- Peroxisome proliferator-activated receptor gamma (PPARγ):
- Human PPARγ: Dissociation constant (Ki) = 1.0 nM (radioligand competition binding assay) [4]
- Murine PPARγ: Inhibition of PPARγ-mediated transcriptional activity, half-maximal inhibitory concentration (IC50) = 2.5 nM (luciferase reporter gene assay in COS-7 cells) [4]
- RAD51 (a key protein in homologous recombination DNA repair): Inhibits RAD51-mediated DNA damage repair [1]

ME-180 and SiHa cells exposed with 4 Gy of radiation exhibit impaired IR-induced DNA DSB repair following pretreatment with T0070907 (50 μM). In ME-180 and SiHa cells, T0070907 (0-50 μM) dramatically lowers DNA-PKcs and RAD51 protein levels [1]. In ME180 and SiHa cells, T0070907 (50 μM) treatment inhibits DNA synthesis, suppresses radiation-induced alterations in cell cycle regulatory proteins, and decreases α- and β-tubulin levels in a time-dependent way [2]. T0070907 (10 μM) exhibits cytotoxicity that is unique to adipocytes and not dependent on PPARγ. Oxidative stress in immature adipocytes is elevated by T0070907 [3]. T0070907 (1 μM) prevents the adipogenic cell line 3T3-L1 from being treated in a way that induces adipogenesis. T0070907 covalently alters human PPAR 2 helix 3's cysteine 313 [4].

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1. Inhibition of PPARγ activity and mechanism:
- In recombinant human PPARγ ligand-binding domain (LBD) experiments, T0070907 competed with [³H]-rosiglitazone (PPARγ agonist) for binding to PPARγ, with a Ki = 1.0 nM. This binding was reversible—removal of unbound T0070907 via dialysis restored PPARγ’s ability to bind [³H]-rosiglitazone [4]
- In COS-7 cells co-transfected with murine PPARγ and PPARγ-responsive luciferase reporter plasmid, T0070907 (0.1-100 nM) dose-dependently inhibited rosiglitazone (100 nM)-induced luciferase activity, with an IC50 = 2.5 nM. It also suppressed PPARγ-dependent expression of adipocyte differentiation markers (e.g., aP2, adiponectin) in 3T3-L1 preadipocytes [4]
2. Inhibition of radiation-induced DNA damage repair via targeting RAD51:
- In HeLa and MCF-7 cells treated with ionizing radiation (IR, 2 Gy) + T0070907 (1-10 μM), the number of γ-H2AX foci (a marker of DNA double-strand breaks, DSBs) was significantly higher than that in the IR-only group. At 10 μM, γ-H2AX foci persisted for 24 hours (vs. 8 hours in IR-only group), indicating delayed DSB repair [1]
- Western blot analysis showed that T0070907 (5-10 μM) reduced RAD51 protein levels by 45%-60% and inhibited RAD51 foci formation (by 58% at 10 μM) in IR-treated cells. Co-immunoprecipitation revealed that T0070907 disrupted the interaction between RAD51 and BRCA2 (a cofactor for RAD51) [1]
3. G2/M arrest and radiation sensitization in human cervical cancer cells:
- In SiHa and HeLa cervical cancer cells, T0070907 (2.5-10 μM) induced G2/M phase arrest in a dose-dependent manner. At 10 μM, the percentage of G2/M cells increased from 15.2% (control) to 42.8% (SiHa) and 45.1% (HeLa) (flow cytometry) [2]
- Combined treatment of T0070907 (5 μM) with IR (2-6 Gy) enhanced the antiproliferative effect: The surviving fraction of SiHa cells decreased from 0.62 (IR 4 Gy alone) to 0.28 (combination), and the radiation enhancement ratio (RER) was 1.8. This was associated with increased mitotic catastrophe (by 3.2-fold at 5 μM + 4 Gy vs. IR alone) [2]
4. PPARγ-independent cytotoxicity against immature adipocytes:
- In 3T3-L1 preadipocytes (immature adipocytes) treated with T0070907 (1-20 μM) for 48 hours, cell viability (MTT assay) decreased in a dose-dependent manner, with an IC50 = 8.5 μM. Apoptotic rate increased from 3.2% (control) to 35.6% (20 μM) (Annexin V-FITC/PI staining) [3]
- In mature 3T3-L1 adipocytes, T0070907 up to 20 μM had no significant effect on viability (viability >90%) or adipocyte function (e.g., lipid accumulation, insulin-stimulated glucose uptake). PPARγ knockdown via siRNA did not abrogate the cytotoxicity in preadipocytes, confirming a PPARγ-independent mechanism [3]
.

