CN114404569A

CN114404569A - Medicine containing lectin family molecule CLEC18A for treating liver injury and liver failure and application thereof

Info

Publication number: CN114404569A
Application number: CN202210052391.6A
Authority: CN
Inventors: 张燕
Original assignee: Jiangsu Kangkede Biotechnology Co ltd
Current assignee: Jiangsu Kangkede Biotechnology Co ltd
Priority date: 2022-01-18
Filing date: 2022-01-18
Publication date: 2022-04-29

Abstract

The invention belongs to the field of biomedicine, and relates to the pharmaceutical application of the lectin family molecule CLEC18A, in particular to a liver injury treatment drug comprising the lectin family molecule CLEC18A and its application. Its active ingredient contains CLEC18A protein. The medicine comprising the CLEC18A protein of the present invention has good preventive and therapeutic effects on liver injury, liver failure and acute liver injury. In vivo injection of mice with preparations containing mouse recombinant CLEC18A demonstrated that recombinant CLEC18A has obvious preventive and therapeutic effects in both the LPS/GalN-induced liver injury (liver failure) model and the CCL4-induced acute liver injury model.

Description

Medicine containing lectin family molecule CLEC18A for treating liver injury and liver failure and application thereof

Technical Field

The invention belongs to the field of biological medicines, relates to the pharmaceutical application of a lectin family molecule CLEC18A, and particularly relates to a liver injury treatment drug containing a lectin family molecule CLEC18A and application thereof.

Background

Acute Liver Injury (ALI) refers to a disease in which sudden abnormalities in liver function occur due to various causes in a short period of time. It is mostly caused by infection, toxic chemicals, viral infection, drugs, ischemia-reperfusion, etc., and in pharmacological studies, models of acute liver injury/liver failure are often established with carbon tetrachloride (CCl4), lipopolysaccharide/D-galactosamine (LPS/GalN). Pathologically, acute liver injury is manifested by severe acute damage to parenchymal liver cells, and glutamic-pyruvic transaminase (ALT) and glutamic-oxaloacetic transaminase (AST)) in liver cells are released into blood in large quantities, resulting in elevation of ALT and AST in blood serum. ALI can induce a series of complications and also exacerbate the progression of other diseases, and is one of the common diseases in emergency medical research. Most ALI patients can recover from drug treatment, but still a subset of patients will progress to high mortality acute liver failure.

The treatment of ALI is currently a combination therapy involving multiple approaches. Clinically, antibiotics, liver-protecting drugs, glucocorticoids, specific antibodies and the like are mainly used. The emergence of drug-resistant bacteria poses a challenge to antibiotic treatment, and the risk of drug-induced liver injury is not completely avoidable: glucocorticoids increase the incidence of complications such as infection; specific antibodies (anti-endotoxin antibodies, TNF α antagonists, IL-1 β antagonists, HMGB1 antagonists, etc.) regulate the further development of endotoxic ALI by inhibiting a certain link of the inflammatory response, but this therapy is expensive and has not been widely validated clinically.

The novel C-type lectin family molecule 18A (C-type lectin family 18 member A, CLEC18A) belongs to the C-type lectin receptor C-type lectin receptors (CTLRs) family members. The family is an evolutionary conserved pattern recognition receptor, and natural ligands of the receptor are polysaccharide, protein, lipid, inorganic molecules and the like, and play an important role in the process of recognizing fungi, tubercle bacillus, viruses, parasites and the like. The molecular structure of human CLEC18A was first confirmed in 2015. Under physiological conditions, human CLEC18A was expressed predominantly in myeloid cells and hepatocytes, and mouse CLEC18A was expressed predominantly in brain, kidney, and heart tissues. Current research on CLEC18A is limited to increased expression in human HBV and HCV infection and to mediating viral infection immune responses as Toll-Like Receptor 3 (TLR 3) co-receptors. As the only exocrine molecule in this family, the expression and role of CLEC18A in other diseases was not reported.

Disclosure of Invention

The technical problem to be solved by the invention is to provide a medicine for treating liver injury and liver failure containing a lectin family molecule CLEC18A, which can utilize the lectin molecule CLEC18A to generate better treatment effect on the liver injury and the liver failure, and an application thereof.

The invention relates to a medicine for treating liver injury and liver failure, which comprises a lectin family molecule CLEC18A, and the active ingredient of the medicine comprises CLEC18A protein.

The medicine consists of an effective component CLEC18A protein and pharmaceutically acceptable auxiliary materials.

The above medicine can be in the form of tablet, capsule, granule, powder, patch, suspension, syrup, oral liquid, injection, suppository or their arbitrary form combination.

The dosage of CLEC18A protein in the above medicine is 1 μ g/kg body weight.

The technical scheme of the invention also comprises the application of the CLEC18A protein in medicaments for treating liver injury or liver failure or acute liver injury.

The medicine containing CLEC18A protein has good prevention and treatment effects on liver injury, liver failure and acute liver injury. The preparation containing the mouse recombinant CLEC18A is injected into mice in vivo, and the recombinant CLEC18A is proved to have obvious prevention and treatment effects in an LPS/GalN induced liver injury (liver failure) model and a CCL4 induced acute liver injury model.

