PPARγ (Peroxisome proliferator-activated receptor γ): IC50 for PPARγ binding = 18.25 μM [1]
- Mitochondrial Pyruvate Carrier (MPC): Interacts with the MPC complex [2]

In primary hepatocytes isolated from diet-induced obese (DIO) mice, treatment with MSDC-0602 (1 μM for 18 h) suppressed the expression of gluconeogenic genes (Pck1 and G6pc) and lipogenic genes (Fasn, Scd1, and Sreb1) at both mRNA and protein levels. This suppression was also observed in hepatocytes from liver-specific PPARγ knockout mice. [1]
- In primary hepatocytes from DIO mice, treatment with MSDC-0602 (5 μM for 24 h) significantly suppressed hepatocyte glucose production. [1]
- In differentiating 3T3-L1 adipocytes, MSDC-0602 (0.1 μM) did not significantly induce the expression of PPARγ target genes (Fabp4, Cidec, Cd36, and Plin4). [1]
- In a BRET-based RESPYR assay using MPC2-knockout HEK293T cells expressing truncated MPC2 (lacking first 16 amino acids), MSDC-0602 (10 μM) caused a dramatic and immediate increase in RESPYR activity, similar to its effect on the wild-type MPC complex. [2]
- In permeabilized HEK293T cells expressing an MPC2 truncation (MPC2^Δ22, the human equivalent of mouse Mpc2^Δ16), MSDC-0602 (20 μM) fully retained its ability to suppress pyruvate-stimulated respiration. In contrast, complete MPC2 knockout cells were insensitive to the inhibitory effects of MSDC-0602 on oxygen consumption. [2]

In ob/ob mice, dietary administration of MSDC-0602 (300 ppm in diet for 4 weeks, resulting in a blood concentration of 2-5 μM) corrected elevated plasma glucose, non-esterified fatty acids, triglycerides, cholesterol, and insulin concentrations. It also significantly improved glucose and insulin tolerance. [1]
- In ob/ob mice, MSDC-0602 treatment improved insulin-stimulated phosphorylation of Ser-473 and Thr-308 of Akt in gastrocnemius muscle. It also corrected the expression of Fabp4, adiponectin, and TNFα in adipose tissue and reduced expression of macrophage cell surface markers (Cd68 and F4/80). [1]
- In diet-induced obese (DIO) mice, MSDC-0602 treatment (300 ppm in high-fat diet for 2 or 4 weeks) significantly lowered plasma glucose and insulin levels and increased plasma adiponectin. Hyperinsulinemic-euglycemic clamp studies showed that MSDC-0602 increased the glucose rate of infusion, enhanced insulin-stimulated glucose uptake into gastrocnemius, adipose tissue, and heart, and improved the suppression of hepatic glucose production by insulin. [1]
- In DIO mice, MSDC-0602 administration (2 or 4 weeks) markedly improved insulin-stimulated phosphorylation of Akt in liver and skeletal muscle. It also reduced the hepatic expression of gluconeogenic enzymes (Pck1, G6pc). [1]
- In DIO mice, MSDC-0602 treatment (4 weeks) significantly increased hepatic mitochondrial oxygen consumption rates (using succinate as a substrate) under basal, ADP-stimulated, and uncoupled conditions. It also reduced hepatic expression of lipogenic genes (Gk, Lpk, Acl, Fasn, Mel, Thrs, Sreb1) and triglyceride synthesis enzymes (Lpin1, Scd1), leading to suppressed rates of de novo fatty acid synthesis and oleate-stimulated TG synthesis in isolated hepatocytes. [1]
- In DIO mice, MSDC-0602 treatment, unlike rosiglitazone, did not induce the hepatic expression of PPARγ target genes that promote lipid storage (Fabp4, Cidec, Cd36, Plin4). [1]
- In both wild-type and Mpc2^Δ16 hypomorphic mice fed a high-fat diet, MSDC-0602 administration (331 ppm in diet for 4 weeks) markedly improved glucose tolerance, reduced hepatic steatosis, lowered plasma triglycerides, cholesterol, free fatty acids, and insulin, and increased plasma adiponectin. It also suppressed adipose tissue expression of proinflammatory markers (Cd68, Emr1, Mcp1, Tnfa) and stimulated the expression of browning markers (Ucp1, Prdm16, Ppargc1a, Cidea, Cox2, Acadm) in epididymal white adipose tissue. The efficacy of MSDC-0602 was not attenuated in the Mpc2^Δ16 mice. [2]

