Use of pcsk9 inhibitor alirocumab in prevention and treatment of gallbladder stones

Abstract

The application discloses application of a PCSK9 inhibitor Alirocumab in prevention and treatment of gallbladder stones. The application is characterized in that, through pharmacological experiments in a mouse gallbladder stone model, the PCSK9 inhibitor Alirocumab can effectively reduce the content of cholesterol in mouse bile, and can prevent the formation of mouse gallbladder stones and dissolve the formed gallbladder stones. After administration, under ultrasonic monitoring, the diameter of the stones gradually decreases with the increase of the administration time. Therefore, the application provides a new drug for effectively preventing and treating cholesteric gallbladder stones. The PCSK9 inhibitor Alirocumab provided by the application has good clinical application value and application prospect as a drug for preventing and treating cholesteric gallbladder stones.

Description

Technical Field

[0001] This invention relates to the application of the PCSK9 inhibitor Alirocumab in the prevention and treatment of gallstones, and belongs to the field of biomedical technology. Background Technology

[0002] Gallstones are a common digestive system disease, with 80% of them being cholesterol gallstones. Their pathogenesis is complex, related to factors such as bile cholesterol supersaturation, impaired gallbladder contraction and emptying, intestinal factors, and genetics. Bile cholesterol supersaturation is the main cause of cholesterol crystal formation. The main components of bile, including cholesterol, bile acids, and phospholipids, are primarily secreted by hepatocytes. Under physiological conditions, cholesterol dissolves in micelles composed of bile acids and phospholipids, maintaining a dynamic equilibrium. Under certain conditions, this balance is disrupted, leading to the precipitation and accumulation of excess cholesterol into cholesterol stones. Currently, the main treatment for gallstones is surgery, which is invasive and carries potential complications. Clinically, drug treatment for gallstones has limited efficacy and a high recurrence rate, necessitating the exploration of new treatment options and effective drugs.

[0003] The liver is the center of cholesterol metabolism in the body, including cholesterol synthesis, uptake, breakdown, and excretion. The proprotein convertase subtilisin 9 (PCSK9) plays a crucial role in regulating cholesterol metabolism. It is known to bind to and mediate the degradation of low-density lipoprotein receptor (LDLR) on the surface of hepatocytes, thereby increasing plasma LDL-C levels. The PCSK9 inhibitor alirocumab inhibits LDLR receptor degradation, lowering blood cholesterol, and is currently approved for the treatment of familial hyperlipidemia and cardiovascular diseases such as atherosclerosis. However, the roles of PCSK9 and its inhibitors in regulating cholesterol metabolism in the liver itself and in the formation of cholesterol gallstones remain unclear. Summary of the Invention

[0004] The purpose of this invention is to address the technical problem of poor efficacy of current clinical drugs for treating gallstones. This invention provides the application of the PCSK9 inhibitor Alirocumab in the prevention and treatment of gallstones, which has the advantages of good efficacy, low price, and few side effects.

[0005] To achieve the above objectives, the present invention provides the use of the PCSK9 inhibitor Alirocumab in the preparation of a medicament for the prevention and treatment of gallstones.

[0006] Preferably, the gallstones include cholesterol gallstones.

[0007] Preferably, the drug prevents gallstones by reducing cholesterol levels in bile, increasing bile acid levels, and inhibiting gallstone formation.

[0008] Preferably, the drug treats gallstones by reducing cholesterol content in bile, increasing bile acid content, and dissolving gallstones.

[0009] This invention also provides the use of the PCSK9 inhibitor Alirocumab in the preparation of drugs that reduce liver and bile cholesterol, increase liver and bile bile acids and / or inhibit gallstone formation.

[0010] Compared with the prior art, the present invention has the following beneficial effects:

[0011] This invention utilizes the PCSK9 inhibitor Alirocumab in vivo in a mouse model of gallstones. The results show that Alirocumab effectively breaks down cholesterol in bile, increases bile acid content, and effectively prevents the formation of gallstones and dissolves existing gallstones. Under ultrasound monitoring, the diameter of the gallstones gradually decreased with increasing treatment time after administration. Therefore, this invention provides a novel drug for the effective prevention and treatment of cholesterol gallstones. The PCSK9 inhibitor Alirocumab provided by this invention has good clinical application value and promising prospects as a drug for the prevention and treatment of cholesterol gallstones. Attached Figure Description

[0012] Figure 1 The preventive effect of PCSK9 inhibitor Alirocumab on gallstones in mice: A: Schematic diagram of mouse experimental procedure; B: Macroscopic appearance of gallbladder and representative images of gallbladder bile under polarized light microscopy; C: Incidence of gallstone formation in mice after 8 weeks of feeding with a litholytic diet; D: Analysis of gallbladder bile composition (cholesterol, bile acids (BAs), phospholipids (PLs), cholesterol saturation index (CSI)); E: Concentration triphase diagram of gallbladder bile, showing relative lipid composition: one phase micelle region is surrounded by a solid curve, and above the micelle region, two solid lines and two dashed lines divide the phase diagram into regions A to E with different crystallization sequences; F: Analysis of liver lipid composition (cholesterol, bile acids, phospholipids); ns: No statistical difference; *: P < 0.05; **: P < 0.01; ***: P < 0.001);

