DN203316
DN203316 (PPARδ agonist 11) is a potent, selective, orally active PPARδ agonist (EC50 = 20 nM) with high selectivity over PPARα and PPARγ. DN203316 suppresses NF-κB-mediated inflammatory signaling, inhibits ferroptosis by upregulating xCT and GPX4, and attenuates STING-TBK1-IRF3-driven fibrogenic responses. DN203316 is useful for research on inflammatory disorders, metabolic dysfunction-associated steatohepatitis (MASH), and liver fibrosis.
For research use only. We do not sell to patients.
- CAS No.: 2982696-04-6
- Formula: C19H15F3N2O3S2
- Molecular Weight:440.46
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Storage:Powder -20°C, 3 years , 4°C, 2 years ; In solvent -80°C, 6 months , -20°C, 1 month
Biological Activity
Description
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hPPARδ 0.02 μM (EC50) |
hPPARα 8 μM (EC50) |
h-PPARγ >10 μM (EC50) |
GPX4 |
TBK1 |
IKKα |
IKKβ |
In Vitro
DN203316 exhibits potent and highly selective PPARδ agonistic activity in CV-1 cells (EC50 = 0.02 μM), with >400-fold selectivity over PPARα (EC50 = 8 μM) and PPARγ (EC50 > 10 μM), showing activity comparable to GW501516 (HY-10838)[1][2].
DN203316 (Compound 11) demonstrates excellent in vitro ADMET properties, including good metabolic stability in human, dog, rat, and mouse liver microsomes, minimal inhibition of major CYP and UGT enzymes, high plasma protein binding, good Caco-2 permeability, and no significant hERG cardiotoxicity liability[1].
DN203316 (12-50 μM; 24 h) does not affect viability or proliferation of Raw264.7 macrophages, indicating no cytotoxicity[1].
DN203316 (12, 25, 50 μM; pre-treated for 2 h, then co-treated with LPS (HY-D1056) for 10-24 h) dose-dependently inhibits IKKα/β phosphorylation, suppresses NF-κB signaling, reduces NF-κB luciferase activity, decreases NO production, and downregulates pro-inflammatory cytokines TNF-α and IL-6 in Raw264.7 macrophages[1].
DN203316 (1 μM; pre-treated for 1 h, then co-treated with cholesterol for 24 h) protects AML12 and HepG2 hepatocytes from cholesterol-induced cell death without cytotoxicity, significantly reduces lipid peroxidation (oxC11-BODIPY signal), and suppresses ferroptosis, an effect reversed by PPARδ knockdown[2].
DN203316 (1 μM; pre-treated for 1 h, then co-treated with cholesterol for 24 h) restores xCT and GPX4 expression at both mRNA and protein levels, increases intracellular GSH levels, and activates xCT promoter activity in a PPARδ-dependent manner, which is abolished by mutation of the PPARδ-binding motif[2].
DN203316 (1 μM; pre-treated for 1 h, then co-treated with cholesterol for 24 h) regulates gene expression in hepatocytes and HSCs, restoring antioxidant genes xCT and GPX4 in hepatocytes while suppressing fibrotic markers α-SMA, COL1A1, and fibronectin in HSCs exposed to ferroptotic hepatocyte conditioned medium[2].
DN203316 (1 μM; pre-treated for 1 h, then co-treated with cholesterol for 24 h) attenuates inflammatory and chemotactic signaling by reducing TNF-α, IL-6, and IL-1β secretion in Kupffer cells and suppressing IL-6 and CXCL12 upregulation in HSCs[2].
DN203316 (1 μM, pre-treated for 1 h, then co-treated with cholesterol for 24 h) inhibits release of exosomal dsDNA from ferroptotic hepatocytes and blocks activation of the STING-TBK1-IRF3 signaling pathway in HSCs; Rab27a knockdown and DNase I treatment further confirm the dependence on the exosomal dsDNA axis[2].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only. Further protocols information, click here.
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Cell Line:AML12 and HepG2 hepatocytes
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Concentration:1 µM
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Incubation Time:1 h, then co-treated with cholesterol for 24 h
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Result:Suppressed cholesterol-induced hepatocyte death without detectable toxicity.
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Cell Line:AML12 and HepG2 hepatocytes
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Concentration:1 µM
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Incubation Time:1 h, then co-treated with cholesterol for 24 h
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Result:Restored expression of xCT and GPX4 at protein and mRNA levels; increased GSH levels (PPARδ-dependent).
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Cell Line:AML12, HepG2 (hepatocytes), primary mouse/human HSCs (LX2), and primary cells isolated from treated mice
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Concentration:1 µM
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Incubation Time:1 h, then co-treated with cholesterol for 24 h
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Result:Restored mRNA levels of xCT and GPX4 in cholesterol-treated hepatocytes in a PPARδ dependent manner
Suppressed mRNA expression of inflammatory and fibrotic genes (α-SMA, COL1A1, fibronectin) in HSCs exposed to ferroptotic hepatocyte conditioned medium.
