| Size | Price | Stock | Qty |
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| 5mg |
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| 10mg |
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| Other Sizes |
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| Targets |
Matrix metalloproteinase 12 (MMP-12)
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| ln Vitro |
Aderamastat is a highly selective oral MMP-12 inhibitor targeting inflammatory and fibrotic diseases. MMP-12 plays a role in asthma pathophysiology and is associated with disease severity.
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| ln Vivo |
Aderamastat showed sustained anti-inflammatory effects and attenuated allergen-induced histopathology in a house dust mite (HDM) mouse model. FP‐025 in a dose‐dependent manner significantly attenuated AHR, BAL inflammatory cell numbers and lung pathohistology in a mouse model of persistent HDM‐allergic asthma. FP‐025 markedly reduced MMP‐12 expression in airway epithelial cells and levels of MMP‐12 in lung parenchyma and airways, indicating that the enhanced expression in allergic asthma depends on an autocrine process. FP‐025 did not negatively affect anti‐viral responses, contributing to its safety profile. These findings warrant interventions with FP‐025 in allergic asthma, also as FP‐025 showed safety, tolerability, and good pharmacokinetics characteristic in a randomized, placebo‐controlled, single, and multiple ascending dose study in healthy subjects.
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| Enzyme Assay |
MMP-12 quantification[2]
Levels of MMP-12 in BAL and lung homogenates were measured by ELISA (MMP-12, LSBio, LS-F2819) according to manufacturers’ specifications. |
| Cell Assay |
Sampling and primary cell isolation[2]
Bronchoalveolar lavage (BAL) was collected as previously described. Briefly, mice were cannulated through the trachea and the lavage was performed by instilling and retrieving 1 mL of sterile NaCl 0.9% into the right lung. BAL recovered was centrifuged, the supernatant was stored, whereas the pellet was immediately suspended in sterile PBS. Cell counts were then determined for each BAL sample and the cellular subset phenotypes were determined by FACS analysis. To remove the intravascular pool of cells, lungs were perfused with 5 ml of sterile NaCl 0.9% via the pulmonary circulation. The left lungs (not lavaged) were removed and minced using iridectomy scissors, thereafter enzymatically digested and grinded, as previously described2,3. Briefly, to prepare a lung homogenate, lungs were incubated for 1 hour at 37C with Collagenase type III (200 U/mL Worthington Biochemical, Lakewood, NJ) and DNAse I (100 U/mL; grade II from bovine pancreas, Sigma) in RPMI 1640. Pulmonary cell suspensions were obtained by grinding the tissue through 70-mm nylon sieves, and red blood cells were lysed with ammonium chloride buffer. Cell counts were then performed before staining for flow cytometric analysis. The cell suspension was lysed with Greenberger buffer (75 mmol/L NaCl, 7.5 mmol/L Tris, 0.5 mmol/L MgCl2, 0.5 mmol/L CaCl2, 0.5% Triton X-100, 4 mg/mL 4-[2-aminoethyl] benzenesulfonyl fluoride, 50 mg/mL Na2-EDTA, 10 ng/mL Pepstatin A, and 10 ng/mL leupeptin, pH 7.4) for 30 minutes on ice, followed by centrifugation at 680g for 10 minutes. Supernatants were stored at -80°C until further use. Cell counts were then performed before staining for flow cytometry analysis. Supernatants were stored at -80°C until further use. |
| Animal Protocol |
Male 8-week-old C57BL/6J mice purchased from Charles River Laboratories (France) were housed under specific pathogen-free conditions and maintained on a 12-hour light – dark cycle with access to food and water ad libitum. The sensitization schedule to elicit a severe and persistent allergic inflammation of the airways was based on a well-characterized model. In short, groups of C57BL/6J mice were randomly assigned to be treated for 5 days a week for five consecutive weeks with either the aeroallergen house dust mite (HDM) (Greer Laboratories, Lenoir, NC, USA) or NaCl through intranasal (i.n.) administration. Lyophilized HDM extract was resuspended in sterile PBS at a concentration of 5 mg (protein)/ml, and 25 µg (5 µl) protein per dose per mouse was administered i.n. in a final volume of 20 µl (in NaCl) in mice lightly anesthetized with isofluorane.
The H3N2 influenza A virus HK X31 stocks and viral titers were obtained by infecting Madin–Darby canine kidney (MDCK) as described before. The viral stock suspension was diluted in NaCl and animals (lightly anesthetized with isofluorane) received i.n. 20 TCID50 in 50 µl or not.
The MMP-12 inhibitor FP-025 (Foresee Pharmaceuticals, Co., Ltd. Taiwan), was dissolved in sterile NaCl 0.5% methyl cellulose containing 0.2% Tween-80 and administered orally, daily, for 7 consecutive days, at a dose of 10 mg/kg or 30 mg/kg or 100 mg/kg in a volume of 50 µl per mouse.
Prednisone (Sigma-Aldrich) was diluted in sterile NaCl (5 mg/kg) in 100 µl and administered daily via the intraperitoneal (i.p) route [2].
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| References |
[1]. Li, Yuhua, et al. Safe administration of MMP-12 inhibitor. Patent. WO2022261624 A1.
[2]. Efficacy of FP-025: A novel matrix metalloproteinase-12 (MMP-12) inhibitor in murine allergic asthma. Allergy. 2023 Feb;78(2):559-562. |
| Molecular Formula |
C21H18N2O4S
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|---|---|
| Molecular Weight |
394.44
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| CAS # |
877176-23-3
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| Appearance |
White to light yellow solid powder
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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.) |
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| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.5352 mL | 12.6762 mL | 25.3524 mL | |
| 5 mM | 0.5070 mL | 2.5352 mL | 5.0705 mL | |
| 10 mM | 0.2535 mL | 1.2676 mL | 2.5352 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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