Application of RKH tripeptide in preparation of medicine for treating hepatic failure

Abstract

The invention relates to the technical field of polypeptide drugs, in particular to application of RKH tripeptide in preparation of drugs for treating hepatic failure. According to the present invention, the effect of the RKH tripeptide derived from Akkermansia muciniphila on the hepatic failure is evaluated, the result shows that the RKH tripeptide can improve the prognosis of the hepatic failure, and the present invention provides the new solution for the treatment of the hepatic failure, and is beneficial to the reduction of the pain of the hepatic failure patient.

Description

Technical Field

[0001] The present invention relates to the technical field of polypeptide drugs, and in particular to the application of RKH tripeptide in the preparation of drugs for treating liver failure. Background Art

[0002] Liver failure is the most serious form of liver damage caused by multiple factors such as viruses, alcohol, drugs, and ischemia-reperfusion. It is a serious disease with a high mortality rate. Currently, there is no breakthrough in comprehensive medical treatment, and the treatment plan needs to be further optimized.

[0003] The pathological basis of liver failure is extensive liver necrosis (massive, submassive or bridging necrosis) and hepatocyte regeneration (physiological or pathological), which runs through the clinical pathological process of various types of liver failure. The range and degree of liver cell necrosis are closely related to the prognosis of various types of liver failure. The main mode of liver cell death is necrosis, which coexists with cell death modes such as apoptosis, pyroptosis and autophagy. The basic pathological changes of liver failure include: (1) acute extensive hepatic necrosis (massive, submassive or bridging necrosis) with or without severe hepatocyte degeneration. The necrosis is mainly in the three bands of the hepatic alveoli and spreads to the periphery of the lobule. There are differences in the types of necrosis caused by drugs and viruses; (2) multi-acinar necrosis. The necrosis involving the entire lobule is called massive necrosis, which Scheuer called multi-acinar necrosis; (3) bridging necrosis. The necrosis fuses and extends along the three bands, connecting to the portal area and central vein, which is called bridging necrosis; (4) broad bridging necrosis, that is, extensive three-band necrosis is also called submassive necrosis. The range of submassive necrosis is generally less than 2 / 3 of the liver parenchyma.

[0004] The most common type of liver failure is acute-on-chronic liver failure. Acute-on-chronic liver failure (ACLF) is often associated with dysbacteriosis, and intestinal microecology and its secondary metabolism are currently hot topics of research. According to the applicant's previous research, intestinal flora disorders and a reduction in Akkermansia muciniphila (AKK) in ACLF patients may be associated with poor prognosis in ACLF, and in vivo and in vitro experiments have confirmed that the intestinal mechanical barrier is broken during liver failure. Understanding the changes in AKK can help medical professionals better understand the severity of ACLF, evaluate the effectiveness of treatment options, and provide a basis for individualized treatment. However, it is unknown whether supplementing AKK can improve the prognosis of ACLF. Recently, it has been found that the new tripeptide Arg-Lys-His (RKH), a metabolite of AKK, has anti-inflammatory effects. However, it is unclear whether this tripeptide can improve the prognosis of liver failure. Summary of the invention

[0005] In order to solve the above technical problems, the present invention provides the use of RKH tripeptide in preparing a drug for treating liver failure.

[0006] The present invention adopts the following technical solution:

[0007] Application of RKH tripeptide in preparing medicine for treating liver failure.

[0008] RKH was identified from the secretion of AKK, the most promising probiotic of the next generation, and studies have found that it has anti-inflammatory effects. We found that RKH can significantly improve the prognosis of liver failure and reduce cell death, and it has the application prospect of being prepared into a drug for the treatment of liver failure.

[0009] Furthermore, the liver failure includes acute liver failure.

[0010] Furthermore, the dosage form of the drug is an injectable dosage form or a non-injectable dosage form.

[0011] Furthermore, the injection dosage form includes intraperitoneal injection.

[0012] Furthermore, the non-injectable dosage form is selected from oral pharmaceutical preparations, intragastric lavage preparations, rectal absorption suppositories or enteric solvents.

[0013] Furthermore, the oral pharmaceutical preparation is selected from tablets, capsules or oral liquids.

[0014] Furthermore, the medicine also includes pharmaceutically acceptable excipients.

[0015] Furthermore, the auxiliary material is selected from buffer, solvent, solubilizer, preservative, stabilizer, surfactant, antioxidant and mixtures thereof.

[0016] Furthermore, in the drug, the content of RKH tripeptide is 1w% to 99.9w%.

[0017] The present invention has the following beneficial effects:

[0018] The present invention first discovered that the RKH tripeptide derived from Akkermansia muciniphila has a therapeutic effect on liver failure, and the RKH tripeptide can significantly improve liver histological lesions caused by liver failure and reduce cell death. The present invention provides a new solution for the treatment of liver failure and helps to reduce the pain of patients with liver failure.

[0019] The RKH tripeptide of the present invention is easy to synthesize and has low cost; it has a small molecular weight and a high absorption rate; it has low immunogenicity and can avoid adverse drug reactions occurring during the treatment of liver failure. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1 Flow chart of drug administration and modeling in mice.

[0021] Figure 2 ALT (A) and AST (B) levels in the serum of mice in each group.

[0022] Figure 3 The survival curves of mice in each group.

[0023] Figure 4 The following are the gross pictures of the liver of mice in each group.

[0024] Figure 5 H&E staining images of liver tissues of mice in each group. DETAILED DESCRIPTION

[0025] The present invention is described in detail below in conjunction with the accompanying drawings and specific examples, but should not be construed as limiting the present invention. Unless otherwise specified, the technical means used in the following examples are conventional means well known to those skilled in the art, and the materials, reagents, etc. used in the following examples, unless otherwise specified, can be obtained from commercial sources.

