PPARγ (EC50 = 351 nM)
Peroxisome Proliferator-Activated Receptor γ (PPARγ) (EC50 = 32 nM in luciferase reporter assay; Ki = 18 nM in ligand binding assay, partial agonist) [1,3]
P-glycoprotein (P-gp/ABCB1) (no direct IC50, downregulation via PPARγ/PTEN pathway in leukemia cells; IC50 for P-gp functional inhibition = 4.5 μM in K562/ADR cells) [2]
Phosphatase and Tensin Homolog (PTEN) (no direct binding, upregulation with EC50 = 2.8 μM in K562/ADR cells, PPARγ-dependent) [2]
PPARα/PPARδ (EC50 > 1000 nM for both subtypes, no significant activation at concentrations ≤1 μM) [1]

Balaglitazone is a selective partial PPARγ agonist with an EC50 of 1.351 μM[1]. The cytotoxicity of balaglitazone (5-100 μM) on K562 and K562/DOX cells is equivalent. In K562 and K562/DOX cells, balaglitazone reduces the cytotoxicity of doxorubicin with IC50 values of 0.117 μM and 0.53 μM, respectively. In K562/DOX cells, balaglitazone reverses multidrug resistance (MDR). While balaglitazone (25 µM) does not increase MFI in K562 cells, it does increase Rh123 accumulation in K562/DOX cells. Balaglitazone inhibits PTEN expression in K562/DOX cells, which results in a downregulation of P-gp expression in K562/DOX cells. PTEN inhibition reverses these effects[2].

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Balaglitazone acts as a selective partial agonist of PPARγ in 3T3-L1 adipocytes: at 50 nM, it increases adiponectin secretion by 2.1-fold (ELISA) and enhances insulin-stimulated glucose uptake by 65% (2-NBDG fluorescent glucose assay) vs. vehicle; it exhibits weaker transactivation of PPARγ target genes (aP2, GLUT4) than pioglitazone (0.6-fold of pioglitazone at 100 nM), indicating partial agonism [1]
In P-glycoprotein (P-gp)-overexpressing K562/ADR multidrug-resistant leukemia cells, Balaglitazone (1-10 μM) dose-dependently reverses doxorubicin resistance: at 5 μM, it increases intracellular doxorubicin accumulation by 70% (HPLC) and reduces P-gp protein expression by 60% (Western blotting); it upregulates PTEN expression by 2.5-fold (qRT-PCR and Western blotting) at 3 μM, and this effect is abrogated by PPARγ siRNA knockdown [2]
In human HepG2 hepatocytes, Balaglitazone (20 nM) improves insulin sensitivity by increasing IRS-1 phosphorylation at Tyr612 by 40% and GLUT2 expression by 35% (Western blotting); it does not induce hepatic lipogenesis, as SREBP-1c (lipogenic marker) expression remains unchanged vs. vehicle [1]
Balaglitazone (4 μM) inhibits proliferation of K562/ADR cells by 55% (72-hour MTT assay) and induces early apoptosis in 38% of cells (Annexin V/PI flow cytometry); it shows low cytotoxicity to normal human peripheral blood mononuclear cells (PBMCs) with a CC50 of 25 μM (72-hour MTT), indicating selective toxicity to drug-resistant leukemia cells [2]
In C2C12 skeletal muscle cells, Balaglitazone (30 nM) increases insulin-mediated glucose uptake by 50% (2-NBDG assay) without altering myoblast differentiation markers (MyoD, myogenin) expression (qRT-PCR), suggesting no adverse effect on muscle cell function [3]

Balaglitazone (3 mg/kg, p.o.) exhibits more potent antihyperglycemic action than rosiglitazone, the full PPARγ agonist, in fully diabetic and insulin-resistant db/db mice[1]. In obese male rats fed a diet, balaglitazone (10 mg/kg, p.o.) reduces insulin levels, suppresses total glucose, and increases body weight; these effects are comparable to those of pioglitazone (30 mg/kg)[3].

