Nodinitib-1 (ML130)

Catalog No.S2863 Batch:S286301

Technical Data

Formula	C₁₄H₁₃N₃O₂S
Molecular Weight	287.34	CAS No.	799264-47-4
Solubility (25°C)*	In vitro	DMSO	57 mg/mL (198.37 mM)
		Ethanol	2 mg/mL (6.96 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

Nodinitib-1(ML130) is a potent and selective inhibitor of NOD1 with IC50 of 0.56 μM, inhibits NF-κB activation, exhibits 36-fold selectivity over NOD2.

Targets

NOD1 ^[1]

0.56 μM

In vitro

ML130 has shown selective inhibition of NOD1-induced NF-κB activation in HEK293 cells with no cytotoxicity and is selected as a probe candidate molecule. ML130 is also confirmed in secondary assays by selectively inhibiting NOD1-dependent IL-8 secretion and also selectively inhibiting the NOD1-dependent pathway to NF-κB activation. ^[1] In another research, ML130 is proved to cause conformational changes of NOD1 in vitro and alter NOD1 subcellular targeting in cells, providing chemical probes for interrogating mechanisms regulating NOD1 activity and tools for exploring the roles of NOD1 in various infectious and inflammatory diseases. ^[2]

Protocol (from reference)

Kinase Assay:^{^[1]}	NOD1 Dose Response assay Day 1 Procedure 1) Harvest HEK-293-T NFKB-Luc at 100% confluency at 100% confluency. 2) Add NOD assay media with Multidrop. 3) Spin down plates at 1000 rpm for 1 min in centrifuge. 4) Serial compound dilutions. 5) Add gamma-tri-DAP to cell suspension at 0.75 ug/mL. 6) Seed 13000 cells/well in 4 uL/well to full plate HEK-293-T NFKB-Luc to TC-treated plate. 7) Spin down plates 500 RPM for 5 min on centrifuge. 8) Lid Plates. Sandwich 4 plates between 2 lidded 384 plates filled with H2O. 9) Wrap plates securely in single layer of Plastic Wrap. 10) Incubate overnight (14 hours) in 37 ℃ and 5% CO₂ incubator. Day 2 Procedure 1) Add 3 ul/well of SteadyGlo solution with Multidrop. 2) Shake plates on a plate shaker for 20 min. 3) Spin plates 1000 RPM for 1 min using centrifuge. 4) Read luminescence.IC50 values are calculated using GraphPad Prism 5.0.The average Z
Cell Assay:^{^[1]}	Cell lines Fa2N-4 cells Concentrations 0.01 μM-50 μM Incubation Time 24 h Method Hepatic toxicity of compounds is determined with Fa2N-4 immortalized human hepatocytes using the ATP-lite 1-step assay. Fa2N-4 cells are seeded at 50,000 cells/well, and incubated with a range of concentrations of the test compound (0.01 µM-50 µM) in MFE support medium for 24 h at 37 ℃, 5% CO₂. At the end of the experiment D-luciferin and luciferase are added. The emitted luminescent signal produced as a result of the reaction with cellular ATP is captured on the Infinite M200 plate reader. The concentration of each compound that killed 50% of the cells (LC50) is calculated by non-linear regression analysis using a log (inhibitior) vs response equation with a variable slope, using the statistic software package Prism4.

Kinase Assay:^{^[1]}

NOD1 Dose Response assay
Day 1 Procedure 1) Harvest HEK-293-T NFKB-Luc at 100% confluency at 100% confluency. 2) Add NOD assay media with Multidrop. 3) Spin down plates at 1000 rpm for 1 min in centrifuge. 4) Serial compound dilutions. 5) Add gamma-tri-DAP to cell suspension at 0.75 ug/mL. 6) Seed 13000 cells/well in 4 uL/well to full plate HEK-293-T NFKB-Luc to TC-treated plate. 7) Spin down plates 500 RPM for 5 min on centrifuge. 8) Lid Plates. Sandwich 4 plates between 2 lidded 384 plates filled with H2O. 9) Wrap plates securely in single layer of Plastic Wrap. 10) Incubate overnight (14 hours) in 37 ℃ and 5% CO₂ incubator. Day 2 Procedure 1) Add 3 ul/well of SteadyGlo solution with Multidrop. 2) Shake plates on a plate shaker for 20 min. 3) Spin plates 1000 RPM for 1 min using centrifuge. 4) Read luminescence.IC50 values are calculated using GraphPad Prism 5.0.The average Z