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Cell Line:Kupffer cells (KCs) and HSCs treated with hepatocyte-derived conditioned medium
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Concentration:1 µM
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Incubation Time:24 h
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Result:Significantly reduced secretion of pro-inflammatory cytokines TNF-α, IL-6, IL-1β from KCs.
Attenuated CM-induced upregulation of IL-6 and CXCL12 in HSCs, with effects comparable to Fer-1 treatment.
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Cell Line:AML12 and HepG2 hepatocytes
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Concentration:1 µM
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Incubation Time:1 h, then co-treated with cholesterol for 24 h
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Result:Significantly reduced cholesterol-induced lipid peroxidation (oxC11-BODIPY signal) in hepatocytes, abolished by PPARδ siRNA knockdown.
Parmacokinetics
In Vivo
DN203316 (3 mg/kg; i.p.; once daily for 5 weeks, week 9-14) in HFHC diet-induced MASH mice (C57BL/6) improves liver morphology and serum ALT/AST; reduces ferroptosis (MDA, 4-HNE, C11-BODIPY), restores redox balance (GSH, xCT, GPX4), decreases fibrosis (Sirius Red, α-SMA, COL1A1), and inhibits STING-TBK1-IRF3 signaling with reduced α-SMA/p-STING colocalization[2].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
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Animal Model:Carrageenan-induced Acute Inflammation and Macrophage Tracking Mice Model (6 weeks old, male, 18-20 g,C57BL/6; Carrageenan, subcutaneous injection into footpads; DiD-labeled primary peritoneal macrophages, intravenous injection (i.v.) via tail vein)[1]
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Dosage:50 mg/kg, single dose
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Administration:oral gavage (p.o.)
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Result:Showed aggressive infiltration of labeled macrophages into the carrageenan-injected paw in vehicle-treated mice.
Significantly reduced macrophage migration and infiltration into inflamed sites, resulting in markedly diminished fluorescence signals in the paw.
Significantly reduced paw edema and hind paw thickness in carrageenan-treated mice.
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Animal Model:HFHC Diet-induced MASH Mice Model (5-6 weeks old, male, C57BL/6 MASH model)[2]
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Dosage:3 mg/kg, once daily for 5 weeks (week 9-14)
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Administration:Intraperitoneal injection (i.p.)
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Result:Improved liver morphology with reduced liver weight and decreased serum ALT and AST levels, without affecting body weight or food intake.
Attenuated hepatic ferroptosis, as indicated by reduced MDA levels, 4-HNE accumulation, and C11-BODIPY fluorescence.
Restored hepatic redox balance by increasing GSH levels and upregulating antioxidant genes xCT and GPX4 in primary hepatocytes.
Reduced hepatic fibrosis, as evidenced by decreased Sirius Red staining and downregulated α-SMA and COL1A1 expression.
Inhibited activation of the STING–TBK1–IRF3 signaling pathway in hepatic stellate cells and reduced co-localization of α-SMA and p-STING in liver tissues.
Chemical Information
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CAS No. 2982696-04-6
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Appearance Solid
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Molecular Weight 440.46
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Formula C19H15F3N2O3S2
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Color White to off-white
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SMILES
CC1=CC(SCC(S2)=NN=C2C3=CC=C(C(F)(F)F)C=C3)=CC=C1OCC(O)=O
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Synonyms
PPARδ agonist 11
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Shipping
Room temperature in continental US; may vary elsewhere.
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Storage
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month
Protocols
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Fibrosis/Collagen Morphometry
Fibrosis and collagen morphometry is based on the quantitative visualization of fibrillar collagen deposition in tissue sections using histochemical stains such as Sirius Red (Picrosirius Red) or Masson's trichrome, followed by image-based or polarization-enhanced analysis to estimate collagen proportional area as a surrogate of extracellular matrix accumulation during fibrotic remodeling. Sirius Red combined with polarized light microscopy enhances detection of collagen fibers due to birefringence properties, enabling more specific visualization of collagen type I and III fibrils compared to conventional bright-field histology, while whole-section or region-restricted digital morphometry reduces field-selection bias in fibrosis assessment. Alternative quantitative approaches include second harmonic generation (SHG) and two-photon excited fluorescence microscopy, which enable label-free detection of fibrillar collagen and have been validated against histological staining and biochemica
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Collagen: Sirius Red Staining
Sirius Red or picrosirius red staining is a histochemical method for visualizing collagen-rich extracellular matrix in tissue sections, and collagen fibers are detected as red-stained structures under bright-field microscopy with enhanced birefringence under polarized light. Picrosirius red is useful for assessing total collagen organization, distribution, and fibrosis burden, but polarized color should not be interpreted as a definitive collagen type I versus type III readout because color is affected by fiber orientation, thickness, and packing.
