受注:045-509-1970 |
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Synonyms | N/A | Storage (From the date of receipt) |
3 years -20°C powder 1 years -80°C in solvent |
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化学式 | C17H11Cl3N2O3 |
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分子量 | 397.64 | CAS No. | 668467-91-2 | ||||||||
Solubility (25°C)* | 体外 | DMSO | 11 mg/mL (27.66 mM) | ||||||||
Water | Insoluble | ||||||||||
Ethanol | Insoluble | ||||||||||
体内 (毎回新しく調製した物を用意してください) |
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* <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. |
製品説明 | LDN-57444 is a reversible, competitive proteasome inhibitor for Uch-L1 with IC50 of 0.88 μM, 28-fold selectivity over isoform Uch-L3. |
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in vitro | Treatment with 50 μM LDN-57444 for 24 h leads to 70% inhibition of the proteasome activity. LDN-57444 causes a significant and concentration-dependent decrease in cell viability at concentrations above 25 μM and the cell viability reduced to 61.81% at 50 μM. LDN-57444 is able to cause cell death through the apoptosis pathway by decreasing the activity of ubiquitin proteasome system and increasing the levels of highly ubiquitinated proteins, both of which can activate unfolded protein response. The apoptosis induced by LDN-57444 may be triggered by the activation of endoplasmic reticulum stress (ERS). [4] |
in vivo | LDN-57444 causes dramatic alterations in synaptic protein distribution and spine morphology in vivo. Treatment with LDN also results in a rapid fall of Uch-L1 activity, but proteasome inhibition has no effect on cAMP levels over a period of several hours. [3] |
キナーゼアッセイ | HTS screen | |
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To start an assay, 0.5 μL of 5 mg/mL test compound (about 50 μM final reaction concentration) or DMSO control is aliquoted into each well. Both enzyme and substrate are prepared in UCH reaction buffer (50 mM Tris-HCl [pH 7.6], 0.5 mM EDTA, 5 mM DTT, and 0.5 mg/mL ovalbumin). 25 μL of 0.6 nM UCH-L1 is then added to each well except substrate control wells, followed by plate shaking for 45–60 s on an automatic shaker. The enzyme/compound mixture is incubated at room temperature for 30 min before 25 μL of 200 nM Ub-AMC is added to initiate the enzyme reaction. The reaction mixture (300 pM UCH-L1, 100 nM Ubiquitin-AMC with 2.5 μg test compound) is incubated at room temperature for 30 additional minutes prior to quenching the reaction by the addition of 10 μL 500 mM acetic acid per well. The fluorescence emission intensity is measured on a LJL Analyst using a coumarin filter set (ex = 365 nm, em = 450 nm) and is subtracted by the intrinsic compound fluorescence to reveal the enzyme activity. A DMSO control (0.5 μL of DMSO, 25 μL of UCH-L1, 25 μL of ubiquitin-AMC, 10 μL of acetic acid), enzyme control (25 μL of UCH-L1, 25 μL of buffer, 10 μL of acetic acid), substrate control (25 μL of buffer, 25 μL of ubiquitin-AMC, 10 μL of acetic acid), and inhibitor control (0.5 μL of ubiquitin aldehyde [100 nM stock], 25 μL of UCH-L1, 25 μL of ubiquitin-AMC, 10 μL of acetic acid) are also performed in each assay plate to ensure quality and reproducibility. Potential UCH-L1 inhibitors are selected if the compounds demonstrated greater than 60% inhibition compared to the controls. The UCH-L1 enzymatic reactions are manually repeated twice using the same protocol to confirm the results for the hit compounds from the primary robot-assisted screen. | ||
細胞アッセイ | 細胞株 | H1299 |
濃度 | ~5 μM | |
反応時間 | 24 h | |
実験の流れ | MTT assay | |
動物実験 | 動物モデル | wild type and a-syn transgenic mice |
投薬量 | 0.5 mg/kg | |
投与方法 | intraperitoneally |
Data from [Data independently produced by , , J Cell Mol Med, 2017, 1-16]
Stem Cell Properties of Gastric Cancer Stem-Like Cells under Stress Conditions Are Regulated via the c-Fos/UCH-L3/β-Catenin Axis [ Mol Cells, 2023, 46(8):476-485] | PubMed: 37460253 |
Stem Cell Properties of Gastric Cancer Stem-Like Cells under Stress Conditions Are Regulated via the c-Fos/UCH-L3/β-Catenin Axis [ Mol Cells, 2023, 46(8):476-485] | PubMed: 37460253 |
UCHL1 regulates inflammation via MAPK and NF-κB pathways in LPS-activated macrophages [ Cell Biol Int, 2021, 10.1002/cbin.11662] | PubMed: 34288216 |
Long-term inhibition of UCHL1 decreases hypertension and retinopathy in spontaneously hypertensive rats [ J Int Med Res, 2021, 49(6):3000605211020641] | PubMed: 34130526 |
The Deubiquitinating Enzyme UCHL1 Promotes Resistance to Pemetrexed in Non-Small Cell Lung Cancer by Upregulating Thymidylate Synthase [ Theranostics, 2020, 15;10(13):6048-6060] | PubMed: 32483437 |
Deubiquitination of CD36 by UCHL1 promotes foam cell formation [ Cell Death Dis, 2020, 11(8):636] | PubMed: 32801299 |
Blockage of UCHL1 Activity Attenuates Cardiac Remodeling in Spontaneously Hypertensive Rats [ Hypertens Res, 2020, 10.1038/s41440-020-0486-1] | PubMed: 32541849 |
UCH-L1 inhibitor LDN-57444 hampers mouse oocyte maturation by regulating oxidative stress and mitochondrial function and reducing ERK1/2 expression [ Biosci Rep, 2020, 40(10)BSR20201308] | PubMed: 33030206 |
The deubiquitinating enzyme UCHL1 is a favorable prognostic marker in neuroblastoma as it promotes neuronal differentiation. [ J Exp Clin Cancer Res, 2018, 37(1):258] | PubMed: 30359286 |
The deubiquitinating enzyme UCHL1 negatively regulates the immunosuppressive capacity and survival of multipotent mesenchymal stromal cells [ Cell Death Dis, 2018, 9(5):459] | PubMed: 29686406 |
長期の保管のために-20°Cの下で製品を保ってください。
人間や獣医の診断であるか治療的な使用のためにでない。
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