PPARα (EC50 = 6 nM); Chk2 (IC50 = 697.4 nM); PPARγ (EC50 = 1.1 μM); PPARδ (EC50 = 6.2 μM)
Peroxisome proliferator-activated receptor alpha (PPARα) agonist. [1]

GW7647 (1 μM) causes both in the presence and absence of IL-1β a significant increase in PDZK1 protein expression in Caco2BBE cells, reaching 129.7 ± 6.5% of vehicle treated control[1]. GW7647 also attenuates the PDZK1 expression decrease mediated by IL-1β.
GW7647(50 nM) increases the amounts of NO released by stimulating the phosphorylation of PI3K and then Akt (Ser473) in the stripped antral mucosa. In antral mucous cells, GW7647 (50 nM) amplifies the first stage of Ca2+-regulated exocytotic events triggered by ACh, but GW7647 by itself does not cause any exocytotic events. In antral mucous cells, GW7647 plus ACh increases the effects of wortmannin (50 nM) and AKT-inh (100 nM) on exocytotic events[2].
GW7647 (100 nM) decreases the AQP9 protein abundance by 43%; however, at 10 and 1,000 nM, it has no discernible effec in WIF-B9 hepatocytes. HepG2 cells exposed to GW7647 (100 nM) exhibit a 24% decrease in AQP9 protein abundance; however, L-FABP protein abundance in HepG2 hepatocytes is not significantly increased[3].

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Effect of the PPARα agonist GW7647 on PDZK1 expression [1]
The transcription factor peroxisome proliferator-activated receptor alpha (PPARα) has been identified as a strong activator of the PDZK1 promoter (Tachibana et al., 2008). The peroxisome proliferator response element, to which the heterodimer PPARα and RXRα binds, is not located in the 5′ end of the PDZK1 promoter, but close to the start of transcription site at 98–86 bp, thus, not in the region sensitive to IL-1ß. Interestingly, IL-1β incubation also decreased PPARα expression in Caco2BBE cells to 65.5 ± 9.6% at 24 h incubation (Supplementary Figure 4), whereas a significant effect on PDZK1 mRNA expression was seen earlier (Figure 3A). It is nevertheless feasible that a PPARα agonist is able to restore PDZK1 expression in pro-inflammatory settings, offering a window for pharmacological intervention. We therefore incubated Caco2BBE cells with the PPARα agonist GW7647, in the absence and presence of IL-1β. At 24 h after GW7647 application, resulted in a significant increase of PDZK1 protein expression to 129.7 ± 6.5% of vehicle treated control (Figures 8A,B), and GW7647 was able to attenuate the IL-1β-mediated decrease in PDZK1 expression (61.1 ± 3.4% in the absence and 33.9 ± 2.3% in the presence of GW7647).

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A PPARα (peroxisome proliferation activation receptor α) agonist (GW7647) activates nitric oxide synthase 1 (NOS1) to produce NO leading to cGMP accumulation in antral mucous cells. In this study, we examined how PPARα activates NOS1. The NO production stimulated by GW7647 was suppressed by inhibitors of PI3K (wortmannin) and Akt (AKT 1/2 Kinase Inhibitor, AKT-inh), although it was also suppressed by the inhibitors of PPARα (GW6471) and NOS1 (N-PLA). GW7647 enhanced the ACh (acetylcholine)-stimulated exocytosis (Ca(2+)-regulated exocytosis) mediated via NO, which was abolished by GW6471, N-PLA, wortmannin, and AKT-inh. The Western blotting revealed that GW7647 phosphorylates NOS1 via phosphorylation of PI3K/Akt in antral mucous cells. The immunofluorescence examinations demonstrated that PPARα existing with NOS1 co-localizes with PI3K and Akt in the cytoplasm of antral mucous cells. ACh alone and AACOCF3, an analogue of arachidonic acid (AA), induced the NOS1 phosphorylation via PI3K/Akt to produce NO, which was inhibited by GW6471. Since AA is a natural ligand for PPARα, ACh stimulates PPARα probably via AA. In conclusion, PPARα activates NOS1 via PI3K/Akt phosphorylation to produce NO in antral mucous cells during ACh stimulation. [2]