[0013] Figure 2The therapeutic effect of PCSK9 inhibitor Alirocumab on gallstones in mice; A: Schematic diagram of mouse experimental procedure; B: Representative ultrasound images of gallstones in the Alirocumab treatment group; C: Changes in the maximum diameter of gallstones; D: Macroscopic appearance of gallbladder and representative images of gallbladder bile under polarized light microscopy in the Alirocumab treatment group; E: Analysis of gallbladder bile components (cholesterol, bile acids (BAs), phospholipids (PLs), cholesterol saturation index (CSI)); F: Analysis of liver lipid components (cholesterol, bile acids, phospholipids) ns: No statistically significant difference; *: P < 0.05; **: P < 0.01; ***: P < 0.001. Detailed Implementation

[0014] To make the present invention more apparent and understandable, preferred embodiments are described in detail below with reference to the accompanying drawings.

[0015] Unless otherwise specified, the experimental methods used in the following examples are conventional methods; the materials and reagents used are commercially available unless otherwise specified.

[0016] Example

[0017] 1. Construction of the gallbladder-integrated model

[0018] The litholytic diet, administered for 8 weeks, is an effective animal model simulating cholesterol gallstones. Mice in all groups were 8 weeks old at the time of the study, with a weight difference of less than 2g between mice. The purchase, feeding, and experimental procedures of the mice were all completed at the Department of Laboratory Animal Science, Fudan University. The litholytic diet, Dyets ASHF4 (40% fat, 1.25% cholesterol, 0.5% sodium cholate), was purchased from Shenzhen Aopu Biotechnology Co., Ltd.

[0019] 2. The preventive effect of the PCSK9 inhibitor Alirocumab on gallstones in mice.

[0020] To confirm the preventive effect of the PCSK9 inhibitor alirocumab on gallstones in mice, 12 mice (n=12) were sacrificed in each group after 8 weeks. The groups were divided into: (1) negative control group, fed with normal diet and subcutaneously injected with 0.1 ml / mouse / week of physiological saline; (2) control group, fed with stone-inducing diet and subcutaneously injected with 0.1 ml / mouse / week of physiological saline; (3) experimental group, fed with stone-inducing diet and subcutaneously injected with 10 mg / kg / week of alirocumab injection; and (4) positive control group, fed with normal diet and subcutaneously injected with 10 mg / kg / week of alirocumab injection. The differences in stone formation rate, bile composition, and liver lipid composition among the four groups were assessed. The results are as follows: Figure 1As shown in A-F, the stone formation rate in mice fed a litholytic diet and treated with Alirocumab was significantly lower than that in the control group. Macroscopic observation showed that the bile was clear, and analysis under a polarized light microscope showed no crystal formation in the bile. Further analysis of the lipid components in the liver and bile showed that, compared with the control group, the cholesterol content was significantly lower, the bile acid content was significantly higher, and the cholesterol saturation index (CSI) was significantly lower.

[0021] 3. The therapeutic effect of the PCSK9 inhibitor Alirocumab on cholesterol stones in mice.

[0022] To confirm the therapeutic effect of the PCSK9 inhibitor Alirocumab on cholesterol stones in mice, a cholesterol stone model was first established by feeding mice with a stone-inducing diet for 8 weeks. A total of 72 mice were used. Stone formation was confirmed by ultrasound in the mice. The 72 mice were randomly divided into 3 groups of 24 mice each for 8 weeks. The treatment groups were: (1) experimental group, fed with stone-inducing diet and subcutaneously injected with Alirocumab injection at 10 mg / kg / week; (2) control group, fed with stone-inducing diet and subcutaneously injected with physiological saline at 0.1 ml / mouse / week; (3) negative control group, fed only with stone-inducing diet. The size of the stones in each group was assessed by ultrasound every week. Three mice in each group were randomly selected and sacrificed every week. Liver, bile, and blood samples were collected to detect the corresponding indicators. The condition of the stones, changes in bile composition and liver lipid composition in the three groups of mice were evaluated. The results are as follows: Figure 2 As shown in A-F, under ultrasound observation, the stones in the Alirocumab-treated mice gradually decreased in size with increasing treatment time, and the maximum diameter of the stones decreased significantly over time. Macroscopic observation showed that the bile gradually became clearer with increasing treatment time, and analysis under a polarizing microscope showed that the bile crystals gradually became smaller. Further analysis of the lipid components in the liver and bile showed that, compared with the control group, cholesterol content decreased significantly, bile acid content increased significantly, and cholesterol saturation index (CSI) decreased significantly with increasing treatment time.

[0023] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any form or substance. It should be noted that those skilled in the art can make several improvements and additions without departing from the present invention, and these improvements and additions should also be considered within the scope of protection of the present invention.

Claims

1. Application of the PCSK9 inhibitor Alirocumab in the preparation of drugs for the prevention and treatment of cholesterol gallstones.

2. The application as described in claim 1, characterized in that, The drug prevents cholesterol gallstones by reducing cholesterol levels in the liver and bile, increasing bile acid levels, and inhibiting gallstone formation.

3. The application as described in claim 1, characterized in that, The drug treats cholesterol gallstones by reducing cholesterol levels in the liver and bile, increasing bile acid levels, and dissolving gallstones.

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