| Size | Price | Stock | Qty |
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| 1mg |
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| 5mg |
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| 10mg |
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| 25mg |
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| 50mg |
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| 100mg | |||
| Other Sizes |
Purity: ≥98%
| Targets |
ATG4B (Autophagin-1), a cysteine protease that processes ATG8 family proteins (such as LC3) during autophagosome formation.
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| ln Vitro |
ATG4B, also known as autophagin-1, is a cysteine protease that cleaves proteins in the ATG8 family. ATG4B is essential to the autophagy pathway and the production of autophagosomes. Using IC50 values of 80 and 73 nM in TR-FRET and cell-based LRA assays, respectively, FMK 9a shows significant inhibitory effects on ATG4B. According to LC-MS/MS investigations, FMK 9a creates an irreversible covalent link with Cys74, a highly reactive thiol group found in the catalytic region of ATG4B, which inactivates ATG4B's proteolytic activity [1].
FMK-9a inhibits ATG4B with IC50 values of 80 µM in a TR-FRET biochemical assay and 73 µM in a cell-based LC3 recruitment assay (LRA). It forms an irreversible covalent bond with Cys74, a highly reactive thiol in the catalytic site of ATG4B, thereby inactivating its proteolytic activity. |
| ln Vivo |
FMK 9a has high clearance from human and mouse liver microsomes, 13.9 and 70 mL/kg per minute, respectively, and intermediate solubility (LYSA: 41 μg/mL) [1].
In vivo, FMK-9a has high clearance from human and mouse liver microsomes, with values of 13.9 and 70 mL/kg per minute, respectively. It also demonstrates intermediate solubility (LYSA: 41 µg/mL). These properties inform its pharmacokinetic profile and potential utility in animal models for studying autophagy inhibition. |
| Enzyme Assay |
ATG4B inhibition is assessed using a time-resolved fluorescence resonance energy transfer (TR-FRET) assay. Recombinant ATG4B enzyme is incubated with a fluorogenic peptide substrate in the presence of varying concentrations of FMK-9a. The cleavage of the substrate by ATG4B results in a fluorescence signal, and the inhibition of this activity is measured to calculate the IC50 value.
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| Cell Assay |
FMK-9a's cellular activity is evaluated using a cell-based LC3 recruitment assay (LRA). Cells are treated with the compound, and the recruitment of LC3 to autophagosomes, a process dependent on ATG4B activity, is monitored. The inhibition of LC3 recruitment is used to determine the cellular potency of the compound, yielding an IC50 of 73 µM.
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| Animal Protocol |
In vivo studies for FMK-9a are typically conducted in rodent models to assess its pharmacokinetic properties and efficacy in modulating autophagy. The compound is administered via intravenous or oral routes, and plasma and tissue samples are collected to measure compound concentration and pharmacodynamic markers of autophagy inhibition. Microsomal stability assays are used to predict in vivo clearance.
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| ADME/Pharmacokinetics |
FMK-9a has a molecular weight of 392.42 g/mol and a molecular formula of C23H21FN2O3. It has high clearance from human and mouse liver microsomes (13.9 and 70 mL/kg/min, respectively) and intermediate solubility (41 µg/mL). The compound has a LogP of 3.83 and is supplied as a white to off-white solid powder.
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| Toxicity/Toxicokinetics |
Specific toxicological data for FMK-9a are not extensively published. As a covalent inhibitor targeting a cysteine protease, it should be handled with standard laboratory safety precautions. The compound is for research use only and not for human therapeutic applications. Appropriate personal protective equipment should be used during handling.
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| References | |
| Additional Infomation |
FMK-9a is a research tool compound for studying the autophagy pathway. It is not approved for clinical use. The compound is a covalent, irreversible inhibitor of ATG4B, making it a valuable probe for understanding the role of this protease in autophagy and related diseases. It is also known as FMK 9a and has the IUPAC name (S)-N-(1-((3-fluoro-2-oxopropyl)amino)-1-oxo-3-phenylpropan-2-yl)-1-naphthamide.
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| Molecular Formula |
C23H21FN2O3
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|---|---|
| Molecular Weight |
392.4228
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| Exact Mass |
392.153
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| CAS # |
1955550-51-2
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| PubChem CID |
127020966
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| Appearance |
White to off-white solid powder
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| Density |
1.2±0.1 g/cm3
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| Boiling Point |
724.6±60.0 °C at 760 mmHg
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| Flash Point |
392.0±32.9 °C
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| Vapour Pressure |
0.0±2.4 mmHg at 25°C
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| Index of Refraction |
1.607
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| LogP |
3.83
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| Hydrogen Bond Donor Count |
2
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| Hydrogen Bond Acceptor Count |
4
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| Rotatable Bond Count |
8
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| Heavy Atom Count |
29
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| Complexity |
577
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| Defined Atom Stereocenter Count |
1
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| SMILES |
C1=CC=C(C=C1)C[C@@H](C(=O)NCC(=O)CF)NC(=O)C2=CC=CC3=CC=CC=C32
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| InChi Key |
VMVOHHPQIAPYHG-NRFANRHFSA-N
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| InChi Code |
InChI=1S/C23H21FN2O3/c24-14-18(27)15-25-23(29)21(13-16-7-2-1-3-8-16)26-22(28)20-12-6-10-17-9-4-5-11-19(17)20/h1-12,21H,13-15H2,(H,25,29)(H,26,28)/t21-/m0/s1
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| Chemical Name |
N-[(1S)-1-Benzyl-2-[(3-fluoro-2-oxo-propyl)amino]-2-oxo-ethyl]naphthalene-1-carboxamide
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| Synonyms |
FMK-9a FMK 9a FMK9a
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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 : ~210 mg/mL (~535.14 mM)
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| Solubility (In Vivo) |
Solubility in Formulation 1: ≥ 2.5 mg/mL (6.37 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 (6.37 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. View More
Solubility in Formulation 3: ≥ 2.5 mg/mL (6.37 mM) (saturation unknown) in 10% DMSO + 90% Corn Oil (add these co-solvents sequentially from left to right, and one by one), clear solution. |
| Preparing Stock Solutions | 1 mg | 5 mg | 10 mg | |
| 1 mM | 2.5483 mL | 12.7415 mL | 25.4829 mL | |
| 5 mM | 0.5097 mL | 2.5483 mL | 5.0966 mL | |
| 10 mM | 0.2548 mL | 1.2741 mL | 2.5483 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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