Leupeptin Ac-LL hydrochloride
Based on 1 Customer Validation
Leupeptin Ac-LL hydrochloride is a broad-spectrum protease inhibitor. By inhibiting the activation of the PTEN/PI3K/Akt/NF-κB/ERK1/2/p38 signaling pathway, Leupeptin Ac-LL hydrochloride significantly reduces LPS-induced NO and ROS production, mitochondrial membrane potential hyperpolarization, phagocytic activity, pro-inflammatory cytokine release, and M1 polarization in mouse peritoneal macrophages, and reverses autophagic flux impairment. It also decreases Concanavalin A (HY-P2149)-induced proliferation index of mouse splenic lymphocytes and the Th1/IL-10 and Th2/IL-10 cytokine ratios, thereby modulating innate and adaptive immune responses. Leupeptin Ac-LL hydrochloride inhibits blood coagulation and tumorigenesis in mouse skin. Leupeptin Ac-LL hydrochloride can be used in research on chronic inflammatory diseases and skin tumorigenesis.
For research use only. We do not sell to patients.
- Purity : 95.0%
- CAS No.: 24125-16-4
- Formula: C20H39ClN6O4
- Molecular Weight:463.01
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Storage:Powder -20°C, 3 years ; In solvent -80°C, 6 months , -20°C, 1 month
To place orders, for customer services and technical support, please contact: MedChemExpress USA
Tel: 609-228-6898 E-mail: [email protected] [email protected]
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Biological Activity
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Chemical Information
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Solvent & Solubility
- Purity & Documentation
- References
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Help & FAQs
All Cathepsin Isoforms
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Biological Activity
Description
|
SARS-CoV-2 Mpro 127.2 μM (IC50) |
Kallikrein 70-75 μg/mL (IC50) |
Plasmin 6-100 μg/mL (IC50) |
Thrombokinase 15-18 μg/mL (IC50) |
papain 0.03-0.51 μg/mL (IC50) |
trypsin 0.1-136 μg/mL (IC50) |
PI3K |
ERK1 |
NF-κB |
iNOS |
Akt |
HTRA1 |
COX-2 |
Threonine Protease |
Cysteine Protease |
ERK2 |
LC3B |
Beclin-1 |
p62 |
Arg 1 |
Msr 1 |
Mrc−1 |
p-PTEN |
In Vitro
Leupeptin (50-200 µM; 3.5 h) Ac-LL hydrochloride significantly reduces LPS-induced NO and ROS production, mitochondrial membrane potential hyperpolarization, phagocytic activity, pro-inflammatory cytokine release, and M1 polarization in mouse peritoneal macrophages, and reverses autophagic flux impairment, with its mechanism associated with the inhibition of PTEN/PI3K/Akt/NF-κB/ERK1/2/p38 signaling pathway activation[1].
Leupeptin (50-200 µM; 48 h) Ac-LL hydrochloride reduces the Con-A (Concanavalin A) (HY-P2149)-induced proliferation index of mouse splenic lymphocytes and significantly decreases the Th1/IL-10 and Th2/IL-10 cytokine ratios, thereby modulating adaptive immune responses[1].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
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Cell Line:Peritoneal macrophages
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Concentration:50 μM; 100 μM; 200 μM
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Incubation Time:3.5 h
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Result:Reduced LPS-induced intracellular ROS accumulation.
Reversed LPS-induced mitochondrial membrane potential hyperpolarization.
Reduced LPS-enhanced macrophage phagocytic activity.
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Cell Line:Peritoneal macrophages
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Concentration:50 μM; 100 μM; 200 μM
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Incubation Time:3.5 h
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Result:Reduced the LPS-induced increases in TNF-α, IL-1β, IFN-γ, and IL-6.
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Cell Line:Peritoneal macrophages
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Concentration:50 μM; 100 μM; 200 μM
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Incubation Time:3.5 h
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Result:Reduced the LPS-induced elevation of iNOS, Cox-2, IL-17, IL-15, and IL-5 mRNA levels.
Reduced LPS-induced mRNA expression of MMP-1, MMP-3, and MMP-9.
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Cell Line:Peritoneal macrophages
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Concentration:50 μM; 100 μM; 200 μM
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Incubation Time:3.5 h
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Result:Reduced the LPS-induced elevations of p-PTEN, p-PI3K, p-Akt, p-NF-κB, p-ERK1/2, and p-p38.
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Cell Line:Splenic lymphocytes
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Concentration:50 μM; 100 μM; 200 μM
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Incubation Time:48 h
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Result:Reduced the Con-A-induced lymphocyte proliferation index.
