CA-074 methyl ester (CA-074 Me)

Catalog No.S7420 Batch:S742001

Technical Data

Formula

C₁₉H₃₁N₃O₆

Molecular Weight

397.47

CAS No.

147859-80-1

Solubility (25°C)*

In vitro

DMSO

79 mg/mL (198.75 mM)

Ethanol

79 mg/mL (198.75 mM)

Water

2.5 mg/mL (6.28 mM)

In vivo (Add solvents to the product individually and in order)

Clear solution	5%DMSO 1 40%PEG300 2 5%Tween80 3 50%ddH2O	5.0mg/ml	Taking the 1 mL working solution as an example, add 50 μL of 100 mg/ml clarified DMSO stock solution to 400 μL of PEG300, mix evenly to clarify it; add 50 μL of Tween80 to the above system, mix evenly to clarify; then continue to add 500 μL of ddH2O to adjust the volume to 1 mL. The mixed solution should be used immediately for optimal results.
Clear solution	5% DMSO 1 95% Corn oil	2.5mg/ml	Taking the 1 mL working solution as an example, add 50 μL of 50 mg/ml clear DMSO stock solution to 950 μL of corn oil and mix evenly. The mixed solution should be used immediately for optimal results.

* <1 mg/ml means slightly soluble or insoluble.
* Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations.
* Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

Preparing Stock Solutions

Biological Activity

Description

CA-074 methyl ester (CA-074 Me, Cathepsin B Inhibitor IV) is a membrane-permeable derivative of CA-074 and acts as an irreversible cathepsin B inhibitor.

Targets

Cathepsin B ^[1]

In vitro

CA-074Me is able to inhibit cathepsin L within living HL-60 cells and bloodstream forms of Trypanosoma brucei brucei, but does not inhibit cathepsin L in the absence of thiols. CA-074Me is converted into CA-074 by intracellular esterasesand is, therefore, able to inhibit cathepsin B in GSH-depleted HL-60 cell. It is suggested that CA-074 rather than CA-074Me should be used to selectively inhibit cathespsin B within living cells^[1]. Myotube size and number and induced levels of fusion-related creatine phosphokinase activity and myosin heavy-chain protein are reduced from 30 to 50% in CA074Me-treated myoblasts. These reductions are also dose related. Micromolar concentrations of CA074Me inhibit catB activity in differentiating myoblasts^[2].

In vivo

Administration of CA074Me to the London APP mice results in significant improvement in memory deficit, reduces amyloid plaque load in brain, reduces levels of Aβ40 and Aβ42 in brain, and reduces C-terminal β-secretase fragment (CTFβ) derived from APP by β-secretase when compared with untreated control animals. It has no effect on any of these parameters in mice expressing the Swe mutant β-secretase site of APP (in Swedish/London APP mice). In vivo icv administration of CA074Me to normal guinea pigs results in substantial reduction of brain Aβ levels and inhibits β-secretase activity^[3].

Protocol (from reference)

Cell Assay:^{^[1]}	Cell lines HL-60 cells Concentrations 100 μM Incubation Time 2 h Method HL-60 cells are pre-treated for 24 h with 200 μM BSO followed by 60 min with 1 mM DEM. Thereafter, the cells(1×10_⁶/ml) are incubated with 100 μM CA-074 or CA-074Me, in the presence of 1% DMSO, 200 μM BSO and 1 mM DEM at 37°C. Control cultures are treated with 1% DMSO alone or with 100 μM Z-FA-DMK in the presence of 200 μM BSO and 1 mM DEM. Untreated HL-60 cells are incubated with 100 μM CA-074 or CA-074Me, in the presence of 1% DMSO at 37°C. Untreated control cultures are incubated with 1% DMSO alone or with 100 μM Z-FA-DMK. After 2 h incubation, cells are washed three times with PBS/1% glucose and lysed in 100 mM citrate, pH 5.0, 2% Chaps (10_⁶ cells/100 μl). Subsequently, the lysate is centrifuged and the clarified supernatant used to assay for proteolytic activity.
Animal Study:^{^[3]}	Animal Models London APP mice Dosages 1 mg/ml Administration An ALZET osmotic minipump was implanted subcutaneously in the midscapular area of the back of each animal. A catheter was connected to the osmotic minipump and inserted into the lateral ventricle of the brain (icv).

