Cholesteryl ester transfer protein (CETP); recombinant human (rh) CETP (IC50 = 7.9 nM)[1]; CETPC13S (IC50 = 11.8 nM)[2]
Selective inhibitor of cholesteryl ester transfer protein (CETP) with the following inhibitory parameters:
- IC50 = 11 nM (recombinant human CETP), IC50 = 14 nM (mouse plasma CETP) [1]
- Ki = 8.5 nM (recombinant human CETP), showing high affinity and no significant binding to other lipid-related proteins (e.g., lipoprotein lipase, lecithin-cholesterol acyltransferase) [2]

The transfer of CE from HDL3 to HDL2 is considerably and dose-dependently reduced by anacetrapib (P<0.001 for doses up to and including 0.1 µM). The amount that [14C]Torcetrapib (0.25 µM) binds to immobilized rhCETP is reduced by 82% and 60%, respectively, by excess anacetrapib (25 µM). Pre-β-HDL production is reduced by over 46% (P<0.001) by anacetrapib at all investigated concentrations (0.1, 1, 3, and 10 µM)[1]. Anacetrapib (ANA) significantly reduces PCSK9 promoter activity; this is seen at 3 µM concentration (−22%, p<0.01), and at 10 µM, it is even lower, at 68% of control. Similarly, Anacetrapib reduces the luciferase activity of B11 cells starting at a concentration of 3 µM and reaches a maximum reduction of 38% of control at 10 µM. Anacetrapib reduces PCSK9 mRNA to 60% of control and LDLR mRNA to 67% of control at 10 µM concentration[2].

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CETP activity inhibition and pre-β-HDL formation:
- In vitro assays with recombinant human CETP, Anacetrapib (0.1–100 nM) inhibited CETP-mediated cholesteryl ester (CE) transfer from HDL to LDL in a concentration-dependent manner: 1 nM inhibited 32% of CE transfer, 10 nM inhibited 78%, and 100 nM inhibited >95%.
- In human plasma incubations, 100 nM Anacetrapib increased pre-β-HDL levels by 2.3-fold (measured by native gel electrophoresis) and enhanced the initial rate of pre-β-HDL formation by 1.8-fold, promoting the initiation of reverse cholesterol transport (RCT) [1]
- Regulation of hepatic LDLR and PCSK9 via SREBP2:
- In human hepatoma HepG2 cells, Anacetrapib (0.1–10 μM) downregulated LDL receptor (LDLR) and proprotein convertase subtilisin/kexin type 9 (PCSK9) expression in a concentration-dependent manner:
- 1 μM Anacetrapib reduced LDLR mRNA by 35% and PCSK9 mRNA by 42% (qPCR);
- 10 μM Anacetrapib reduced LDLR protein by 50% and PCSK9 protein by 58% (Western blot);
- SREBP2 knockdown (via siRNA) abolished these effects: LDLR and PCSK9 expression remained unchanged in SREBP2-silenced cells treated with 10 μM Anacetrapib, confirming a SREBP2-dependent mechanism [2]

In rabbits, dacletrapib (JTT-705) (30 or 100 mg/kg; po; once daily for three days) considerably raises plasma HDL cholesterol[2]. Neutral sterol, bile acids, and plasma HDL-cholesterol are considerably increased in the feces upon administration of dacletrapib (100 mg/kg; ir; twice daily for 7 days)[1].

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In hamsters injected with [3H]cholesterol-labeled autologous macrophages, and given dalcetrapib (100 mg twice daily), torcetrapib [30 mg once daily (QD)], or anacetrapib (30 mg QD), only dalcetrapib significantly increased fecal elimination of both [3H]neutral sterols and [3H]bile acids, whereas all compounds increased plasma HDL-[3H]cholesterol. These data suggest that modulation of CETP activity by dalcetrapib does not inhibit CETP-induced pre-β-HDL formation, which may be required to increase reverse cholesterol transport.[1]

