The only reported target of GSK3787 is peroxisome proliferator-activated receptor delta (PPARδ, also named PPARβ), a selective and irreversible antagonist. Key parameters and selectivity data are as follows:
- Human PPARδ:
- Dissociation constant (Ki) = 1.8 nM (radioligand competition binding assay with recombinant human PPARδ ligand-binding domain, LBD) [1]
- Inhibition of PPARδ-mediated transcriptional activity: Half-maximal inhibitory concentration (IC50) = 3.2 nM (luciferase reporter gene assay in CV-1 cells transfected with human PPARδ) [2]
- Selectivity over other PPAR isoforms:
- Human PPARα: Ki > 1000 nM (no significant binding at concentrations up to 1000 nM) [1,2]
- Human PPARγ: Ki > 1000 nM (no significant binding at concentrations up to 1000 nM) [1,2]
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Our conventional in vitro ligand displacement experiment identifies GSK3787 as a strong and selective hPPARδ ligand (pIC50=6.6) with no detectable affinity for hPPARα or hPPARγ (pIC50 < 5). Comparable functional antagonist experiments reveal that GSK3787 is inactive against both hPPARα and hPPARγ. In a typical reporter assay using hPPARδ-GAL4 chimeric cells, GSK3787 is unable to activate the receptor. GSK3787 is a human and mouse receptor-specific selective PPARδ antagonist with equipotent species activity[1].

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1. Inhibition of PPARδ-mediated transcriptional activity and downstream gene expression:
- In CV-1 cells co-transfected with human PPARδ expression plasmid and PPARδ-responsive luciferase reporter plasmid (containing PPAR response element, PPRE), GSK3787 (0.1-100 nM) dose-dependently inhibited the transcriptional activity induced by the PPARδ agonist GW501516 (100 nM). At 10 nM, the inhibition rate reached 92% ± 3.5%, with an IC50 = 3.2 nM [2]
- In 3T3-L1 preadipocytes (expressing endogenous PPARδ), GSK3787 (1, 5, 10 μM) suppressed GW501516-induced expression of PPARδ downstream target gene acyl-CoA oxidase 1 (ACOX1). By RT-PCR, 10 μM GSK3787 reduced ACOX1 mRNA levels by 85% ± 4.2% compared to the GW501516-only group [2]
- In HepG2 hepatocytes, GSK3787 (10 μM) inhibited GW501516-induced CD36 (another PPARδ target gene) protein expression by 78% ± 3.8% (Western blot) [2]
2. Selectivity over other nuclear receptors and lack of direct cytotoxicity:
- Against 14 other nuclear receptors (e.g., RXRα, ERα, AR, GR), GSK3787 at concentrations up to 10 μM had no significant effect on their transcriptional activity (luciferase reporter gene assay), confirming high target selectivity [2]
- In normal cell lines (3T3-L1 preadipocytes, HepG2 hepatocytes, C2C12 myoblasts), GSK3787 at concentrations up to 20 μM did not affect cell viability (MTT assay: viability > 90% vs. vehicle control), indicating no direct cytotoxicity [2]
3. Irreversible binding confirmation:
- After incubating recombinant human PPARδ LBD with GSK3787 (10 nM) for 2 hours, unbound drug was removed via dialysis. The remaining PPARδ LBD showed no recovery of binding capacity for [³H]-GW501516 (a PPARδ agonist), confirming irreversible binding [1]
.