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In Caco-2BBE cells, pretreatment with 1 μM GW7647 for 1 hour followed by co-incubation with IL-1β (10 ng·mL⁻¹) for 48 hours significantly attenuated the IL-1β-mediated decrease in PDZK1 protein expression. The inhibitory effect of IL-1β on PDZK1 expression was reduced from 61.1 ± 3.4% inhibition (in the absence of GW7647) to 33.9 ± 2.3% inhibition (in the presence of GW7647). Furthermore, GW7647 (1 μM, 24 hours) alone significantly increased basal PDZK1 protein expression to 129.7 ± 6.5% of vehicle-treated control levels. [1]

GW7647 (3 mg/kg per day) inhibits the decrease in left ventricular ejection fraction but does not stop the development of cardiac hypertrophy in vivo[4].

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Volume-overload cardiac hypertrophy was produced in 7-day-old rabbits via an aorto-caval shunt, after which, the rabbits were treated with or without GW7647 (3 mg/kg per day) for 14 days. Biventricular working hearts were subjected to 35 minutes of aerobic perfusion, 25 minutes of global no-flow ischemia, and 30 minutes of aerobic reperfusion. GW7647 treatment did not prevent the development of cardiac hypertrophy, but did prevent the decline in left ventricular ejection fraction in vivo. GW7647 treatment increased cardiac fatty acid β-oxidation rates before and after ischemia, which resulted in a significant increase in overall ATP production and an improved in vitro post-ischemic functional recovery. A decrease in post-ischemic proton production and endoplasmic reticulum stress, as well as an activation of sarcoplasmic reticulum calcium ATPase isoform 2 and citrate synthase, was evident in GW7647-treated hearts.
Conclusions: Stimulating fatty acid β-oxidation in neonatal hearts may present a novel cardioprotective intervention to limit post-ischemic contractile dysfunction. [4]

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GW7647 Treatment Improves In Vivo and In Vitro Contractile Function in Hypertrophied Neonatal Rabbit Hearts. GW7647 Treatment Prevents the Hypertrophy-Induced Shift in Myocardial Energy Metabolism. GW7647 Treatment Enhances Rates of ATP Production and Tricarboxylic Acid Cycle Activity in Hypertrophied Hearts. GW7647 Treatment Increases Mitochondrial Biogenesis and Fatty Acid Oxidative Capacity. GW7647 Treatment Reduces Myocardial Triacylglycerol Content by Enhancing Its Turnover. GW7647 Treatment Activates Calcium-Handling Proteins, Reduces Ceramide Synthesis, Endoplasmic Reticulum Stress, and Reduces Inflammation. [4]

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Administration of GW7647 (3 mg/kg po bid) to cholesterol/cholic acid-fed rats5 for 4 days resulted in a 60% increase in HDL-cholesterol, a 60% decrease in triglycerides, and a 40% decrease in serum apolipoprotein CIII [5].

