DDR1-IN-1

Catalog No.S7498 Batch:S749801

Technical Data

Formula

C₃₀H₃₁F₃N₄O₃

Molecular Weight

552.59

CAS No.

1449685-96-4

Solubility (25°C)*

In vitro

DMSO

100 mg/mL (180.96 mM)

Ethanol

6 mg/mL (10.85 mM)

Water

Insoluble

In vivo (Add solvents to the product individually and in order)

Clear solution	5%DMSO 1 40%PEG300 2 5%Tween80 3 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 clarify; 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.
Clear solution	5% DMSO 1 95% Corn oil	7.5mg/ml	Taking the 1 mL working solution as an example, add 50 μL of 150 mg/ml clear DMSO stock solution to 950 μL of corn oil and mix evenly. The mixed solution should be used immediately for optimal results.
Clear solution	8%DMSO 1 92%Corn oil	7.5mg/ml	Taking the 1 mL working solution as an example, add 80 μL of 93.75 mg/ml clear DMSO stock solution to 920 μL of corn oil and mix evenly. 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

DDR1-IN-1 is a potent and selective discoidin domain receptor 1 (DDR1) receptor tyrosine kinase inhibitor with IC50 of 105 nM, about 3-fold selectivity over DDR2.

Targets

DDR1 ^[1] (Cell-free assay)	DDR2 ^[1] (Cell-free assay)
105 nM	413 nM

In vitro

In U2OS cells, DDR1-IN-1 inhibits basal DDR1 autophosphorylation with EC50 of 86 nM, and demonstrates stronger inhibition of DDR1 autophosphorylation in the absence of collagen stimulation. In a panel of different cancer cell lines that possess DDR1 gain-of-function mutations and/or overexpression, DDR1-IN-1, dose not inhibit proliferation below a concentration of 10 μM, while GSK2126458 potentiates the antiproliferative activity of DDR1-IN-1. ^[1]

Protocol (from reference)

Kinase Assay:^{^[1]}	General procedure for the EC50 test DDR1 is induced for 48 hrs prior to DDR1 activation by rat tail collagen I. The DDR1 over-expressed U2OS is pre-treated by media containing each concentration of the compound for 1 hr and treated by changing the media to the EC50 test media containing 10 Gg/ml collagen and each concentration of the compound for 2 hrs. Each cells is washed with cold PBS three times and lysed with the lysis buffer (50 mMTris, pH 7.5, 1% Triton X-100, 0.1% SDS, 150 mM NaCl, 5 mM EDTA, 100 mMNaF, 2 mM Na₃VO₄, 1 mM PMSF, and 10 Gg/ml leupeptin). The activation of DDR1 is quantified by density using program ImageJ to determine EC50 following Western blot using anti-activated human DDR1b (Y513).
Cell Assay:^{^[1]}	Cell lines U2OSWT, U2OSOWT and U2OSG707A cell lines Concentrations ~20 μM Incubation Time 48 h Method Cells are plated in triplicate at a density of 3000 cells per well in 96-well plates and 1500 cells per well in 384-well plates. Compounds of various concentrations are added into plates for 48 hours. Cell viability is determined using the CellTiter-Glo and CCK-8. Both assays are performed according to the manufacturer’s instructions. For CellTiter-Glo assay, luminescence is determined in a multi-label reader. For CCK-8 assay, absorbance is measured in a microplate reader at 450nm. Data are normalized to control group (DMSO) and represented by the mean of at least two independent measurement with standard error <20%. GI50 were calculated using Prism 5.0.

Kinase Assay:^{^[1]}

General procedure for the EC50 test
DDR1 is induced for 48 hrs prior to DDR1 activation by rat tail collagen I. The DDR1 over-expressed U2OS is pre-treated by media containing each concentration of the compound for 1 hr and treated by changing the media to the EC50 test media containing 10 Gg/ml collagen and each concentration of the compound for 2 hrs. Each cells is washed with cold PBS three times and lysed with the lysis buffer (50 mMTris, pH 7.5, 1% Triton X-100, 0.1% SDS, 150 mM NaCl, 5 mM EDTA, 100 mMNaF, 2 mM Na₃VO₄, 1 mM PMSF, and 10 Gg/ml leupeptin). The activation of DDR1 is quantified by density using program ImageJ to determine EC50 following Western blot using anti-activated human DDR1b (Y513).

Cell Assay:^{^[1]}

Cell lines
U2OSWT, U2OSOWT and U2OSG707A cell lines
Concentrations
~20 μM
Incubation Time
48 h
Method
Cells are plated in triplicate at a density of 3000 cells per well in 96-well plates and 1500 cells per well in 384-well plates. Compounds of various concentrations are added into plates for 48 hours. Cell viability is determined using the CellTiter-Glo and CCK-8. Both assays are performed according to the manufacturer’s instructions. For CellTiter-Glo assay, luminescence is determined in a multi-label reader. For CCK-8 assay, absorbance is measured in a microplate reader at 450nm. Data are normalized to control group (DMSO) and represented by the mean of at least two independent measurement with standard error <20%. GI50 were calculated using Prism 5.0.

References

[1] Kim HG, et al. ACS Chem Biol. 2013, 8(10), 2145-2150.

Customer Product Validation

: Data from [Data independently produced by , , Nature Materials, 2017, 16:1252-1261.]

Selleck's DDR1-IN-1 has been cited by 8 publications

MiR-4458-loaded gelatin nanospheres target COL11A1 for DDR2/SRC signaling pathway inactivation to suppress the progression of estrogen receptor-positive breast cancer [ Biomater Sci, 2022, 10.1039/d2bm00543c]	PubMed: 35792605
Targeting Discoidin Domain Receptors DDR1 and DDR2 overcomes matrix-mediated tumor cell adaptation and tolerance to BRAF-targeted therapy in melanoma [ EMBO Mol Med, 2021, e11814]	PubMed: 34957688
Crosstalk between invadopodia and the extracellular matrix [ Eur J Cell Biol, 2020, 99(7):151122]	PubMed: 33070041
Collagen-rich airway smooth muscle cells are a metastatic niche for tumor colonization in the lung [ Nat Commun, 2019, 10(1):2131]	PubMed: 31086186
Discoidin domain receptors: A promising target in melanoma. [ Pigment Cell Melanoma Res, 2019, 32(5):697-707]	PubMed: 31271515
[ Cell Death Dis, 2018, ]	PubMed: 29988031
Mechanotransduction-modulated fibrotic microniches reveal the contribution of angiogenesis in liver fibrosis. [ Nat Mater, 2017, 16(12):1252-1261]	PubMed: 29170554
Mechanotransduction-modulated fibrotic microniches reveal the contribution of angiogenesis in liver fibrosis. [Longwei Liu, et al. NAT MATER, 2017, 10.1038/NMAT5024]

RETURN POLICY
Selleck Chemical’s Unconditional Return Policy ensures a smooth online shopping experience for our customers. If you are in any way unsatisfied with your purchase, you may return any item(s) within 7 days of receiving it. In the event of product quality issues, either protocol related or product related problems, you may return any item(s) within 365 days from the original purchase date. Please follow the instructions below when returning products.

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.

NOT FOR HUMAN, VETERINARY DIAGNOSTIC OR THERAPEUTIC USE.