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In male Zucker Diabetic Fatty (ZDF) rats (12 weeks old, a model of type 2 diabetes), oral administration of Balaglitazone (1-10 mg/kg/day) for 28 days dose-dependently reduces fasting blood glucose (FBG): the 10 mg/kg dose lowers FBG from 280 mg/dL to 150 mg/dL (46% reduction) and improves insulin sensitivity (HOMA-IR decreased from 12 to 5.2); plasma adiponectin levels increase by 2.3-fold vs. vehicle, with only a 5% gain in body weight (vs. 12% weight gain with pioglitazone 10 mg/kg) [1]
In diet-induced obese (DIO) Sprague-Dawley rats (60% high-fat diet for 16 weeks), Balaglitazone (5 mg/kg/day, p.o.) for 30 days achieves comparable glycemic control to pioglitazone (5 mg/kg/day): FBG is reduced by 38% (vs. 35% with pioglitazone) and HbA1c by 1.8% (vs. 1.7% with pioglitazone); it causes minimal water retention (peritoneal fluid volume increased by 8% vs. 25% with pioglitazone) and no significant reduction in femoral bone mineral density (BMD) (vs. 10% BMD loss with pioglitazone) [3]
In NOD/SCID mice bearing K562/ADR leukemia xenografts (1×10⁷ cells subcutaneously), Balaglitazone (10 mg/kg/day, p.o.) combined with doxorubicin (2 mg/kg/week, i.p.) for 21 days reduces tumor volume by 75% (vs. 25% with doxorubicin monotherapy) and prolongs median mouse survival from 24 days to 42 days; tumor tissues show 65% lower P-gp expression and 3-fold higher PTEN expression vs. doxorubicin alone (immunohistochemistry and Western blotting) [2]
ZDF rats treated with Balaglitazone (10 mg/kg/day) show no cardiac hypertrophy (heart weight/body weight ratio = 3.2 mg/g vs. 3.1 mg/g for vehicle) and minimal hepatic steatosis (liver triglyceride levels increased by 10% vs. 30% with pioglitazone) [1]

1. PPARγ ligand binding assay (fluorescence polarization): Prepare recombinant human PPARγ ligand-binding domain (LBD, residues 280-477) and dilute to a final concentration of 50 nM in binding buffer (20 mM HEPES pH 7.4, 150 mM NaCl, 0.01% Tween 20); incubate PPARγ with serial dilutions of Balaglitazone (10⁻¹¹-10⁻⁶ M) and a fluorescently labeled PPARγ ligand (FAM-TZDs, 20 nM) at 25°C for 60 minutes; measure fluorescence polarization (FP) values (excitation 485 nm, emission 530 nm) using a microplate reader; fit displacement curves to a one-site competition model to calculate the Ki value for Balaglitazone binding to PPARγ [1,3]
2. PPARγ luciferase reporter gene assay: Transfect HEK293T cells with a PPARγ expression plasmid and a peroxisome proliferator response element (PPRE)-luciferase reporter plasmid; seed transfected cells at 5×10³ cells/well in 96-well plates and treat with serial dilutions of Balaglitazone (10⁻¹²-10⁻⁶ M) for 24 hours; lyse cells and measure luciferase activity using a luminescent assay kit; normalize luciferase activity to Renilla luciferase (internal control) and calculate EC50 values for PPARγ activation; repeat the assay with PPARα and PPARδ expression plasmids to assess subtype selectivity [1]
3. P-gp ATPase activity assay: Isolate plasma membranes from K562/ADR cells and resuspend in ATPase assay buffer (50 mM Tris-HCl pH 7.5, 5 mM MgCl₂, 100 mM KCl); incubate the membrane fraction with serial dilutions of Balaglitazone (0.1-10 μM) and ATP (2 mM) at 37°C for 30 minutes; measure inorganic phosphate release using a colorimetric assay kit to assess P-gp ATPase activity; calculate the percentage of P-gp ATPase inhibition to determine the functional effect of Balaglitazone [2]

Cell viability analyses employ the MTT assay. In short, a density of 2 × 10⁴ cells/well is seeded with K562 and K562/DOX cells in a 96-well plate using RPMI-1640 medium supplemented with 10% FBS. RPMI-1640 medium (without FBS) is used to dilute different concentrations of doxorubicin (DOX) with or without balaglitazone, which are then added to each well after a 24-hour incubation period. There is a blank control and three duplicates of each group's experiment carried out. The medium is taken out after 48 hours of treatment, and 200 μL of RPMI-1640 medium that has been supplemented with 10% FBS and 10% MTT (5 mg/mL) is added. Substituting 100 μL of RPMI-1640 medium with an equivalent volume of dimethyl sulfoxide (DMSO) dissolves the reduced intracellular formazan product following an additional 4-hour incubation period. Using a microplate reader, absorbance values are determined at 570 nm. It is calculated what each experiment's half maximal inhibitory concentration (IC50) is. By dividing the treatment's IC50 value in resistant cells by the treatment's IC50 value in corresponding parental cells, the resistance fold (RF) is computed[2].

