ATP-citrate lyase (ACL); AMPK
AMP-activated protein kinase (AMPK) (activation via free acid form, LKB1-dependent) [1]
ATP-citrate lyase (ACL) (inhibition via CoA thioester form, IC50 = 17.8 μM under substrate-saturated conditions; IC50 = 4.01 μM under apparent Km CoASH conditions) [1]

Bempedoic acid (ETC-1002) activates AMP-activated protein kinase in a Ca2+/calmodulin-dependent kinase β-independent and liver kinase β 1-dependent manner, without detectable changes in adenylate energy charge.It has been demonstrated that bempedoic acid quickly converts to a CoA thioester in the liver, which directly inhibits ATP-citrate lyase[1]. Increased AMP-activated protein kinase (AMPK) phosphorylation is associated with decreased MAP kinase activity and decreased production of proinflammatory cytokines and chemokines in cells treated with bempedoic acid (ETC-1002)[2].

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ETC-1002 is an investigational drug currently in Phase 2 development for treatment of dyslipidemia and other cardiometabolic risk factors. In dyslipidemic subjects, ETC-1002 not only reduces plasma LDL cholesterol but also significantly attenuates levels of hsCRP, a clinical biomarker of inflammation. Anti-inflammatory properties of ETC-1002 were further investigated in primary human monocyte-derived macrophages and in in vivo models of inflammation. In cells treated with ETC-1002, increased levels of AMP-activated protein kinase (AMPK) phosphorylation coincided with reduced activity of MAP kinases and decreased production of proinflammatory cytokines and chemokines. AMPK phosphorylation and inhibitory effects of ETC-1002 on soluble mediators of inflammation were significantly abrogated by siRNA-mediated silencing of macrophage liver kinase B1 (LKB1), indicating that ETC-1002 activates AMPK and exerts its anti-inflammatory effects via an LKB1-dependent mechanism. In vivo, ETC-1002 suppressed thioglycollate-induced homing of leukocytes into mouse peritoneal cavity. Similarly, in a mouse model of diet-induced obesity, ETC-1002 restored adipose AMPK activity, reduced JNK phosphorylation, and diminished expression of macrophage-specific marker 4F/80. These data were consistent with decreased epididymal fat-pad mass and interleukin (IL)-6 release by inflamed adipose tissue. Thus, ETC-1002 may provide further clinical benefits for patients with cardiometabolic risk factors by reducing systemic inflammation linked to insulin resistance and vascular complications of metabolic syndrome.[2]

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In primary rat hepatocytes, Bempedoic acid is rapidly taken up and converted to its CoA thioester. This conversion is associated with immediate inhibition (≤5 min) of de novo lipid synthesis, as measured by [14C]acetate incorporation, and transient increases in phosphorylation of AMPK (T172), ACC (S79), and HMGR (S872). [1]
Treatment of primary rat hepatocytes with Bempedoic acid results in concentration-dependent reductions in acetyl-CoA, malonyl-CoA, and HMG-CoA, with concomitant increases in citrate and the formation of ETC-1002-CoA. These effects occur within 5 minutes of treatment. [1]
Triacsin C, an inhibitor of long-chain acyl-CoA synthetases, reduces the intracellular concentration of Bempedoic acid-CoA and attenuates the compound's effects on metabolic intermediates (acetyl-CoA, malonyl-CoA, HMG-CoA) and de novo lipid synthesis, demonstrating that CoA thioester formation is required for these inhibitory activities. [1]
In HepG2 cells, which do not form significant amounts of Bempedoic acid-CoA, the free acid form of the compound causes a sustained and concentration-dependent increase in AMPK (T172) and ACC (S79) phosphorylation, comparable to the effect of metformin (1000 μM). This activation is not inhibited by the CaMKKβ inhibitor STO-609 or the AMP analog Compound C. [1]
Bempedoic acid treatment does not alter adenylate energy charge (AEC) or ATP levels in HepG2 cells, even when grown in galactose medium to force mitochondrial oxidative phosphorylation, indicating AMPK activation is independent of energy depletion. [1]
In primary rat hepatocytes, Bempedoic acid (IC50 = 3.6 μM) reduces glucagon-stimulated glucose production. This effect is associated with decreased protein expression of PEPCK and G6Pase, as well as reduced basal levels of FOXO1. [1]
Knockdown of LKB1 via siRNA in HepG2 cells abolishes Bempedoic acid-dependent phosphorylation of ACC (S79) and reduction in HNF-4α protein levels, demonstrating that AMPK activation by the compound is LKB1-dependent. [1]