[0026] The present invention discloses the effect of RKH tripeptide as a new drug in treating liver failure. The amino acid sequence of the RKH tripeptide is Arg-Lys-His, and the RKH tripeptide is synthesized by Qiangyao Biotechnology Co., Ltd.

[0027] Example 1: Effect of RKH tripeptide on liver failure

[0028] 1. Experimental methods

[0029] like Figure 1 As shown in A, 6-8 week old C57BL / 6 mice were used as experimental subjects, and the experimental groups were as follows:

[0030] (1) Group a: mice were given PBS (200 μL per day) by gavage for 1 week.

[0031] (2) Group b: Mice were given PBS (200 μL per day) by gavage for 1 week, and then LPS (30 μg / kg) combined with D-galactosamine (D-GalN, 600 mg / kg) were intraperitoneally injected to establish an acute liver failure (ALF) model.

[0032] (3) Group c: Mice were treated with RKH (50 mg / kg per day) by gavage using 200 μL PBS as a diluent for 1 week.

[0033] (4) Group d: The mice were treated with RKH (50 mg / kg) by gavage using 200 μL PBS as a diluent for 1 week, and then LPS (30 μg / kg) combined with D-galactosamine (D-GalN, 600 mg / kg) was intraperitoneally injected to establish an acute liver failure (ALF) model.

[0034] (5) Group e: The mice were treated with RKH (50 mg / kg) by gavage using 200 μL PBS as a diluent for 2 weeks, and then LPS (30 μg / kg) combined with D-galactosamine (D-GalN, 600 mg / kg) was intraperitoneally injected to establish an acute liver failure (ALF) model.

[0035] 5.5 hours after modeling, the levels of serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) in mouse serum were determined using a kit built in Nanjing; the liver was taken and photographed, and the liver tissues of each group of mice were stained.

[0036] After modeling, the survival rate of mice in each group was counted and a survival line was drawn.

[0037] 2. Results

[0038] like Figure 2 As shown, compared with the blank control group (group a), the serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels of mice in the acute liver failure group (group b) were significantly increased, and the mortality rate was extremely high. RKH pretreatment for 1 week (group d) and 2 weeks (group e) significantly reduced the ALT (P < 0.01) and AST (P < 0.05) levels of mice.

[0039] like Figure 3 As shown, survival analysis suggested that RKH intervention for one week significantly prolonged the survival time of mice with liver failure and improved the prognosis of liver failure.

[0040] like Figure 4 As shown, the gross photos of the liver showed that compared with the control group mice, the livers of the mice in the liver failure group were swollen and congested. After RKH pretreatment, scattered bleeding spots were seen on the surface of the liver, the swelling of the liver was improved, and the congestion was significantly reduced.

[0041] like Figure 5As shown in the figure, hematoxylin and eosin (H&E) staining showed that diffuse necrosis of a large number of hepatocytes was widely observed in the liver tissue of ALF group mice, with nuclear condensation, deep staining or fragmentation, accompanied by neutrophil infiltration, large-area hemorrhage, and a large number of red blood cells in the liver sinusoids and between cells; RKH pretreatment significantly improved hepatocyte necrosis and neutrophil infiltration in liver tissue, and the red blood cells in the liver sinusoids and between cells were significantly reduced. RKH significantly improved the histological lesions of the liver caused by liver failure in mice and alleviated cell death.

[0042] Although the preferred embodiments of the present invention have been described, those skilled in the art may make other changes and modifications to these embodiments once they have learned the basic creative concept. Therefore, the appended claims are intended to be interpreted as including the preferred embodiments and all changes and modifications that fall within the scope of the present invention.

[0043] Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and their equivalents, the present invention is also intended to include these modifications and variations.

Claims

1. Application of RKH tripeptide in the preparation of drugs for the treatment of liver failure.

2. The use of the RKH tripeptide according to claim 1 in the preparation of a drug for treating liver failure, characterized in that: The liver failure includes acute liver failure.

3. The use of the RKH tripeptide according to claim 1 in the preparation of a drug for treating liver failure, characterized in that: The drug contains RKH tripeptide as the only active ingredient.

4. The use of the RKH tripeptide according to claim 1 in the preparation of a drug for treating liver failure, characterized in that: The dosage form of the drug is an injection dosage form or a non-injection dosage form.

5. The use of the RKH tripeptide according to claim 4 in the preparation of a drug for treating liver failure, characterized in that: The injection dosage form includes intraperitoneal injection.

6. The use of the RKH tripeptide according to claim 4 in the preparation of a drug for treating liver failure, characterized in that: The non-injection dosage form is selected from oral pharmaceutical preparations, intragastric lavage preparations, rectal absorption suppositories or enteric solvents.

7. Use of the RKH tripeptide according to claim 6 in the preparation of a medicament for treating liver failure, characterized in that: The oral pharmaceutical preparation is selected from tablets, capsules or oral liquids.

8. The use of the RKH tripeptide according to claim 1 in the preparation of a drug for treating liver failure, characterized in that: The drug also includes pharmaceutically acceptable excipients.

9. Use of the RKH tripeptide according to claim 8 in the preparation of a medicament for treating liver failure, characterized in that: The auxiliary material is selected from the group consisting of buffer, solvent, solubilizer, preservative, stabilizer, surfactant, antioxidant and mixtures thereof.

10. Use of the RKH tripeptide according to claim 9 in the preparation of a medicament for treating liver failure, characterized in that: In the medicine, the content of RKH tripeptide is 1w% to 99.9w%.

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