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Cell Line:Splenic lymphocytes
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Concentration:50 μM; 100 μM; 200 μM
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Incubation Time:48 h
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Result:Reduced Con-A-induced levels of TNF-α and IFN-γ, as well as the TNF-α/IL-10 and IFN-γ/IL-10 ratios.
In Vivo
Leupeptin (40 mg/kg; i.p.; single dose; sacrificed 3 h post-injection) Ac-LL hydrochloride detects a 45% increase in macroautophagic flux in calorie-starved mouse liver and significant suppression of flux in refed mouse liver, with a 3-hour flux value of 53.94 ng/mg total protein in starved mice[3].
Leupeptin (40 mg/kg; i.p.; single dose; sacrificed 60 min or 180 min post-injection) Ac-LL hydrochloride detects a significant reduction in p62 flux (but not statistically significant reduction in LC3b-II flux) in beclin 1+/- mouse liver compared to wild-type littermates, with an 180-minute p62 flux value of 18 ng/mg total protein in heterozygous mice[3].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
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Animal Model:C57BL/6NCrl (male, 6-8 weeks old, 20-25 g)[3]
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Dosage:9 mg/kg; 18 mg/kg; 20 mg/kg; 36 mg/kg; 40 mg/kg
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Administration:i.p.; single dose; sacrificed 1 h, 4 h, 60 min, 90 min, 120 min, 180 min, or 240 min post-injection; administered 60 min prior to cycloheximide injection, sacrificed 60 min post-cycloheximide injection
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Result:Produced a dose-dependent increase in LC3b-II levels in mouse liver total extracts and lysosome-enriched (LE) fractions, with the 36 mg/kg dose inducing the strongest signal.
Led to time-dependent LC3b-II accumulation across multiple organs, with liver reaching 69.1 ng/mg total protein in LE fractions at 60 min, a flux of 42.1 ng/mg total protein, and spleen reaching 37.1 ng/mg total protein in LE fractions at 60 min, a flux of 11.0 ng/mg total protein.
Increased LC3a-II and p62 levels in liver LE fractions, but did not increase levels of GABARAP or GATE-16.
Induced a 6.88-fold increase in cross-sectional area of late autophagosome/lysosomal compartments in hepatocytes (from 0.36% to 2.50% of total cytoplasmic area; p = 0.013), with this increase correlating with LC3b-II levels in LE fractions (R2 = 0.7741).
Increased LC3b-II and LC3a-II content in dense, lysosome-enriched fractions of lung homogenates.
Stabilized LC3b-II and LC3a-II levels in liver LE fractions (but not cytosolic fractions) following cycloheximide-induced translational inhibition.
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Animal Model:C57BL/6NCrl (male, 6-8 weeks old, 20-25 g)[3]
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Dosage:40 mg/kg
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Administration:i.p.; single dose; sacrificed 1 h or 3 h post-injection
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Result:Induced an LC3b-II flux of 37.21 ng/mg total protein at 3 h post-injection in ad libitum-fed mice.
Induced an LC3b-II flux of 53.94 ng/mg total protein at 3 h post-injection in calorie-starved mice, representing a 45% increase compared to ad libitum-fed mice (p = 0.04).
Induced an LC3b-II flux of only 3.00 ng/mg total protein at 3 h post-injection in refed mice, significantly suppressed compared to ad libitum-fed mice (p < 0.001).
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Animal Model:Beclin 1+/+; beclin 1+/- (male, 6-8 weeks old)[3]
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Dosage:40 mg/kg
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Administration:i.p.; single dose; sacrificed 60 min or 180 min post-injection
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Result:Showed an LC3b-II flux of ~25 ng/mg total protein in beclin 1+/- mice at 60 min post-injection, compared to ~38 ng/mg total protein in beclin 1+/+ mice (no statistical significance reported).
Showed an LC3b-II flux of ~56 ng/mg total protein in beclin 1+/- mice at 180 min post-injection, compared to ~73 ng/mg total protein in beclin 1+/+ mice (no statistical significance reported).
Showed a p62 flux of ~4 ng/mg total protein in beclin 1+/- mice at 60 min post-injection, compared to ~2 ng/mg total protein in beclin 1+/+ mice.
Showed a p62 flux of ~18 ng/mg total protein in beclin 1+/- mice at 180 min post-injection, compared to ~35 ng/mg total protein in beclin 1+/+ mice (p < 0.05).