Bempedoic acid (ETC-1002) treatment for two weeks causes a noticeable and long-lasting increase in AMPK and ACC phosphorylation in the livers of rats. In rat liver, Bempedoic acid is more prevalent >100-fold than CoA thioester and is connected to AMPK activation[1]. Leukocytes' ability to home into the mouse peritoneal cavity is suppressed by bempedoic acid (ETC-1002). In a mouse model of diet-induced obesity, Bempedoic acid improves adipose AMPK activity, lowers JNK phosphorylation, and decreases expression of the macrophage-specific marker 4F/80[2].

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ETC-1002 (8-hydroxy-2,2,14,14-tetramethylpentadecanedioic acid) is a novel investigational drug being developed for the treatment of dyslipidemia and other cardio-metabolic risk factors. The hypolipidemic, anti-atherosclerotic, anti-obesity, and glucose-lowering properties of ETC-1002, characterized in preclinical disease models, are believed to be due to dual inhibition of sterol and fatty acid synthesis and enhanced mitochondrial long-chain fatty acid β-oxidation. However, the molecular mechanism(s) mediating these activities remained undefined. Studies described here show that ETC-1002 free acid activates AMP-activated protein kinase in a Ca(2+)/calmodulin-dependent kinase β-independent and liver kinase β 1-dependent manner, without detectable changes in adenylate energy charge. Furthermore, ETC-1002 is shown to rapidly form a CoA thioester in liver, which directly inhibits ATP-citrate lyase. These distinct molecular mechanisms are complementary in their beneficial effects on lipid and carbohydrate metabolism in vitro and in vivo. Consistent with these mechanisms, ETC-1002 treatment reduced circulating proatherogenic lipoproteins, hepatic lipids, and body weight in a hamster model of hyperlipidemia, and it reduced body weight and improved glycemic control in a mouse model of diet-induced obesity. ETC-1002 offers promise as a novel therapeutic approach to improve multiple risk factors associated with metabolic syndrome and benefit patients with cardiovascular disease.[1]

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In chow-fed Wistar rats, oral administration of Bempedoic acid (30 mg/kg/day for 14 days) significantly increases hepatic AMPK (T172) (358.3% ± 48.14; P = 0.0007) and ACC (S79) phosphorylation (164.7% ± 12.39; P = 0.001). This is associated with a 70% reduction in hepatic triglycerides, a 51% increase in plasma β-hydroxybutyrate (β-HBA), and no change in hepatic adenylate energy charge. [1]
In nutritionally staged (fasted/refed high-carbohydrate diet) Wistar rats, a single oral dose of Bempedoic acid (30 mg/kg) leads to the formation of ETC-1002-CoA in the liver and is associated with reduced hepatic acetyl-CoA, malonyl-CoA, and HMG-CoA, and increased citrate levels 2 hours post-dose. [1]
In a hamster model of hyperlipidemia (high-fat, high-cholesterol diet), treatment with Bempedoic acid (30 mg/kg/day for 3 weeks) results in a 14.4% decrease in body weight gain, a 20% increase in plasma β-HBA, a 34% reduction in plasma NEFA, a 35% reduction in epididymal fat mass, a 64% reduction in hepatic TG, a 67% reduction in hepatic cholesteryl ester (CE), a 31% reduction in hepatic free cholesterol (FC), a 41% reduction in plasma TG, a 41% reduction in plasma total cholesterol, a 64% reduction in LDL-C, and a 62% reduction in VLDL-C. No significant changes in food consumption or plasma ALT/AST were observed. [1]
In a mouse model of diet-induced obesity (DIO) (60% kcal fat diet for 12 weeks), treatment with Bempedoic acid (30 mg/kg/day for 14 days) results in a 9% reduction in body weight without affecting food consumption. Fasting plasma glucose is reduced by 13%, and fasting plasma insulin is reduced by 42%. [1]

Glucose production assay[1]
Glucose production was measured in primary rat hepatocyte cultures. Cells were cultured in glucose- and phenol red-free DMEM, containing 10 mM lactate, 1 mM pyruvate, and nonessential amino acids (glucose production buffer, GPB). To assess the effects of ETC-1002 on glucagon-stimulated glucose production, cells were incubated with and without 0.3 μM glucagon with various concentrations of ETC-1002 (0.1 to 100 μM). Media was sampled over time. Following specified treatments, cells were washed twice in GPB. Cells were then incubated for an additional hour to assess glucose production by adding GPB containing equivalent glucagon concentrations without ETC-1002. Cells were incubated for 1 h, and the concentration of glucose in the media was determined using a glucose oxidase assay kit.

