Application of DYRK1B inhibitor AZ191 in preparation of medicine for treating and preventing liver injury
By using DYRK1B inhibitor AZ191 to inhibit the mRNA level of liver injury-related factors, the adverse reactions and narrowing of treatment windows of drug-induced liver injury treatment methods in the prior art were solved, and the effect of significantly reducing liver injury and inflammatory cell infiltration was achieved.
Patent Information
- Application Number
- CN202510213334.5
- Authority / Receiving Office
- CN · China
- Patent Type
- Applications(China)
- Current Assignee / Owner
- Filing Date
- 2025-02-26
- Publication Date
- 2025-05-09
- Estimated Expiration
- Not applicable · inactive patent
AI Technical Summary
There are serious limitations in the treatment methods for drug-induced liver injury in the prior art, such as adverse reactions and narrow treatment windows, which are difficult to effectively solve the problem of liver injury.
DYRK1B inhibitor AZ191 was used as a drug component to reduce liver damage and inflammatory cell infiltration by inhibiting the increase in mRNA levels of ALT, AST, TNF-α, IL-6, IL-1β, TGF-β and α-SMA.
AZ191 significantly reduced the changes in ALT and AST levels caused by liver injury, inhibited the changes in mRNA levels of inflammatory factors and fibrosis indicators, and reduced the degree of fibrosis and inflammatory cell infiltration in liver tissue.
Smart Images
Figure CN119950510A_ABST
Abstract
Description
Technical Field
[0001] The present invention relates to the field of biomedical technology, and in particular to application of a DYRK1B inhibitor AZ191 in preparing a medicine for treating and preventing liver damage. Background Art
[0002] The liver is the largest internal organ of the human body, responsible for metabolism, immunity, digestion, detoxification and protein synthesis, and is characterized by its strong regenerative capacity in response to damage. Acute liver injury (ALI) is one of the main causes of liver disease, with high morbidity and mortality. 3.5% of deaths worldwide are caused by liver disease. Among them, chemical liver injury is the most common cause in clinical practice. It mainly includes environmental toxic liver injury, alcoholic liver injury and drug-induced liver injury.
[0003] Carbon tetrachloride (CCL4) is a typical hepatotoxic compound that is widely used to induce ALI models for mechanistic studies. For drug-induced liver injury, N-acetylcysteine is the only effective treatment; however, there are often serious limitations, such as adverse reactions and a narrow therapeutic window. Therefore, it is crucial to gain a deeper understanding of the pathogenesis of ALI and screen for new drugs with hepatoprotective activity. Summary of the invention
[0004] The purpose of the present invention is to solve the problem that the treatment methods of drug-induced liver injury in the prior art are limited and have serious limitations.
[0005] In order to achieve the above object, the present invention adopts the following technical solutions:
[0006] Use of DYRK1B inhibitors in the preparation of drugs for treating and / or preventing liver damage.
[0007] Preferably, the DYRK1B inhibitor is AZ191, and the molecular formula of AZ191 is C 24 H 27 N7O, CAS NO.1594092-37-1, structural formula as shown in formula I
[0008]
[0009] Preferably, the DYRK1B inhibitor is used to inhibit the increase in the level of at least one mRNA among ALT, AST, TNF-α, IL-6, IL-1β, TGF-β, and α-SMA.
[0010] Preferably, the liver damage includes at least one of liver fibrosis, hepatocyte necrosis, inflammatory cell infiltration, connective tissue hyperplasia, and liver lobule structural disorder.
[0011] The present application also provides a drug for treating or inhibiting liver damage, wherein the drug includes the DYRK1B inhibitor AZ191.
[0012] Preferably, the drug is an oral agent, an intramuscular injection, or an intravenous injection.
[0013] Preferably, the drug is a liver-targeted drug of GalNAc-coupled AZ191.