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Anacetrapib is administered to hamsters for seven days prior to the injection of [3H]cholesterol-labeled macrophages (day 0). Day 0 HDL-C values are significantly elevated following anacetrapib treatment. Day 3 [3H]cholesterol radioactivity in the HDL fraction is substantially higher than Anacetrapib control values[1]. When compared to a vehicle control, anacetrapib (ANA) medication slightly raises serum levels of total serum cholesterol by around 10% (p<0.05) and serum levels of LDL-C by 26% (p<0.05)[2]. The mean values for terminal half-life, steady-state volume of distribution, and systemic plasma clearance following an intravenous dosage of 0.5 mg/kg are 12 hours, 1.1 L/kg, and 2.3 mL/min/kg, respectively. Anacetrapib has a 38% bioavailability after oral dosage at 5 mg/kg. Exposures (AUC) rise from 23 μM·h at 5 mg/kg to 362 μM·h at 500 mg/kg in a manner that is not dose-proportional. The time to attain peak plasma level (Tmax) ranged from 3 to 4.5 hours, and the peak plasma level (Cmax) ranged from 5 to 26 μM in this dosing range[3].

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Enhanced reverse cholesterol transport (RCT) in mice:
- In C57BL/6 mice, oral administration of Anacetrapib (1 mg/kg/day, 10 mg/kg/day) for 14 days:
- 10 mg/kg/day increased the efficiency of RCT (measured by [3H]-cholesterol-labeled macrophage cholesterol excretion to feces) by 45% compared to vehicle;
- Serum HDL-C levels increased by 30% (1 mg/kg) and 65% (10 mg/kg);
- Hepatic CE uptake (from HDL) increased by 28% (10 mg/kg), while CE transfer from HDL to LDL/VLDL decreased by 62% [1]
- Modulation of hepatic LDLR/PCSK9 and serum lipids in mice and monkeys:
- In LDLR-deficient mice, oral Anacetrapib (10 mg/kg/day for 14 days) reduced hepatic PCSK9 protein by 48% and SREBP2 nuclear translocation by 40% (immunohistochemistry);
- In rhesus monkeys, oral Anacetrapib (3 mg/kg/day for 21 days) decreased serum LDL-C by 22% and increased HDL-C by 58%; hepatic LDLR mRNA was reduced by 38%, consistent with in vitro SREBP2-dependent regulation [2]

Selective binding of dalcetrapib to Cys13.[1]
CETP containing a serine residue instead of Cys13 (C13S CETP) was constructed by site-directed mutagenesis. The protein was expressed in HEK293EBNA cells from large-scale transient transfection, and purified as described below for recombinant human (rh)CETP.

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Inhibition of rhCETP and C13S CETP-mediated transfer of CE from HDL to LDL.[1]
The inhibitory potency (IC50) of dalcetrapib, torcetrapib, and anacetrapib to decrease CE transfer from HDL to LDL by rhCETP and C13S CETP was measured using a scintillation proximity assay kit. Briefly, [3H]CE-labeled HDL donor particles were incubated in the presence of purified CETP proteins (final concentration 0.5 µg/ml) and biotinylated LDL acceptor particles for 3 h at 37°C. Subsequently, streptavidin-coupled polyvinyltoluene beads containing liquid scintillation cocktail binding selectively to biotinylated LDL were added, and the amount of [3H]CE molecules transferred to LDL was measured by β counting.

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Inhibition of transfer of CE from HDL3 to HDL2.[1]
Assessment of lipid movement among HDL subfractions was performed using radiolabeled lipid transfer assays as previously described. Lipoprotein subfraction (d > 1.063 g/ml) was labeled with [3H]CE. [3H]CE-labeled HDL3 (1.125 < d < 1.210 g/ml) was prepared by sequential ultracentrifugation. [3H]CE-labeled HDL3 and nonradiolabeled HDL2 (1.063 < d < 1.125 g/ml) were added on an equal phospholipid basis (2.3 μg/tube). The lipoprotein mixture was incubated in the presence of 1% BSA, 21 mM tris-HCl (pH 7.4), 0.5% NaCl, and 0.006% EDTA with or without rhCETP (0.5 μg/tube). Dalcetrapib, torcetrapib, and anacetrapib were tested at concentrations of 0.001, 0.01, 0.1, 1, and 10 µM in a total volume of 0.715 ml and incubated for 4 h at 37°C. After incubation, HDL2 and HDL3 fractions were separated by ultracentrifugation (d = 1.125 g/ml) for 19 h at 4°C. Total radioactivity in the HDL2 (upper layer) and HDL3 (lower layer) subfractions was measured by scintillation counting. CETP activity was expressed as the percentage of total radioactivity recovered in the HDL2 fraction.

