| Size | Price | Stock | Qty |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg |
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| 250mg |
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| 500mg | |||
| Other Sizes |
Purity: ≥98%
| Targets |
Endothelial lipase (EL) [1]
EL: IC50 = 0.237 μM (human, cell-free FELA mixed micelle assay) [1] EL: IC50 = 0.11 μM (human, functional HDL particle assay) [1] EL: IC50 = 0.25 μM (human, cell-based assay using EL-transfected HEK cells and PED-A substrate) [1] LPL: IC50 = 209.5 μM (human) [1] |
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| ln Vitro |
XEN445 shows good in vitro inhibitory potency against EL, and high selectivity over LPL and HL.
Cell Assay: In vitro, XEN445 has been reported to selectively inhibit EL with IC50 values of 0.237μM, 20μM and 9.5μM for hEL, hLPL and hHL, respectively. In addition, XEN445 has been revealed to suppress EL in EL-transfected HEK cells with an IC50 value of 0.25μM which is very similar to that tested in cell-free assay. Besides, XEN445 has shown significantly high oral bioavailability and low clearance rates in C57BL6 mice. XEN445 exhibited potent inhibitory activity against human endothelial lipase (EL) in a cell-free fluorogenic phosphatidylcholine mixed micelle assay (FELA, bis-BODIPY-FL C11-PC), with an IC50 of 0.237 μM [1]. In a more physiological functional HDL particle assay using purified human HDL as substrate and measuring free fatty acid release, XEN445 showed an IC50 of 0.11 μM [1]. In a cell-based assay using EL-transfected HEK cells and the soluble EL fluorogenic substrate PED-A, after 30 min preincubation of cells with XEN445, an IC50 value of 0.25 μM was obtained, similar to the cell-free assay [1]. Selectivity: XEN445 demonstrated high selectivity over human lipoprotein lipase (LPL, IC50 = 209.5 μM) with >880-fold selectivity; selectivity over human hepatic lipase (HL) was not determined but compounds in this class generally showed good selectivity [1]. |
| ln Vivo |
In vivo, XEN445 displays good ADME and PK properties. In mice, XEN445 (30 mg/kg, p.o.) raises plasma HDL-cholesterol concentration.
Oral administration of XEN445 at 30 mg/kg twice daily (b.i.d.) for 3 days to wild-type C57BL/6 mice raised total plasma cholesterol by 18% and plasma HDL-cholesterol (HDLc) by 16% compared to vehicle-treated wild-type mice, with average plasma levels of XEN445 at termination (16 h post final dose) of 9.9 μM [1]. Dosing XEN445 at 10 mg/kg b.i.d. for 3 days to wild-type mice resulted in approximately 10% increase in plasma HDLc [1]. To confirm on-target efficacy, XEN445 was dosed to EL knockout (LIPG KO) mice at 30 mg/kg b.i.d. for 3 days; no significant change in total plasma cholesterol or HDLc was observed compared to vehicle-treated EL knockout mice, while vehicle-treated EL knockout mice had 121% higher HDLc than wild-type controls [1]. Extended treatment of wild-type mice with XEN445 at 30 mg/kg b.i.d. for 9 days resulted in a 30% increase in plasma HDLc (vs. 16% at 3 days) [1]. |
| Enzyme Assay |
The activity of endothelial lipase (EL) was measured using a mixed micelle fluorogenic phosphatidylcholine/Triton X-100 assay. Recombinant human or mouse EL was diluted in PBS containing 14% DMEM, 2% glycerol. 40 μL of diluted enzyme (1.4 nM final concentration) was added to each well of a 96-well plate containing inhibitors or vehicle (5 μL in 10% DMSO). The enzyme-inhibitor mixture was incubated at room temperature for 30 min, then 5 μL of 10 μM bis-BODIPY-FL C11-PC in 0.1% Triton X-100 was added. Fluorescence was read at excitation 490 nm and emission 520 nm. Human LPL (4 nM final) assay required addition of ApoCII (1 μg/μL final). Human HL (6 nM final) was assayed in 50 mM Tris pH 8.5, 1 M NaCl. Background activity was determined using mock-transfected HEK-293 cell media [1].
