Sacubitrilat (formerly known as LBQ-657; LBQ657) is the activated form of Sacubitril (AHU-377; AHU-377; Entresto), which is a novel and potent NEP (neutral endopeptidase 24.11) inhibitor used in combination with valsartan (LCZ696) for treating heart failure. Sacubitril is a prodrug that requires esterases to de-ethylate in order to be converted into the active form LBQ657. Atrial and brain natriuretic peptides are two peptides that lower blood pressure primarily by decreasing blood volume. LBQ657 inhibits the enzyme neprilysin, which is in charge of breaking these peptides down.

Physicochemical Properties

Molecular Formula	C22H25NO5
Molecular Weight	383.4376
Exact Mass	383.173
Elemental Analysis	C, 68.91; H, 6.57; N, 3.65; O, 20.86
CAS #	149709-44-4
Related CAS #	Sacubitrilat-d4
PubChem CID	10430040
Appearance	White to off-white solid powder
LogP	4.196
Hydrogen Bond Donor Count	3
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	10
Heavy Atom Count	28
Complexity	521
Defined Atom Stereocenter Count	2
SMILES	C[C@H](C[C@@H](CC1=CC=C(C=C1)C2=CC=CC=C2)NC(=O)CCC(=O)O)C(=O)O
InChi Key	DOBNVUFHFMVMDB-BEFAXECRSA-N
InChi Code	InChI=1S/C22H25NO5/c1-15(22(27)28)13-19(23-20(24)11-12-21(25)26)14-16-7-9-18(10-8-16)17-5-3-2-4-6-17/h2-10,15,19H,11-14H2,1H3,(H,23,24)(H,25,26)(H,27,28)/t15-,19+/m1/s1
Chemical Name	(2R,4S)-4-(3-carboxypropanoylamino)-2-methyl-5-(4-phenylphenyl)pentanoic acid
Synonyms	Sacubitrilat; LBQ657; LBQ-657; LBQ 657
HS Tariff Code	2934.99.9001
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)