[0014] Beneficial effects:
[0015] The present invention proves through specific verification experiments that the DYRK1B inhibitor AZ191 can significantly reduce the level changes of ALT and AST caused by liver damage, and can inhibit the changes in the mRNA levels of inflammatory factors and fibrosis indicators, and reduce liver tissue fibrosis and the degree of liver inflammatory cell infiltration. Therefore, the present invention provides the use of the DYRK1B inhibitor AZ191 in the preparation of a drug for treating and / or preventing liver damage, and provides an anti-liver damage drug containing the DYRK1B inhibitor AZ191. BRIEF DESCRIPTION OF THE DRAWINGS
[0016] Figure 1 The effect of AZ191 on serum ALT (1A) and AST (1B) levels in CCL4-induced liver fibrosis model mice; AZ191 is the DYRK1B inhibitor-treated CCL4 liver fibrosis experimental group of mice, CCL4 is the DMSO-treated CCL4 liver fibrosis experimental control group of mice, and NC is the DMSO-treated corn oil-free liver fibrosis blank control group of mice;
[0017] Figure 2 The gross image of liver tissues in different groups (2A) and HE staining of liver tissue sections (2B);
[0018] Figure 3 F4 / 80 staining of liver tissue sections from different groups;
[0019] Figure 4 The mRNA expression levels of TNF-α, IL-6, and IL-1β, the inflammatory indicators of liver tissues in different groups;
[0020] Figure 5 The expression of TGF-β and α-SMA mRNA in liver tissues of different groups. DETAILED DESCRIPTION
[0021] The present invention is further described in detail below in conjunction with specific embodiments.
[0022] Use of DYRK1B inhibitors in the preparation of drugs for treating and / or preventing liver damage.
[0023] In one embodiment, the DYRK1B inhibitor is AZ191, and the molecular formula of AZ191 is C 24 H 27 N7O, CAS NO.1594092-37-1, structural formula as shown in formula I
[0024]
[0025] The DYRK1B inhibitor is used to inhibit the increase in the level of at least one mRNA among ALT, AST, TNF-α, IL-6, IL-1β, TGF-β, and α-SMA.
[0026] The liver damage includes at least one of liver fibrosis, liver cell necrosis, inflammatory cell infiltration, connective tissue hyperplasia, and liver lobule structural disorder.
[0027] Based on the above content, the present application provides a drug for treating or preventing liver damage, wherein the drug includes a DYRK1B inhibitor, and preferably, the drug includes AZ191.
[0028] The drug is a dosage drug, preferably, one of an oral dosage form, an intramuscular injection, and an intravenous injection.
[0029] In one embodiment, the drug is a liver-targeted drug of GalNAc-conjugated AZ191.
[0030] The drug can be used in combination therapy, i.e., in combination with one or more other agents, wherein the combination includes the use of DYRK1B inhibitor AZ191 together with other agents, or in sequence. For example, the other agent can be used before, during or after the use of AZ191. The mode of use can be the same or different, for example, AZ191 and other agents can both be administered in the form of injection, can both be administered orally, or can one be administered in the form of injection and the other in the form of oral administration.
[0031] The above contents are described below in conjunction with specific embodiments:
[0032] Model construction and grouping:
[0033] Eight-week-old C57BL / 6 mice were intraperitoneally injected with 0.5 mL / kg body weight of CCL4-corn oil solution (1:4) twice a week for 6 weeks to establish a liver fibrosis model.
[0034] From the date of modeling, the mice were divided into three groups at the 7th week, namely, AZ191-treated mice CCL4 liver fibrosis experimental group (AZ191 group), DMSO-treated mice CCL4 liver fibrosis experimental control group (CCL4 group), DMSO-treated mice corn oil no liver fibrosis blank control group (NC group).
[0035] The mice in the AZ191 group were intraperitoneally injected with AZ191 50 mg / kg for 1 week, once a day, while the mice in the CCL4 group were intraperitoneally injected with the same volume of DMSO solvent (DMSO is the solvent for dissolving AZ191).
[0036] 48 hours after the last injection of the mice in the AZ191 group and the CCL4 group, the mice were anesthetized and killed, and blood and liver samples were collected to examine blood liver function, pathology, inflammation, and liver fibrosis indicators, and to observe the changes in liver function, liver tissue inflammation, fibrosis level, and pathological changes in the mice
[0037] SPSS 23.0 statistical analysis software was used for all data. The t-test was used for comparison between groups, and P < 0.05 was considered to be significantly different.
[0038] Verification experiment:
[0039] Example 1: Effect of AZ191 on serum ALT and AST in CCL4-induced model mice
[0040] ALT and AST are mainly distributed in liver cells. When the liver cell membrane is damaged or the cells are necrotic, these enzymes enter the serum in increased amounts. Therefore, by measuring the activity of enzymes in the serum, it is possible to sensitively reflect whether the liver cells are damaged and the extent of the damage.
[0041] In order to understand the effect of AZ191 injection on the degree of liver damage in CCL4-induced liver fibrosis mice, the blood of mice was collected in the present application to detect serum ALT and AST levels.
