PPARγ (IC50 = 19 nM); PPARα (IC50 = 38 nM)
PPARα (IC50 = 0.038 μM; EC50 = 0.050 μM, 156% effect relative to Farglitazar)
PPARγ (IC50 = 0.019 μM; EC50 = 0.021 μM, 67% effect relative to Edaglitazone)
PPARδ (EC50 = 0.053 μM, 22% effect relative to GW501516)[1]

Aleglitazar demonstrates species selectivity towards PPARα, as evidenced by its EC50 values of 50 nM, 2.26 µM, and 2.34 µM for PPARα in humans, rat PPARα, and mouse PPARα, respectively[1].
Aleglitazar (0.01-40 µM; 12-48 hours) significantly increases lactate dehydrogenase (LDH) release at concentrations of 30 µM and 40 µM, but not at concentrations of 0.1 µM to 20 µM[2].
Aleglitazar (0.01–20 µM; 48 hr) reduces caspase-3 activity, cytochrome-C release, and apoptosis induced by hyperglycemia (HG, glucose 25 mM)[2].
Aleglitazar increases the viability of cells exposed to high blood sugar levels[2].

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Aleglitazar attenuated hyperglycaemia-induced apoptosis, caspase-3 activity and cytochrome-C release and increased viability in human cardiomyocyte, cardiomyocytes from cardiac-specific peroxisome proliferator-activated receptor-γ knockout and wild-type mice. Hyperglycaemia reduced the antioxidant capacity and Aleglitazar significantly blunted this effect. Hyperglycaemia-induced reactive oxygen species production was attenuated by Aleglitazar in both human cardiomyocyte and wild-type mice cardiomyocytes. Aleglitazar improved cell viability in cells exposed to hyperglycaemia. The protective effect was partially blocked by short interfering RNA against peroxisome proliferator-activated receptor-α alone and short interfering RNA against peroxisome proliferator-activated receptor-γ alone and completely blocked by short interfering RNA to both peroxisome proliferator-activated receptor-α and peroxisome proliferator-activated receptor-γ.Conclusion: Aleglitazar protects cardiomyocytes against hyperglycaemia-induced apoptosis by combined activation of both peroxisome proliferator-activated receptor-α and peroxisome proliferator-activated receptor-γ in a short-term vitro model.[2]

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Aleglitazar showed potent agonistic activity at both PPARα and PPARγ receptors in binding and transactivation assays. The X‑ray crystal structure of the human PPARα ligand binding domain in complex with (S)-2Aa and a SRC‑1 coactivator fragment was solved at 2.2 Å resolution, revealing four strong hydrogen bonds between the ligand carboxylate and Ser, His, Tyr residues (all H‑bond distances ≤3.0 Å). Similarly, the ternary complex with human PPARγ ligand binding domain was solved at 2.3 Å, showing an analogous binding mode. The central benzothiophene linker fits snugly into the protein cavity.[1]
The structure–activity relationship indicated that increasing length and size of the R1 side chain enhances potency at PPARγ but reduces affinity for PPARα due to a smaller cleft in PPARα (Phe363↔Ile354 difference). (R)-isomers were almost inactive. Ortho-substitution on the phenyl-oxazole abolished activity completely.[1]

Aleglitazar (0.3–3.0 mg/kg; intraperitoneal; daily; for 7 days) improves the anatomical and functional results of mild brain ischemia[3].
Aleglitazar inhibits the production of NO, the release of proinflammatory cytokines, migration, and phagocytosis, among other important aspects of microglia activation[3].
Aleglitazar reduces post-ischemic brain inflammatory responses[3].

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In db/db mice, oral administration of Aleglitazar at 0.3 or 3 mg/kg/d for 12 days reduced non‑fasted plasma glucose levels and improved oral glucose tolerance. At ten times lower doses, it compared favorably with Rosiglitazone.[1]
In a hyperinsulinemic‑euglycemic clamp study in Zucker fa/fa rats treated for 7 days, (S)-2Aa significantly improved insulin resistance (HOMA‑IR index) compared to Rosiglitazone (3 mg/kg/d) and Farglitazar (10 mg/kg/d).[1]
In human ApoAI‑transgenic mice treated for 12 days, (S)-2Aa increased HDLc levels and induced a shift in HDL particle size. The effect was less pronounced than that of Fenofibrate, but Fenofibrate was used at 100‑1000 times higher doses. In contrast, the pure PPARγ agonist Farglitazar had no effect on HDLc.[1]
In (pre)diabetic rhesus monkeys, (S)-2Aa showed extraordinary efficacy, which is anticipated to translate into a stronger pharmacodynamic effect due to higher potency in primates.[1]