PPARγ competitive binding assay: The ability of MSDC-0602 to bind to the ligand binding domain of PPARγ was assessed using a TR-FRET-based LanthaScreen competitive binding assay. The IC50 value for MSDC-0602 was determined to be 18.25 μM. [1]
- Gal4-PPARγ transactivation assay: HepG2 cells were co-transfected with expression vectors for Gal4-PPARγ (ligand binding domain only), a firefly luciferase reporter construct driven by Gal4 response elements, and a renilla luciferase reporter construct. Transfected cells were treated with MSDC-0602 for 24 hours. Even at a concentration of 50 μM, MSDC-0602 only minimally activated Gal4-PPARγ. [1]
- Gal4-PPARα transactivation assay: Using a similar co-transfection system with a Gal4-PPARα construct, MSDC-0602 did not activate PPARα at physiological concentrations. The EC50 for PPARα activation was greater than 100 μM. [1]
- Mitochondrial binding assay: A radiolabeled probe (125I-labeled MSDC-1101) was used in a cross-linking reaction with mitochondrial membranes. Unlabeled MSDC-0602 competed with the probe for binding to mitochondrial membranes, demonstrating that MSDC-0602 binds to mitochondrial membranes with affinity equivalent to that of rosiglitazone and pioglitazone. [1]

Primary mouse hepatocyte isolation and treatment: Hepatocytes were isolated from diet-induced obese (DIO) mice or liver-specific PPARγ knockout mice. Cells were plated and treated with MSDC-0602 (1 μM or 5 μM for 18-24 h) or vehicle (DMSO). [1]
- Fatty acid synthesis assay in hepatocytes: After treatment, hepatocytes were incubated with [14C] acetate to assess rates of de novo fatty acid synthesis. [1]
- Triglyceride synthesis assay in hepatocytes: Hepatocytes were incubated with [3H] glycerol in the presence of 300 μM oleate to quantify triglyceride synthesis rates. [1]
- Glucose production assay in hepatocytes: After an 18-hour pretreatment with MSDC-0602 or vehicle, hepatocytes were washed and incubated in buffer containing lactate and glucagon with or without insulin. Glucose content in the media was determined enzymatically. [1]
- 3T3-L1 adipocyte differentiation: Preadipocytes were grown to confluence and differentiation was initiated with a hormone mixture. Cells were treated with vehicle, rosiglitazone (0.1 μM), pioglitazone (0.1 μM), or MSDC-0602 (0.1 μM). RNA was collected after 48 hours for analysis of PPARγ target genes. [1]
- RESPYR (Reporter Sensitive to Pyruvate) BRET assay: HEK293T cells lacking endogenous MPC2 were transiently transfected with MPC1-Venus and either wild-type MPC2-RLuc8 or a mutant MPC2-RLuc8 lacking the first 16 amino acids. After 48 hours, cells were stimulated with vehicle, 5 mM pyruvate, 5 μM UK-5099, or 10 μM MSDC-0602, and BRET was measured. [2]
- Oxygraph respiration in permeabilized cells: HEK293T cells (wild-type, MPC2 knockout, or expressing MPC2^Δ22) were suspended in respiration buffer and permeabilized with digitonin. After establishing maximal pyruvate/malate-stimulated respiration with ADP, 20 μM MSDC-0602 was injected to test its acute effect on pyruvate-mediated respiration. [2]

Ob/ob mouse study: 6-week-old male ob/ob mice were fed a diet containing MSDC-0602 (300 ppm) for 4 weeks. The dose was chosen to achieve a blood concentration of 2-5 μM. Mice were sacrificed after a 4-hour fast. In a subset, insulin (10 milliunits/g body weight) was injected intraperitoneally 5 minutes before sacrifice. [1]
- Diet-induced obesity (DIO) mouse study: Male C57BL/6 mice were fed a 60% high-fat diet from 6 weeks of age. After 8 weeks on the high-fat diet, mice were randomized to continue the high-fat diet or receive the high-fat diet containing MSDC-0602 (300 ppm, providing ~30 mg/kg/day) for an additional 2 or 4 weeks. [1]
- Hyperinsulinemic-euglycemic clamp in DIO mice: DIO mice, untreated or treated with MSDC-0602 for 3 weeks, were surgically catheterized in the jugular vein and carotid artery. After recovery, conscious mice were subjected to a hyperinsulinemic-euglycemic clamp. Insulin was infused at 2.5 milliunits/kg/min. A bolus of 2-[1-14C]deoxyglucose was given to estimate tissue-specific glucose uptake. [1]
- High-fat diet feeding in Mpc2^Δ16 mice: 6-8 week-old male wild-type and Mpc2^Δ16 mice were fed a 60% high-fat diet for 10 weeks. Mice were then randomized to remain untreated or receive high-fat chow containing pioglitazone (300 ppm) or MSDC-0602 (331 ppm) for an additional 4 weeks. Body weight was monitored weekly. Glucose tolerance tests were performed after 2 weeks of treatment. [2]

