DMH1 1206711-16-1

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Molecular Weight:

380.44 DMH1 is a selective BMP receptor inhibitor with IC50 of 107.9 nM for ALK2, exhibiting no inhibition on AMPK, ALK5, KDR (VEGFR-2) or PDGFR.

Biological Activity

DMH1 inhibits BMP signaling with IC50 of 100 nM, and selectively inhibits the BMP-induced Smad1/5/8 activation. DMH1 increases cardiomyocyte progenitors and promotes cardiac differentiation in mouse embryonic stem cells. In addition, DMH1 as a BMP inhibitor, significantly reduces NSCLC cell growth, migration and invasion.


DMH1 dorsalizes the embryonic axis without disrupting the angiogenic process in Zebrafish embryos. In proepicardial explants, DMH1 results in the greatest inhibition of epithelial sheet migration. DMH1 (5 mg/kg i.p.)attenuates xenograft lung tumor growth in mice bearing A549 xenograft.






Kinase assay


All kinase assays are conducted by Reaction Biology Corp. In brief,
compounds are tested at 10 concentrations by 3-fold serial dilutions
starting at 30 μM, using nonspecific kinase inhibitor staurosporine as
control. In vitro kinase reactions are carried out in the presence of 10
μM (33P)γATP.


Five kinases tested are the human BMP type-I
receptor activin receptor-like kinase 2 (ALK-2/ACVR1), the human TGFβ
type-I receptor activin receptor-like kinase 5 (Alk5/TGFβR1), the human
VEGF type-II receptor (KDR/Flk-1/VEGFR2), the human AMP-activated
protein kinase (AMPK/A1/B1/G1) and the human platelet-derived growth
factor receptor-β (PDGFRβ).






Method


About 10,000 A549 cells per well are seeded in 96-well plates and
incubated for overnight. The culture medium is then changed to fresh
medium containing DMSO or DMH1 at various concentrations. The cells are
then incubated for 48 hours and 96 hours before treatment termination by
replacing the medium with 100 μL of 10% trichloroacetic acid in 1× PBS,
followed by incubation at 4°C for at least 1 hour.


Subsequently, the
plates are washed with water and air dried. The plates are stained with
50 μL 0.4% sulphorhodamine assay in 1% acetic acid for 30 minutes at
room temperature. Unbound dye is washed off with 1% acetic acid. After
air drying and solubilization of the protein-bound dye in 10 mM Tris
solution, absorbance is read in a microplate reader at 565 nm.

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Product Categories : TGF-beta/Smad