Aderamastat (FP-025) is an orally bioactive matrix metalloproteinase 12 (MMP-12) inhibitor. FP‐025 is a novel, potent, and highly selective non‐hydroxamate inhibitor of MMP‐12 with 90‐fold selectivity over the closest family member (MMP‐2) and two to three orders of magnitude over the seven other MMP family members. Aderamastat may be utilized in the research of asthma, chronic obstructive pulmonary disease (COPD) and pulmonary fibrosis. The key role of MMP-12 in disease is supported by single-nucleotide polymorphisms/genetic evidence related to inflammatory-fibrotic diseases, including asthma and COPD. MMP-12 is a key immune-fibrotic modulator secreted by macrophages and a key regulator of macrophage, neutrophil, and lung epithelial cell biology and is becoming increasingly recognized as a key marker of inflammatory exacerbations and fibrosis. Importantly, a Phase 2 allergen challenge asthma proof-of-concept study has been successfully completed.

Physicochemical Properties

CAS #	877176-23-3
PubChem CID	67177374
Appearance	White to light yellow solid powder
Hydrogen Bond Donor Count	2
Hydrogen Bond Acceptor Count	5
Rotatable Bond Count	6
Heavy Atom Count	28
Complexity	567
Defined Atom Stereocenter Count	0
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	Matrix metalloproteinase 12 (MMP-12)
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.
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.
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 37C 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].
References	[1]. Safe administration of MMP-12 inhibitor. Patent. WO2022261624 A1.

Solubility Data

Solubility (In Vitro)

May dissolve in DMSO (in most cases), if not, try other solvents such as H2O, Ethanol, or DMF with a minute amount of products to avoid loss of samples

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 (e.g. IP/IV/IM/SC) Injection Formulation 1: DMSO : Tween 80： Saline = 10 : 5 : 85 (i.e. 100 μL DMSO stock solution → 50 μL Tween 80 → 850 μL Saline) *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). 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)] *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. Injection Formulation 5: 2-Hydroxypropyl-β-cyclodextrin : Saline = 50 : 50 (i.e. 500 μL 2-Hydroxypropyl-β-cyclodextrin → 500 μL Saline) Injection Formulation 6: DMSO : PEG300 : castor oil : Saline = 5 : 10 : 20 : 65 (i.e. 50 μL DMSO → 100 μLPEG300 → 200 μL castor oil → 650 μL Saline) Injection Formulation 7: Ethanol : Cremophor : Saline = 10： 10 : 80 (i.e. 100 μL Ethanol → 100 μL Cremophor → 800 μL Saline) Injection Formulation 8: Dissolve in Cremophor/Ethanol (50 : 50), then diluted by Saline Injection Formulation 9: EtOH : Corn oil = 10 : 90 (i.e. 100 μL EtOH → 900 μL Corn oil) Injection Formulation 10: EtOH : PEG300：Tween 80 : Saline = 10 : 40 : 5 : 45 (i.e. 100 μL EtOH → 400 μLPEG300 → 50 μL Tween 80 → 450 μL 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). Oral Formulation 3: Dissolved in PEG400 Oral Formulation 4: Suspend in 0.2% Carboxymethyl cellulose Oral Formulation 5: Dissolve in 0.25% Tween 80 and 0.5% Carboxymethyl cellulose Oral Formulation 6: Mixing with food powders Note: Please be aware that the above formulations are for reference only. InvivoChem strongly recommends customers to read literature methods/protocols carefully before determining which formulation you should use for in vivo studies, as different compounds have different solubility properties and have to be formulated differently.  (Please use freshly prepared in vivo formulations for optimal results.)