ameliorates cancer-related anemia [ Stem Cell Res Ther, 2021, 12(1):65] PubMed: 33461597

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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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