| Size | Price | Stock | Qty |
|---|---|---|---|
| 1mg |
|
||
| 5mg |
|
||
| 10mg |
|
||
| Other Sizes |
| Targets |
Lonodelestat TFA targets human neutrophil elastase (hNE), a serine protease released by activated neutrophils. NE plays a crucial role in host defense but also contributes to tissue damage and inflammation in chronic lung diseases.
|
|---|---|
| ln Vitro |
Lonodelestat TFA is a potent and selective peptide inhibitor of human neutrophil elastase (hNE). As a protein epitope mimetic (PEM), it binds to the active site of NE and blocks its proteolytic activity. It is designed for oral or aerosol delivery.
|
| ln Vivo |
In HNE-treated mice, lonodelestat (POL6014) dramatically and effectively decreased the inflammatory processes of ALI[1]. The amount of macrophages, epithelial cells, neutrophils, and lymphocytes recovered in BAL was significantly and dose-dependently decreased by lonsodelestat (POL6014, 0.1, 0.5, 2 and 10 mg/kg, delivered intranasally). The reduction of neutrophils by 65% (p<0.001), epithelial cells by 68% (p<0.001), macrophages by 33% (p<0.001), and lymphocytes by 77% (p<0.001) was the greatest inhibition attained at 2 mg/kg[1].
In HNE-treated mice, lonodelestat dramatically and effectively decreased the inflammatory processes of acute lung injury (ALI). The number of macrophages, epithelial cells, neutrophils, and lymphocytes recovered in bronchoalveolar lavage (BAL) was significantly and dose-dependently decreased. The reduction of neutrophils by 65% (p<0.001), epithelial cells by 68% (p<0.001), macrophages by 33% (p<0.001), and lymphocytes by 77% (p<0.001) was the greatest inhibition attained at 2 mg/kg. |
| Enzyme Assay |
Non-cell enzyme assays for lonodelestat TFA are performed using purified human neutrophil elastase (hNE). A fluorogenic peptide substrate (e.g., MeOSuc-Ala-Ala-Pro-Val-AMC) is commonly used. The assay buffer consists of 50 mM Tris-HCl (pH 7.5), 150 mM NaCl, 0.01% Tween-20. Serial dilutions of lonodelestat TFA (0.001-1000 nM) are prepared in 96-well plates. The fluorogenic substrate is added to each well at a final concentration of 50-200 uM. The reaction is initiated by adding 0.5-5 nM purified hNE. The increase in fluorescence (excitation 380 nm, emission 460 nm) is monitored kinetically over 5-30 minutes at 37degC in a fluorescence plate reader. Initial velocities (v0) are calculated. IC50 values are determined from dose-response curves (log[inhibitor] vs. % activity). For Ki determination, the assay is performed at varying substrate concentrations (10-500 uM) with and without fixed concentrations of lonodelestat. Kinetic data are fit to the Michaelis-Menten equation. For selectivity assays, lonodelestat is tested against other serine proteases (e.g., cathepsin G, proteinase 3, trypsin, chymotrypsin) using their specific fluorogenic substrates.
|
| Cell Assay |
Cellular assays for lonodelestat TFA are performed using human neutrophils isolated from peripheral blood. Neutrophils are isolated by density gradient centrifugation (Ficoll-Paque) followed by dextran sedimentation. Isolated neutrophils (1×10⁶ cells/mL) are pre-incubated with lonodelestat TFA at concentrations of 0.1-1000 nM for 15-30 minutes at 37degC. Neutrophil elastase release can be stimulated by adding phorbol 12-myristate 13-acetate (PMA, 1 uM) or the chemotactic peptide fMLF (1 uM) with cytochalasin B (5 ug/mL). After 30-60 minutes, the cells are pelleted by centrifugation, and the supernatant is collected. NE activity in the supernatant is measured by adding the fluorogenic substrate (as above) and monitoring fluorescence. Alternatively, a chromogenic substrate (e.g., MeOSuc-Ala-Ala-Pro-Val-pNA) is used, and the release of p-nitroaniline (pNA) is measured at 405 nm. The percent inhibition is calculated compared to the vehicle control. For cytotoxicity assays, neutrophils are treated with lonodelestat (0.1-10 uM) for 4-24 hours, and viability is assessed using trypan blue exclusion or a luminescence-based assay (CellTiter-Glo).