Biological Activity

Targets	NEP Sacubitrilat (LBQ657) has a distinct stereocenter and is a single diastereomer. Sacubitrilat exhibits a high inhibitory efficacy of 5 nM upon binding to the active site of NEP via a complicated response network including all of the compound's functional groups [1].
ln Vitro	Sacubitrilat (LBQ657) has a distinct stereocenter and is a single diastereomer. Sacubitrilat exhibits a high inhibitory efficacy of 5 nM upon binding to the active site of NEP via a complicated response network including all of the compound's functional groups [1].
ln Vivo	The present study summarizes the pharmacokinetics of sacubitrilat, sacubitrilat la (LBQ657), and the insecticide Sartan after a single sidewall dose of LCZ696 at 400 or 1200 mg during fasting. With a median Tmax of 0.52 h for the 400 mg dose and 1.05 h for the 1200 mg dose, Sacubitril's mean intraperitoneal concentration rose quickly. Sacubitril was next, with corresponding Tmax values of 2.07 and 3.05 h, respectively. The median brasartan Tmax for the 400 mg and 1200 mg dosages of LCZ696 was 2.07 hours. Sacubitril's Cmax shown a dose-proportional increase, while the Cmaxes of both Sacubitril and Sacubitril demonstrated an increase that was not proportionate to the dosages. Arithmetic tools AUC0-24 h and AUClast exhibited less of a dose-proportional increase for Sartan, but increased about dose-proportionally for Sacubitril and Sacubitrilat [2].
Enzyme Assay	Sacubitril is an ethyl ester prodrug of LBQ657, the active neprilysin (NEP) inhibitor, and a component of LCZ696 (sacubitril/valsartan). We report herein the three-dimensional structure of LBQ657 in complex with human NEP at 2 Å resolution. The crystal structure unravels the binding mode of the compound occupying the S1, S1' and S2' sub-pockets of the active site, consistent with a competitive inhibition mode. An induced fit conformational change upon binding of the P1'-biphenyl moiety of the inhibitor suggests an explanation for its selectivity against structurally homologous zinc metallopeptidases [1].
Animal Protocol	Purpose: Sacubitril/valsartan (LCZ696) is a first-in-class angiotensin receptor neprilysin inhibitor (ARNI) indicated to reduce the risk of cardiovascular death and hospitalization for heart failure in patients with chronic heart failure (NYHA class II-IV) and reduced ejection fraction. This study was aimed to evaluate the effect of single oral therapeutic (400 mg) and supratherapeutic (1200 mg) doses of LCZ696 on cardiac repolarization.[2] Method: This randomized double-blind crossover study in healthy male subjects compared the effect of therapeutic and supratherapeutic doses of LCZ696 with placebo and moxifloxacin 400 mg (open-label treatment) as positive control. The primary assessment was mean baseline- and placebo-corrected QTcF (∆∆QTcF; Fridericia correction). Additional assessments included the ∆∆QTcB (Bazett's correction), PR interval, QRS duration, heart rate (HR), LCZ696 pharmacokinetics, pharmacokinetic/pharmacodynamic relationships, and safety.[2] Results: Of the 84 subjects enrolled, 81 completed the study. The maximum upper bound of the two-sided 90 % confidence interval for ∆∆QTcF for LCZ696 400 mg and 1200 mg were <10 ms, and assay sensitivity was confirmed with moxifloxacin. No relevant treatment-emergent changes were observed in any of the ECG-derived parameters with LCZ696 or placebo, and the incidence of adverse events was comparable among the treatment groups.[2]
ADME/Pharmacokinetics	This study provides pharmacokinetic (PK) parameters for LBQ657 following single oral doses of LCZ696 400 mg (therapeutic) and 1200 mg (supratherapeutic) in healthy male subjects under fasting conditions. [2] After LCZ696 400 mg dose (N=81-82): Arithmetic mean AUC₀₋₂₄ was 122,000 h·ng/mL (SD 19,700, CV% 16.2); Arithmetic mean AUCₗₐₛₜ was 121,000 h·ng/mL (SD 22,200, CV% 18.3); Arithmetic mean Cₘₐₓ was 13,700 ng/mL (SD 2,490, CV% 18.2); Median Tₘₐₓ was 2.07 hours (range 1.05-5.07). [2] After LCZ696 1200 mg dose (N=82): Arithmetic mean AUC₀₋₂₄ was 364,000 h·ng/mL (SD 62,300, CV% 17.1); Arithmetic mean AUCₗₐₛₜ was 364,000 h·ng/mL (SD 62,400, CV% 17.1); Arithmetic mean Cₘₐₓ was 40,700 ng/mL (SD 6,990, CV% 17.2); Median Tₘₐₓ was 3.05 hours (range 2.05-5.07). [2] The Cₘₐₓ of LBQ657 showed a dose-proportional increase between the 400 mg and 1200 mg doses of LCZ696. The AUC increased approximately dose-proportionally. The half-life is mentioned to be short, but no specific t₁/₂ value is provided. The document also states that all LCZ696 analytes (including LBQ657) have high plasma protein binding, but no specific percentage is given. [2]
Toxicity/Toxicokinetics	The safety assessment focused on the combination drug LCZ696. In this study of healthy subjects, single doses of LCZ696 400 mg and 1200 mg (which delivers LBQ657) were generally safe and well tolerated. The overall incidence of adverse events (AEs) was comparable to placebo (14.6% for each LCZ696 dose vs. 12.2% for placebo). Common AEs included headache, nasopharyngitis, postural dizziness, and nausea. Most AEs were mild and resolved without treatment. No deaths, serious AEs, or AEs leading to discontinuation were reported. No clinically significant changes in lab parameters, vital signs, or safety ECGs (beyond the primary QTc assessment) were observed. The study concluded that LCZ696 did not affect cardiac repolarization (QTc interval), indicating a low risk for drug-induced proarrhythmia related to its components, including LBQ657. [2]
References	[1]. Structure of neprilysin in complex with the active metabolite of sacubitril. Sci Rep. 2016 Jun 15;6:27909. [2]. Single therapeutic and supratherapeutic doses of sacubitril/valsartan (LCZ696) do not affect cardiac repolarization. Eur J Clin Pharmacol. 2016 Aug;72(8):917-24.
Additional Infomation	A metabolite of LCZ696 with neprilysin inhibitory activity. Sacubitrilat is a Neprilysin Inhibitor. The mechanism of action of sacubitrilat is as a Neprilysin Inhibitor. Sacubitrilat (LBQ657) is the active neprilysin (NEP) inhibitor metabolite generated from the prodrug sacubitril upon oral administration of LCZ696 (sacubitril/valsartan). LCZ696 is an angiotensin receptor neprilysin inhibitor (ARNI) approved to reduce the risk of cardiovascular death and hospitalization in chronic heart failure patients with reduced ejection fraction. This dedicated thorough QT (TQT) study demonstrated that single therapeutic (400 mg) and supratherapeutic (1200 mg) oral doses of LCZ696 did not prolong the QTc interval in healthy subjects, meeting the criteria set by ICH E14 guidelines. The mean baseline- and placebo-corrected QTcF (ΔΔQTcF) for both LCZ696 doses remained below 5 ms, with the upper bound of the 90% confidence interval below 10 ms at all time points. Assay sensitivity was confirmed using moxifloxacin (positive control). The study supports the cardiac repolarization safety profile of LCZ696 and, by extension, suggests no significant QT-prolonging risk from its components, including the NEP inhibitor LBQ657. [2]

Solubility Data

Solubility (In Vitro)

DMSO: ~77 mg/mL (~200.8 mM) Ethanol: ~77 mg/mL (~200.8 mM)

Solubility (In Vivo)

Solubility in Formulation 1: ≥ 2.5 mg/mL (6.52 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. Solubility in Formulation 2: ≥ 2.5 mg/mL (6.52 mM) (saturation unknown) 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. Solubility in Formulation 3: ≥ 2.5 mg/mL (6.52 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.  (Please use freshly prepared in vivo formulations for optimal results.)

Preparing Stock Solutions		1 mg	5 mg	10 mg
	1 mM	2.6080 mL	13.0398 mL	26.0797 mL
	5 mM	0.5216 mL	2.6080 mL	5.2159 mL
	10 mM	0.2608 mL	1.3040 mL	2.6080 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.