In this study, researchers incubated human cardiomyocytes, cardiomyocytes from cardiac-specific peroxisome proliferator-activated receptor-γ knockout or wild-type mice in normoglycaemic or hyperglycaemic conditions (glucose 25 mM). Cells were treated with different concentrations of Aleglitazar for 48 h. Researchers measured viability, apoptosis, caspase-3 activity, cytochrome-C release, total antioxidant capacity and reactive oxygen species formation in the treated cardiomyocytes. Human cardiomyocytes were transfected with short interfering RNA against peroxisome proliferator-activated receptor-α or peroxisome proliferator-activated receptor-γ[2].

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Radioligand binding assays were performed to determine IC50 values for human PPARα and PPARγ using standard protocols as described in Binggeli et al. Functional transactivation was measured using a luciferase transcriptional reporter gene assay in cells expressing the respective human PPAR receptors; effects were reported relative to Farglitazar (100% for PPARα) and Edaglitazone (100% for PPARγ).[1]
Co‑crystals of human PPARα‑LBD with (S)-2Aa and an SRC‑1 coactivator fragment were obtained using a protocol similar to Burgermeister et al. (Mol. Endocrinol. 2006, 20, 809). Data were collected in‑house on a rotating anode (λ=1.5418 Å) to a maximum resolution of 2.2 Å. Crystals belonged to orthorhombic space group P212121 with cell axes a=42.4, b=76.1, c=98.5 Å. The structure was determined by molecular replacement using chain A of PDB entry 1K71 and refined. Difference electron density was used to place the ligand by real space refinement.[1]
Co‑crystals of human PPARγ‑LBD with (S)-2Aa and SRC‑1 were obtained similarly, with data collected to 2.3 Å resolution. Crystals belonged to orthorhombic space group P212121 with cell axes a=53.9, b=70.0, c=88.5 Å. The structure was determined by molecular replacement using chain A of PDB entry 1PRG and refined.[1]

Cell Line: human cardiomyocytes (HCM), wild-type mice cardiomyocytes (mCM-WT)
Concentration: 0.01 µM, 0.05 µM, 0.1 µM, 0.5 µM, 1 µM, 5 µM, 10 µM, 20 µM, 30 µM, 40 µM
Incubation Time: 12 hours, 24 hours, 48 hours
Result: Increased LDH release at concentrations of 30 µM and 40 µM.

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Functional transactivation assays were performed using a luciferase reporter gene system in cells transfected with human PPARα, PPARγ, or PPARδ receptors. Cells were treated with the test compound, and luciferase activity was measured. EC50 values were calculated, and effects were expressed as percentage of the maximal response induced by reference agonists (Farglitazar for PPARα, Edaglitazone for PPARγ, GW501516 for PPARδ).[1]

Male 129S6/SvEv mice (24-30 g), middle cerebral artery occlusion (MCAo) models[3]
0.3 mg/kg, 3.0 mg/kg
Intraperitoneal injection, daily, for 7 days

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db/db mice: (S)-2Aa was administered orally once daily at doses of 0.3 or 3 mg/kg/d for 12 days. An oral glucose tolerance test (OGTT) was performed after overnight fasting on the day following the last drug administration (given by gavage on the prior day). Non‑fasted plasma glucose was measured in the morning after the dark/light cycle switch.[1]
Zucker fa/fa rats: Animals were treated with (S)-2Aa for 7 days, then subjected to a hyperinsulinemic‑euglycemic clamp study. HOMA‑IR index was calculated from fasting plasma insulin and fasting blood glucose (FBG) levels measured at time 0 of the OGTT (prior to glucose challenge) using the formula: HOMA‑IR = (fasting insulin (mU/ml) × FBG (mM)) / 22.5.[1]
Human ApoAI‑transgenic mice: (S)-2Aa was administered for 12 days. Plasma lipid levels were measured by FPLC from pooled plasma (N=8 per group). HDL particle size shift was also assessed.[1]
Rhesus monkeys: (S)-2Aa was tested in (pre)diabetic rhesus monkeys, showing extraordinary efficacy (specific dose regimen not detailed).[1]

Total clearance: 6.2 ml/min/kg in rat, 1.6 ml/min/kg in cynomolgus monkey.
Bioavailability: 70% in rat, 68% in cynomolgus monkey.
Half‑life: 4 h in rat, 12.9 h in cynomolgus monkey.[1]

In cytotoxicity assessment using LDH release, Aleglitazar at concentrations of 0.1 to 20 μM did not significantly increase LDH release in human cardiomyocytes and wild-type mouse cardiomyocytes after 12, 24, or 48 h. However, at 30 and 40 μM, there was a significant increase in LDH release. [2]
A phase III clinical trial (AleCardio) was halted due to safety signals and lack of efficacy in reducing cardiovascular events and mortality. The manufacturer terminated the AleCardio trial and all other trials involving Aleglitazar. [2]

[1]. Aleglitazar, a new, potent, and balanced dual PPARalpha/gamma agonist for the treatment of type II diabetes. Bioorg Med Chem Lett. 2009 May 1;19(9):2468-73.