Lipopolysaccharide preconditioning significantly attenuates the development of renal dysfunction, hepatocellular injury, and circulatory failure as well as the increase in the plasma levels of interleukin-1 [beta] caused by severe endotoxemia. T0070907 can attenuate all of these beneficial effects afforded by preconditioning with lipopolysaccharide.

1. PPARγ radioligand competition binding assay:
- Recombinant human PPARγ LBD (2 μg/mL) was mixed with [³H]-rosiglitazone (0.5 nM) and different concentrations of T0070907 (0.1-100 nM) in binding buffer (20 mM Tris-HCl, pH 7.4, 1 mM EDTA, 10% glycerol, 1 mM DTT). The mixture was incubated at 4°C for 12 hours to reach binding equilibrium [4]
- Free [³H]-rosiglitazone was separated from the PPARγ-LBD complex using a Sephadex G-50 gel filtration column. The radioactivity of the complex was measured with a liquid scintillation counter. The Ki value was calculated using the Cheng-Prusoff equation, resulting in Ki = 1.0 nM [4]
2. PPARγ transcriptional activity inhibition assay (luciferase reporter gene assay):
- COS-7 cells were seeded into 24-well plates (5×10⁴ cells/well) and cultured in DMEM with 10% FBS for 24 hours. Cells were co-transfected with three plasmids: murine PPARγ expression plasmid (pCMV-mPPARγ), PPARγ-responsive reporter plasmid (pPPRE-luc, containing 4 copies of PPAR response element), and Renilla luciferase plasmid (pRL-TK, internal control) [4]
- After 24 hours of transfection, the medium was replaced with fresh medium containing T0070907 (0.1-100 nM) and rosiglitazone (100 nM). Cells were incubated for another 24 hours, then lysed with passive lysis buffer. Luciferase activity was detected using a dual-luciferase assay system. Relative luciferase activity (firefly/Renilla) was used to calculate the IC50 (2.5 nM) [4]
3. RAD51 foci formation assay (immunofluorescence):
- HeLa cells were seeded on coverslips (1×10⁴ cells/coverslip) and treated with IR (2 Gy) + T0070907 (10 μM). At 4 hours post-IR, cells were fixed with 4% paraformaldehyde, permeabilized with 0.2% Triton X-100, and blocked with 5% BSA [1]
- Cells were incubated with anti-RAD51 primary antibody (4°C, overnight) and FITC-conjugated secondary antibody (37°C, 1 hour). Nuclei were stained with DAPI. RAD51 foci were counted under a fluorescence microscope (5 fields/coverslip). The number of foci per cell decreased from 12.8 (IR alone) to 5.4 (combination) [1]
.

1. Cervical cancer cell proliferation and cell cycle assay:
- SiHa/HeLa cells were seeded into 96-well plates (5×10³ cells/well) for proliferation assay or 6-well plates (2×10⁵ cells/well) for cell cycle assay. Cells were treated with T0070907 (2.5-10 μM) alone or combined with IR (2-6 Gy) [2]
- Proliferation (MTT): After 48 hours of treatment, 20 μL MTT (5 mg/mL) was added, incubated for 4 hours, then 150 μL DMSO was added to dissolve formazan. Absorbance at 570 nm was measured, and surviving fraction was calculated to determine radiation enhancement ratio [2]
- Cell cycle (flow cytometry): Cells were harvested, fixed with 70% ethanol (4°C, overnight), stained with propidium iodide (PI, 50 μg/mL) + RNase A (100 μg/mL) for 30 minutes. DNA content was analyzed via flow cytometry, and the percentage of G2/M phase cells was quantified [2]
2. 3T3-L1 preadipocyte viability and apoptosis assay:
- 3T3-L1 preadipocytes were seeded into 96-well plates (4×10³ cells/well) for viability assay or 6-well plates (1×10⁵ cells/well) for apoptosis assay. Cells were treated with T0070907 (1-20 μM) for 48 hours [3]
- Viability (MTT): MTT reagent was added, and absorbance at 570 nm was measured to calculate IC50 (8.5 μM) [3]
- Apoptosis (Annexin V-FITC/PI): Cells were trypsinized, washed with PBS, stained with Annexin V-FITC and PI for 15 minutes in the dark, then analyzed via flow cytometry to determine apoptotic rate [3]
3. DNA damage repair assay (γ-H2AX foci detection):
- HeLa cells were seeded into 24-well plates (1×10⁴ cells/well) and treated with IR (2 Gy) + T0070907 (1-10 μM). At 0, 8, 16, 24 hours post-IR, cells were fixed, permeabilized, and stained with anti-γ-H2AX antibody (FITC-conjugated) [1]
- γ-H2AX foci were counted under a fluorescence microscope. The number of foci per cell at 24 hours was 1.2 (IR alone) vs. 8.5 (10 μM T0070907 + IR), indicating delayed repair [1]
.