Cell Assay:^{^[1]}

Cell lines
Fa2N-4 cells
Concentrations
0.01 μM-50 μM
Incubation Time
24 h
Method
Hepatic toxicity of compounds is determined with Fa2N-4 immortalized human hepatocytes using the ATP-lite 1-step assay. Fa2N-4 cells are seeded at 50,000 cells/well, and incubated with a range of concentrations of the test compound (0.01 µM-50 µM) in MFE support medium for 24 h at 37 ℃, 5% CO₂. At the end of the experiment D-luciferin and luciferase are added. The emitted luminescent signal produced as a result of the reaction with cellular ATP is captured on the Infinite M200 plate reader. The concentration of each compound that killed 50% of the cells (LC50) is calculated by non-linear regression analysis using a log (inhibitior) vs response equation with a variable slope, using the statistic software package Prism4.

References

Customer Product Validation

: , , Gut, 2016, 66(6):1060-1073

: Data from [Data independently produced by , , Vet Immunol Immunopathol, 2015, 168(1-2):68-76]

Selleck's Nodinitib-1 (ML130) has been cited by 9 publications

Adh Promotes Actinobacillus pleuropneumoniae Survival in Porcine Alveolar Macrophages by Inhibiting CHAC2-Mediated Respiratory Burst and Inflammatory Cytokine Expression [ Cells, 2023, 12(5)696]	PubMed: 36899832
NOD1/RIP2 signalling enhances the microglia-driven inflammatory response and undergoes crosstalk with inflammatory cytokines to exacerbate brain damage following intracerebral haemorrhage in mice [ J Neuroinflammation, 2020, 17(1):364]	PubMed: 33261639
A Specific Strain of Lactic Acid Bacteria, Lactobacillus paracasei, Inhibits Inflammasome Activation In Vitro and Prevents Inflammation-Related Disorders [ J Immunol, 2020, 205(3):811-821]	PubMed: 32591398
Ginsenoside Rg3 protects against iE-DAP-induced endothelial-to-mesenchymal transition by regulating the miR-139-5p-NF-κB axis. [ J Ginseng Res, 2020, 44(2):300-307]	PubMed: 32148412
Suppression of αvβ6 Integrin Expression by Polymicrobial Oral Biofilms in Gingival Epithelial Cells. [Bi J, et al. Sci Rep, 2017, 30;7(1):4411]	PubMed: 28667248
The Helicobacter pylori type IV secretion system promotes IL-8 synthesis in a model of pediatric airway epithelium via p38 MAP kinase [Dela Pena-Ponce MG, et al. PLoS One, 2017, 12(8):e0183324]	PubMed: 28813514
Impaired antibacterial autophagy links granulomatous intestinal inflammation in Niemann–Pick disease type C1 and XIAP deficiency with NOD2 variants in Crohn’s disease [Schwerd T, et al. Gut, 2016, 10.1136/gutjnl-2015-310382]	PubMed: 26953272
Role of the NOD1/NF-κB pathway on bovine neutrophil responses to crude lipopolysaccharide [Wei LJ, et al. Vet J, 2016, 214:24-31]	PubMed: 27387722
Effector responses of bovine blood neutrophils against Escherichia coli: Role of NOD1/NF-κB signalling pathway [Tan X, et al. Vet Immunol Immunopathol, 2015, 168(1-2):68-76]	PubMed: 26321220

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