Drawings

FIG. 1 is a graph showing the results of comparative experiments of LPS/GalN-induced liver injury in Wild-type mice (Wild-type, WT) and CLEC 18A-deficient mice (Knockout, KO) in example 1, wherein A is a graph showing the results of ALT and AST concentration measurements in the serum of the two groups of mice, and B is a graph showing the results of HE staining of liver tissues of the two groups of mice;

FIG. 2 is a graph of the results of the model that the recombinant mouse CLEC18A protein in example 1 of the present invention can inhibit the liver damage of C57bl/6 mice induced by LPS/GalN, specifically, the serum ALT and AST concentrations of C57bl/6 mice were examined;

FIG. 3 is a graph showing the results of comparative tests of CCL4 induced liver injury in wild type mice (WT) and mice lacking CLEC18A (KO) during example 2 of the present invention, and showing the results of measurement of serum ALT and AST concentrations in these two groups of mice;

FIG. 4 is a diagram showing the serum ALT of the recombinant mouse CLEC18A protein capable of inhibiting the liver injury of C57bl/6 mice induced by CCL4 in example 2 of the invention, and the AST concentration detection result.

Detailed Description

Example 1:

1. after mice liver was specifically knocked out of CLEC18A, it was verified that LPS/GalN induced liver damage aggravation.

A liver specific knockout CLEC18A mouse (KO) is obtained by using a Cre-Loxp technology, and is constructed, bred and identified by a southern model animal center.

KO mice 8-10 weeks old were selected as a test group, while littermates wild type mice (WT) were selected as a control group, 8 mice per group. LPS (10mg/kg)/GalN (750. mu.g/kg) was intraperitoneally injected in a total volume of 200. mu.l. After 6 hours, serum of the mouse is taken, ALT and AST concentrations are detected, liver tissue sections are cut, and hematoxylin and acid staining solution eosin (HE) staining is utilized to detect the liver injury condition.

Liver damage was significantly exacerbated in liver knockout CLEC18A mice (KO) compared to littermate control Wild Type (WT) mice. The verification results are shown in fig. 1.

2. The recombinant mouse CLEC18A protein is used for treating C57bl/6 mice with LPS/GalN induced liver injury.

The test process comprises the following steps: 8-10 week-old C57bl/6 mice were randomly selected into 2 groups, labeled as treatment group and control group, respectively, and 6 mice were each group, and all mice were intraperitoneally injected with LPS (10mg/kg)/GalN (750. mu.g/kg) in a total volume of 200. mu.l. After 1 hour, 100. mu.l PBS was intraperitoneally injected into the control mice, and 100. mu.l CLEC18A (1. mu.g/kg) was intraperitoneally injected into the treatment mice. After 6 hours, all mice were serum tested for ALT and AST concentrations.

As a result: as shown in FIG. 2, the recombinant mouse CLEC18A protein significantly reduced liver damage in C57bl/6 mice induced by LPS/GalN.

Example 2:

1. after mouse liver specific knockout of CLEC18A, CCl4 induced liver injury aggravation was verified.

8-10 week-old liver-specific knockout CLEC18A mice (KO) were selected as test groups, and littermate wild-type mice (WT) were selected as controls, 6 mice per group. CCL 41 mg/kg was intraperitoneally injected in a total volume of 200. mu.l. After 24 hours, serum of the mice was taken and ALT and AST concentrations were measured.

Liver damage was significantly exacerbated in liver knockout CLEC18A (KO) mice compared to littermate control Wild Type (WT) mice. The verification results are shown in fig. 3.

2. The recombinant mouse CLEC18A protein is used for treating C57bl/6 mice with CCl4 induced liver injury.

The test process comprises the following steps: 8-10 week-old C57bl/6 mice were randomly divided into 2 groups, i.e., a treatment group and a control group, wherein each group had 8 mice, and all mice were injected with CCL 41 mg/kg in the abdominal cavity in a total injection volume of 200. mu.l. After 4 hours, the mice in the control group were intraperitoneally injected with 100. mu.l of PBS, and the mice in the treatment group were intraperitoneally injected with 100. mu.l of CLEC18A (2. mu.g/kg) in a total volume of 200. mu.l. After 24 hours, all mice were serum tested for ALT and AST concentrations.

As a result: as shown in FIG. 4, the recombinant mouse CLEC18A protein significantly reduced the C57bl/6 mouse liver damage induced by CCl 4.

Claims

1. A drug for treating liver injury and liver failure comprising a lectin family molecule CLEC18A, wherein the active ingredient comprises CLEC18A protein.

2 . The medicine for treating liver injury and liver failure according to claim 1 , wherein the medicine is composed of the active ingredient CLEC18A protein and pharmaceutically acceptable excipients. 3 .

3. The medicine for treating liver injury and liver failure comprising lectin family molecule CLEC18A according to claim 1, characterized in that: the dosage form of the medicine is tablet, capsule, granule, powder, patch, suspension, One or more combinations of syrups, oral liquids, injections, and suppositories.

4 . The drug for treating liver injury and liver failure according to claim 1 , wherein the dosage of CLEC18A protein is more than 1 μg per kilogram of body weight. 5 .

5. Application of CLEC18A protein in the treatment of liver injury or liver failure or acute liver injury.