|
| Animal Protocol |
In vivo animal studies have been conducted in mouse models of lung inflammation. For the acute lung injury (ALI) model, 6-8 week old C57BL/6 mice are treated with human neutrophil elastase (hNE) intratracheally (0.5-2 U/kg) to induce ALI. Lonodelestat TFA is delivered intranasally at doses of 0.1, 0.5, 2, and 10 mg/kg, either prophylactically (e.g., 30 minutes before hNE) or therapeutically (1 hour after hNE). At 6-24 hours after hNE challenge, mice are euthanized, and bronchoalveolar lavage (BAL) fluid is collected by cannulating the trachea and instilling and aspirating PBS. BAL fluid is centrifuged, and the total cell count is determined. Differential cell counts (neutrophils, macrophages, lymphocytes, epithelial cells) are performed on cytospin slides stained with Wright-Giemsa. The protein concentration in the BAL fluid is measured as an indicator of vascular leakage. Lung tissue is fixed in 10% formalin, embedded in paraffin, sectioned, and stained with H&E for histopathological analysis. For a sub-chronic tobacco smoke (TS) model, mice are exposed to cigarette smoke (4 cigarettes/day, 5 days/week) for 2-4 weeks, with lonodelestat (0.5-2 mg/kg) administered intranasally daily. At the end of the study, BAL fluid and lung tissue are analyzed as above. Cytokine levels (KC, MIP-2, IL-6, TNF-alpha) in BAL fluid are measured by ELISA.
|
| ADME/Pharmacokinetics |
Lonodelestat TFA (POL6014, MW ~1592.8 Da) is a peptide inhibitor formulated for aerosol delivery via nebulization (e.g., using the Pari eFlow system). Preclinical pharmacokinetic studies have been performed in animal models and humans. After inhalation, the compound is delivered directly to the lung, where it achieves high local concentrations with minimal systemic exposure. In healthy volunteers and subjects with cystic fibrosis, inhaled lonodelestat was well-tolerated, and systemic absorption was low (as expected for a peptide). The elimination half-life in the lungs is likely prolonged due to the local retention and slow clearance from the epithelial lining fluid. The TFA salt form improves solubility and stability. Detailed PK parameters (e.g., Cmax, Tmax, AUC) in humans are not publicly available.
|
| Toxicity/Toxicokinetics |
In preclinical toxicology studies, lonodelestat was well-tolerated in animals and healthy human volunteers. In single-dose escalation studies with inhaled POL6014 in healthy volunteers and subjects with cystic fibrosis, the compound was generally safe and well-tolerated. No serious adverse events were reported. In animal models, intranasal administration of lonodelestat at doses up to 10 mg/kg did not cause overt toxicity (no weight loss, no clinical signs). The TFA counterion may be irritating at high doses. No genotoxicity or carcinogenicity studies have been published. As an inhibitor of host defense mechanisms (neutrophil elastase), there is a theoretical risk of increased susceptibility to infection; however, this was not observed in clinical studies.
|
| References |
|
| Additional Infomation |
Lonodelestat (POL6014) is a protein epitope mimetic (PEM) that was in clinical development for the treatment of cystic fibrosis (CF). Neutrophil elastase (NE) activity in the airways is a key driver of inflammation and tissue damage in CF and other chronic lung diseases. Lonodelestat acts as a potent and selective NE inhibitor, reducing lung inflammation. Phase I clinical trials in healthy volunteers and CF subjects demonstrated safety and tolerability. However, further clinical development status may be discontinued or inactive. The compound has not received regulatory approval (FDA/EMA) for marketing. The TFA salt is used in research; formulations for clinical use were likely the free base. The product is for research use only.
|
| Molecular Formula |
C73H112F3N15O21
|
|---|---|
| Molecular Weight |
1592.75
|
| Related CAS # |
Lonodelestat;906547-89-5
|
| Appearance |
White to off-white solid powder
|
| 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 Note: Please store this product in a sealed and protected environment (e.g. under nitrogen), avoid exposure to moisture and light. |
| Shipping Condition |
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
|
| Solubility (In Vitro) |
H2O :~50 mg/mL (~31.39 mM)
|
|---|---|
| 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 | 0.6278 mL | 3.1392 mL | 6.2784 mL | |
| 5 mM | 0.1256 mL | 0.6278 mL | 1.2557 mL | |
| 10 mM | 0.0628 mL | 0.3139 mL | 0.6278 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.
Products are for research use only; We do not sell to patients
Copyright 2020 InvivoChem LLC | All Rights Reserved