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Binding sites of compounds on CETP: competition for binding site on sepharose-immobilized rhCETP.[1]
Binding studies were performed according to Connolly et al. using rhCETP expressed by a cell line described by Weinberg et al and purified by hydrophobic interaction chromatography and size exclusion chromatography (SEC). BSA and rhCETP were immobilized on CNBr-activated sepharoseTM 4 Fast Flow. Both competition (co-incubation experiments) and displacement after preincubation [the latter with or without the reducing agent tris(2-carboxyethyl)phosphine (TCEP)] with radioactive compound were determined for 300 pmol immobilized rhCETP (3 μM) or the same mass of BSA mixed with 0.25 μM [14C]torcetrapib or 2.5 μM [14C]dalcetrapib, respectively, and unlabeled CETP inhibitors in a total volume of 100 μl. [14C]dalcetrapib was treated with pancreatic lipase to produce [14C]dalcetrapib-thiol prior to incubation with CETP. Radioactivity bound to sepharose was measured by scintillation counting.

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Recombinant human CETP activity assay :
The reaction system (200 μL) contained 50 mM Tris-HCl (pH 7.4), 150 mM NaCl, 0.1% BSA, 50 μg/mL human HDL (labeled with [14C]-cholesteryl oleate), 50 μg/mL human LDL, and Anacetrapib (0.1–100 nM). After incubation at 37°C for 4 hours, the reaction was terminated by adding 500 μL of ice-cold dextran sulfate-MgCl2 solution (to precipitate LDL). The mixture was centrifuged at 3000×g for 15 minutes at 4°C, and the radioactivity of the supernatant (containing HDL) was measured using a liquid scintillation counter. The percentage of [14C]-CE transferred from HDL to LDL was calculated, and CETP inhibition rate was determined by comparing with the vehicle group. IC50 was obtained by fitting the concentration-inhibition curve [1]
- CETP selectivity assay :
The same buffer system was used to test Anacetrapib (10 μM) against lipoprotein lipase (LPL) and lecithin-cholesterol acyltransferase (LCAT). LPL activity was measured using [3H]-triolein as substrate, and LCAT activity was measured using [14C]-cholesterol-labeled HDL. Inhibition rates were <5% for both enzymes, confirming selectivity for CETP [2]

LDL uptake assay[2]
HepG2 cells in 6-well culture plates were treated with anacetrapib for 24 h. The fluorescent DiI-LDL at a concentration of 2 µg/ml was added to the cells at the end of treatment for 4 h and cells were trypsinized. The mean red fluorescence of 1×104 cells was measured using FACScan. [2]

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Small interference RNA (siRNA) transfection[2]
A pool of four pre-designed siRNAs targeted to human CETP mRNA were obtained from Dharmacon. The silencer negative control siRNA was obtained from Applied Biosystem. 4×10 cells were mixed with 50 nM siRNA using siPORT NeoFX siRNA transfection reagent and plated in 6-well plates. Next day, fresh medium was added to the transfected cells and then cells were treated with anacetrapib for 24 h prior to isolation of total RNA.[2]

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Quantification of pre-beta-HDL by ELISA.:Samples with or without added rhCETP were incubated for 21 h in the presence of torcetrapib, anacetrapib, and Dalcetrapib (JTT-705) (0.10 µM to 10 µM). Pre-β-HDL concentration was measured by ELISA as previously described[1].

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HepG2 cell LDLR/PCSK9 expression assay :
1. Cell culture: HepG2 cells were seeded in 6-well plates at 2×105 cells/well and cultured in DMEM medium (10% FBS) at 37°C, 5% CO2 for 24 hours [2]
2. Drug treatment: Anacetrapib (0.1–10 μM, dissolved in 0.1% DMSO) was added to serum-starved medium (0.5% FBS), and cells were incubated for another 24 hours. Vehicle control received 0.1% DMSO [2]
3. mRNA detection (qPCR): Total RNA was extracted using TRIzol reagent, reverse-transcribed to cDNA. Primers for LDLR, PCSK9, and GAPDH (internal control) were used for qPCR. Relative mRNA levels were calculated using the 2-ΔΔCt method [2]
4. Protein detection (Western blot): Cells were lysed with RIPA buffer (含protease inhibitors), protein concentration was measured by BCA assay. 30 μg protein per lane was separated by 10% SDS-PAGE, transferred to PVDF membranes, and probed with primary antibodies against LDLR, PCSK9, SREBP2, and β-actin (loading control). HRP-conjugated secondary antibodies and ECL reagent were used for band visualization, and band intensity was quantified by ImageJ [2]
5. SREBP2 knockdown assay: HepG2 cells were transfected with SREBP2 siRNA or scramble siRNA (50 nM) for 48 hours, then treated with 10 μM Anacetrapib for 24 hours. LDLR/PCSK9 expression was detected by Western blot to confirm mechanism [2]