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ETC-1002 formulation / ETC-1002-CoA synthesis[2]
For in vitro assays, ETC-1002 was formulated using aseptic technique at 30 and 100 mM in sterile dimethylsulfoxide (DMSO) and stored in sterile microcentrifuge tubes at 4°C for up to four weeks (stability was assessed). Working solutions of ETC-1002 were prepared in serum-free RPMI 1640 containing 12 mM HEPES, 10,000 U/ml penicillin, and 100 μg/ml streptomycin. ETC-1002-CoA was synthesized using rat liver microsomes as described.

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7.5× compounds are added to a 96-well PolyPlate containing 60 μL of Buffer per well with substrates CoA (200 μM), ATP (400 μM), and [14C]citrate. Reaction is started with 4 μL (300 ng/well) ACL, and the plate is incubated at 37°C for 3 h.

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The activity of recombinant human ATP-citrate lyase (ACL) was measured using a cell-free assay. The reaction mixture contained 87 mM Tris (pH 8.0), 20 μM MgCl2, 10 mM KCl, 10 mM dithiothreitol, 200 μM CoA, 400 μM ATP, and 150 μM [14C]citrate (specific activity: 2 μCi/μmol). The reaction was initiated by adding 300 ng of ACL and incubated at 37°C for 3 hours. The reaction was terminated with EDTA, and the [14C]acetyl-CoA product was detected by liquid scintillation counting. Under these substrate-saturated conditions, Bempedoic acid-CoA inhibited ACL with an IC50 of 17.8 μM. When the assay was performed with a CoA concentration comparable to the apparent Km for ACL, the potency increased to an IC50 of 4.01 μM, indicating the inhibition is competitive with CoASH. The free acid form of the compound showed no inhibition. [1]

Glucose production is measured in primary rat hepatocyte cultures. It contains nonessential amino acids, 10 mM lactate, 1 mM pyruvate, and is free of glucose and phenol red. Cells are cultured in this mixture. Bempedoic acid (0.1 to 100 μM) is incubated with the cells in a variety of concentrations[1].

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Protein arrays[2]
At the end of differentiation period, macrophages were washed with PBS and switched to RPMI 1640 containing 5% autologous serum and supplemented with 14 mM HEPES, 100 U/ml penicillin, 50 U/ml streptomycin, and 2 mM L-glutamine. ETC-1002 at various concentrations (50 μM and 100 μM) was added to the media 1 h prior to stimulation with 100 ng/ml of lipopolysaccharide (LPS) from Escherichia coli 0111:B4. Media conditioned by MDMs was collected 12 h following LPS stimulation and assayed with Proteome Profiler Human Cytokine Array Kit, Panel A, and Human Matrix Metalloproteinase Array, according to the manufacturer's instructions. Data for cytokine and matrix metalloproteinase (MMP) arrays were captured and analyzed with a Kodak 4000MM Image Station. Net signal intensity for each analyte was expressed as percentage of the internal reference standard for each individual array membrane. Data are presented as mean ± SEM. Comparisons between groups were performed by one-way ANOVA. A Bonferroni's post hoc multiple comparison test was used to assess significant differences revealed by the ANOVA. Significance was accepted at P ≤ 0.05.[2]