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Connective Tissue: Masson's Trichrome/Collagen Trichrome Staining
Masson’s Trichrome (collagen/trichrome staining) is a histological technique that differentially stains tissue compartments using sequential acidic dyes to distinguish collagen from muscle and cytoplasmic components based on dye affinity and tissue permeability differences, enabling visualization of fibrosis and connective tissue architecture in histological sections. The classical formulation typically uses Weigert's iron hematoxylin for nuclear staining, Biebrich scarlet-acid fuchsin for cytoplasm and muscle, and aniline blue (or light green variants) for collagen, producing a characteristic blue/green collagen signal contrasted against red cytoplasm and dark nuclei. The staining principle relies on selective displacement of smaller dye molecules by larger anionic dyes in collagen-rich regions under controlled acidified conditions, which enhances collagen-specific dye retention. This property makes the method widely used for fibrosis assessment in organs such as heart, liver, lung, a
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Research Protocol for Inflammation-related Diseases
The NLRP3 inflammasome is a cytosolic innate immune signaling platform that integrates priming signals and danger-signal activation to promote caspase-1 activation, maturation of IL-1β and IL-18, and gasdermin D-mediated pyroptotic cell death. The core experimental logic is to determine whether inflammatory disease phenotypes are driven by increased NLRP3 expression, ASC-containing inflammasome assembly, caspase-1 cleavage, GSDMD cleavage, and extracellular release of IL-1β/IL-18 rather than by nonspecific cell injury alone. The pathway is strongly linked to inflammation-related disease phenotypes because monosodium urate crystals activate NALP3/NLRP3 inflammasome signaling in gout-like crystal inflammation, cholesterol crystals activate NLRP3 inflammasomes in atherogenesis models, and DSS-induced intestinal inflammation has been reported to involve NLRP3 inflammasome activity. However, experimental colitis studies also show context-dependent protective effects of NLRP3 inflammasome co
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Ferroptosis Solutions
Ferroptosis is an iron-dependent, non-apoptotic form of regulated cell death characterized by lethal lipid peroxidation and sensitivity to suppression by iron chelators or lipophilic radical-trapping antioxidants. The core pathway links cystine uptake through system Xc−, glutathione availability, GPX4-dependent detoxification of phospholipid hydroperoxides, iron-dependent oxidative reactions, and polyunsaturated-phospholipid metabolism into a cell-death program that is biochemically and morphologically distinct from apoptosis, necrosis, and autophagy. The ferroptosis pathway is experimentally linked to phenotype through chemical and genetic perturbation. Erastin induces ferroptosis by inhibiting cystine uptake through system Xc− and weakening antioxidant defenses, while GPX4 inhibition or depletion causes lipid peroxide accumulation and ferroptotic cancer-cell death. ACSL4 and oxidizable arachidonoyl- or adrenoyl-containing phosphatidylethanolamines shape ferroptosis sensitivity by con
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Liver Histomorphometry
Liver histomorphometry is a quantitative histological approach used to measure structural alterations in hepatic tissue, including parenchymal loss, steatosis, fibrosis, and vascular remodeling, by combining stained tissue section analysis with stereological or computerized image-based measurements. Classical morphometric frameworks quantify volume fractions of liver compartments and fibrotic regions using systematic sampling and image analysis, enabling objective comparison of pathological changes across experimental groups. These approaches are widely applied in liver cirrhosis and fibrosis studies to reduce subjectivity in histological scoring and improve reproducibility of tissue evaluation. Recent methodological advances integrate automated image analysis and radiomics-based extraction of histological features from standard liver stains (e. g. , H&E and fibrotic stains), enabling quantitative correlation between morphometric features and fibrosis stages in non-alcoholic fatty live
Purity & Documentation
References
[1]. Kim J, et al., Discovery of the therapeutic potential of PPARδ agonist bearing 1,3,4- thiadiazole in inflammatory disorders. Eur J Med Chem. 2024 Sep 12;279:116856. [Content Brief]
[2]. Kim YJ, et al. DN203316, a novel PPARδ agonist, suppresses ferroptotic signaling and fibrogenesis in metabolic dysfunction-associated steatohepatitis. Exp Mol Med. 2026 Jun 5. [Content Brief]
Calculators
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
Keywords
- DN203316
- 2982696-04-6
- PPARδ agonist 11
- DN 203316
- DN-203316
- PPARδ agonist11
- PPARδ agonist-11
- PPAR
- Ferroptosis
- Glutathione Peroxidase
- STING
- IKK
- NF-κB
- TNF Receptor
- PPARδ agonist
- orally active
- NF-κB inhibitor
- anti-inflammatory
- ferroptosis inhibitor
- xCT/GPX4 axis
- RAW264.7 cells
- AML12 cells
- HepG2 cells
- LX2 cells
- primary hepatocytes
- hepatic stellate cells
- carrageenan-induced paw edema
- carrageenan acute inflammation model
- cholesterol-induced ferroptosis
- HFHC diet MASH model
- metabolic dysfunction-associated steatohepatitis
- MASH
- liver fibrosis
- macrophage infiltration
- Inhibitor
- inhibitor
- inhibit