LDN-214117

Catalog No.S7627 Batch:S762702

Technical Data

Formula	C₂₅H₂₉N₃O₃
Molecular Weight	419.52	CAS No.	1627503-67-6
Solubility (25°C)*	In vitro	DMSO	84 mg/mL (200.22 mM)
		Ethanol	21 mg/mL (50.05 mM)
		Water	Insoluble
* <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.)

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Biological Activity

Description

LDN-214117 is a potent and selective BMP type I receptor kinase ALK2 inhibitor with IC50 of 24 nM.

Targets

ALK2 ^[1]
(Cell-free assay)

24 nM

In vitro

The kinase most highly inhibited by LDN-214117 is ALK2 (IC50, 24 nM), followed by TNIK, RIPK2, and ABL1. LDN-214117 demonstrates selective inhibition of ALK2 and ALK1 in preference to ALK3 kinase activity. LDN-214117 exhibits relatively selective inhibition of BMP6 versus BMP2 or BMP4. In a cell-based assay, LDN-214117 exhibits a selective inhibition on BMP6 with IC50 of approximately 100 nM, and 164-fold selectivity for BMP6 versus TGF-β1. ^[1]

Protocol (from reference)

Kinase Assay:^{^[1]}	ALK2 kinase assay Puriﬁed recombinant ALK2 proteins, ATP, ATP[γ-³²P], and dephosphorylated casein at ﬁnal concentrations of 2.5 nM, 6 μM, 0.05 μCi/μL , and 0.5 mg/mL, respectively, are aliquoted in kinase buﬀer containing 0.2% BSA supplemented with 10 mM MnCl₂ into 96-microwell plates, in combination with inhibitor compounds diluted at varying concentrations (0.01 nM to 100 μM). Positive control samples lacking inhibitor compounds, and negative controls lacking recombinant kinase, are also measured. The mixture is reacted at RT for 45 min, quenched with a ﬁnal concentration of 2% phosphoric acid. The reaction mixture is transferred to 96-well P81 phosphocellulose ﬁlter plates and bound for 5 min. The plates are washed 20 times with 150 μL of 1% phosphoric acid solution per well by vacuum manifold. Plates are dried at RT for 1 h, sealed, and assayed with Microscint 20 scintillation ﬂuid using a Spectramax L luminometer. Data is normalized to positive controls at 100% enzyme activity, with negative controls being subtracted as background.
Cell Assay:^{^[1]}	Cell lines HepG2 hepatocarcinoma cells Concentrations 100 μM Incubation Time 24 h Method Cells are seeded at 25000 cells per well in 96-well plates and incubated for 2 h at 37℃ and 5% CO₂. Compounds of LDN-214117 or DMSO are diluted in DMEM and added at ﬁnal compound concentrations of 1, 10, and 100 μM. Cells are incubated for 4 and 24 h, after which the media is discarded. Cells are lysed by adding 30 μL of passive lysis buﬀer and shaken at RT for 15 min. Cell viability is determined by quantifying the ATP present in each well by adding 10 μL of Cell Titer Glo per well and measuring the light output by Spectramax L luminometer. Data is normalized to 100% viability for cells receiving only DMSO.

Kinase Assay:^{^[1]}

ALK2 kinase assay
Puriﬁed recombinant ALK2 proteins, ATP, ATP[γ-³²P], and dephosphorylated casein at ﬁnal concentrations of 2.5 nM, 6 μM, 0.05 μCi/μL , and 0.5 mg/mL, respectively, are aliquoted in kinase buﬀer containing 0.2% BSA supplemented with 10 mM MnCl₂ into 96-microwell plates, in combination with inhibitor compounds diluted at varying concentrations (0.01 nM to 100 μM). Positive control samples lacking inhibitor compounds, and negative controls lacking recombinant kinase, are also measured. The mixture is reacted at RT for 45 min, quenched with a ﬁnal concentration of 2% phosphoric acid. The reaction mixture is transferred to 96-well P81 phosphocellulose ﬁlter plates and bound for 5 min. The plates are washed 20 times with 150 μL of 1% phosphoric acid solution per well by vacuum manifold. Plates are dried at RT for 1 h, sealed, and assayed with Microscint 20 scintillation ﬂuid using a Spectramax L luminometer. Data is normalized to positive controls at 100% enzyme activity, with negative controls being subtracted as background.

Cell Assay:^{^[1]}

Cell lines
HepG2 hepatocarcinoma cells
Concentrations
100 μM
Incubation Time
24 h
Method
Cells are seeded at 25000 cells per well in 96-well plates and incubated for 2 h at 37℃ and 5% CO₂. Compounds of LDN-214117 or DMSO are diluted in DMEM and added at ﬁnal compound concentrations of 1, 10, and 100 μM. Cells are incubated for 4 and 24 h, after which the media is discarded. Cells are lysed by adding 30 μL of passive lysis buﬀer and shaken at RT for 15 min. Cell viability is determined by quantifying the ATP present in each well by adding 10 μL of Cell Titer Glo per well and measuring the light output by Spectramax L luminometer. Data is normalized to 100% viability for cells receiving only DMSO.

References

[1] Mohedas AH, et al. J Med Chem. 2014, 57(19), 7900-7915.

Selleck's LDN-214117 has been cited by 4 publications

Live-Cell Invasive Phenotyping Uncovers ALK2 as a Therapeutic Target in LKB1-Mutant Lung Cancer [ Cancer Res, 2024, 10.1158/0008-5472.CAN-23-2631]	PubMed: 39207369
Live-cell invasive phenotyping uncovers the ALK2/BMP6 iron homeostasis pathway as a therapeutic vulnerability in LKB1-mutant lung cancer [ bioRxiv, 2023, 2023.06.14.544941]	PubMed: 37398244
Fluid shear stress generates a unique signaling response by activating multiple TGFβ family type I receptors in osteocytes [ FASEB J, 2021, 35(3):e21263]	PubMed: 33570811
Dual Inhibition of BMP and WNT Signals Promotes Pancreatic Differentiation from Human Pluripotent Stem Cells. [ Stem Cells Int, 2019, 2019:5026793]	PubMed: 31885612

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