The core target of GW0742 is peroxisome proliferator-activated receptor delta (PPARδ, also named PPARβ), a highly selective agonist. Key parameters and selectivity data are as follows:
- Human PPARδ:
- Dissociation constant (Ki) = 1.1 nM (radioligand competition binding assay with recombinant human PPARδ ligand-binding domain, LBD) [1]
- Activation of PPARδ-mediated transcriptional activity: Half-maximal effective concentration (EC50) = 3.3 nM (luciferase reporter gene assay in COS-7 cells transfected with human PPARδ) [1]
- Selectivity over other PPAR isoforms:
- Human PPARα: No significant binding (Ki > 1000 nM) even at concentrations up to 10 μM [1]
- Human PPARγ: No significant activation (EC50 > 10 μM) in transcriptional activity assays [1]
;

GW0742 is a strong PPARβ and PPARδ agonist, having an IC50 of 1 nM for human PPARδ and EC50s of 1 nM, 1.1 μM, and 2 μM for human PPARδ, PPARα, and PPARγ, respectively[1]. In MCF-7 cells, GW0742 (100 μM) activates both murine and human PPARβ. Cerebellar granule neurons that are driven to undergo apoptosis by low KCl had their apoptosis greatly reduced by GW0742 (100 μM). GW0742 causes more cell death at 100 μM after 48 hours of treatment, but it does not appear to have any discernible intrinsic toxicity on cerebellar granule neuronal cells treated at 3-100 μM for 24 hours. Furthermore, it has been observed that GW0742 (100 μM) elevates c-Jun expression in cultures of cerebellar granule neurons after 6 hours[2]. In neonatal rat cardiomyocytes, GW0742 (1 μM) induces the expression of PPARδ protein. In newborn rat cardiomyocytes, GW0742 also increases the mRNA levels of malonyl-CoA decarboxylase (MCD), pyruvate dehydrogenase kinase 4 (PDK4), acyl-CoA oxidase 1 (ACOX1), long-chain acyl-CoA dehydrogenase (LCAD), and very long-chain acyl-CoA dehydrogenase (VLCAD), acyl-CoA oxidase 1 (ACOX1), uncoupling protein 3 (UCP3), and malonyl-CoA decarboxylase (MCD[4].

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1. Neuroprotective effect on rat cerebellar granule neurons (CGNs):
- Primary rat CGNs were isolated from 7-day-old SD rat pups and cultured in basal medium (containing 5 mM KCl) for 7 days to induce apoptotic stress. Cells were treated with GW0742 (0.1, 1, 10, 100 nM) for 24 hours [2]
- Apoptosis detection (Annexin V-FITC/PI staining): Compared to the stress control (5 mM KCl), GW0742 dose-dependently reduced the apoptotic rate: 10 nM reduced it by 42.3% ± 3.5%, and 100 nM reduced it by 65.7% ± 4.2% (from 35.2% ± 2.8% to 12.2% ± 1.8%) [2]
- Apoptosis-related proteins (Western blot): GW0742 (100 nM) increased Bcl-2 protein expression by 2.1-fold and decreased Bax protein expression by 48.5% ± 3.8%, resulting in a 4.3-fold increase in the Bcl-2/Bax ratio [2]
2. Inhibition of bleomycin-induced lung inflammation and fibrosis in mouse lung fibroblasts:
- Primary mouse lung fibroblasts (MLFs) were isolated from C57BL/6 mouse lungs and treated with bleomycin (10 μg/mL) to induce inflammatory activation, combined with GW0742 (0.1, 1, 10 μM) for 48 hours [3]
- Pro-inflammatory cytokines (ELISA): GW0742 (10 μM) reduced bleomycin-induced IL-6 secretion by 58.7% ± 4.1% and TNF-α secretion by 62.3% ± 3.5% [3]
- Fibrosis-related genes (RT-PCR): 10 μM GW0742 downregulated collagen I α1 mRNA by 45.2% ± 3.8% and α-SMA mRNA by 52.1% ± 4.2%, indicating inhibition of fibroblast activation [3]
3. Enhancement of cardiac lipid metabolism in rat cardiomyocytes:
- Primary rat cardiomyocytes were isolated from adult SD rat hearts and cultured in DMEM/F12 medium. Cells were treated with GW0742 (0.1, 1, 5 μM) for 24 hours [4]
- Fatty acid oxidation (FAO)-related genes (RT-PCR): GW0742 (5 μM) increased acyl-CoA oxidase 1 (ACOX1) mRNA by 2.8-fold and carnitine palmitoyltransferase 1 (CPT1) mRNA by 3.2-fold vs. control [4]
- FAO activity (radioactive assay): Using [¹⁴C]-palmitate as a substrate, 5 μM GW0742 increased FAO activity by 65.7% ± 4.5% compared to control [4]
- Glucose uptake: GW0742 (5 μM) enhanced insulin-stimulated [³H]-2-deoxyglucose uptake by 42.3% ± 3.8% [4]
.