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In neonatal rabbits with volume-overload cardiac hypertrophy induced by aorto-caval shunt, treatment with GW7647 (3 mg/kg/day, i.p., for 14 days) prevented the hypertrophy-induced decline in left ventricular ejection fraction in vivo, restoring it to levels comparable with sham-operated hearts. [4]
In ex vivo isolated biventricular working heart perfusions, hearts from GW7647-treated hypertrophied rabbits showed markedly improved recovery of cardiac function during reperfusion after 25 minutes of global no-flow ischemia, recovering to similar levels as sham hearts, whereas vehicle-treated hypertrophied hearts had significantly worse recovery. [4]
GW7647 treatment increased cardiac fatty acid β-oxidation rates both before and after ischemia in hypertrophied hearts. [4]
This led to a significant increase in overall ATP production rates and an increase in tricarboxylic acid cycle activity (measured as acetyl-CoA production) in GW7647-treated hypertrophied hearts compared to vehicle-treated ones. [4]
GW7647 treatment attenuated the hypertrophy-induced increase in glycolysis rates, especially during the post-ischemic period. [4]
Consequently, myocardial proton production (which increases when glycolysis is uncoupled from glucose oxidation) was significantly decreased in GW7647-treated hypertrophied hearts, especially post-ischemia. [4]
GW7647 treatment increased the expression of malonyl-CoA decarboxylase and decreased malonyl-CoA levels in hypertrophied hearts. [4]
It increased the incorporation of radiolabeled palmitate into the myocardial triacylglycerol (TG) pool, decreased total TG content, and increased the expression of adipose triglyceride lipase (ATGL), suggesting increased TG turnover. [4]
GW7647 treatment increased glycerol-3-phosphate dehydrogenase (GPD) activity in hypertrophied hearts. [4]
It prevented the hypertrophy-mediated decrease in sarcoplasmic reticulum calcium ATPase isoform 2 (SERCA2) protein expression and increased phosphorylated phospholamban (p-PLN) at Ser-16 in hypertrophied hearts. [4]
GW7647 treatment prevented the hypertrophy-induced increase in the endoplasmic reticulum (ER) stress marker glucose-regulated protein 78 (GRP78) and the pro-apoptotic protein BNIP3 in the left ventricle. It also attenuated the hypertrophy-induced increase in serine-palmitoyltransferase (SPT1 and SPT2) expression in the right ventricle. [4]
GW7647 treatment reduced the nuclear retention of nuclear factor-κB subunit p65 (NF-κB p65) in the right ventricle of hypertrophied hearts and increased cytosolic expression of its inhibitor, IκBα. [4]
GW7647 treatment increased citrate synthase activity in the left ventricle of hypertrophied hearts. [4]

Cell culture, seeding density, and cytokine treatment [1]
\nCaco-2BBE cells were grown at 37°C in a humidified atmosphere containing 5% CO2 and 95% O2, in Dulbecco's Modified Eagle Medium containing 4.5 g·L−1 D-glucose and sodium pyruvate supplemented with 10% FCS, 50 units penicillin/streptomycin, and 1% non-essential amino acids. Huh-7 cells were grown in Dulbecco's Modified Eagle Medium containing 4.5 g·L−1 D-glucose and sodium pyruvate supplemented with 10% FCS, 50 units penicillin/ streptomycin. 0.25 × 106 cells were plated in 6 well-plates and grown for 24 h, serum starved (0.5% FCS in medium) for 24 h and then treated with IL-1β (0.1, 1, or 10 ng·mL−1), TNF-α (up to 20 ng·mL−1) and IFN-γ (up to 30 ng·mL−1) for the times indicated in the respective figures. Concentration response curves for these cytokines in Caco-2BBE cells had been tested in a previous study (Yeruva et al., 2008), and were applied accordingly in this study. For each experiment, two (qPCR) or three (Western Analysis) individual dishes were studied for each experimental condition, and the results pooled for n = 1. Each experiment was repeated three times in different cell passages. n indicates the number of repeats in different cell passages. Due to the homogeneity of a cell line, and the duplicate or triplicate measurements within the same experiment, we believe that three repeats are acceptable for a statistically valid conclusion.\n

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\n\nTransient transfections and luciferase assays [1]
\n3 × 104 cells were seeded in 24 wells and grown overnight. Two hundred and fifty nanograms of plasmid per well was mixed with 10 ng of renilla luciferase plasmid and transfections were done using a Jet prime® polyplus transfection reagent from Peqlab according manufacturer's protocol. Cells were serum starved overnight before adding cytokines and then treated with cytokines for the time periods indicated in the text. After the treatment cells were lysed in 1X passive lysis buffer by shaking at room temperature for 15 min. Luciferase assay was performed as described previously.\n

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\n\nPatients selection [1]
\nThe details of the patients who provided the biopsies of UC patients are given in detail in a previous report (Yeruva et al., 2015).\n