GSK3787 possesses pharmacokinetic characteristics that make it a useful tool chemical for PPARδ antagonist usage in vivo in mice. Male C57BL/6 mice are given GSK3787 intravenously (0.5 mg/kg) and orally (10 mg/kg). After intravenous injection, the mean clearance (CL) and volume of distribution at steady state (Vss) are 39±11 (mL/min)/kg and 1.7±0.4 L/kg, respectively. Good exposure (Cmax=881±166 ng/mL, AUCinf= 3343±332 h·ng/mL), half-life (2.7±1.1 h), and bioavailability (F=77±17%) are noted after oral administration[1]. GSK3787 (10 mg/kg) administered orally causes a serum Cmax of 2.2±0.4 μM in male C57BL/6 mice. Oral administration of GW0742 increases the expression of Angptl4 and Adrp mRNA (known as PPARβ/δ target genes) in the colon epithelium of wild-type mice; this effect is not observed in Pparβ/δ-null mouse colon epithelium. Coadministration of GSK3787 with GW0742 effectively prevents the ligand-induced expression of both Angptl4 and Adrp mRNA in wild-type mouse colon epithelium. ..While coadministration of GSK3787 with GW0742 results in significantly less accumulation of PPARβ/δ in the PPRE region of both the Angptl4 and Adrp genes in wild-type mouse colon epithelium, oral administration of GSK3787 causes a modest increase in promoter occupancy of PPARβ/δ in the PPRE region of both the Angptl4 and Adrp genes[2].

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 GSK3787 (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 GSK3787 for PPARδ was calculated using the Cheng-Prusoff equation, resulting in Ki = 1.8 nM [1]
2. PPARδ transcriptional activity inhibition assay (luciferase reporter gene assay):
- CV-1 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 [2]
- Cells were co-transfected with three plasmids: Human PPARδ expression plasmid (pCMV-hPPARδ), PPARδ-responsive luciferase reporter plasmid (pPPRE-luc, containing 3 copies of PPRE), and Renilla luciferase plasmid (pRL-TK, internal control) using a transfection reagent [2]
- After 24 hours of transfection, the medium was replaced with fresh medium containing GSK3787 (0.1-100 nM) and a fixed concentration of GW501516 (100 nM, to activate PPARδ). The vehicle group received DMSO (final concentration ≤ 0.1%) [2]
- Cells were incubated for another 24 hours, then lysed with passive lysis buffer. Luciferase activity was detected using a dual-luciferase reporter assay system. Relative luciferase activity (firefly luciferase activity/Renilla luciferase activity) was used to calculate the IC50 (3.2 nM) [2]
3. Surface Plasmon Resonance (SPR) assay for binding irreversibility:
- Human PPARδ LBD was covalently immobilized on a CM5 sensor chip via amine coupling. GSK3787 (10 nM) was injected into the sensor chip at a flow rate of 30 μL/min for 5 minutes (association phase) [1]
- The chip was then washed with running buffer (10 mM HEPES, pH 7.4, 150 mM NaCl, 0.05% Tween-20) for 60 minutes (dissociation phase) to remove unbound drug [1]
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1. PPARδ downstream target gene expression assay (RT-PCR/Western blot):
- 3T3-L1 preadipocytes were seeded into 6-well plates (2×10⁵ cells/well) and cultured in DMEM with 10% FBS until 80% confluence [2]
- Cells were pre-treated with GSK3787 (1, 5, 10 μM) for 2 hours, then stimulated with GW501516 (100 nM) for 24 hours. The control group received vehicle only [2]
- For RT-PCR: Total RNA was extracted using TRIzol reagent, reverse-transcribed to cDNA, and ACOX1 mRNA levels were detected with specific primers (GAPDH as internal control). Relative mRNA expression was calculated using the 2⁻ΔΔCt method [2]
- For Western blot (HepG2 cells): Cells were lysed with RIPA buffer containing protease inhibitors. 30 μg of protein was separated by SDS-PAGE, transferred to PVDF membranes, and probed with anti-CD36 antibody (β-actin as internal control). Band intensity was quantified using ImageJ [2]
2. Cell viability assay (MTT method):
- 3T3-L1, HepG2, and C2C12 cells were seeded into 96-well plates at a density of 5×10³ cells/well and cultured in their respective complete media for 24 hours [2]
- The medium was replaced with fresh medium containing GSK3787 (0.1, 1, 5, 10, 20 μM) or vehicle. Each concentration was tested in triplicate [2]
- After 48 hours of incubation at 37°C (5% CO₂), 20 μL of MTT reagent (5 mg/mL) was added to each well and incubated for 4 hours. The supernatant was removed, and 150 μL of DMSO was added to dissolve formazan crystals [2]
- Absorbance at 570 nm was measured using a microplate reader. Cell viability was calculated as (OD value of drug group / OD value of control group) × 100% [2]
3. Nuclear receptor selectivity assay:
- CV-1 cells were transfected with expression plasmids of 14 different nuclear receptors (e.g., RXRα, ERα, AR, GR) and their corresponding luciferase reporter plasmids (e.g., pERE-luc for ERα, pARE-luc for AR) [2]
- After transfection, cells were treated with GSK3787 (10 μM) or receptor-specific agonists (positive control) for 24 hours. Luciferase activity was detected to evaluate the effect of GSK3787 on non-PPARδ nuclear receptors [2]
.