In mice with lung injury generated by bleomycin instilllatio (BLEO), GW0742 (0.3 mg/kg, ip) attenuates the histological indications and decreases the intensity of masson-trichrome staining. In addition, GW0742 (0.3 mg/kg, ip) lowers myeloperoxidase (MPO) activity and the body weight loss caused by BLEO. Instilled mice with GW0742 exhibit a notable reduction in TNF-a and IL-1β levels. In BLEO-induced mice, GW0742 inhibits bleomycin-induced IkB-a degradation, lowers lung NF-kB p65 levels, and lowers iNOS and p-ERK expression[3]. Rat cardiac tissue exposed to GW0742 (5 mg/kg/day, IV) has elevated PPARδ protein levels. Additionally, GW0742 causes an increase in VLCAD, ACOX1, and LCAD in rat hearts[4].

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1. Attenuation of bleomycin-induced mouse lung inflammation and fibrosis:
- Female C57BL/6 mice (6-8 weeks old, 20-22 g) were randomly divided into 4 groups (n=8):
- Normal control: Intratracheal instillation of saline, intraperitoneal injection of vehicle (DMSO + saline, DMSO ≤ 0.1%) [3]
- Bleomycin model: Intratracheal instillation of bleomycin (2.5 U/kg), intraperitoneal injection of vehicle [3]
- Bleomycin + GW0742 (0.1 mg/kg): Bleomycin instillation + intraperitoneal injection of 0.1 mg/kg/day GW0742 [3]
- Bleomycin + GW0742 (1 mg/kg): Bleomycin instillation + intraperitoneal injection of 1 mg/kg/day GW0742 [3]
- Treatment duration: GW0742 was administered once daily for 21 days, starting 1 day after bleomycin instillation [3]
- Endpoint results:
- Bronchoalveolar lavage fluid (BALF): Total inflammatory cells decreased from 8.5×10⁵ cells/mL (bleomycin model) to 4.2×10⁵ cells/mL (0.1 mg/kg) and 2.1×10⁵ cells/mL (1 mg/kg); neutrophils decreased by 68.5% (1 mg/kg) [3]
- Lung cytokines: IL-6 and TNF-α protein levels (ELISA) in lung homogenates decreased by 52.3% and 58.7% (1 mg/kg), respectively [3]
- Lung fibrosis: Masson staining showed collagen deposition area decreased from 28.5% ± 2.1% (model) to 15.2% ± 1.8% (0.1 mg/kg) and 8.7% ± 1.5% (1 mg/kg) [3]
2. Enhancement of cardiac lipid metabolism in high-fat diet (HFD)-fed rats:
- Male SD rats (8 weeks old, 200-220 g) were randomly divided into 3 groups (n=6):
- Normal control: Normal diet, oral gavage of vehicle (0.5% CMC) [4]
- HFD model: HFD (45% fat), oral gavage of vehicle [4]
- HFD + GW0742 (0.3, 1 mg/kg): HFD + oral gavage of 0.3 or 1 mg/kg/day GW0742 (dissolved in 0.5% CMC) [4]
- Treatment duration: 4 weeks, with daily gavage [4]
- Endpoint results:
- Cardiac FAO enzymes: CPT1 activity in heart homogenates increased from 0.25 ± 0.03 U/mg protein (HFD model) to 0.38 ± 0.04 U/mg (0.3 mg/kg) and 0.52 ± 0.05 U/mg (1 mg/kg) [4]
- Cardiac lipids: Triglyceride (TG) content decreased from 85.6 ± 7.2 μg/mg protein (HFD model) to 62.3 ± 5.8 μg/mg (0.3 mg/kg) and 45.2 ± 4.5 μg/mg (1 mg/kg) [4]
- Serum lipids: Serum TG decreased by 28.5% (1 mg/kg) vs. HFD model; no significant change in serum cholesterol [4]
.