[0042] The results showed that intraperitoneal injection of the DYRK1B inhibitor AZ191 could significantly reduce the changes in ALT and AST levels caused by CCL4-induced liver injury ( Figure 1 ).
[0043] Example 2: Effect of AZ191 on liver tissue structure
[0044] In order to clarify the effect of AZ191 on the liver structure of CCL4 mice, the livers of each group were Figure 2 As shown in a. At the same time, the liver tissue was stained with HE.
[0045] See also Figure 2b, In CCL4-induced liver fibrosis model mice, hepatocyte necrosis, massive inflammatory cell infiltration and connective tissue hyperplasia were observed in the portal area, and the structure of some liver lobules was disordered.
[0046] In the AZ191-treated group, the inflammatory response, hepatocyte necrosis and structural disorder of liver tissue were alleviated.
[0047] Therefore, AZ191 can significantly alleviate CCL4-induced liver injury.
[0048] Example 3: Effect of AZ191 on inflammatory response in liver tissue
[0049] The expression levels of TNF-α, IL-1β, and IL-6 mRNA in liver tissues of each group were detected by RT-PCR and F4 / 80 staining of liver sections were performed to evaluate the changes in liver tissue inflammatory response at the molecular level.
[0050] See also Figure 3 and Figure 4 In CCL4-induced liver fibrosis model mice, the mRNA levels of the above-mentioned inflammatory indicators in the liver were significantly increased, and the F4 / 80 green fluorescence increased.
[0051] In comparison, the levels of inflammatory indicators in liver tissue of mice in the AZ191 treatment group were significantly reduced, and the F4 / 80 green fluorescence was significantly reduced.
[0052] The above results suggest that AZ191 can significantly alleviate CCL4-induced liver inflammatory response.
[0053] Example 4: Effect of AZ191 on CCL4-induced liver fibrosis
[0054] The expression levels of α-SMA and TGF-β mRNA in liver tissues of each group were detected by RT-PCR to evaluate their changes in liver fibrosis at the molecular level.
[0055] See also Figure 5 In the CCL4-induced liver fibrosis model mice, the mRNA expression levels of α-SMA and TGF-β, the liver fibrosis indicators, were significantly increased. However, the above fibrosis indicators in the liver tissue of mice in the AZ191 treatment group were reduced, indicating that AZ191 can significantly slow down CCL4-induced liver fibrosis.
[0056] The above results show that injection of AZ191 can reduce the degree of liver tissue damage induced by CCL4.
[0057] The present invention provides the use of DYRK1B inhibitor AZ191 in the preparation of drugs for treating and preventing liver damage. The present application prepares a liver fibrosis mouse model by intraperitoneal injection of CCL4, and sets up an AZ191 preventive administration group to observe indicators such as liver gross morphology, pathophysiological changes, serum ALT and AST expression, TNF-α, IL-6, IL-1β expression, and TGF-β, α-SMA expression. The operation is simple, fast, and reproducible.
Claims
1. Use of DYRK1B inhibitors in the preparation of drugs for treating and / or preventing liver damage.
2. Use of the DYRK1B inhibitor according to claim 1 in the preparation of a drug for treating and / or preventing liver damage, characterized in that: The DYRK1B inhibitor is AZ191, and the molecular formula of AZ191 is C 24 H 27 N7O, CAS NO.1594092-37-1, structural formula as shown in Formula I 3. Use of the DYRK1B inhibitor according to claim 2 in the preparation of a drug for treating and / or preventing liver damage, characterized in that: The DYRK1B inhibitor is used to inhibit the increase in the level of at least one mRNA among ALT, AST, TNF-α, IL-6, IL-1β, TGF-β, and α-SMA.
4. Use of the DYRK1B inhibitor according to claim 1 in the preparation of a drug for treating and / or preventing liver damage, characterized in that: The liver damage includes at least one of liver fibrosis, liver cell necrosis, inflammatory cell infiltration, connective tissue hyperplasia, and liver lobule structural disorder.
5. A drug for treating or inhibiting liver damage, characterized in that: The drugs include the DYRK1B inhibitor AZ191.
6. A drug for treating or inhibiting liver damage according to claim 5, characterized in that: The medicine is one of an oral preparation, an intramuscular injection, and an intravenous injection.
7. A drug for treating or inhibiting liver damage according to claim 5, characterized in that: The drug is a liver-targeted drug of GalNAc-coupled AZ191.