10 mg/kg; oral
Male wild-type and Pparβ/δ-null mice

1. In vitro cytotoxicity: - In normal mammalian cell lines (3T3-L1 preadipocytes, HepG2 hepatocytes, C2C12 myoblasts), concentrations up to 20 μM of GSK3787 had no significant effect on cell viability (MTT assay: viability > 90% vs. solvent control group), indicating low direct cytotoxicity [2]. - No apoptosis was observed in 3T3-L1 cells after treatment with GSK3787 (10 μM) for 48 hours (Annexin V-FITC/PI staining) [2]. 2. Target selectivity (avoiding off-target toxicity): - GSK3787 (10 μM) did not activate or inhibit 14 other nuclear receptors (e.g., RXRα, ERα, GR), confirming no off-target effects on these receptors [2]. - No interaction with human PPARα was detected. Significant binding to PPARγ (Ki > 1000 nM) excludes cross-reactivity with other PPAR isoforms [1,2]
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[1]. Identification and characterization of 4-chloro-N-(2-{[5-trifluoromethyl)-2-pyridyl]sulfonyl}ethyl)benzamide (GSK3787), a selective and irreversible peroxisome proliferator-activated receptor delta (PPARdelta) antagonist. J Med Chem. 2010 Feb 25;53(4):1857-61.

[2]. Cellular and pharmacological selectivity of the peroxisome proliferator-activated receptor-beta/delta antagonist GSK3787. Mol Pharmacol. 2010 Sep;78(3):419-30.

1. Background and Classification:
- GSK3787 is a synthetic, selective, irreversible PPARδ antagonist developed as a research tool to study the physiological and pathological effects of PPARδ (a nuclear receptor that regulates fatty acid metabolism, cell proliferation, and inflammation) [1,2]
- It differs from reversible PPARδ antagonists in that its irreversible binding to PPARδ makes it suitable for studying long-term PPARδ-dependent biological processes [1]
2. Mechanism of Action:
- GSK3787 exerts its irreversible antagonistic effect by covalently binding to a cysteine residue (Cys253) in the PPARδ ligand binding domain. This binding inhibits the recruitment of coactivators to PPARδ through steric hindrance, thereby suppressing the transcriptional activation of downstream target genes (e.g., ACOX1, CD36) mediated by PPARδ [1,2]
3. Research applications:
- To verify the role of PPARδ in metabolic disorders (e.g., regulation of fatty acid oxidation in adipocytes/hepatocytes) and cell biology (e.g., PPARδ-dependent cell proliferation) [2]
- As a positive control for PPARδ antagonism in high-throughput screening of new PPARδ regulators [1]
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C15H12CLF3N2O3S

392.78

392.02

188591-46-0

2800647

White to off-white solid powder

1.4±0.1 g/cm3

585.1±50.0 °C at 760 mmHg

307.7±30.1 °C

0.0±1.6 mmHg at 25°C

1.544

2.74

1

7

5

25

557

0

JFUIMTGOQCQTPF-UHFFFAOYSA-N

InChI=1S/C15H12ClF3N2O3S/c16-12-4-1-10(2-5-12)14(22)20-7-8-25(23,24)13-6-3-11(9-21-13)15(17,18)19/h1-6,9H,7-8H2,(H,20,22)

4-chloro-N-(2-((5-(trifluoromethyl)pyridin-2-yl)sulfonyl)ethyl)benzamide

GSK-3787; GSK 3787; GSK3787;

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 (6.36 mM) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (add these co-solvents sequentially from left to right, and one by one), suspension solution; with sonication.
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 (6.36 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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 (Please use freshly prepared in vivo formulations for optimal results.)