Functional HDL particle assay: Potency of EL inhibitors was determined using purified human HDL particles as a physiologically relevant substrate. Release of free fatty acids was measured as readout of lipase activity [1]. Cell-based EL activity assay: EL-transfected HEK cells were preincubated with XEN445 for 30 min, then the soluble EL fluorogenic substrate PED-A was added. The IC50 was determined [1]. Recombinant human and mouse EL, human LPL, and human HL were obtained by transient transfection of HEK-293 cells with codon-optimized sequences in pcDNA3.1. Cells were grown in DMEM +10% FBS to 50-80% confluency in 225 cm² flasks and transfected with 95 μg plasmid DNA using Lipofectamine. After 5 h, media was replaced with Opti-MEM and cells cultured for 72 h. Cell-associated lipase was displaced by adding heparin to 50 U/mL for 30 min. Media was removed and concentrated 100-fold using concentrators (30 kDa cutoff). Active site concentration of each lipase was determined using orlistat titration [1]. |
| Cell Assay |
XEN445 was tested in a cell-based assay using EL-transfected HEK cells. After 30 min preincubation of EL-expressing HEK cells with XEN445, the soluble EL fluorogenic substrate PED-A was added, and an IC50 value of 0.25 μM was obtained, similar to that determined in the cell-free assay [1].
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| Animal Protocol |
Formulated in 0.2% Tween-20/1% carboxymethyl-cellulose; 30 mg/kg bid; p.o. Male EL knockout mice and age-matched wild-type male C57BL6 mice
Mouse pharmacokinetics: Male C57BL/6 mice (non-fasted, n=3) were administered XEN445 orally (po) at 10 mg/kg in 50% PEG 400 and 50% D5W (dextrose 5% in water), or intravenously (iv) at 1 mg/kg in 10% ethanol, 25% PEG 400 and 65% D5W. Pharmacokinetic parameters were determined [1]. In vivo efficacy study: Male C57BL/6 wild-type or EL knockout (LIPG KO) mice were orally dosed with XEN445 at 30 mg/kg or 10 mg/kg twice daily (b.i.d.) for 3 or 9 days. Vehicle groups received the same dosing regimen without compound. Blood was collected on the morning of day 4 (or day 10) at 16 h post final dose. Plasma total cholesterol and HDLc concentrations were measured [1]. |
| ADME/Pharmacokinetics |
Mouse pharmacokinetic parameters of XEN445 (iv 1 mg/kg, po 10 mg/kg in male C57BL/6 mice, n=3): Cmax (po) = 6.2 μM; Tmax (po) = 0.5 h; Cmin at 24 h (po) = 4.5 μM; AUC last (po) = 47 μM·h; AUC last (iv) = 25.3 μM·h; terminal half-life (t1/2, po) = 11.9 h; clearance (CL, iv) = 0.06 L/h/kg; oral bioavailability (F) = 54% [1].
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| Toxicity/Toxicokinetics |
Plasma protein binding (PPB): XEN445 showed high affinity for rat plasma proteins (97.4% bound at 2.5 μM compound in 10% rat plasma/90% PBS by equilibrium dialysis for 4 h at 37°C), mouse plasma proteins (88.8% bound), and human plasma proteins (95.4% bound) [1].
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| References |
Bioorg Med Chem.2013 Dec 15;21(24):7724-34.
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| Additional Infomation |
XEN445 (compound 13) is a potent and selective endothelial lipase (EL) inhibitor belonging to the anthranilic acid class. It was discovered through optimization of a high-throughput screening hit (compound 1) with focus on safety, potency, selectivity, and protein binding. EL hydrolyzes phosphatidylcholine in HDL particles, reducing HDL particle size. EL activity is inversely correlated with plasma HDLc levels. XEN445 demonstrated in vivo efficacy in raising plasma HDLc in wild-type mice, and the effect was confirmed to be on-target using EL knockout mice. This compound is useful for further defining catalytic and non-catalytic roles of EL in HDL metabolism and as a starting point for developing EL-directed therapeutics for dyslipidemia-related cardiovascular disease [1].