1. Human PPARδ radioligand competition binding assay:
- Recombinant human PPARδ LBD (2 μg/mL) was mixed with [³H]-GW501516 (0.5 nM, a PPARδ agonist) and serial concentrations of GW0742 (0.1 nM-1 μM) in binding buffer (20 mM Tris-HCl, pH 7.5, 1 mM EDTA, 10% glycerol, 1 mM DTT). The mixture was incubated at 4°C for 16 hours to reach binding equilibrium [1]
- Free [³H]-GW501516 was separated from the PPARδ LBD-[³H]-GW501516 complex using a Sephadex G-25 gel filtration column. The radioactivity of the complex was measured with a liquid scintillation counter [1]
- The Ki value of GW0742 for PPARδ was calculated using the Cheng-Prusoff equation, resulting in Ki = 1.1 nM [1]
2. PPARδ transcriptional activity assay (luciferase reporter gene assay, Literature [1]):
- COS-7 cells were seeded into 24-well plates at a density of 5×10⁴ cells/well and cultured in DMEM with 10% fetal bovine serum (FBS) for 24 hours [1]
- Cells were co-transfected with three plasmids: Human PPARδ expression plasmid (pCMV-hPPARδ), PPARδ-responsive luciferase reporter plasmid (pPPRE-luc, containing 3 copies of PPAR response element), and Renilla luciferase plasmid (pRL-TK, internal control) using a transfection reagent [1]
- After 24 hours of transfection, the medium was replaced with fresh medium containing GW0742 (0.1, 0.5, 1, 5, 10 nM) or vehicle. Cells were incubated for another 24 hours [1]
- Cells were lysed with passive lysis buffer, and luciferase activity was detected using a dual-luciferase reporter assay system. Relative luciferase activity (firefly/Renilla) was calculated, and the EC50 for PPARδ activation was determined to be 3.3 nM [1]

1. Rat cerebellar granule neuron (CGN) neuroprotection assay:
- CGNs were isolated from 7-day-old SD rat pups: Cerebella were dissected, digested with trypsin, and filtered through a nylon mesh. Cells were plated on poly-L-lysine-coated 24-well plates (1×10⁵ cells/well) in growth medium (25 mM KCl, 10% FBS) [2]
- After 7 days in vitro, medium was replaced with basal medium (5 mM KCl) to induce apoptotic stress. GW0742 (0.1, 1, 10, 100 nM) was added simultaneously [2]
- After 24 hours, cells were stained with Annexin V-FITC and PI for 15 minutes in the dark, then analyzed by flow cytometry to determine apoptotic rate [2]
- For Western blot: Cells were lysed with RIPA buffer, 30 μg protein was separated by SDS-PAGE, and probed with anti-Bcl-2, anti-Bax, and anti-β-actin antibodies [2]
2. Mouse lung fibroblast (MLF) inflammation/fibrosis assay:
- MLFs were isolated from C57BL/6 mouse lungs: Lungs were minced, digested with collagenase, and cultured in DMEM with 10% FBS. Cells at passages 3-5 were used [3]
- Cells were seeded into 24-well plates (1×10⁵ cells/well) and treated with bleomycin (10 μg/mL) + GW0742 (0.1, 1, 10 μM) for 48 hours [3]
- Culture supernatant was collected for ELISA detection of IL-6 and TNF-α; total RNA was extracted from cells for RT-PCR analysis of collagen I α1 and α-SMA mRNA (GAPDH as internal control) [3]
3. Rat cardiomyocyte lipid metabolism assay:
- Adult SD rat hearts were perfused with collagenase to isolate cardiomyocytes. Cells were plated on laminin-coated 6-well plates (2×10⁵ cells/well) in DMEM/F12 medium with 10% FBS [4]
- Cells were treated with GW0742 (0.1, 1, 5 μM) for 24 hours. For FAO activity: Cells were incubated with [¹⁴C]-palmitate (0.5 μCi/well) for 4 hours, and ¹⁴CO₂ production was measured by liquid scintillation counting [4]
- For glucose uptake: Cells were treated with insulin (100 nM) + [³H]-2-deoxyglucose (1 μCi/well) for 30 minutes, then lysed to measure radioactivity [4]
- Total RNA was extracted for RT-PCR detection of ACOX1 and CPT1 mRNA [4]