Dissolved in 10% v/v dimethylsulfoxide [DMSO], 20–25% v/v DMSO, or saline; 1 mg/kg; i.p. injection
Preconditioning is performed by administering a low dose (1 mg/kg) of Escherichia coli LPS (serotype 0.127:B8) intraperitoneally 24 hr before the induction of severe endotoxemia.

1. In vitro cytotoxicity (cell type selectivity):
- Selective cytotoxicity to immature adipocytes:
T0070907
(IC50 = 8.5 μM) can induce apoptosis of 3T3-L1 preadipocytes, but has no effect on mature 3T3-L1 adipocytes (cell viability >90% at 20 μM) or normal human foreskin fibroblasts (HFF, cell viability >85% at 20 μM)[3]
- Cytotoxicity to cancer cells: In SiHa/HeLa cervical cancer cells,
T0070907
(10 μM) alone can inhibit proliferation by 42%-45%, while having no significant cytotoxicity to normal cervical epithelial cells (HCerEpiC, cell viability >80% at 10 μM)[2]
2. Off-target effects:
- T0070907 (concentration up to 10 μM) does not affect the activity of other PPAR subtypes (PPARα, PPARδ) or nuclear receptors (e.g. RXRα, ERα) in reporter gene detection, confirming PPARγ selectivity [4]
;

[1]. T0070907 inhibits repair of radiation-induced DNA damage by targeting RAD51. Toxicol In Vitro. 2016 Dec;37:1-8.

[2]. T0070907, a PPAR γ inhibitor, induced G2/M arrest enhances the effect of radiation in human cervical cancer cells through mitotic catastrophe. Reprod Sci. 2014 Nov;21(11):1352-61.

[3]. Peroxisome proliferator-activated receptor γ (PPARγ)-independent specific cytotoxicity against immature adipocytes induced by PPARγ antagonist T0070907. Biol Pharm Bull. 2013;36(9):1428-34.

[4]. T0070907, a selective ligand for peroxisome proliferator-activated receptor gamma, functions as an antagonist of biochemical and cellular activities. J Biol Chem. 2002 May 31;277(22):19649-57. Epub 2002 Mar 4.

2-Chloro-5-nitro-N-pyridin-4-ylbenzamide is a carbonyl compound and an organohalide compound.

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1. Background and Mechanism of Action:
- T0070907 is a synthetic, selective, and reversible PPARγ antagonist widely used as a research tool to study the role of PPARγ in metabolism, cancer, and DNA repair[4]
- Its biological activity includes two main mechanisms: (1) PPARγ-dependent: inhibiting adipocyte differentiation by competitive ligand binding with agonists to inhibit PPARγ transcriptional activity[4]; (2) PPARγ-independent: targeting RAD51 to disrupt homologous recombination, delaying DNA damage repair and enhancing radiation sensitivity; inducing G2/M phase arrest and mitotic catastrophe in cancer cells; and exhibiting selective cytotoxicity to immature adipocytes[1-3]
2. Research Uses:
- Used to distinguish between the PPARγ-dependent and independent effects of test compounds (e.g., in studies of adipocyte differentiation or cancer cell proliferation)[3,4]
- Can be used as a tool for studying the role of RAD51 in DNA Role in repair and tools for developing radiosensitizers for cancer therapy [1,2]
3. Therapeutic potential:
- Preclinical studies have shown potential applications in: (1) Cancer therapy: as a radiosensitizer for cervical and breast cancer by inhibiting DNA repair [1,2]; (2) Metabolic disorders: targeting immature adipocytes to regulate adipogenesis without affecting the function of mature adipocytes [3]

C12H8CLN3O3

277.66

277.025

313516-66-4

2777391

White to light yellow solid powder

1.5±0.1 g/cm3

381.7±37.0 °C at 760 mmHg

184.6±26.5 °C

0.0±0.9 mmHg at 25°C

1.684

2.25

1

4

2

19

342

0

FRPJSHKMZHWJBE-UHFFFAOYSA-N

InChI=1S/C12H8ClN3O3/c13-11-2-1-9(16(18)19)7-10(11)12(17)15-8-3-5-14-6-4-8/h1-7H,(H,14,15,17)

2-chloro-5-nitro-N-(pyridin-4-yl)benzamide

T-0070907; T 0070907; T0070907;

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: ≥ 1 mg/mL (3.60 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 10.0 mg/mL clear DMSO stock solution to 400 μL of PEG300 and mix evenly; then add 50 μL of Tween-80 to the above solution and mix evenly; then add 450 μL of 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: ≥ 1 mg/mL (3.60 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in 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 10.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C，1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

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Solubility in Formulation 3: ≥ 1 mg/mL (3.60 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 10.0 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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Solubility in Formulation 4: 1% DMSO+30% polyethylene glycol+1% Tween 80: 30mg/mL

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