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| Molecular Formula |
C18H17F3N2O3
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| Molecular Weight |
366.33
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| Exact Mass |
366.119
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| Elemental Analysis |
C, 59.02; H, 4.68; F, 15.56; N, 7.65; O, 13.10
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| CAS # |
1515856-92-4
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| Related CAS # |
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| PubChem CID |
72736173
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| Appearance |
White to off-white solid powder
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| Density |
1.4±0.1 g/cm3
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| Boiling Point |
490.9±45.0 °C at 760 mmHg
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| Flash Point |
250.7±28.7 °C
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| Vapour Pressure |
0.0±1.3 mmHg at 25°C
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| Index of Refraction |
1.583
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| LogP |
3.3
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| Hydrogen Bond Donor Count |
1
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| Hydrogen Bond Acceptor Count |
8
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| Rotatable Bond Count |
5
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| Heavy Atom Count |
26
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| Complexity |
489
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| Defined Atom Stereocenter Count |
1
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| SMILES |
FC(C1C([H])=C([H])C(=C(C(=O)O[H])C=1[H])N1C([H])([H])C([H])([H])[C@@]([H])(C1([H])[H])OC([H])([H])C1=C([H])C([H])=C([H])C([H])=N1)(F)F
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| InChi Key |
NBGRERFNOKZQLO-AWEZNQCLSA-N
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| InChi Code |
InChI=1S/C18H17F3N2O3/c19-18(20,21)12-4-5-16(15(9-12)17(24)25)23-8-6-14(10-23)26-11-13-3-1-2-7-22-13/h1-5,7,9,14H,6,8,10-11H2,(H,24,25)/t14-/m0/s1
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| Chemical Name |
(S)-2-(3-(pyridin-2-ylmethoxy)pyrrolidin-1-yl)-5-(trifluoromethyl)benzoic acid
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| Synonyms |
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| HS Tariff Code |
2934.99.9001
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| Storage |
Powder -20°C 3 years 4°C 2 years In solvent -80°C 6 months -20°C 1 month |
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| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
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| Solubility (In Vitro) |
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| Solubility (In Vivo) |
Note: Listed below are some common formulations that may be used to formulate products with low water solubility (e.g. < 1 mg/mL), you may test these formulations using a minute amount of products to avoid loss of samples.
Injection Formulations
Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline)(e.g. IP/IV/IM/SC) *Preparation of saline: Dissolve 0.9 g of sodium chloride in 100 mL ddH ₂ O to obtain a clear solution. Injection Formulation 2: DMSO : PEG300 ：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL DMSO → 400 μLPEG300 → 50 μL Tween 80 → 450 μL Saline) Injection Formulation 3: DMSO : Corn oil = 10 : 90 (i.e. 100 μL DMSO → 900 μL Corn oil) Example: Take the Injection Formulation 3 (DMSO : Corn oil = 10 : 90) as an example, if 1 mL of 2.5 mg/mL working solution is to be prepared, you can take 100 μL 25 mg/mL DMSO stock solution and add to 900 μL corn oil, mix well to obtain a clear or suspension solution (2.5 mg/mL, ready for use in animals). View More
Injection Formulation 4: DMSO : 20% SBE-β-CD in saline = 10 : 90 [i.e. 100 μL DMSO → 900 μL (20% SBE-β-CD in saline)] Oral Formulations
Oral Formulation 1: Suspend in 0.5% CMC Na (carboxymethylcellulose sodium) Oral Formulation 2: Suspend in 0.5% Carboxymethyl cellulose Example: Take the Oral Formulation 1 (Suspend in 0.5% CMC Na) as an example, if 100 mL of 2.5 mg/mL working solution is to be prepared, you can first prepare 0.5% CMC Na solution by measuring 0.5 g CMC Na and dissolve it in 100 mL ddH2O to obtain a clear solution; then add 250 mg of the product to 100 mL 0.5% CMC Na solution, to make the suspension solution (2.5 mg/mL, ready for use in animals). View More
Oral Formulation 3: Dissolved in PEG400 (Please use freshly prepared in vivo formulations for optimal results.) |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.7298 mL | 13.6489 mL | 27.2978 mL | |
| 5 mM | 0.5460 mL | 2.7298 mL | 5.4596 mL | |
| 10 mM | 0.2730 mL | 1.3649 mL | 2.7298 mL |
*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.
Calculation results
Working concentration： mg/mL；
Method for preparing DMSO stock solution： mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.
Method for preparing in vivo formulation:：Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O，mix and clarify.
(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
(2) Be sure to add the solvent(s) in order.
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