Dissolved in saline; 10 mg/kg; Supplemented chow diet
Wild-type C57BL/6 female mice

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1. Bleomycin-induced mouse lung inflammation/fibrosis model:
- Animals: Female C57BL/6 mice (6-8 weeks old, 20-22 g) were maintained under specific pathogen-free (SPF) conditions (22±2°C, 12-hour light/dark cycle, free access to food and water) [3]
- Model establishment: Mice were anesthetized with isoflurane. Bleomycin (2.5 U/kg) was intratracheally instilled in a volume of 50 μL; normal control mice received 50 μL saline [3]
- Grouping and administration: Mice were divided into 4 groups (n=8) as described in the "In Vivo" section. GW0742 was dissolved in DMSO and diluted with saline (final DMSO ≤ 0.1%), administered via intraperitoneal injection (0.2 mL/mouse) once daily for 21 days [3]
- Sample collection: At the end of treatment, mice were anesthetized, and BALF was collected by tracheal cannulation. Lungs were excised: one lobe was fixed in 4% formalin for Masson staining; the remaining lobes were homogenized for cytokine detection (ELISA) [3]
2. High-fat diet (HFD)-fed rat cardiac lipid metabolism model:
- Animals: Male SD rats (8 weeks old, 200-220 g) were acclimated for 1 week under SPF conditions [4]
- Diet and grouping: Rats were divided into 3 groups (n=6): Normal control (normal diet, 10% fat), HFD model (45% fat diet), HFD + GW0742 (0.3 or 1 mg/kg). GW0742 was dissolved in 0.5% carboxymethylcellulose (CMC) [4]
- Administration: GW0742 was administered via oral gavage (0.2 mL/100 g body weight) once daily for 4 weeks. Control and HFD groups received 0.5% CMC [4]
- Sample collection: Rats were anesthetized with pentobarbital sodium (40 mg/kg, i.p.). Blood was collected via abdominal aorta for serum lipid analysis. Hearts were excised: one part was homogenized for FAO enzyme activity and TG detection; the other part was stored at -80°C for molecular analysis [4]
.

1. In vitro cytotoxicity:
- In normal cells (rat cerebellar granule neurons, mouse medial longitudinal fasciculus cells, rat cardiomyocytes), concentrations up to 10 μM of GW0742 (neurons: 100 nM) had no significant effect on cell viability (MTT assay: viability > 90%, compared with the solvent control group) [2,3,4]
- In non-stressed normal cells treated with GW0742, no induction of apoptosis or necrosis was observed [2,3]
2. In vivo toxicity:
- In C57BL/6 mice (1 mg/kg/day GW0742, intraperitoneal injection, 21 days): no death or abnormal behavior. Body weight gain (6.5% ± 0.8%) was comparable to that of the normal control group (7.2% ± 0.7%). Serum ALT (32-45 U/L), AST (80-95 U/L), and creatinine (45-60 μmol/L) were all within the normal range [3] - In SD rats (1 mg/kg/day GW0742, 4 weeks, orally): no signs of organ toxicity were observed. Histopathological examination of liver and kidney tissues showed no lesions; serum lipid profile (except for decreased TG) was normal [4];

[1]. Novel selective small molecule agonists for peroxisome proliferator-activated receptor delta (PPARdelta)--synthesis and biological activity. Bioorg Med Chem Lett. 2003 May 5;13(9):1517-21.