Blood concentration: In obese mice, dietary administration of MSDC-0602 at 300 ppm resulted in a blood concentration of 2-5 μM. [1]
- Dosage: The dosage selected for mouse studies (300 ppm in diet) provides approximately 30 mg/kg/day of MSDC-0602. [1]

Unlike PPARγ agonists, MSDC-0602 did not cause weight gain. In DIO mice, MSDC-0602 treatment actually led to a notable amount of weight loss, which was not associated with increased locomotor activity or an overt reduction in food intake. [1][2]
- MSDC-0602 did not induce the hepatic expression of PPARγ target genes that promote lipid storage (e.g., Fabp4, Cidec, Cd36, Plin4), suggesting a reduced risk of PPARγ-mediated side effects such as hepatic steatosis. [1]

[1]. Resistance and metabolic derangements in obese mice are ameliorated by a novel peroxisome proliferator-activated receptor γ-sparing thiazolidinedione. J Biol Chem. 2012 Jul 6;287(28):23537-48.

[2]. The beneficial metabolic effects of sensitizers are not attenuated by mitochondrial pyruvate carrier 2 hypomorphism. Exp Physiol. 2017 Aug 1;102(8):985-999.

[3]. Loss of Mitochondrial Pyruvate Carrier 2 in the Liver Leads to Defects in Gluconeogenesis and Compensation via Pyruvate-Alanine Cycling. Cell Metab. 2015 Oct 6;22(4):682-94.

[4]. Targeting the mitochondrial pyruvate carrier attenuates fibrosis in a mouse model of nonalcoholic steatohepatitis. Hepatology. 2017 May;65(5):1543-1556.

[5]. MSDC-0160 and MSDC-0602 binding with human mitochondrial pyruvate carrier (MPC) 1 and 2 heterodimer: PPARγ activating and sparing TZDs as therapeutics. Int. J. Knowl. Knowl. Bioinform.2017, 7, 43–67.

[6]. Insulin resistance and metabolic derangements in obese mice are ameliorated by a novel peroxisome proliferator-activated receptor γ-sparing thiazolidinedione. J Biol Chem. 2012 Jul 6;287(28):23537-48.

Azeglitazone is being investigated in the clinical trial NCT01280695 (a randomized, double-blind, controlled, placebo-controlled, multi-dose study designed to evaluate the safety, tolerability, and efficacy of three dose levels of MSDC-0602 in patients with type 2 diabetes).

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MSDC-0602 is a novel thiazolidinedione (TZD) analog designed with modifications to the carbon backbone to impair binding to PPARγ, making it a PPARγ-sparing TZD (PsTZD). [1]
- The beneficial pharmacology of MSDC-0602 on insulin sensitivity suggests that PPARγ-sparing TZDs can be effective for treating type 2 diabetes with a potentially reduced risk of PPARγ-mediated side effects. [1]
- In a Phase 2a clinical trial with a first-generation PsTZD (MSDC-0160), beneficial effects on glucose, insulin, circulating lipids, and blood pressure were observed without causing weight gain or blood volume expansion. [1]
- The interaction between TZDs and the mitochondrial pyruvate carrier (MPC) is a proposed alternative mechanism of action. MSDC-0602 has been shown to bind to mitochondrial membranes and interact with the MPC complex. [1][2]
- In cells with a truncated MPC2 protein (lacking first 16 amino acids), the ability of MSDC-0602 to interact with the MPC and suppress pyruvate-mediated respiration was fully retained, indicating that the N-terminus of MPC2 is not required for these effects. [2]

C19H17NO5S

371.4

371.082

1133819-87-0

Azemiglitazone potassium;1314533-27-1

25230266

White to off-white solid powder

1.3±0.1 g/cm3

608.1±40.0 °C at 760 mmHg

321.6±27.3 °C

0.0±1.7 mmHg at 25°C

1.617

2.82

1

6

7

26

532

0

YAUMOGALQJYOJQ-UHFFFAOYSA-N

InChI=1S/C19H17NO5S/c1-24-15-4-2-3-13(10-15)16(21)11-25-14-7-5-12(6-8-14)9-17-18(22)20-19(23)26-17/h2-8,10,17H,9,11H2,1H3,(H,20,22,23)

5-(4-(2-(3-methoxyphenyl)-2-oxoethoxy)benzyl)thiazolidine-2,4-dione

MSDC-0602 MSDC0602 Azemiglitazone MSDC 0602

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 : ~125 mg/mL (~336.56 mM)
H2O : < 0.1 mg/mL

Solubility in Formulation 1: ≥ 2.08 mg/mL (5.60 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 20.8 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: ≥ 2.08 mg/mL (5.60 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 20.8 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: ≥ 2.08 mg/mL (5.60 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 20.8 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.)