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Primary rat hepatocytes were isolated from male Sprague-Dawley rats and cultured in collagen-coated plates. For de novo lipid synthesis, cells were treated with Bempedoic acid (10 or 100 μM) or AICAR (500 μM) for 5-30 minutes, then pulsed with [14C]acetate for 15 minutes. Saponified (fatty acids) and nonsaponified (sterols) lipids were extracted and counted. Results showed immediate inhibition of lipid synthesis by Bempedoic acid. [1]
For Western blot analysis of AMPK pathway activation, primary rat hepatocytes were treated with 30 μM Bempedoic acid for various times (5-120 min). Cell lysates were prepared, proteins separated by SDS-PAGE, transferred to PVDF membranes, and probed with antibodies against phospho-AMPKα (T172), total AMPKα, phospho-ACC (S79), total ACC, and phospho-HMGR (S872). Transient increases in phosphorylation were observed. [1]
For glucose production assays, primary rat hepatocytes were cultured in glucose- and phenol red-free DMEM containing 10 mM lactate, 1 mM pyruvate, and nonessential amino acids. Cells were stimulated with 0.3 μM glucagon with or without various concentrations of Bempedoic acid (0.1 to 100 μM). Media glucose concentration was measured using a glucose oxidase assay. Bempedoic acid inhibited glucagon-stimulated glucose production with an IC50 of 3.6 μM. Cell lysates from this assay were also analyzed by Western blot for PEPCK, G6Pase, and FOXO1, showing reduced protein levels. [1]
In HepG2 cells, reverse transfection with LKB1 or negative control siRNA was performed using Lipofectamine 2000. After 48 hours, cells were treated with vehicle, 100 μM Bempedoic acid, or 1000 μM metformin for 24 hours. Cell lysates were analyzed by Western blot for LKB1, phospho-ACC (S79), total ACC, and HNF-4α. Intracellular ATP, ADP, and AMP were measured by LC-MS/MS, and total cholesterol and triglycerides were quantified. LKB1 knockdown abolished the effects of Bempedoic acid on ACC phosphorylation and HNF-4α reduction, as well as its lipid-lowering effects. [1]

Rats: Male Wistar Han rats are fasted for 48 hours and then given a single dose of bempedoic acid before receiving a second 48-hour feeding of a high-carbohydrate diet. Rats are kept on a standard chow diet and given oral gavage doses of bempedoic acid for a two-week assessment. The dose is 30 mg/kg/day given in the morning. Food is discontinued two hours before the final oral dose of engine control or bempedoic acid after nutritional staging and/or dosing[1].

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For in vivo experiments, ETC-1002 dosing solutions were formulated by preparing a disodium salt aqueous solution using 2:1 molar ratio of NaOH to ETC-1002 in water. Carboxymethyl cellulose (CMC) and Tween-20 were added to make a final solution containing 0.5% CMC and 0.025% Tween with a final pH 7–8. Compound concentrations in dosing solutions were administered at a volume of 10 ml/kg body.[1],2]

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Wistar rats.[1]
Male Wistar Han [Crl:WI] rats weighing 225–250 g were acclimated to the laboratory environment for seven days, housed 2–3 per cage in a temperature controlled room, and maintained on a 12 h light and dark cycle with ad libitum access to food and water. Prior to single-dose ETC-1002 administration, rats were fasted for 48 h and refed a high-carbohydrate diet for an additional 48 h. For two-week assessment, rats were maintained on standard chow diet (Purina 5001) and dosed by oral gavage with ETC-1002 at 30 mg/kg/day for two weeks in the morning. Following nutritional staging and/or dosing, food was withdrawn 2 h prior to last the oral dose of vehicle control or ETC-1002. Blood and liver were collected from isofluorane-anesthetized animals 2 or 8 h after the last dose, blood was collected from the subclavian vein, and liver tissue was harvested by freeze clamp. The freeze-clamped liver samples were held frozen in liquid nitrogen immediately following excision and stored at −70°C. Plasma triglycerides, β-hydroxybutyrate (β-HBA), and total cholesterol levels were measured with commercially available kits (Wako Diagnostics, Richmond, VA) adapted to a 96-well format.