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\n\nInhibition of NF-κB and MAPKs pathways [1]
\nFor NF-κB and MAPK pathway inhibition experiments, Caco-2BBE cells were pretreated with a NF-κB inhibitor BAY11-7082 (10 μM), a p38MAPK inhibitor BIRB-796 (μM), a JNK inhibitor SP600125 (25 μM), MEK1/2 inhibitor PD98059 (30 μM), and U0126 (10 μM) for 1 h, followed by exposure to IL-1β (10 ng·mL−1) for 48 h. Cells were lysed for Western blot analysis as described below.\n

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\n\n9-cis retinoic acid treatment experiments [1]
\nFor PDZK1 mRNA measurements, Caco-2BBE cells were pretreated for 30 mins with 9-cis retinoic acid (RA) or vehicle at a concentration of 1 μM and then treated with IL-1β for 3, 6, 12, and 24 h. For PDZK1 protein assessment, the cells were pretreated with 1 μM RA or vehicle for 30 min before addition of IL-1β (10 ng·mL−1) and the cells were harvested after 48 h.\n

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\n\nRNA isolation and real-time PCRs [1]
\nRNA isolation from cells was done using Qiagen RNA isolation kit and real time PCRs were performed as explained previously (Yeruva et al., 2015).\n

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\n\nImmunoblot analysis [1]
\nAfter the treatment, cells were lysed in lysis buffer and protein concentration was estimated with Bio-rad Bradford assay. Twenty to forty micrograms of total cellular proteins were separated on 8–10% SDS-poly acryl amide gels and transferred to polyvinylene difluoride membranes. Antibodies were diluted in TBST containing 5% non-fat dry milk and blots were incubated overnight at 4°C, washed with TBST and incubated with secondary antibodies conjugated to horseradish peroxidase, washed with TBST and then developed using enhanced chemiluminescence kit from GE health sciences.\n

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\n\nWST-1 cell viability assay [1]
\nThe reagent WST-1 was used to determine cell viability according to the manufacturer's instructions. In brief, Caco-2BBE cells were seeded at a density of 1 × 104 cells in each well of a 96 well-plate and were grown and treated as stated in the section “Cell culture, seeding density, and cytokine treatment.” Cells were incubated with respective cytokines for 24 h. At the end of treatment, 10 μl WST-1 were added to each well and 1 h later absorbance was measured at 450 and 630 nm using the BioTek® Epoch Reader. No decrease in viability was detected during exposure of any of the tested cytokines or their combination (Supplementary Figure 5).

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The effect of GW7647 on PDZK1 expression was assessed in the human intestinal epithelial cell line Caco-2BBE. Cells were grown in Dulbecco's Modified Eagle Medium supplemented with 10% FCS, penicillin/streptomycin, and non-essential amino acids. For the experiments, cells were plated and after 24 hours, serum-starved (0.5% FCS) for another 24 hours. Cells were then pretreated with 1 μM GW7647 (or vehicle) for 1 hour. Following pretreatment, cells were treated with or without IL-1β (10 ng·mL⁻¹) for an additional 48 hours. After treatment, cells were lysed for protein isolation. Total cellular proteins were separated by SDS-PAGE, transferred to membranes, and subjected to immunoblot analysis using a PDZK1-specific antibody. Protein expression was normalized to a loading control (β-actin) and quantified using image analysis software. [1]

New Zealand newborn Seven-day-old white rabbits weighing between 90 and 200 grams are anesthetized with 2% isofluorane inhaled, and they undergo an aorto-caval shunt to cause volume-overload cardiac hypertrophy. When using a color flow doppler on postoperative days 7 and 13, it is possible to see a physical shunt between the inferior vena cava and the abdominal aorta in both the axial and transverse planes, indicating the presence of a successful fistula. The enlarged inferior vena cava serves as additional confirmation of this. Following validation, animals in the shunt group are randomized to receive either GW7647 (3 mg/kg per day; EC50=6 nM for PPARα) or the vehicle (dimethyl sulfoxide, the solvent of GW7647) twice daily for 14 days via intraperitoneal injection. Animals that have surgery to create a shunt are not allowed to continue in the study if the shunt closes or does not exhibit. Transthoracic echocardiography is used to measure the left ventricular ejection fraction (%) and other cardiac parameters on postoperative days 7 and 13. All animals are put to sleep with Na⁺ pentobarbital at age 21 (14 days after surgery), and their hearts are removed for isolated biventricular working heart perfusions.[4]