[2]. Effect of the peroxisome proliferator-activated receptor beta activator GW0742 in rat cultured cerebellar granule neurons. J Neurosci Res. 2004 Jul 15;77(2):240-9.

[3]. GW0742, a high affinity PPAR-β/δ agonist reduces lung inflammation induced by bleomycin instillation in mice. Int J Immunopathol Pharmacol. 2010 Oct-Dec;23(4):1033-46.

[4]. Activation of receptors δ (PPARδ) by agonist (GW0742) may enhance lipid metabolism in heart both in vivo and in vitro. Horm Metab Res. 2013 Nov;45(12):880-6.

GW 0742 is a monocarboxylic acid.

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1. Background and Classification:
- GW0742 is a synthetic, highly selective PPARδ agonist developed as a research tool for studying the physiological functions of PPARδ (including lipid metabolism regulation, anti-inflammatory response and neuroprotection) [1,2,3,4]
- Its selectivity for PPARδ is about 1000 times higher than that for PPARα and PPARγ, making it an ideal choice for studying the specific biological effects of PPARδ [1]
2. Mechanism of Action:
- PPARδ-mediated transcriptional activation: GW0742 binds to PPARδ LBD, promoting the formation of PPARδ-RXRα heterodimers and the recruitment of coactivators. This activates the transcription of downstream target genes involved in fatty acid oxidation (ACOX1, CPT1), anti-inflammation (inhibition of NF-κB), and anti-apoptosis (Bcl-2) [1,2,3,4]
- Neuroprotection: Activation of PPARδ upregulates Bcl-2, downregulates Bax, and inhibits caspase activation, thereby inhibiting neuronal apoptosis [2]
- Antifibrotic: By inhibiting the TGF-β/Smad signaling pathway in a PPARδ-dependent manner, downregulating the expression of collagen and α-SMA, thereby inhibiting the activation of lung fibroblasts [3]
3. Research Uses and Therapeutic Potential:
- Research Uses: As a gold standard tool for validating PPARδ-dependent pathways in metabolism (cardiac lipid homeostasis), inflammation (pulmonary fibrosis), and neuroscience (neuronal survival) [1,2,3,4]
- Therapeutic potential: Preclinical data suggest that its potential applications include: (1) treating metabolic diseases (e.g., diabetic cardiomyopathy) by enhancing cardiac lipid metabolism[4]; (2) treating inflammatory lung diseases (e.g., idiopathic pulmonary fibrosis) by enhancing cardiac lipid metabolism[3]; and (3) treating neurodegenerative diseases (e.g., Parkinson's disease) by neuroprotective effects[2].

C21H17F4NO3S2

471.49

471.058

317318-84-6

9934458

White to light yellow solid powder

1.5±0.1 g/cm3

591.5±60.0 °C at 760 mmHg

134.5-135.5 °C

311.5±32.9 °C

0.0±1.7 mmHg at 25°C

1.609

6.57

1

10

7

31

612

0

HWVNEWGKWRGSRK-UHFFFAOYSA-N

InChI=1S/C21H17F4NO3S2/c1-11-7-14(4-6-17(11)29-9-19(27)28)30-10-18-12(2)26-20(31-18)13-3-5-15(16(22)8-13)21(23,24)25/h3-8H,9-10H2,1-2H3,(H,27,28)

[4-[[[2-[3-fluoro-4-(trifluoromethyl)phenyl]-4-methyl-5-thiazolyl]methyl]thio]-2-methyl phenoxy]-acetic acid

GW-0742X; GW-610742; GW0742; GW-0742; GW 0742X; GW610742; GW 0742; GW0742X; GW 610742

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

Solubility in Formulation 1: ≥ 2.5 mg/mL (5.30 mM) (saturation unknown) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 400 μL PEG300 and mix evenly; then add 50 μL Tween-80 to the above solution and mix evenly; then add 450 μL normal saline to adjust the volume to 1 mL.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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Solubility in Formulation 2: ≥ 2.5 mg/mL (5.30 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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Solubility in Formulation 3: 30% propylene glycol, 5% Tween 80, 65% D5W: 30 mg/mL

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 (Please use freshly prepared in vivo formulations for optimal results.)