S1PR4-targeting composition for preventing or treating non-alcoholic steatohepatitis
A steatohepatitis, non-alcoholic technology, applied in the direction of active ingredients of phosphorus compounds, drug combinations, active ingredients of heterocyclic compounds, etc.
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Embodiment 1
[0119] Example 1. Selection of sphingolipid candidate substances
[0120] In order to derive lead candidates for the development of therapeutic agents for non-alcoholic steatohepatitis, from more than 600 kinds of sphingolipid compounds possessed by the sphingolipid library, we provided the combination of myriocin and FTY720 (fingolimod) 15 sphingolipid compounds with similar structures.
[0121] The results of measuring the agonistic activity related to sphingosine1-phosphate receptors using the GPCR activity measurement method of Discovex Corporation confirms that 2-amino-2-indicated by the following chemical formula 2 (2-(1-decyl-1H-1,2,3-triazol-4-yl)ethyl)propane-1,3-diol (hereinafter, referred to as SLB736) is susceptible to sphingosine 1-phosphate Body 4 has a specific agonistic effect.
[0122] Chemical formula 2:
Embodiment 2
[0123] Example 2. NLRP3 Inflammatory Body Inhibition Ability
[0124] It is well known that if the macrophages (Raw cells) are treated with lipopolysaccharide (LPS), the inflammasome (inflammasome) will be activated, and the production of interleukin-1β (IL-1β) will increase (Mariathasanet al ., Nat Rev Immunol. 2007; 17186029). In order to confirm the inhibitory effect of the inflammasome (NLRP3 inflammasome) activity, which is a representative inflammation-mediated protein, when lipopolysaccharide is processed on macrophages, the production amount of interleukin-1β as a candidate substance is measured and evaluated.
[0125] First, the raw cells (cell line) of the mouse macrophage cell line were treated with lipopolysaccharide (100ng / ml, Sigma-aldrich). After 12 hours, the in the culture medium could be observed. The concentration of interleukin-1β increased significantly.
[0126] As a control group, when macrophages were treated with myriocin at the same concentration and treat...
Embodiment 3
[0128] Example 3. Confirmation of the tissue and intracellular expression inhibitory effects related to the sphingosine 1-phosphate receptor 4 of SLB736
[0129] In order to confirm whether the compound SLB736 of the present invention acts as a functional inhibitor related to sphingosine 1-phosphate receptor 4, after administration of the candidate substance to non-alcoholic steatohepatitis model mice and mouse macrophages, The changes in sphingosine 1-phosphate receptor 4 protein expression were confirmed.
[0130] It is well known that if a methionine choline-deficinet diet (MCDD) is administered to mice, non-alcoholic steatohepatitis will occur. The experimental mice (C57BL / 6N, Orient Bio, Inc., Korea) were divided into three groups, and each group was fed only with methionine choline deficiency diet, methionine choline deficiency diet, and FTY720, 1mg / kg methionine choline deficiency diet After six weeks with 1 mg / kg of SLB736, the expression changes of sphingosine 1-phosphate...
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