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Assessment of Myocardial Function [4]
Newborn New Zealand White rabbits of either sex (7 days old, 90–200 g) were anesthetized with inhaled isofluorane (2%), and were subjected to an aorto-caval shunt to induce volume-overload cardiac hypertrophy as described previously.18 The presence of a successful fistula was verified at postsurgical days 7 and 13 by color flow doppler that visualizes a physical shunt between the abdominal aorta and the inferior vena cava in both an axial and transverse plane. This is further validated by an enlarged inferior vena cava (an expanded Methods section is available in the Online Data Supplement). After validation, the animals in shunt group were randomly assigned to receive an intraperitoneal injection of vehicle (dimethyl sulfoxide, the solvent of GW7647) or GW7647 (3 mg/kg per day; EC50=6 nmol/L for PPARα) twice a day for 14 days. Animals that underwent surgery to create shunt, but consequently the shunt either did not exhibit or closed, were excluded from the study. Left ventricular ejection fraction (%) and other cardiac parameters were assessed by transthoracic echocardiography at postsurgical days 7 and 13 as described previously.18 At 21 days of age (14 days post surgery), all animals were euthanized with Na+ pentobarbital, and hearts were removed for isolated biventricular working heart perfusions.22

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Volume-overload cardiac hypertrophy was induced in 7-day-old New Zealand White rabbits via surgical creation of an aorto-caval shunt. After verification of a successful shunt, animals were randomly assigned to receive intraperitoneal injections of either GW7647 (3 mg/kg per day) or vehicle (dimethyl sulfoxide, DMSO) twice daily for 14 days. Left ventricular function was assessed by transthoracic echocardiography at postsurgical days 7 and 13. At 21 days of age (14 days post-surgery), animals were euthanized, and hearts were removed for ex vivo isolated biventricular working heart perfusions. [4]

[1]. IL-1β-Induced Downregulation of the Multifunctional PDZ Adaptor PDZK1 Is Attenuated by ERK Inhibition, RXRα, or PPARα Stimulation in Enterocytes. Front Physiol. 2017 Feb 7;8:61.

[2]. PPARα induced NOS1 phosphorylation via PI3K/Akt in guinea pig antral mucous cells: NO-enhancement in Ca(2+)-regulated exocytosis. Biomed Res. 2016;37(3):167-78.

[3]. Hepatic AQP9 expression in male rats is reduced in response to PPARα agonist treatment. Am J Physiol Gastrointest Liver Physiol. 2015 Feb 1;308(3):G198-205.

[4]. Activating PPARα prevents post-ischemic contractile dysfunction in hypertrophied neonatal hearts. Circ Res. 2015 Jun 19;117(1):41-51.

[5]. Identification of a subtype selective human PPARalpha agonist through parallel-array synthesis. Bioorg Med Chem Lett. 2001 May 7;11(9):1225-7.

GW 7647 is a monocarboxylic acid, 2-(phenylthio)isobutyric acid, in which the para-position of the phenyl group is replaced by a 3-aza-7-cyclohexylhept-1-yl group, the nitrogen atom of which is acylated by a (cyclohexylamino)carbonyl group. It is a PPARα agonist. It belongs to the urea class, aryl thioether class, and monocarboxylic acid class.