[2]. Aleglitazar, a dual peroxisome proliferator-activated receptor-α and -γ agonist, protects cardiomyocytes against the adverse effects of hyperglycaemia. Diab Vasc Dis Res. 2017 Mar; 14(2): 152–162.

[3]. Dual PPARα/γ agonist aleglitazar confers stroke protection in a model of mild focal brain ischemia in mice. J Mol Med (Berl). 2019; 97(8): 1127-1138.

Aleglitazar, an investigational drug developed by Hoffmann-La Roche, is currently undergoing a Phase III clinical trial called ALECARDIO. This drug is being investigated for its use in treating patients with type 2 diabetes to reduce cardiovascular mortality and morbidity. Aleglitazar is a peroxisome proliferator-activated receptor (PPAR) agonist that simultaneously activates both PPARα and PPARγ receptor subtypes. In the Phase II clinical trial SYNCHRONY, Aleglitazar demonstrated synergistic control of blood lipid and blood glucose levels in patients with type 2 diabetes, with fewer side effects and toxicity. Aleglitazar is a dual peroxisome proliferator-activated receptor (PPAR) agonist with hypoglycemic activity. Aleglitazar binds to both PPARα and PPARγ, lowering plasma glucose, low-density lipoprotein cholesterol (LDL-C), and triglyceride (TG) levels, while raising high-density lipoprotein cholesterol (HDL) levels. This drug can be used to treat diabetes and cardiovascular disease. Drug Indications Alogliptin has been studied for the treatment of type 2 diabetes to reduce cardiovascular mortality and morbidity. Mechanism of Action Alogliptin is rationally designed as a peroxisome proliferator-activated receptor (PPAR) agonist, acting on both PPARα and PPARγ receptor subtypes. PPARα agonists control lipid levels, thereby improving dyslipidemia; PPARγ agonists control blood glucose levels, thereby improving insulin sensitivity in diabetic patients.

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Aleglitazar is a balanced dual PPARα/γ agonist designed to address both hyperglycemia and the enhanced cardiovascular risk of diabetic patients. It is an α-alkoxy-β-arylpropionic acid derivative with a benzothiophene core. The compound exhibited species selectivity for PPARα, with functional affinity dropping almost two orders of magnitude from human to rodent receptors, which explains the moderate lipid‑lowering effect in mice despite robust HDLc increase. The coordinates of (S)-2Aa co‑crystallized with human PPARα and PPARγ ligand binding domains have been deposited in the Protein Data Bank under PDBids 3G8I (PPARα) and 3G9E (PPARγ). The compound successfully completed phase II clinical development under the USAN name Aleglitazar.[1]

C24H23NO5S

437.51

437.13

C, 65.89; H, 5.30; N, 3.20; O, 18.28; S, 7.33

475479-34-6

475479-34-6

10274777

White to yellow solid powder

1.29

665.151ºC at 760 mmHg

356.071ºC

1.627

5.128

1

7

9

31

586

1

CC1=C(CCOC2=CC=C(C[C@@H](C(O)=O)OC)C3=C2C=CS3)N=C(C4=CC=CC=C4)O1

DAYKLWSKQJBGCS-NRFANRHFSA-N

InChI=1S/C24H23NO5S/c1-15-19(25-23(30-15)16-6-4-3-5-7-16)10-12-29-20-9-8-17(14-21(28-2)24(26)27)22-18(20)11-13-31-22/h3-9,11,13,21H,10,12,14H2,1-2H3,(H,26,27)/t21-/m0/s1

(2S)-2-methoxy-3-[4-[2-(5-methyl-2-phenyl-1,3-oxazol-4-yl)ethoxy]-1-benzothiophen-7-yl]propanoic acid

RG-1439; RO-0728804; RG1439; R 1439; Aleglitazar (USAN); Aleglitazar [USAN]; R1439; RO0728804; RO 0728804; RG 1439; R-1439; Aleglitazar

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: ≥ 50 mg/mL (~114.3 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (5.71 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 (5.71 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.)