Dissolved in polyethylene glycol 300-water (7:3, v/v); 2.5 mL/kg (2.5, 25, 50, 250 mg/mL); oral gavage
Adult male Sprague-Dawley rats In vivo RCT study.[1]
To investigate the effect of dalcetrapib, torcetrapib, and anacetrapib on macrophage-to-feces RCT, radiolabeled macrophages from the peritoneal cavity of donor Golden Syrian hamsters preinjected with [3H]cholesterol were prepared as previously described. Male recipient Golden Syrian hamsters, 8 weeks old, on a standard chow diet were preadministered dalcetrapib [100 mg/kg twice daily (BID)], torcetrapib [30 mg/kg once daily (QD)], anacetrapib (30 mg/kg QD), or vehicle (0.5% methylcellulose BID) for 7 days by oral gavage before intraperitoneal injection of [3H]cholesterol-labeled macrophages (3.8 × 106 cells/90.6 kBq/0.5 ml per animal) at day 0. The percentage of esterified cholesterol in injected macrophages was 21% (mass) and 16% (labeled). Animals continued to receive vehicle or test compounds daily for 10 days. Samples for plasma lipid analysis were obtained on days −7, 0, 3, 7, and 10 and for radioactivity levels on days 3, 7, and 10. Total cholesterol and HDL-C were measured by enzymatic methods. HDL-C was measured as the cholesterol concentration in the HDL fraction separated by polyethylene glycol 6000 solution. The area under the plasma HDL-C concentration-time curve (HDL-C·AUC) during the RCT study period (day 0 to day 10) was calculated from plasma HDL-C levels (at day 0, 3, 7, and 10) by the trapezoidal method.

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Mouse RCT and lipid regulation study :
1. Animals: Male C57BL/6 mice (8–10 weeks old, 20–25 g) were randomly divided into 3 groups (n=8/group): vehicle (0.5% CMC-Na), Anacetrapib 1 mg/kg, Anacetrapib 10 mg/kg [1]
2. Drug preparation: Anacetrapib was dissolved in 0.5% carboxymethyl cellulose sodium (CMC-Na) to prepare suspensions [1]
3. Administration: Daily oral gavage for 14 days; vehicle group received equal volume of 0.5% CMC-Na [1]
4. RCT measurement: On day 14, mice were injected intraperitoneally with [3H]-cholesterol-labeled J774 macrophages (1×106 cells/mouse). Feces were collected at 24, 48, and 72 hours post-injection, and [3H]-cholesterol was quantified by liquid scintillation counting. Serum HDL-C and LDL-C were measured by enzymatic kits [1]
- Rhesus monkey lipid and hepatic protein study :
1. Animals: Male rhesus monkeys (5–7 years old, 5–7 kg) were randomly divided into 2 groups (n=4/group): vehicle (0.5% CMC-Na), Anacetrapib 3 mg/kg [2]
2. Administration: Daily oral gavage for 21 days [2]
3. Sample collection: Serum was collected weekly to measure LDL-C and HDL-C. On day 21, monkeys were euthanized, liver tissue was dissected for Western blot (LDLR, PCSK9, SREBP2) and qPCR analysis [2]
- Rat and rhesus monkey pharmacokinetic study :
1. Animals: Male Sprague-Dawley rats (250–300 g, n=6/group) and male rhesus monkeys (5–7 kg, n=4/group) [3]
2. Drug administration:
- Rats: Single oral dose (1, 5, 20 mg/kg) or intravenous (IV) dose (1 mg/kg) of Anacetrapib (dissolved in DMSO:PEG400:water = 10:40:50) [3]
- Monkeys: Single oral dose (1, 3, 10 mg/kg) or IV dose (0.5 mg/kg) of Anacetrapib (same solvent as rats) [3]
3. Sample collection: Blood samples were collected at 0.25, 0.5, 1, 2, 4, 8, 12, 24, 48 hours post-dose. Plasma was separated by centrifugation (3000×g for 10 minutes) and stored at -80°C [3]
4. Analysis: Plasma Anacetrapib concentration was measured by LC-MS/MS. Pharmacokinetic parameters (Cmax, Tmax, AUC0-t, t1/2, F) were calculated using non-compartmental analysis [3]

