| Size | Price | |
|---|---|---|
| 500mg | ||
| 1g | ||
| Other Sizes |
Leupeptin is a novel and potent protease inhibitor as well as a lysosome inhibitor with potential antioxidant and anti-inflammatory activity. It inhibits Trypsin, Papain, Plasmin, Thrombokinase, Kallikrein and Cathepsin B with IC50s ranging from 0.5 to 75 μg/ml, depending on the enzyme and the substrate. Leupeptin does not inhibit Chymotrypsin, Elastase, Renin, or Pepsin.
| Targets |
protease: Cathepsin B; Cathepsin L; Cathepsin H; Ser/Thr Protease; Mpro
|
|---|---|
| ln Vitro |
Leupeptin (0.06-200 µM; 72 h) dramatically reduces the SARS-CoV-2 viral RNA (vRNA) replication number in Vero cells [1]. Leupeptin has an EC50 value of 42.34 and inhibits SARS-CoV-2's RNA level in Vero cells. Leupeptin's IC50 value of 127.2 µM indicates that it exhibits some inhibitory effect against Mpro [1]. Leupeptin's half-life against the human coronavirus 229E strain is 0.4 µg/mL, or roughly 1 µM[1].
|
| ln Vivo |
Leupeptin (0, 9, 18, 36 mg/kg; i.p.; single dose) produced strong, dose-dependent increases in LC3b-II fraction and was well tolerated by animals in both total tissue extracts and lysosome- and autophagosome-enriched pellets [2].
|
| Enzyme Assay |
Mpro enzyme activity inhibition test. [2]
A total of 20 mM leupeptin hemisulfate in deionized water was diluted to 2 mM to 31.25 μM with 25 mM Tris buffer (pH 8.0). A 30-μl inhibitor solution with a series of concentrations in 25 mM Tris buffer (pH 8.0) was first mixed with 10 μl 100 μM peptide substrate (Dabcyl-TSAVLQ↓SGFRKMK-Edans; GenScript). Next, 10 μl of a final concentration of 200 nM Mpro was added to the plate. The relative fluorescence unit (RFU) value was measured with an excitation wavelength of 360 nm and an emission wavelength of 490 nm at 37°C for 1 h by using a SpectraMax Paradigm multimode detection platform (Molecular Devices, USA). Experiments were performed in triplicate. The enzyme activity reaction rate and inhibition rate were calculated by using MS Excel. The inhibition curve was plotted by using GraphPad Prism 8.0.[2] In vitro antiviral assays.[2] A total of 20 mM leupeptin hemisulfate in deionized water was diluted to 200 μM to 0.06 μM with DMEM containing 1% FBS. Vero cells cultured overnight in 96-well plates were infected by virus at a multiplicity of infection (MOI) of 0.01 for 2 h. The medium was removed, and fresh drug-containing medium was then added to the cells. After 48 h, the cells were lysed in lysis buffer. The viral RNA in 100 μl of the cell supernatant was quantified by reverse transcription-PCR (RT-PCR). Seventy-two hours later, the changes of cytopathic effect were also observed by microscopy. Experiments were performed in triplicate. The experimental results were processed using MS Excel and GraphPad Prism 8.0. |
| Cell Assay |
RT-PCR[1]
Cell Types: Vero Cell Tested Concentrations: 0.06-200 µM Incubation Duration: 72 hrs (hours) Experimental Results: Significant reduction in copy number of SARS-CoV-2 viral RNA (vRNA). |
| Animal Protocol |
Animal/Disease Models: C57BL/6NCrl male mice (6-8 weeks old, 20-25g) [1].
Doses: 0, 9, 18, 36 mg/kg Route of Administration: intraperitoneal (ip) injection; single Experimental Results:promotes the accumulation of LC3b-II in mouse liver. |
| References | |
| Additional Infomation |
Leupeptin is a tripeptide composed of N-acetylleucyl, leucyl and argininal residues joined in sequenceby peptide linkages. It is an inhibitor of the calpains, a family of calcium-activated proteases which promote cell death. It has a role as a serine protease inhibitor, a bacterial metabolite, a cathepsin B inhibitor, a calpain inhibitor and an EC 3.4.21.4 (trypsin) inhibitor. It is a tripeptide and an aldehyde. It is a conjugate base of a leupeptin(1+).
Leupeptin has been reported in Streptomyces lavendulae, Streptomyces exfoliatus, and other organisms with data available. |
| Molecular Formula |
C20H38N6O4
|
|---|---|
| Molecular Weight |
426.55352
|
| Exact Mass |
950.558
|
| Elemental Analysis |
C, 56.32; H, 8.98; N, 19.70; O, 15.00
|
| CAS # |
55123-66-5
|
| Related CAS # |
Leupeptin hemisulfate;103476-89-7;Leupeptin Ac-LL;24365-47-7; 39740-82-4 (HCl); 55123-66-5; 1082207-96-2 (hemisulfate hydrate); 103476-89-7 (hemisulfate)
|
| PubChem CID |
72429
|
| Sequence |
Ac-Leu-Leu-Arg-al
|
| SequenceShortening |
LLR
|
| Appearance |
Typically exists as solid at room temperature
|
| LogP |
5.185
|
| Hydrogen Bond Donor Count |
5
|
| Hydrogen Bond Acceptor Count |
5
|
| Rotatable Bond Count |
14
|
| Heavy Atom Count |
30
|
| Complexity |
602
|
| Defined Atom Stereocenter Count |
3
|
| SMILES |
CC(C[C@H](NC(C)=O)C(N[C@H](C(N[C@H](C=O)CCCNC(N)=N)=O)CC(C)C)=O)C
|
| InChi Key |
GDBQQVLCIARPGH-ULQDDVLXSA-N
|
| InChi Code |
InChI=1S/C20H38N6O4/c1-12(2)9-16(24-14(5)28)19(30)26-17(10-13(3)4)18(29)25-15(11-27)7-6-8-23-20(21)22/h11-13,15-17H,6-10H2,1-5H3,(H,24,28)(H,25,29)(H,26,30)(H4,21,22,23)/t15-,16-,17-/m0/s1
|
| Chemical Name |
(2S)-2-acetamido-N-[(2S)-1-[[(2S)-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-4-methyl-1-oxopentan-2-yl]-4-methylpentanamide
|
| Synonyms |
Leupeptin; 55123-66-5; Leupeptin hemisulfate; CHEBI:6426; Ac-Leu-Leu-Arg-H; Leupeptin Ac-LL; 24365-47-7; 103476-89-7;
|
| 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)
|
| 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
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 | 2.3444 mL | 11.7220 mL | 23.4439 mL | |
| 5 mM | 0.4689 mL | 2.3444 mL | 4.6888 mL | |
| 10 mM | 0.2344 mL | 1.1722 mL | 2.3444 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