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Golden Syrian hamsters.[1]
Male golden Syrian hamsters were obtained from Charles River (Montreal, QC) at 8–10 weeks of age and weighed 100–120 g. Animals were maintained on Prolab RMH 1000 standard rodent chow diet during a seven-day quarantine period. Following randomization into treatment groups (n = 6), hyperlipidemia was induced by feeding high-fat, high-cholesterol (HFHC) Prolab RMH 1000 diet containing: 11.5% coconut oil, 11.5% corn oil, 5% fructose, and 0.5% cholesterol. During the study, animals were individually housed in an environmentally controlled room with a 12 h light and dark cycle. Following two weeks on HFHC diet, hamsters were dosed by oral gavage once daily with vehicle (0.5% carboxymethyl cellulose and 0.025% Tween-20, pH 7–8) or vehicle plus ETC-1002 (30 mg/kg) for three weeks. Body weights were recorded every two days at the beginning of dosing, and food consumption was measured every four days. Blood samples were collected by administering isoflurane anesthesia and bleeding from the orbital venous plexus in lithium heparinized tubes during the study and by cardiac puncture under anesthesia at the end of the study. Plasma samples were analyzed for triglycerides, total cholesterol, nonesterified fatty acids, and β-hydroxybutyrate on an automated chemistry analyzer. Liver and epididymal fat were collected, weighed, frozen in liquid nitrogen, and stored at −80°C until processing. All hamster procedures were conducted in accordance with the current guidelines for animal welfare at the Hospital for Sick Children and were in compliance with National Institutes of Health Publication 86-23, 1985; Animal Welfare act, 1966, as amended in 1970, 1976, and 1985, 9 CFR Parts 1, 2, and 3.

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Diet-induced obesity in mice.[1]
Male C57BL/6N mice were obtained from Taconic at 8 weeks of age and singly housed on α-dri paper bedding on a normal 12 h light and dark cycle (6 AM to 6 PM). Upon arrival mice, were fed a high-fat diet (HFD) containing 60% kcal fat for 12 weeks. Mice were randomized into two treatment arms at 20 weeks of age based on 4 h fasted blood glucose and body weight and received oral dosing of either CMC/Tween vehicle or 30 mg/kg/day ETC-1002 q.d in the morning for an additional two weeks. Body weight and food consumption were monitored throughout the study. Following the two-week dosing period, food was removed at 8 AM, and bedding was changed 2 h prior to oral administration of ETC-1002. Two hours post dose, fasting samples were collected. Fasting blood glucose levels were measured immediately prior to anesthesia using a hand-held Alphatrak glucometer (Abbott, Chicago, IL), with blood collected by unrestrained tail snip. For insulin determinations, blood was collected under isoflurane anesthesia via retro-orbital sinus into EDTA-coated tubes, and plasma was isolated by centrifugation. Plasma insulin levels were measured with a commercially available ELISA

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Rat Studies:** Male Wistar Han rats (225-250 g) were used. For the 2-week efficacy study, rats were maintained on standard chow and dosed daily by oral gavage with vehicle (0.5% carboxymethyl cellulose and 0.025% Tween-20, pH 7-8) or 30 mg/kg Bempedoic acid for 14 days. Food was withdrawn 2 hours prior to the last dose. Blood and freeze-clamped liver were collected 2 or 8 hours post-dose. For the nutritional staging study, rats were fasted for 48 hours and then refed a high-carbohydrate diet for 48 hours prior to a single oral dose of 30 mg/kg Bempedoic acid or vehicle. Liver was collected 2 and 8 hours post-dose. [1]
* **Hamster Study:** Male golden Syrian hamsters (8-10 weeks old) were fed a high-fat, high-cholesterol diet (11.5% coconut oil, 11.5% corn oil, 5% fructose, 0.5% cholesterol) for two weeks to induce hyperlipidemia. They were then dosed orally once daily with vehicle or 30 mg/kg Bempedoic acid for three weeks while continuing on the same diet. Body weight and food consumption were monitored. At the end of the study, blood, liver, and epididymal fat were collected. [1]
* **Mouse Study:** Male C57BL/6N mice (8 weeks old) were fed a high-fat diet (60% kcal fat) for 12 weeks to induce obesity. They were then randomized and dosed orally once daily with vehicle or 30 mg/kg/day Bempedoic acid for an additional two weeks while on the HFD. Body weight and food consumption were monitored. After a 4-hour fast, blood glucose and plasma insulin were measured. [1]