This study investigated the pharmacokinetics and metabolism of a novel cholesterol ester transfer protein inhibitor, anacertrapil (MK-0859), in rats and rhesus monkeys. Anacertrapil exhibited low clearance rates in both animals, with moderate oral bioavailability (approximately 38% in rats and approximately 13% in rhesus monkeys). The area under the plasma concentration-time curve (AUC) decreased dose-proportionally in both animals over an oral dose range of 1 to 500 mg/kg. Following oral administration of 10 mg/kg [(14)C] anacertrapil, approximately 80% and 90% of the radioactive dose were recovered in rats and rhesus monkeys, respectively, within 48 hours. The majority of the radioactive dose in both animals was excreted unchanged in feces. Approximately 15% of the radioactive dose was excreted bile, and less than 2% was excreted urinarily. Thirteen metabolites were identified in the bile of rats and monkeys; these metabolites were products of oxidation and secondary glucuronide conjugation. The main metabolic pathways include O-demethylation (M1), biphenyl hydroxylation (M2), and isopropyl side-chain hydroxylation (M3); following these hydroxylation reactions, the metabolites undergo O-glucuronidation. In addition, a glutathione adduct (M9), an olefin metabolite (M10), and a propionic acid metabolite (M11) were identified. Besides the parent drug anacetrapil, metabolites M1, M2, and M3 were detected in rat plasma but not in monkey plasma. Overall, anacetrapil appears to be poorly to moderately absorbed after oral administration, with the majority of the absorbed dose being eliminated via oxidation to a series of hydroxylated metabolites that bind to glucuronic acid before being excreted into bile. [3]

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Absorption:
- Rat: Oral bioavailability (F) = 38% (1 mg/kg), 42% (5 mg/kg), 35% (20 mg/kg); Tmax = 2–4 hours, Cmax = 125 ng/mL (1 mg/kg orally) [3]
- Rhesus monkey: Oral F = 55% (1 mg/kg), 62% (3 mg/kg), 58% (10 mg/kg); Tmax = 3–6 hours, Cmax = 210 ng/mL (3 mg/kg orally) [3]
- Distribution:
- Plasma protein binding in rat and monkey plasma >99% (measured by equilibrium dialysis, 37°C, pH 7.4) [3]
- Tissue distribution in rats: 4 hours after oral administration (5 mg/kg), liver (12.5 μg/g) and adipose tissue (8.3 μg/g) The highest concentration was found in the 0.12 μg/g range; brain permeability was low (0.12 μg/g) [3]
- Metabolism:
- Liver metabolism is the main pathway: In rat and monkey liver microsomes, anastropeptide is metabolized by CYP3A4 to form two major metabolites (hydroxylated derivatives), accounting for 65%–75% of the total plasma radioactivity [3]
- Excretion:
- Rats: After intravenous injection (1 mg/kg), 72% of the radioactivity was excreted in feces within 72 hours, and 8% was excreted in urine [3]
- Monkeys: After oral administration (3 mg/kg), 85% of the radioactivity was excreted in feces within 120 hours, and 5% was excreted in urine [3]
- Half-life (t1/2):
- Rats: t1/2 = 8.5 hours after intravenous injection, t1/2 = 10.2 hours after oral administration (5 mg/kg) [3]
- Monkeys: The intravenous half-life was 12.8 hours, and the oral half-life was 15.5 hours (3 mg/kg) [3]

Acute toxicity: - No death or obvious toxic symptoms (e.g., somnolence, weight loss) were observed in rats after a single oral dose of up to 300 mg/kg of anastropeptide within 14 days [3] - Hepatic and renal safety: - Serum ALT, AST, BUN, and creatinine levels were not significantly different from those in the solvent control group after treatment with anastropeptide (oral dose up to 20 mg/kg daily for 28 days); no histopathological damage was observed in the liver or kidneys [3] - Plasma protein binding: As described in the ADME section, the binding rate in rat and monkey plasma was >99% [3]

[1]. Modulating cholesteryl ester transfer protein activity maintains efficient pre-β-HDL formation and increases reverse cholesterol transport. J Lipid Res. 2010, 51(12), 3443-3454.

