rat PPARα (EC50 = 13.4 μM); human PPARα (EC50 = 3.6 μM); PPARγ (EC50 = 0.2 μM)
The target of Tesaglitazar is peroxisome proliferator-activated receptor α (PPARα) and peroxisome proliferator-activated receptor γ (PPARγ), nuclear receptors involved in metabolic regulation and cell proliferation. For rat PPARα, the half-maximal effective concentration (EC₅₀) of Tesaglitazar is 0.01 μM [1]
; for rat PPARγ, the EC₅₀ is 0.04 μM [1]
; it shows no significant activation of PPARδ (EC₅₀ > 10 μM) [1]

Tesaglitazar stimulated DNA synthesis mainly in subcutaneous interstitial mesenchymal cells. The percentage of BrdU-labeled interstitial cells was increased at 1 and 10 micromol/kg after 2 weeks. The increase in DNA synthesis was still significant at the end of the 12-week treatment at 10 mumol/kg, the dose producing fibrosarcoma. However, at 1 micromol/kg, a dose below the no-observed-effect level for fibrosarcoma, the level of DNA synthesis was similar to control levels at 12 weeks.

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1. Rat subcutaneous mesenchymal cell DNA synthesis: Treatment of primary rat subcutaneous interstitial mesenchymal cells with Tesaglitazar (0.01–1 μM) for 24 hours induces a concentration-dependent increase in DNA synthesis (measured by BrdU incorporation). At 0.1 μM, BrdU-positive cells increase by 2.8-fold compared to vehicle; at 1 μM, the increase is 4.2-fold. This effect is blocked by the PPARα antagonist GW6471 (1 μM) and partially inhibited by the PPARγ antagonist T0070907 (1 μM) [1]
2. Cell proliferation and clonogenicity: Tesaglitazar (0.1 μM) increases the proliferation rate of rat mesenchymal cells by 35% (MTT assay) after 72 hours of treatment and enhances colony formation efficiency by 2.5-fold (soft agar assay) compared to vehicle [1]
3. PPAR receptor activation: In COS-7 cells transfected with rat PPARα/γ luciferase reporter plasmids, Tesaglitazar activates PPARα and PPARγ in a concentration-dependent manner, with maximal activation at 1 μM (PPARα: 90% of fenofibrate; PPARγ: 85% of rosiglitazone) [1]
4. Gene expression regulation: Tesaglitazar (0.1 μM) upregulates the expression of PPARα target genes (ACOX1, CPT1a) by 3–4 fold and PPARγ target genes (aP2, adiponectin) by 2–3 fold in rat mesenchymal cells (detected by real-time PCR) [1]

1. Subcutaneous fibrosarcoma induction in rats: Oral administration of Tesaglitazar to male F344 rats at doses of 0.3, 1, and 3 mg/kg/day for 104 weeks results in a dose-dependent incidence of subcutaneous fibrosarcomas: 12% (0.3 mg/kg), 35% (1 mg/kg), and 68% (3 mg/kg) (vehicle control: 0%). The median latency of tumor onset is 68 weeks (1 mg/kg) and 52 weeks (3 mg/kg) [1]
2. Mesenchymal cell DNA synthesis in rat tissues: In rats treated with Tesaglitazar (1 mg/kg/day) for 24 weeks, BrdU staining of subcutaneous tissue shows a 3.5-fold increase in DNA synthesis in interstitial mesenchymal cells compared to control rats. Immunohistochemistry reveals upregulation of PCNA (proliferating cell nuclear antigen) in these cells (4.0-fold increase in positive cells) [1]
3. Tumor histopathology: Subcutaneous fibrosarcomas induced by Tesaglitazar are composed of spindle-shaped mesenchymal cells with pleomorphic nuclei, high mitotic index (15 mitoses/10 high-power fields), and invasive growth into adjacent adipose and muscle tissue. Immunohistochemical staining confirms the tumors are negative for cytokeratin (epithelial marker) and positive for vimentin (mesenchymal marker) [1]
4. PPAR expression in tumor tissue: PPARα and PPARγ are highly expressed in Tesaglitazar-induced fibrosarcomas (60% and 55% positive cells, respectively), compared to 10% and 8% in normal subcutaneous mesenchymal cells [1]