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Rat Studies: Male Wistar Han rats (225-250 g) were used. For the 2-week efficacy study, rats were maintained on standard chow and dosed daily by oral gavage with vehicle (0.5% carboxymethyl cellulose and 0.025% Tween-20, pH 7-8) or 30 mg/kg Bempedoic acid for 14 days. Food was withdrawn 2 hours prior to the last dose. Blood and freeze-clamped liver were collected 2 or 8 hours post-dose. For the nutritional staging study, rats were fasted for 48 hours and then refed a high-carbohydrate diet for 48 hours prior to a single oral dose of 30 mg/kg Bempedoic acid or vehicle. Liver was collected 2 and 8 hours post-dose. [1]
Hamster Study: Male golden Syrian hamsters (8-10 weeks old) were fed a high-fat, high-cholesterol diet (11.5% coconut oil, 11.5% corn oil, 5% fructose, 0.5% cholesterol) for two weeks to induce hyperlipidemia. They were then dosed orally once daily with vehicle or 30 mg/kg Bempedoic acid for three weeks while continuing on the same diet. Body weight and food consumption were monitored. At the end of the study, blood, liver, and epididymal fat were collected. [1]
Mouse Study: Male C57BL/6N mice (8 weeks old) were fed a high-fat diet (60% kcal fat) for 12 weeks to induce obesity. They were then randomized and dosed orally once daily with vehicle or 30 mg/kg/day Bempedoic acid for an additional two weeks while on the HFD. Body weight and food consumption were monitored. After a 4-hour fast, blood glucose and plasma insulin were measured. [1]

Absorption, Distribution and Excretion
Bepedocoxel is rapidly absorbed in the small intestine. The time to peak concentration (Tmax) for a 180 mg tablet is estimated to be 3.5 hours. Conjugates of bepedocoxel are primarily excreted via urine (70%) and feces (30%). 5% of the total parenteral drug is excreted via urine and feces. The apparent volume of distribution of bepedocoxel is approximately 18 liters. In clinical trials, the steady-state clearance (CL/F) of bepedocoxel was estimated to be 11.2 mL/min. Metabolism/Metabolites The two main metabolites of bepedocoxel are ETC-1002-CoA and ESP15228. Bepedocoxel is primarily excreted via the metabolism of its acyl glucuronide. Based on observations from in vitro studies, the drug is reversibly converted to the active metabolite (ESP15228). Both compounds produced by bepedocoxel metabolism are metabolized by the UGT2B7 enzyme to inactive glucuronide conjugates.
Biological Half-Life
The half-life of bepedocoxel is 15 to 24 hours. Prescription information indicates a clearance rate of 21 hours ± 11 hours.

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Bempedoic acid is rapidly taken up by primary rat hepatocytes and converted to its CoA thioester. The intracellular ratio of free acid to CoA thioester in vitro is approximately 1:1 to 2:1. [1]
In vivo, after oral administration to rats, Bempedoic acid free acid is >100-fold more prevalent than its CoA thioester in the liver. The CoA thioester is detectable in liver extracts of rats and hamsters following oral dosing. [1]
The highly hydrophobic nature of the compound suggests it is not rapidly cleared from tissues. [1]

Effects During Pregnancy and Lactation
◉ Overview of Use During Lactation
Currently, there is no published information regarding the use of bepedioxic acid during lactation. Bepedioxic acid and its metabolites have a plasma protein binding rate as high as 99%, therefore the concentration in breast milk may be very low. However, due to concerns about disrupting the infant's lipid metabolism, it is best to avoid using bepedioxic acid during lactation. The use of other medications is recommended, especially when breastfeeding newborns or premature infants.
◉ Effects on Breastfed Infants
As of the revision date, no relevant published information was found.
◉ Effects on Lactation and Breast Milk
As of the revision date, no relevant published information was found.

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Protein Binding
The plasma protein binding rate of bepedioxic acid and its metabolites is approximately 99%.

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In a 4-week safety study in Wistar rats, oral administration of Bempedoic acid at 30 mg/kg/day resulted in plasma exposures equivalent to clinical exposures with no meaningful drug-related effect on safety parameters, including liver markers of injury ALT and AST. [1]
In the 3-week hamster study, no significant changes in plasma ALT or AST were observed with Bempedoic acid treatment. [1]

[1]. AMP-activated protein kinase and ATP-citrate lyase are two distinct molecular targets for ETC-1002, a novel small molecule regulator of lipid and carbohydrate metabolism. J Lipid Res. 2013 Jan;54(1):134-51.