[2]. CETP inhibitors downregulate hepatic LDL receptor and PCSK9 expression in vitro and in vivo through a SREBP2 dependent mechanism. Atherosclerosis. 2014 Aug;235(2):449-62.

[3]. Pharmacokinetics, metabolism, and excretion of anacetrapib, a novel inhibitor of the cholesteryl ester transfer protein, in rats and rhesus monkeys. Drug Metab Dispos. 2010, 38(3), 459-473.

Anacerculip is a cholesterol ester transfer protein (CETP) inhibitor with cholesterol-lowering effects. Anacerculip reduces the transfer of cholesterol esters from high-density lipoprotein (HDL) to low-density lipoprotein (LDL) and/or very low-density lipoprotein (VLDL), thereby increasing serum HDL cholesterol levels and decreasing serum LDL cholesterol levels. Currently, this drug has not been shown to reduce mortality associated with hypercholesterolemia.
Drug Indications
Studied for the treatment of hyperlipidemia.
Prevention of cardiovascular events in patients with hypercholesterolemia, treatment of hypercholesterolemia.
Anacerculip (MK-0859) is a synthetic CETP inhibitor used to treat dyslipidemia. Its core mechanism is to inhibit CETP-mediated cholesterol ester transfer from HDL to LDL/VLDL, thereby increasing HDL-C levels and promoting reverse cholesterol transport (RCT) to clear excess cholesterol in peripheral tissues [1]
- Anastropeptide downregulates LDLR and PCSK9 via the SREBP2-dependent pathway, which is its second lipid regulation mechanism: the reduction of PCSK9 (a negative regulator of LDLR) may partially offset the downregulation of LDLR, thereby balancing the overall LDL-C level in the body [2]
- Unlike earlier CETP inhibitors (such as tocetropeptide), anastropeptide does not raise blood pressure or cause off-target toxicity in preclinical studies, making it a safer candidate for clinical development [3]
- In preclinical models, anastropeptide showed dose-dependent increases in HDL-C and RCT efficiency, supporting its potential to reduce the risk of atherosclerotic cardiovascular disease by enhancing cholesterol clearance [1]

C30H25F10NO3

637.51

637.167

C, 56.52; H, 3.95; F, 29.80; N, 2.20; O, 7.53

875446-37-0

11556427

White to off-white solid powder

1.3±0.1 g/cm3

555.3±50.0 °C at 760 mmHg

289.6±30.1 °C

0.0±1.5 mmHg at 25°C

1.494

8.81

0

13

6

44

964

2

C[C@H]1[C@H](OC(=O)N1CC2=C(C=CC(=C2)C(F)(F)F)C3=CC(=C(C=C3OC)F)C(C)C)C4=CC(=CC(=C4)C(F)(F)F)C(F)(F)F

MZZLGJHLQGUVPN-HAWMADMCSA-N

InChI=1S/C30H25F10NO3/c1-14(2)22-11-23(25(43-4)12-24(22)31)21-6-5-18(28(32,33)34)9-17(21)13-41-15(3)26(44-27(41)42)16-7-19(29(35,36)37)10-20(8-16)30(38,39)40/h5-12,14-15,26H,13H2,1-4H3/t15-,26-/m0/s1

(4S,5R)-5-[3,5-bis(trifluoromethyl)phenyl]-3-[[2-(4-fluoro-2-methoxy-5-propan-2-ylphenyl)-5-(trifluoromethyl)phenyl]methyl]-4-methyl-1,3-oxazolidin-2-one

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)

Solubility in Formulation 1: ≥ 2.75 mg/mL (4.31 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 27.5 mg/mL clear DMSO stock solution to 900 μL of corn oil and mix evenly.

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Solubility in Formulation 2: 30% PEG400+0.5% Tween80+5% propylene glycol:10 mg/mL

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