Cell Assay:^{^[1]}

Cell lines
HL-60 cells
Concentrations
100 μM
Incubation Time
2 h
Method
HL-60 cells are pre-treated for 24 h with 200 μM BSO followed by 60 min with 1 mM DEM. Thereafter, the cells(1×10_⁶/ml) are incubated with 100 μM CA-074 or CA-074Me, in the presence of 1% DMSO, 200 μM BSO and 1 mM DEM at 37°C. Control cultures are treated with 1% DMSO alone or with 100 μM Z-FA-DMK in the presence of 200 μM BSO and 1 mM DEM. Untreated HL-60 cells are incubated with 100 μM CA-074 or CA-074Me, in the presence of 1% DMSO at 37°C. Untreated control cultures are incubated with 1% DMSO alone or with 100 μM Z-FA-DMK. After 2 h incubation, cells are washed three times with PBS/1% glucose and lysed in 100 mM citrate, pH 5.0, 2% Chaps (10_⁶ cells/100 μl). Subsequently, the lysate is centrifuged and the clarified supernatant used to assay for proteolytic activity.

Animal Study:^{^[3]}

Animal Models
London APP mice
Dosages
1 mg/ml
Administration
An ALZET osmotic minipump was implanted subcutaneously in the midscapular area of the back of each animal. A catheter was connected to the osmotic minipump and inserted into the lateral ventricle of the brain (icv).

References

Selleck's CA-074 methyl ester (CA-074 Me) has been cited by 35 publications

Melatonin alleviates chronic stress-induced hippocampal microglia pyroptosis and subsequent depression-like behaviors by inhibiting Cathepsin B/NLRP3 signaling pathway in rats [ Transl Psychiatry, 2024, 14(1):166]	PubMed: 38538614
Gasdermin D regulates soluble fms-like tyrosine kinase 1 release in macrophages [ Reprod Biol, 2024, 24(1):100857]	PubMed: 38295720
Monocyte-derived macrophages orchestrate multiple cell-type interactions to repair necrotic liver lesions in disease models [ J Clin Invest, 2023, 133(15)e166954]	PubMed: 37338984
The impact of cycling hypoxia on the phenotype of HPV-positive cervical cancer cells [ J Med Virol, 2023, 95(12):e29280]	PubMed: 38054507
Resistance of hypervirulent Klebsiella pneumoniae to cathepsin B-mediated pyroptosis in murine macrophages [ Front Immunol, 2023, 14:1207121]	PubMed: 37457695
Parallel use of human stem cell lung and heart models provide insights for SARS-CoV-2 treatment [ Stem Cell Reports, 2023, 18(6):1308-1324]	PubMed: 37315523
Acid Ceramidase Inhibitor LCL-805 Antagonizes Akt Signaling and Promotes Iron-Dependent Cell Death in Acute Myeloid Leukemia [ Cancers (Basel), 2023, 15(24)5866]	PubMed: 38136410
Effects of HMGB1/RAGE/cathespin B inhibitors on alleviating hippocampal injury by regulating microglial pyroptosis and caspase activation in neonatal hypoxic-ischemic brain damage [ J Neurochem, 2023, 167(3):410-426]	PubMed: 37753942
Trimethylamine N-Oxide (TMAO) Inducing Endothelial Injury: UPLC-MS/MS-Based Quantification and the Activation of Cathepsin B-Mediated NLRP3 Inflammasome [ Molecules, 2023, 28(9)3817]	PubMed: 37175227
Pyroptosis leads to loss of centrosomal integrity in macrophages [ bioRxiv, 2023, 10.1101/2023.11.22.568260]	PubMed: none

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SHIPPING AND STORAGE
Selleck products are transported at room temperature. If you receive the product at room temperature, please rest assured, the Selleck Quality Inspection Department has conducted experiments to verify that the normal temperature placement of one month will not affect the biological activity of powder products. After collecting, please store the product according to the requirements described in the datasheet. Most Selleck products are stable under the recommended conditions.

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