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Background: The PDZ adaptor protein PDZK1 regulates the membrane expression and function of various intestinal receptors and ion/nutrient transporters. PDZK1 expression is significantly reduced in the inflamed intestinal mucosa of mice and patients with inflammatory bowel disease (IBD). Objectives and Methods: We investigated whether the downregulation of inflammation-related PDZK1 is a direct result of the release of pro-inflammatory cytokines by treating intestinal Caco-2BBE cells with TNF-α, IFN-γ, and IL-1β and analyzing PDZK1 promoter activity, mRNA, and protein expression. Results: We found that IL-1β significantly reduced PDZK1 promoter activity, mRNA, and protein expression in Caco-2BBE cells. The distal region of the hPDZK1 promoter is crucial for basal expression and IL-1β responsiveness. This region contains the retinoic acid-responsive element RARE and binding sites for transcription factors involved in downstream IL-1β signaling. Inhibition of ERK1/2 by the specific MEK1/2 inhibitor PD98059/U0126 significantly attenuated IL-1β-mediated PDZK1 downregulation, while NF-κB, p38 MAPK, and JNK inhibitors had no such effect. In the inflamed colonic mucosa of patients with ulcerative colitis and in IL-1β-treated Caco-2-BBE cells, the expression of nuclear receptors RXRα and PPARα was reduced. Furthermore, the RAR/RXR ligand 9-cis-retinoic acid and the PPARα agonist GW7647 stimulated PDZK1 mRNA and protein expression and attenuated IL-1β-mediated inhibition. Conclusion: The significant decrease in PDZK1 expression during intestinal inflammation may be partly due to IL-1β-mediated inhibition of RXRα and PPARα, and can be attenuated by nuclear receptor agonists or ERK1/2 inhibitors. The negative impact of inflammation-induced PDZK1 downregulation on epithelial transport function may be improved by drug treatment. [1]

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Peroxisome proliferator receptor α (PPARα) is a key regulator of the liver’s fasting response and has effects on both lipid and carbohydrate metabolism. Studies have also found that PPARα plays a role in hepatic glycerol metabolism; however, some of the results are contradictory. Aquaporin 9 (AQP9) is a transmembrane protein that forms pores and promotes the uptake of glycerol by the liver. In male rodents, AQP9 expression is negatively regulated by insulin and increases during fasting. Previous studies have shown that PPARα plays a key role in the induction of AQP9 mRNA during fasting. This study investigated the effects of PPARα activation on hepatic AQP9 expression and the abundance of glycerol metabolism-related enzymes using in vivo and in vitro experiments with PPARα agonists. In ad libitum-fed male rats, treatment with the PPARα agonist WY 14643 (3 mg·kg⁻¹·day⁻¹) reduced hepatic AQP9 abundance by 50%, and this effect was limited to AQP9 expressed in periportal hepatocytes. Pharmacological activation of PPARα did not affect GlyK abundance but increased the expression of hepatic GPD1, GPAT1, and L-FABP proteins. In WIF-B9 and HepG2 hepatocytes, both WY 14643 and another PPARα agonist, GW 7647, reduced AQP9 protein abundance. In conclusion, pharmacological activation of PPARα significantly reduces AQP9 abundance in periportal hepatocytes. Combined with the effects on the glycerol metabolism enzyme system, our results suggest that, under feeding conditions, activation of PPARα directs glycerol to glycerol ester synthesis rather than de novo glucose synthesis. [3]