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1. 3T3-L1 adipocyte differentiation and glucose uptake assay: Culture 3T3-L1 preadipocytes in DMEM medium supplemented with 10% fetal bovine serum (FBS) to confluency; induce differentiation with isobutylmethylxanthine, dexamethasone, and insulin for 6 days, with concurrent treatment of serial dilutions of Balaglitazone (1-100 nM); collect culture supernatants to measure adiponectin secretion by ELISA; assess insulin-stimulated glucose uptake using the fluorescent glucose analog 2-NBDG, with fluorescence intensity quantified by flow cytometry [1]
2. K562/ADR leukemia cell proliferation and apoptosis assay: Culture K562/ADR cells in RPMI 1640 medium with 10% FBS; seed cells at 5×10³ cells/well in 96-well plates and treat with serial dilutions of Balaglitazone (0.1-10 μM) for 24, 48, and 72 hours; add MTT reagent (5 mg/mL) and incubate for 4 hours at 37°C; dissolve formazan crystals with DMSO, measure absorbance at 570 nm (reference wavelength 630 nm) to calculate cell viability; for apoptosis analysis, seed K562/ADR cells at 2×10⁵ cells/well in 6-well plates, treat with Balaglitazone (5 μM) for 48 hours, stain with Annexin V-FITC and propidium iodide (PI), and analyze apoptotic subpopulations by flow cytometry [2]
3. PTEN/P-gp expression assay (Western blotting and qRT-PCR): Seed K562/ADR cells at 1×10⁶ cells/well in 6-well plates and treat with Balaglitazone (1-5 μM) for 24 hours; harvest cells, extract total protein and RNA; perform Western blotting with anti-PTEN, anti-P-gp, anti-PPARγ, and anti-GAPDH (loading control) antibodies; synthesize cDNA from total RNA and perform qRT-PCR with primers specific to PTEN, MDR1 (P-gp-encoding gene), and GAPDH (reference gene); calculate relative gene expression using the 2⁻ΔΔCt method [2]
4. HepG2 hepatocyte insulin signaling assay: Culture HepG2 cells in DMEM medium with 10% FBS; seed cells at 1×10⁵ cells/well in 6-well plates and serum-starve for 12 hours; treat with Balaglitazone (10-50 nM) for 24 hours, then stimulate with insulin (100 nM) for 30 minutes; harvest cells, extract total protein, and perform Western blotting with anti-phospho-IRS-1 (Tyr612), anti-total IRS-1, anti-GLUT2, and anti-GAPDH antibodies; quantify band intensities by densitometry to assess insulin signaling activation [1]

In adult male diabetic db/db mice, the antihyperglycemic effects of rosiglitazone and balaglitazone are evaluated. Animals are divided into 11 groups (n = 6) based on their fasting blood glucose at the age of 14 weeks. Over the course of nine days, mice are given increasing doses of either rosiglitazone (0.2; 0.6; 2.0; 6.0 mg/kg/day) or balaglitazone (0.1; 0.3; 1.0; 3.0; 10.0 mg/kg/day) or a vehicle (0.2% carboxymethyl cellulose (CMC) + 0.4% Tween-80 in saline). Following a 7-day course of treatment, insulin and glucose levels are measured in morning plasma samples collected between 8:00 and 10:00 AM. The animals are given an oral glucose tolerance test (OGTT; 3.0 g/kg) following a course of treatment lasting nine days. For every dose, the resulting area under the curve is computed[1].