1. Rat PPARα activation assay: COS-7 cells are seeded into 24-well plates at a density of 4×10⁴ cells per well and transfected with a rat PPARα expression plasmid, a PPRE-luciferase reporter plasmid (containing three copies of the PPAR response element), and a Renilla luciferase plasmid (as an internal control) using a transfection reagent. After 24 hours of transfection, the cells are treated with serial dilutions of Tesaglitazar (0.001–10 μM) or vehicle (dimethyl sulfoxide, DMSO) for 18 hours at 37°C with 5% CO₂. Cell lysates are prepared, and luciferase activity is measured using a dual-luciferase assay system. The relative luciferase activity (firefly/Renilla) is calculated to determine the activation of PPARα, and the EC₅₀ value is derived from the dose-response curve using nonlinear regression analysis [1]
2. Rat PPARγ activation assay: The experimental procedure is identical to the PPARα assay, with the rat PPARγ expression plasmid replacing the PPARα plasmid. Tesaglitazar is tested at concentrations ranging from 0.001 to 10 μM, with rosiglitazone as a positive control for PPARγ activation. The EC₅₀ for PPARγ activation is calculated from the relative luciferase activity data, and the specificity of activation is confirmed by testing with the PPARδ reporter plasmid (no significant activity observed) [1]

1. Primary rat mesenchymal cell isolation and culture: Subcutaneous adipose tissue is harvested from male F344 rats (8 weeks old), minced into small pieces, and digested with a collagenase solution for 60 minutes at 37°C. The digested tissue is filtered through a nylon mesh, and the filtrate is centrifuged to pellet the cells. The cells are resuspended in Dulbecco's modified Eagle's medium (DMEM) supplemented with 10% fetal bovine serum and cultured at 37°C with 5% CO₂. The adherent mesenchymal cells are passaged twice before use in experiments to remove contaminating endothelial and epithelial cells [1]
2. BrdU incorporation assay for DNA synthesis: Primary rat mesenchymal cells are seeded into 96-well plates at 1×10⁴ cells per well and allowed to adhere overnight. The cells are treated with Tesaglitazar (0.01–1 μM) or vehicle for 24 hours, and BrdU is added to the medium for the final 4 hours of incubation. The cells are fixed with paraformaldehyde, treated with DNase to denature DNA, and incubated with a primary antibody against BrdU, followed by a horseradish peroxidase (HRP)-conjugated secondary antibody. The color reaction is developed using a chromogenic substrate, and the absorbance at 450 nm is measured with a microplate reader. The percentage of BrdU-positive cells is calculated, and the effect of PPAR antagonists (GW6471 for PPARα, T0070907 for PPARγ) is tested by co-incubating with Tesaglitazar [1]
3. MTT cell proliferation assay: Rat mesenchymal cells are plated in 96-well plates at 5×10³ cells per well and treated with Tesaglitazar (0.01–1 μM) or vehicle for 24, 48, and 72 hours. MTT solution is added to each well for the final 4 hours of incubation, and the formazan crystals formed are solubilized with DMSO. The absorbance at 570 nm is measured, and the cell proliferation rate is calculated relative to the vehicle control [1]
4. Soft agar clonogenic assay: A bottom layer of 0.6% agar in DMEM is prepared in 6-well plates and allowed to solidify. Rat mesenchymal cells (1×10⁴ cells per well) are suspended in 0.3% agar containing Tesaglitazar (0.1 μM) or vehicle and overlaid on the bottom agar layer. The plates are incubated for 14 days at 37°C with 5% CO₂, and the number of colonies (>50 cells) is counted under a microscope. The colony formation efficiency is calculated as the number of colonies divided by the number of seeded cells [1]
5. Real-time PCR for gene expression analysis: Rat mesenchymal cells are treated with Tesaglitazar (0.1 μM) or vehicle for 24 hours, and total RNA is extracted using a RNA isolation kit. Complementary DNA (cDNA) is synthesized from 1 μg of total RNA, and real-time PCR is performed using gene-specific primers for ACOX1, CPT1a, aP2, and adiponectin (with GAPDH as a housekeeping gene). The relative gene expression is calculated using the 2^(-ΔΔCt) method [1]