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Reason: Post-ischemic systolic dysfunction is one of the factors contributing to morbidity and mortality after surgical correction of congenital heart disease in newborns. Pre-existing hypertrophy of the newborn heart can exacerbate these ischemic injuries, partly due to a low rate of fatty acid β-oxidation leading to reduced cardiac energy supply. Objective: We investigated whether stimulation of fatty acid β-oxidation with the peroxisome proliferation-activating receptor α (PPARα) activator GW7647 could improve energy production and post-ischemic functional recovery in the neonatal heart of volume-overload-induced cardiac hypertrophy. Methods and Results: A volume-overload cardiac hypertrophy model was established in 7-day-old rabbits via aortovesical shunt. The rabbits were then randomly divided into two groups, receiving GW7647 (3 mg/kg/day) or not for 14 days. Biventricular working hearts were subjected to 35 minutes of aerobic perfusion, 25 minutes of total cardiac ischemia without blood flow, and 30 minutes of aerobic reperfusion. GW7647 treatment did not prevent the occurrence of cardiac hypertrophy, but it did prevent the decrease in left ventricular ejection fraction in vivo. GW7647 treatment increased the rate of fatty acid β-oxidation in the heart before and after ischemia, thereby significantly increasing total ATP production and improving functional recovery after in vitro ischemia. In the heart treated with GW7647, proton production and endoplasmic reticulum stress were reduced after ischemia, and sarcoplasmic reticulum calcium ATPase 2 isoform and citrate synthase were activated. Conclusion: Stimulating fatty acid β-oxidation in the neonatal heart may be a new cardioprotective intervention that helps limit systolic dysfunction after ischemia. [4] A series of urea-substituted thioisobutyric acids were synthesized by solid-phase parallel array synthesis and their activity against human PPAR isoforms was determined. GW7647 (3) was identified as a potent human PPARα agonist with approximately 200-fold selectivity for PPARγ and PPARδ and significant lipid-lowering activity in animal models of dyslipidemia. GW7647 (3) will become an important chemical tool for studying PPARα biology in human cells and disease animal models. GW7647 (3) is a potent human PPARα agonist with approximately 200-fold selectivity for other subtypes and lipid-lowering activity in vivo. To further characterize the compound, we determined its activity against three mouse PPAR subtypes. The EC50 values of compound 3 against mouse PPARα, PPARγ and PPARδ were 0.001, 1.3 and 2.9 μM, respectively. [5] GW7647 is a pharmacological tool belonging to the PPARα agonist class for studying the regulation of the PDZ adaptor protein PDZK1 in the context of intestinal inflammation. It can increase PDZK1 expression and attenuate IL-1β-induced downregulation, suggesting that PPARα activation may be a potential therapeutic strategy to combat inflammation-related epithelial dysfunction. The PDZK1 promoter contains a peroxisome proliferative response element (PPRE), to which PPARα can form a heterodimer with RXRα and bind. [1]

C29H46N2O3S

502.75214

502.322

C, 69.28; H, 9.22; N, 5.57; O, 9.55; S, 6.38

265129-71-3

265129-71-3

3392731

White to off-white solid powder

1.1±0.1 g/cm3

693.9±55.0 °C at 760 mmHg

373.5±31.5 °C

0.0±2.3 mmHg at 25°C

1.569

8.59

2

4

12

35

636

0

CC(C)(SC1=CC=C(CCN(C(NC2CCCCC2)=O)CCCCC3CCCCC3)C=C1)C(O)=O

PKNYXWMTHFMHKD-UHFFFAOYSA-N

InChI=1S/C29H46N2O3S/c1-29(2,27(32)33)35-26-18-16-24(17-19-26)20-22-31(28(34)30-25-14-7-4-8-15-25)21-10-9-13-23-11-5-3-6-12-23/h16-19,23,25H,3-15,20-22H2,1-2H3,(H,30,34)(H,32,33)

2-[4-[2-[4-cyclohexylbutyl(cyclohexylcarbamoyl)amino]ethyl]phenyl]sulfanyl-2-methylpropanoic acid

GW 7647; GW-7647; 265129-71-3; 2-[(4-{2-[(4-cyclohexylbutyl)(cyclohexylcarbamoyl)amino]ethyl}phenyl)sulfanyl]-2-methylpropanoic acid; 2-[[4-[2-[[(cyclohexylamino)carbonyl](4-cyclohexylbutyl)amino]ethyl]phenyl]thio]-2-methylpropanoic acid; MFCD06798379; NTL2A9CAZ7; GW7647

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 (~198.9 mM)
Ethanol: ~25 mg/mL (~49.7 mM)

Solubility in Formulation 1: ≥ 2.5 mg/mL (4.97 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 (4.97 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.)