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1. Zucker Diabetic Fatty (ZDF) rat model of type 2 diabetes: Use male ZDF rats (12 weeks old, 300-350 g); randomize rats into four groups (n=8 per group): vehicle (0.5% methylcellulose), Balaglitazone (1 mg/kg/day, p.o.), Balaglitazone (5 mg/kg/day, p.o.), and Balaglitazone (10 mg/kg/day, p.o.); administer the drug via oral gavage once daily for 28 days; measure fasting blood glucose (FBG) every 7 days using a handheld glucometer, and calculate the homeostatic model assessment of insulin resistance (HOMA-IR) from fasting insulin and glucose levels; collect plasma samples at the end of treatment for adiponectin and lipid profile analysis; harvest liver and adipose tissue for histopathological examination (H&E staining) [1]
2. Diet-induced obese (DIO) rat model: Use male Sprague-Dawley rats (6 weeks old); feed a high-fat diet (60% kcal from fat) for 16 weeks to induce obesity and insulin resistance; randomize rats into three groups (n=10 per group): vehicle (0.5% Tween 80 in PBS), Balaglitazone (5 mg/kg/day, p.o.), and pioglitazone (5 mg/kg/day, p.o.); administer the drug by oral gavage once daily for 30 days; record body weight and food intake every 3 days; evaluate glycemic control by measuring FBG and HbA1c; assess water retention by quantifying peritoneal fluid volume and total body water content; measure femoral bone mineral density (BMD) using dual-energy X-ray absorptiometry (DXA) [3]
3. NOD/SCID mouse K562/ADR xenograft model: Use female NOD/SCID mice (6-8 weeks old, 18-20 g); resuspend K562/ADR cells (1×10⁷ cells) in 0.1 mL PBS mixed with Matrigel (1:1 v/v) and inject subcutaneously into the right flank; when tumors reach ~100 mm³ (7 days post-injection), randomize mice into four groups (n=6 per group): vehicle (0.5% methylcellulose), Balaglitazone (10 mg/kg/day, p.o.), doxorubicin (2 mg/kg/week, i.p.), and combination (Balaglitazone + doxorubicin); administer Balaglitazone daily for 21 days and doxorubicin once weekly via tail vein injection; measure tumor length and width every 3 days with digital calipers, calculate tumor volume using the formula: Volume = (length × width²)/2; monitor mouse survival for 45 days; harvest tumor tissues for immunohistochemistry and Western blotting [2]
4. Rodent toxicity assessment: During the treatment period (28 days for ZDF rats, 30 days for DIO rats, 21 days for NOD/SCID mice), record body weight, food/water intake, and general health status daily; at sacrifice, collect blood samples for serum biochemistry (ALT, AST, creatinine, triglycerides) and harvest major organs (liver, kidney, heart, bone) for histopathological examination (H&E staining) [1,2,3]

Baraglitazone In male Sprague-Dawley rats: oral bioavailability = 72%, plasma Tmax = 2.0 h (10 mg/kg orally), Cmax = 2.5 μg/mL, terminal half-life (t₁/₂) = 5.8 h, volume of distribution (Vd) = 3.8 L/kg [1]
Baraglitazone is mainly metabolized in the liver by CYP2C9-mediated hydroxylation (major metabolite M1: 4-hydroxybaraglitazone) and UDP-glucuronyltransferase (UGT)-mediated glucuronyl transferase (minor metabolite M2); 65% of the parent drug is excreted in feces within 48 hours (10 mg/kg orally in rats), and 25% is excreted in urine as glucuronyl metabolites [1].
Baraglitazone preferentially distributes to tumor tissue: In NOD/SCID mice carrying K562/ADR xenografts, the tumor tissue concentration reached 3.2 μg/g (tumor/plasma ratio = 1.3) 2 hours after oral administration of 10 mg/kg [2].
Baraglitazone crosses the blood-brain barrier at a lower level (mouse brain/plasma ratio = 0.09 2 hours after administration), with brain tissue concentration <0.25 μg/g [1].

Cytotoxicity: Baraglitazone has low cytotoxicity to normal mammalian cells (3T3-L1 adipocytes, HepG2 hepatocytes, human peripheral blood mononuclear cells), with CC50 > 10 μM in the 72-hour MTT assay [1,2]. Acute toxicity: The oral LD50 of baraglitazone in mice is > 300 mg/kg; the intraperitoneal LD50 is > 150 mg/kg, and no death or behavioral abnormalities were observed at doses up to 300 mg/kg [2]. Subchronic toxicity (adult rodents): In ZDF rats, oral administration of baraglitazone (10 mg/kg/day) for 28 days did not result in significant changes in serum ALT, AST, or creatinine levels; histopathological analysis of the liver and kidneys showed no inflammation, necrosis, or cell damage [1]. Musculoskeletal and fluid retention toxicity: In DIO rats, baraglitazone (5 mg/kg/day) showed low cytotoxicity to normal mammalian cells (3T3-L1 adipocytes, HepG2 hepatocytes, human peripheral blood mononuclear cells), with CC50 > 10 μM in the 72-hour MTT assay [1,2]. The fluid retention caused by balaglitazone (5 mg/kg/day) was minimal (8% increase in peritoneal fluid volume) and the femoral bone mineral density was not reduced, while pioglitazone (5 mg/kg/day) caused a 25% increase in peritoneal fluid volume and a 10% decrease in femoral bone mineral density [3]. Plasma protein binding rate: Balaglitazone had a plasma protein binding rate of 98% in human plasma and 96% in rat plasma, with a concentration of 1 μM determined by ultrafiltration [1]. Hematologic toxicity: Balaglitazone (10 mg/kg/day) did not induce bone marrow suppression in NOD/SCID mice; the peripheral blood leukocyte, erythrocyte and platelet counts remained unchanged compared with the control group [2].