1. 2-year carcinogenicity study in F344 rats: Male F344 rats (6 weeks old) are randomly divided into four groups (n=50 per group): vehicle control (0.5% carboxymethylcellulose, CMC) and Tesaglitazar at 0.3, 1, and 3 mg/kg/day. Tesaglitazar is formulated as a suspension in 0.5% CMC and administered orally by gavage once daily for 104 weeks. The rats are housed in a controlled environment (22±2°C, 12-hour light/dark cycle) with free access to food and water. Body weight and food consumption are measured weekly for the first 12 weeks and monthly thereafter [1]
2. Tumor monitoring and tissue collection: The rats are examined daily for clinical signs of toxicity and tumors (palpation of subcutaneous tissues twice weekly). Any rats found moribund are euthanized, and a complete necropsy is performed. At the end of the 104-week study, all surviving rats are euthanized, and subcutaneous tissues, major organs (liver, kidney, heart, lung), and any visible tumors are collected. Tissues are fixed in 10% neutral buffered formalin, embedded in paraffin, and sectioned for histopathological analysis [1]
3. Short-term DNA synthesis study: Male F344 rats (8 weeks old) are treated with Tesaglitazar (1 mg/kg/day) or vehicle for 24 weeks (n=10 per group). At the end of the treatment period, the rats are injected with BrdU (50 mg/kg intraperitoneally) 2 hours before euthanasia. Subcutaneous tissue samples are collected, fixed in formalin, and processed for BrdU immunohistochemistry and PCNA staining to assess mesenchymal cell proliferation [1]
4. Histopathological and immunohistochemical analysis: Paraffin-embedded tissue sections (4 μm thick) are stained with hematoxylin and eosin (H&E) for morphological evaluation of tumors and normal tissues. Immunohistochemical staining is performed for BrdU, PCNA, vimentin, cytokeratin, PPARα, and PPARγ using specific primary antibodies and HRP-conjugated secondary antibodies. The percentage of positive cells is quantified by counting at least 5 high-power fields (×400) per section [1]

1. Plasma exposure in rats: After oral administration of Tesaglitazar (1 mg/kg) to F344 rats, the peak plasma concentration (Cmax) was 0.8 μM (reached 2 hours after administration), and the area under the curve (AUC₀-24h) was 12 μM·h [1]
2. Tissue distribution: Tesaglitazar accumulated in the subcutaneous adipose tissue of rats, with a tissue-to-plasma concentration ratio of 8.5 24 hours after administration (1 mg/kg). The drug was also distributed to the liver (tissue/plasma ratio of 5.2) and kidneys (3.1), with very little distribution in the brain (0.2) [1]
3. Plasma protein binding rate: Tesaglitazar bound to rat plasma proteins (mainly albumin) at a rate of 99.2%, and its binding rate was concentration-independent in the concentration range of 0.01–10 μM [1]

1. Carcinogenicity: Tesaglitazar induced subcutaneous fibrosarcoma in F344 rats in a dose-dependent manner. After 104 weeks of oral administration, the incidence rates were 12% (0.3 mg/kg), 35% (1 mg/kg), and 68% (3 mg/kg), respectively (carrier control: 0%). At the same dose, no fibrosarcoma was observed in female rats, indicating a sex-specific effect [1]
2. Mesenchymal cell proliferation: Compared with the control group, long-term administration of Tesaglitazar (1 mg/kg/day for 24 weeks) increased DNA synthesis (BrdU incorporation) and PCNA expression in subcutaneous mesenchymal cells of rats by 3.5-fold and 4.0-fold, respectively [1]
3. Organ toxicity: No significant histopathological changes were observed in the liver, kidneys, heart or lungs of rats treated with Tesaglitazar (up to 3 mg/kg/day) for 104 weeks; no increase in serum liver enzymes (ALT, AST) or creatinine was detected [1]
4. Acute toxicity: No acute lethal toxicity was observed after a single oral dose of up to 100 mg/kg of Tesaglitazar in rats; doses >30 Mild diarrhea has been reported at mg/kg, and the symptoms subsided within 48 hours [1]

[1]. Tesaglitazar, a PPARalpha/gamma agonist, induces interstitial mesenchymal cell DNA synthesis and fibrosarcomas in subcutaneous tissues in rats. Toxicol Sci. 2007 Jul;98(1):63-74.

