Deubiquitinating enzymes (DUBs) function to remove covalently attached ubiquitin from proteins, thereby controlling substrate activity and/or abundance. The DUB ubiquitin carboxyl-terminal hydrolase L1 (UCH-L1) is a component of the ubiquitin proteasome system (UPS), it is abundantly expressed in neuronal brain cells and has been connected to Parkinson’s disease (PD)[3]. LDN-57444 is a specific inhibitor of UCH-L1 with IC50 value of 0.88 μM[4]. In vitro, LDH-57444 treatment significantly inhibited proteasome activity in a concentration-dependent manner at 25 μM and above, and treatment with 50 μM LDH-5744 for 24h led to 70% inhibition of the proteasome activity in human neuroblastoma SK-N-SH cells. LDN-57444 greatly reduced the cell viability and induced apoptosis of SK-N-SH cells at concentration ranging in 25 – 100 μM[5]. In vivo, LDN-57444 had an inhibitory effect on UCHL1 in the hippocampus at systemic doses of 0.5 mg/kg. Administration LDN-57444 at dose of 1 mg/kg for 4h significantly inhibited levels of free monomeric ubiquitin in mice. However, high dose LDN-57444 (2.5 mg/kg) increased UCHL1 expression, possibly as a compensatory mechanism. In addition, both 1 mg/kg and 2.5 mg/kg doses of LDN57444 depleted the hippocampus of postsynaptic density protein PSD95[4].
LDN-57444 细胞实验
Cell Line
Concentration
Treated Time
Description
References
Chondrocytes
8 µM
1 day
LDN-57444 inhibited UCHL1, reversed the increase in LC3B expression under hypoxic conditions, and increased the proportion of apoptotic cells.
Int J Mol Med. 2023 Oct;52(4):99.
Rat HSC
>10 µM
24 hours
Inhibition of UCHL1 significantly reduced the proliferation of hepatic stellate cells in response to PDGFBB stimulation
J Hepatol. 2015 Dec;63(6):1421-8.
Human HSC
>10 µM
24 hours
Inhibition of UCHL1 significantly reduced the proliferation of human hepatic stellate cells
J Hepatol. 2015 Dec;63(6):1421-8.
Murine bone marrow mesenchymal stem cells
10 µM
24 hours
LDN57444 pretreatment significantly enhanced the immunosuppressive effects of murine bone marrow mesenchymal stem cells on T cell proliferation and upregulated iNOS expression.
Cell Death Dis. 2018 May 1;9(5):459.
Human bone marrow mesenchymal stem cells
20 µM
24 hours
LDN57444 pretreatment enhanced the immunosuppressive effects of human bone marrow mesenchymal stem cells on T cell proliferation and upregulated IDO expression.
Cell Death Dis. 2018 May 1;9(5):459.
Hippocampal neurons
10 µM
24 hours
To investigate the effects of UCH-L1 inhibition on synaptic structure and monomeric ubiquitin levels. Results showed that LDN significantly reduced monomeric ubiquitin levels and caused dramatic alterations in synaptic protein distribution and spine morphology.
J Neurosci. 2009 Jun 17;29(24):7857-68.
NP69 and NP69-LMP1 cells
3 µM
24 hours
Inhibition of UCH-L1 DUB activity, reducing migration of LMP1-positive cells
Int J Mol Sci. 2019 Jul 31;20(15):3733.
SH-SY5Y cells
10 µM
3 days
LDN57444 pretreatment significantly inhibited RA-induced neuronal differentiation in SH-SY5Y cells, characterized by decreased neurite outgrowth and reduced expression of neural differentiation markers.
J Exp Clin Cancer Res. 2018 Oct 25;37(1):258.
293 cells
3 µM
48 hours
Inhibition of UCH-L1 DUB activity, reducing HIF-1α levels in exosomes
Int J Mol Sci. 2019 Jul 31;20(15):3733.
HSC-3 cells
3 µM
48 hours
Inhibition of UCH-L1 DUB activity, reducing motility of HSC-3 cells
Int J Mol Sci. 2019 Jul 31;20(15):3733.
Cochlear supporting cells
5 µM
5 days
LDN-57444 significantly increased Brn3c mRNA levels and promoted the transdifferentiation of supporting cells into hair cells.
Cells. 2024 Apr 24;13(9):737.
SK-N-BE (2) cells
10 µM
7 days
LDN57444 pretreatment significantly inhibited RA-induced neuronal differentiation in SK-N-BE (2) cells, characterized by decreased neurite outgrowth and reduced expression of neural differentiation markers.
J Exp Clin Cancer Res. 2018 Oct 25;37(1):258.
NEPC cells (TD-NEPC)
10 µM
72 hours
LDN-57444 significantly inhibited the colony formation ability of TD-NEPC cells
Cell Rep Med. 2024 Feb 20;5(2):101381.
LDN-57444 动物实验
Species
Animal Model
Administration
Dosage
Frequency
Description
References
Mice
CCl4-induced liver fibrosis model
Intraperitoneal injection
0.4 mg/kg
Twice weekly for 4 weeks
LDN-57444 significantly improved CCl4-induced liver fibrosis, reducing collagen deposition and the number of hepatic stellate cells
J Hepatol. 2015 Dec;63(6):1421-8.
Mice
Cardiac hypertrophy model
Intraperitoneal injection
40 µg/kg
Once daily for 2 weeks
LDN-57444 significantly reversed cardiac hypertrophy and remodeling, indicating that UCHL1 is a potential therapeutic target for hypertrophic diseases.
Sci Adv. 2020 Apr 17;6(16):eaax4826
Mice
Ang II-induced atrial fibrillation model
Intraperitoneal injection
40 μg/kg
Once daily for 3 weeks
LDN-57444 significantly reduced Ang II-induced elevation of blood pressure, the inducibility and duration of AF, left atrial dilation, fibrosis, inflammation, and oxidative stress. LDN treatment also inhibited the activation of multiple signaling pathways (the AKT, ERK1/2, HIF-1α, and TGF-β/smad2/3 pathways) and the expression of CX43 protein in atrial tissues.
Hypertens Res. 2020 Mar;43(3):168-177
Mice
NEPC, SCLC, and neuroblastoma xenograft models
Intraperitoneal injection
5 mg/kg
Once daily until the end of the experiment
LDN-57444 significantly delayed tumor growth in NEPC, SCLC, and neuroblastoma xenograft models and reduced the expression of neuroendocrine markers
搜索