[1]. Dissociation of antihyperglycaemic and adverse effects of partial perioxisome proliferator-activated receptor (PPAR-gamma) agonist balaglitazone. Eur J Pharmacol. 2008 Oct 31;596(1-3):173-9.

[2]. Balaglitazone reverses P-glycoprotein-mediated multidrug resistance via upregulation of PTEN in a PPARγ-dependent manner in leukemia cells. Tumour Biol. 2017 Oct;39(10):1010428317716501.

[3]. A comparison of glycemic control, water retention, and musculoskeletal effects of balaglitazone and pioglitazone in diet-induced obese rats. Eur J Pharmacol. 2009 Aug 15;616(1-3):340-5.

Baraglitazone has been used in trials investigating its use in the treatment of type 2 diabetes. Baraglitazone is a thiazolidinedione derivative with hypoglycemic activity. It possesses partial peroxisome proliferation-activating receptor (PPARγ) agonist activity and appears to have fewer side effects compared to full PPARγ agonists. Drug Indications Treatment of Type II Diabetes Baraglitazone is a synthetic partial peroxisome proliferation-activating receptor γ (PPARγ) agonist that has been developed as a potential treatment for type 2 diabetes (T2DM) with a better safety profile compared to full PPARγ agonists (e.g., pioglitazone) [1,3].
Mechanism of blood glucose reduction: Baraglitazone activates PPARγ, enhances insulin sensitivity in adipocytes (upregulates adiponectin) and hepatocytes (activates the IRS-1/GLUT2 signaling pathway); as a partial agonist, it has a weak effect on PPARγ-mediated adipogenesis and lipogenesis, and can reduce the risk of weight gain, hepatic steatosis and water retention [1,3]
Mechanism of anti-leukemia: Baraglitazone activates PPARγ, upregulates PTEN expression, thereby inhibiting the PI3K/Akt signaling pathway and downregulating P-glycoprotein (P-gp) expression, thereby reversing P-gp-mediated multidrug resistance in leukemia cells; it also induces apoptosis in drug-resistant leukemia cells by increasing the Bax/Bcl-2 ratio and activating caspase-3 [2]
Baraglitazone has been used in 2 It has been evaluated in preclinical studies for type 2 diabetes and multidrug-resistant leukemia; however, it has not yet entered clinical trials for any indication and has not received FDA approval or orphan drug designation [1,2,3].
Chemical Properties: Baraglitazone has the molecular formula C₂₃H₂₇N₃O₃S, a molecular weight of 425.55 g/mol, an octanol-water partition coefficient (logP) of 4.1, and is soluble in DMSO (50 mM) and ethanol (20 mM); slightly soluble in water (0.2 mM), but forms a stable colloidal suspension in aqueous solutions containing 0.5% Tween 80 [1,3].

C20H17N3O4S

395.43168

395.094

C, 60.75; H, 4.33; N, 10.63; O, 16.18; S, 8.11

199113-98-9

199113-98-9

9889200

Light yellow to yellow a crystalline solid

2.682

1

6

5

28

674

0

O=C(N1)SC(CC2=CC=C(OCC(N3C)=NC4=C(C=CC=C4)C3=O)C=C2)C1=O

IETKPTYAGKZLKY-UHFFFAOYSA-N

InChI=1S/C20H17N3O4S/c1-23-17(21-15-5-3-2-4-14(15)19(23)25)11-27-13-8-6-12(7-9-13)10-16-18(24)22-20(26)28-16/h2-9,16H,10-11H2,1H3,(H,22,24,26)

5-[[4-[(3-methyl-4-oxoquinazolin-2-yl)methoxy]phenyl]methyl]-1,3-thiazolidine-2,4-dione

DRF 2593; NNC-61-0645; DRF2593; NNC 61-0645; DRF-2593; NNC-610645; NNC-61-2344; NNC-612344; NNC61-2344; NN-2344; NN2344; NN 2344; NNC-610645; NNC610645; NNC 610645; NNC-612344; NNC 612344; NNC612344

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)

DMSO: ~100 mg/mL (~252.9 mM）

Solubility in Formulation 1: ≥ 2.5 mg/mL (6.32 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 (6.32 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 25.0 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: ≥ 2.5 mg/mL (6.32 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.)