Chemical Information
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CAS No. 24125-16-4
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Appearance Solid
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Molecular Weight 463.01
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Formula C20H39ClN6O4
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Color White to off-white
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SMILES
CC(C)C[C@H](NC(C)=O)C(N[C@@H](CC(C)C)C(NC(C=O)CCCNC(N)=N)=O)=O.Cl
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Structure Classification
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Initial Source
Actinomycetes
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Shipping
Room temperature in continental US; may vary elsewhere.
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Storage
Powder -20°C 3 years In solvent -80°C 6 months -20°C 1 month
Solvent & Solubility
In Vitro:
H2O : 125 mg/mL (269.97 mM; Need ultrasonic)
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.
Concentration (start) × Volume (start) = Concentration (final) × Volume (final)
Purity & Documentation
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Data Sheet (284 KB)
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SDS (394 KB)
- English - EN (394 KB)
- Français - FR (394 KB)
- Deutsch - DE (394 KB)
- Norwegian - NO (394 KB)
- Español - ES (394 KB)
- Swedish - SV (394 KB)
- Italian - IT (394 KB)
- Korean - KR (394 KB)
- Portuguese - PT (394 KB)
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Handling Instructions (2659 KB)
References
[1]. Sharma A, et al. Leupeptin maintains redox homeostasis via targeting ROS-autophagy-inflammatory axis in LPS-stimulated macrophages and cytokines dichotomy in Con-A challenged lymphocyte. Peptides. 2023 Oct;168:171066. [Content Brief]
[2]. Hozumi M, et al. Inhibition of tumorigenesis in mouse skin by leupeptin, a protease inhibitor from Actinomycetes. Cancer Res. 1972 Aug;32(8):1725-8. [Content Brief]
[3]. Haspel J, et al. Characterization of macroautophagic flux in vivo using a leupeptin-based assay. Autophagy. 2011 Jun;7(6):629-42. [Content Brief]
[4]. Billinger E, et al. Inhibition properties of free and conjugated leupeptin analogues. FEBS open bio. 2020 Dec;10(12):2605-2615. [Content Brief]
Complete Stock Solution Preparation Table
Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
| Optional Solvent | Concentration Solvent Mass | 1 mg | 5 mg | 10 mg | 25 mg |
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| H2O | 1 mM | 2.1598 mL | 10.7989 mL | 21.5978 mL | 53.9945 mL |
| 5 mM | 0.4320 mL | 2.1598 mL | 4.3196 mL | 10.7989 mL | |
| 10 mM | 0.2160 mL | 1.0799 mL | 2.1598 mL | 5.3995 mL | |
| 15 mM | 0.1440 mL | 0.7199 mL | 1.4399 mL | 3.5996 mL | |
| 20 mM | 0.1080 mL | 0.5399 mL | 1.0799 mL | 2.6997 mL | |
| 25 mM | 0.0864 mL | 0.4320 mL | 0.8639 mL | 2.1598 mL | |
| 30 mM | 0.0720 mL | 0.3600 mL | 0.7199 mL | 1.7998 mL | |
| 40 mM | 0.0540 mL | 0.2700 mL | 0.5399 mL | 1.3499 mL | |
| 50 mM | 0.0432 mL | 0.2160 mL | 0.4320 mL | 1.0799 mL | |
| 60 mM | 0.0360 mL | 0.1800 mL | 0.3600 mL | 0.8999 mL | |
| 80 mM | 0.0270 mL | 0.1350 mL | 0.2700 mL | 0.6749 mL | |
| 100 mM | 0.0216 mL | 0.1080 mL | 0.2160 mL | 0.5399 mL |
* Note: If you choose water as the stock solution, please dilute it to the working solution, then filter and sterilize it with a 0.22 μm filter before use.
Leupeptin Ac-LL hydrochloride Related Classifications
HY-18234C Related Classifications
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Keywords
- Leupeptin Ac-LL
- 24125-16-4
- Ser/Thr Protease
- Cathepsin
- Autophagy
- SARS-CoV
- Atg8/LC3
- NO Synthase
- COX
- p62
- Arginase
- PTEN
- NF-κB
- PI3K
- Akt
- p38 MAPK
- ERK
- Reactive Oxygen Species (ROS)
- kallikrein
- cathepsins B
- L
- and H
- LPS-induced mitochondrial membrane potential hyperpolarization
- mouse skin
- papain
- chymotrypsin
- mouse hepatocytes
- trypsin
- beclin 1+/- mouse liver
- plasmin
- Inhibitor
- inhibitor
- inhibit