| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
| Targets |
The parent compound Sacubitril, when administered as Sacubitril/Valsartan (Entresto®), targets neprilysin. Sacubitril is a prodrug that is activated by carboxylesterase 1 (CES1) in the liver to its active metabolite, sacubitrilat (LBQ657). Sacubitrilat is a potent inhibitor of neprilysin, a neutral endopeptidase that degrades vasoactive peptides including natriuretic peptides (ANP, BNP), bradykinin, and adrenomedullin. By inhibiting neprilysin, the drug increases the levels of these vasoactive peptides, leading to vasodilation, natriuresis, and diuresis, thereby reducing preload and afterload in heart failure. (Z)2S,4R-Sacubitril is an inactive impurity and lacks the correct stereochemistry for neprilysin inhibition.
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| ln Vitro |
As a stereochemical impurity, (Z)2S,4R-Sacubitril has no significant in vitro activity. The (2S,4R) isomer is the active stereoisomer of sacubitril. The (Z) isomer (the specific form of the double bond impurity) and other stereoisomers are expected to have significantly reduced or no biological activity at neprilysin. It is used as a reference standard to control the quality and purity of the sacubitril active pharmaceutical ingredient (API) in pharmaceutical manufacturing.
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| ln Vivo |
(Z)2S,4R-Sacubitril has no in vivo activity; it is an inactive impurity. The active (2S,4R) isomer of sacubitril, when combined with valsartan, alleviates heart failure in animal models and is clinically effective in patients with chronic heart failure. Sacubitril/Valsartan (Entresto®) is superior to enalapril (an ACE inhibitor) in reducing cardiovascular death and hospitalization for heart failure, as demonstrated in the landmark PARADIGM-HF clinical trial. The activity of the drug is dependent on the correct (2S,4R) stereochemistry.
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| Enzyme Assay |
(8Z)2S,4R-Sacubitril is not intended for enzyme/receptor binding assays. The active sacubitril is a prodrug, and its activity is measured by its conversion to sacubitrilat and subsequent inhibition of neprilysin. A non-cell neprilysin inhibition assay is performed using purified recombinant human neprilysin. The active metabolite sacubitrilat (LBQ657) is incubated with a fluorogenic substrate, such as MCA-RPPGFSAFK(Dnp)-OH, in assay buffer (50 mM HEPES, pH 7.4, 100 mM NaCl). The increase in fluorescence (ex 320 nm, em 405 nm) is measured over time. The IC50 for neprilysin inhibition is calculated from the reduction in fluorescence.
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| Cell Assay |
(Z)2S,4R-Sacubitril is not used for cellular assays. The active sacubitril is a prodrug that requires conversion by carboxylesterase 1 (CES1). To test the activity of sacubitril or its impurity, HepG2 cells (which express CES1) can be used. Cells are treated with the test compound, and the production of the active metabolite sacubitrilat in the culture medium is measured by LC-MS/MS. The inactive impurity would show no or minimal conversion to the active metabolite, confirming its lack of biological activity. These assays are used to assess the pharmacokinetic properties of the impurity.
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| Animal Protocol |
(Z)2S,4R-Sacubitril is not used in animal models for efficacy testing. It is a research tool for analytical method development. For the parent drug, in vivo efficacy is tested in the rat model of chronic heart failure (e.g., the coronary ligation-induced heart failure model). Rats are orally administered sacubitril (a combination of the active (2S,4R) isomer) for 4-8 weeks. Endpoints include echocardiography to assess cardiac function (ejection fraction, fractional shortening), measurement of plasma natriuretic peptide levels, and assessment of survival. The impurity is used to test the specificity of analytical methods for quantifying the drug in plasma samples from these animal studies.
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| ADME/Pharmacokinetics |
(Z)2S,4R-Sacubitril is a solid powder with a molecular weight of 409.47. It is soluble in DMSO (~100 mg/mL) and is used as a reference standard for HPLC and LC-MS/MS analysis. The parent drug sacubitril is orally bioavailable and is rapidly hydrolyzed by CES1 in the liver to the active metabolite sacubitrilat. The half-life of sacubitril in humans is approximately 1-2 hours, while the active metabolite sacubitrilat has a half-life of approximately 12 hours. The impurity would be expected to have similar absorption and distribution but no target engagement.
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| Toxicity/Toxicokinetics |
(Z)2S,4R-Sacubitril is a non-toxic stereoisomer and is classified as an impurity. The purity of the active pharmaceutical ingredient is controlled to ensure that the levels of this impurity do not exceed regulatory limits (typically NMT 0.1-0.5%). The active drug Entresto® has a well-established safety profile, with the most common adverse events being hypotension, hyperkalemia, cough, and dizziness. (Z)2S,4R-Sacubitril is not present at levels that would contribute to toxicity.
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| References |
[1]. Shi J, et al. Sacubitril Is Selectively Activated by Carboxylesterase 1 (CES1) in the Liver and the Activation Is Affected by CES1 Genetic Variation. Drug Metab Dispos. 2016 Apr;44(4):554-9.
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| Additional Infomation |
(Z)2S,4R-Sacubitril (Sacubitril-Valsartan Impurity 12) is a high-purity research-grade analytical reference standard. It is used for impurity profiling, method development, and validation in the pharmaceutical quality control (QC) of Sacubitril/Valsartan (Entresto®) drug substance and drug product. Sacubitril is approved by the FDA and other regulatory agencies for the treatment of heart failure with reduced ejection fraction (HFrEF). This product is for research and analytical applications (RUO) only and is not for human use as a therapeutic agent. Key reference: Shi J, et al. Drug Metab Dispos. 2016.
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| Molecular Formula |
C24H27NO5
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| Molecular Weight |
409.47
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| Related CAS # |
Sacubitril;149709-62-6
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| Appearance |
Typically exists as solid at room temperature
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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 |
| 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) |
DMSO :~100 mg/mL (~244.22 mM)
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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 | 2.4422 mL | 12.2109 mL | 24.4218 mL | |
| 5 mM | 0.4884 mL | 2.4422 mL | 4.8844 mL | |
| 10 mM | 0.2442 mL | 1.2211 mL | 2.4422 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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