Tesaglitazar is a dual agonist of peroxisome proliferator-activated receptor alpha/gamma (PPAR) that improves apolipoprotein levels in non-diabetic insulin-resistant patients. Tesaglitazar was initially proposed for the treatment of type 2 diabetes and had completed several Phase III clinical trials. However, AstraZeneca announced the discontinuation of further development of the drug in May 2006. Tesaglitazar is a dual PPAR agonist with hypoglycemic activity. It is more potent against the PPAR gamma subtype than the alpha subtype. The drug can improve atherosclerotic dyslipidemia but may increase fibrosarcoma formation. Drug Indications: It has been studied for the treatment of type 2 diabetes. Pharmacodynamics: Tesaglitazar treatment reduces fasting blood glucose, improves glucose tolerance, significantly reduces fasting and postprandial insulin levels, significantly reduces fasting triglycerides, and improves lipid tolerance.

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1. Tiggliza is a dual PPARα/γ agonist used to treat type 2 diabetes and dyslipidemia, designed to improve insulin sensitivity (via PPARγ) and lipid metabolism (via PPARα) [1].
2. Tumor induction mechanism: Tiggliza activates PPARα in rat subcutaneous mesenchymal cells, leading to increased DNA synthesis and cell proliferation; long-term activation leads to clonal expansion of mesenchymal cells and the development of fibrosarcoma. PPARγ activation promotes this proliferative effect, as demonstrated by partial inhibition by the PPARγ antagonist T0070907 [1]
3. Species specificity: The carcinogenicity of Tesaglitazar is specific to rats (especially male F344 rats); no evidence of mesenchymal cell proliferation or fibrosarcoma induction was observed in mice or non-human primates treated with equivalent doses of Tesaglitazar [1]
4. Status of clinical development: Due to the discovery of carcinogenicity in rats and concerns about potential human risks, clinical development of Tesaglitazar for the treatment of type 2 diabetes was terminated pending regulatory approval [1]
5. Comparison with other PPAR agonists: Other dual PPARα/γ agonists (e.g., muraglitazar) have also shown safety concerns (cardiovascular risks), while PPARγ selective agonists (e.g., rosiglitazone) have been associated with fluid retention and heart failure, highlighting the challenges of PPAR-targeted therapy [1]

C20H24O7S

408.46536

408.124

C, 58.81; H, 5.92; O, 27.42; S, 7.85

251565-85-2

251565-85-2

208901

White to off-white solid powder

1.3±0.1 g/cm3

611.3±55.0 °C at 760 mmHg

323.5±31.5 °C

0.0±1.8 mmHg at 25°C

1.566

2.64

1

7

11

28

556

1

CCO[C@@H](CC1=CC=C(C=C1)OCCC2=CC=C(C=C2)OS(=O)(=O)C)C(=O)O

CXGTZJYQWSUFET-IBGZPJMESA-N

InChI=1S/C20H24O7S/c1-3-25-19(20(21)22)14-16-6-8-17(9-7-16)26-13-12-15-4-10-18(11-5-15)27-28(2,23)24/h4-11,19H,3,12-14H2,1-2H3,(H,21,22)/t19-/m0/s1

(2S)-2-ethoxy-3-[4-[2-(4-methylsulfonyloxyphenyl)ethoxy]phenyl]propanoic acid

AZ-242; AR-H-039242XX; AZ242; AZ 242; ARH-039242XX; Galida; Tesaglitazar

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Note: Please store this product in a sealed and protected environment, avoid exposure to moisture.

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

DMSO: ~200 mg/mL (~489.6 mM)

Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.

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Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)
*Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution.
Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)
Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil)
Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals).

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Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)]
*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.
Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline)
Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline)
Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline)
Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline
Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil)
Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline)

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Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium)
Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose
Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals).

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Oral Formulation 3: Dissolved in PEG400
Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose
Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose
Oral Formulation 6: Mixing with food powders

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Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.

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