[2]. ETC-1002 regulates immune response, leukocyte homing, and adipose tissue inflammation via LKB1-dependent activation of macrophage AMPK. J Lipid Res. 2013 Aug;54(8):2095-108.

Pharmacodynamics
Bepedocoxel inhibits cholesterol synthesis in the liver, thereby lowering low-density lipoprotein cholesterol (LDL-C) levels. This can reduce the formation of atherosclerotic plaques, thus reducing the risk of cardiovascular events. Early clinical trials investigating the effects of bepedocoxel showed that, in addition to a reduction in the number of LDL particles, LDL-C levels also decreased in a dose-dependent manner, along with levels of apolipoprotein B, non-HDL cholesterol, and high-sensitivity C-reactive protein. Due to its unique mechanism of action, bepedocoxel does not cause myositis, a common side effect of statin therapy. Recent trials have shown that this drug significantly reduces LDL-C levels after 12 weeks of treatment, and further reduces LDL-C levels when used in combination with ezetimibe and statins. The impact of bepedocoxel on mortality is currently unknown.

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Bempedoic acid (8-hydroxy-2,2,14,14-tetramethylpentadecanedioic acid), also known as ESP55016 and ETC-1002, is a novel investigational drug being developed for the treatment of dyslipidemia and other cardiometabolic risk factors. It is a first-in-class small molecule that regulates lipid and carbohydrate metabolism. [1]
The compound's mechanism of action is dual and tissue-specific. The free acid form activates AMPK in an LKB1-dependent but energy- and calcium-independent manner. The CoA thioester form, formed in the liver, directly inhibits ATP-citrate lyase (ACL). These two mechanisms are complementary in reducing lipid synthesis (sterol and fatty acid) at both the signal transduction and substrate levels. [1]
Inhibition of ACL by Bempedoic acid-CoA reduces cytosolic acetyl-CoA, the common substrate for both fatty acid and sterol synthesis. This also leads to reductions in malonyl-CoA (which de-inhibits CPT-1, increasing fatty acid oxidation) and HMG-CoA. [1]
The combination of AMPK activation (which phosphorylates and inhibits ACC and HMGR) and ACL inhibition provides a robust effect on hepatic lipid metabolism, leading to reduced hepatic steatosis and lower circulating proatherogenic lipoproteins (LDL-C, VLDL-C) in preclinical models. It also improves glycemic control by reducing hepatic glucose production. [1]

C19H36O5

344.4861

344.256

C, 66.25; H, 10.53; O, 23.22

738606-46-7

Bempedoic acid-d4;2408131-70-2;Bempedoic acid-d5;2408131-71-3

10472693

White to light yellow solid powder

87-92

4.469

3

5

14

24

351

0

OC(CCCCCC(C(=O)O)(C)C)CCCCCC(C(=O)O)(C)C

HYHMLYSLQUKXKP-UHFFFAOYSA-N

InChI=1S/C19H36O5/c1-18(2,16(21)22)13-9-5-7-11-15(20)12-8-6-10-14-19(3,4)17(23)24/h15,20H,5-14H2,1-4H3,(H,21,22)(H,23,24)

8-hydroxy-2,2,14,14-tetramethylpentadecanedioic acid

ETC-1002; ETC 1002; ETC1002; ESP-55016; Bempedoate; 8-Hydroxy-2,2,14,14-tetramethylpentadecanedioic acid; Nexletol; Nilemdo; ESP-55016; Bempedoic acid; ETC-1002ESP55016; ETC1002ESP

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 (~290.3 mM)

Solubility in Formulation 1: ≥ 2.87 mg/mL (8.33 mM) (saturation unknown) in 5% DMSO + 40% PEG300 + 5% Tween80 + 50% Saline (add these co-solvents sequentially from left to right, and one by one), clear solution.
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: 2.87 mg/mL (8.33 mM) in 5% DMSO + 95% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), suspension solution; with ultrasonication.
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 (7.26 mM) 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 4: ≥ 2.5 mg/mL (7.26 mM) 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 5: ≥ 2.5 mg/mL (7.26 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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Solubility in Formulation 6: 0.57 mg/mL (1.65 mM) in 1% DMSO + 99% Saline (add these co-solvents sequentially from left to right, and one by one),suspension solution;clear solution.
Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH₂ O to obtain a clear solution.

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