Use of dronedarone hydrochloride in the preparation of a medicament for the treatment of non-alcoholic fatty liver disease and cholestatic liver disease

Abstract

The application relates to the field of biological medicine, and provides application of hydrochloric acid dinedarong in preparation of a medicine for treating non-alcoholic fatty liver disease and cholestatic liver disease. The application discloses a new medical use of hydrochloric acid dinedarong, which can significantly reduce liver injury, lipid deposition in the liver and liver fibrosis caused by high-fat high-fructose high-cholesterol diet, and can reduce liver injury and liver fibrosis caused by cholestasis. Therefore, the hydrochloric acid dinedarong has the effect of treating non-alcoholic fatty liver disease and cholestatic liver disease.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and in particular to the use of dronedarone hydrochloride in the preparation of medicaments for treating non-alcoholic fatty liver disease and cholestatic liver disease. Background Technology

[0002] Nonalcoholic fatty liver disease (NAFLD) is one of the most common liver diseases. The NAFLD spectrum includes simple fatty liver (NAFL), nonalcoholic steatohepatitis (NASH), and nonalcoholic steatocirrhosis. As the disease progresses, approximately 25% of patients with simple fatty liver develop nonalcoholic steatohepatitis, eventually progressing to cirrhosis and liver cancer, seriously threatening human physical and mental health and quality of life. The pathological feature of NAFLD is the excessive deposition of triglycerides in hepatocytes. According to the "multiple-hit hypothesis," the causes of excessive triglyceride deposition in hepatocytes include: genetic susceptibility, epigenetic alterations, oxidative stress, and endoplasmic reticulum stress. These factors work together on the liver, not only leading to intrahepatic lipid "overload" but also causing persistent hepatocyte damage, inflammatory responses, and liver fibrosis. Currently, there are no marketed drugs with proven efficacy and safety for the treatment of non-alcoholic fatty liver disease.

[0003] Cholestatic liver disease refers to liver lesions caused by biliary tract diseases, infections, genetics, or other factors that lead to impaired bile formation or secretion, resulting in the accumulation of bile acids in the liver. This can cause liver damage, cirrhosis, liver failure, and even death. Except for obstructive cholestasis, which can be surgically cured, other types can only be treated with medication. Currently, only ursodeoxycholic acid and obeticholic acid are available for treatment, and their efficacy varies greatly from person to person. Ursodeoxycholic acid has no significant therapeutic effect in approximately 40% of patients, while obeticholic acid treatment is expensive and can cause adverse reactions such as itching and a decrease in high-density lipoprotein cholesterol. Therefore, there is an urgent need to develop and explore drugs for the treatment of non-alcoholic fatty liver disease and cholestatic liver disease.

[0004] The development of entirely new drugs is often multi-step, lengthy, and costly, while existing drugs generally have better safety profiles and more abundant research and clinical data. Drug repurposing is an effective strategy in drug development, which can shorten the development cycle, reduce costs, decrease risks, and increase output. In the treatment of patients with non-alcoholic fatty liver disease and cholestatic liver disease, it can buy time and offer a significant advantage, demonstrating enormous application potential in the medical field.

[0005] dronedarone hydrochloride is a drug used to treat cardiac arrhythmias. Its chemical name is N-(2-butyl-3-(4-(3-dibutylaminopropoxy)benzoyl)benzofuran-5-yl)methanesulfonamide hydrochloride. Dronedarone hydrochloride is a multi-channel inhibitor, blocking sodium, potassium, and calcium ion channels and β-receptors. It inhibits CYP3A and CYP2D6, but has no significant inhibitory effect on CYP1A2, CYP2C9, CYP2C19, CYP2C8, and CYP2B6. The major metabolite of dronedarone, SR90154, inhibits organic anion transport peptides (OATP1B1, OATP1B3) in vivo.

[0006] Given the limitations of existing treatments, it is essential to explore new uses for drugs used to treat non-alcoholic fatty liver disease and cholestatic liver disease. Summary of the Invention

[0007] The purpose of this invention is to provide a new drug for the treatment of non-alcoholic fatty liver disease and cholestatic liver disease, which can significantly reduce liver damage, liver inflammation and fibrosis.

[0008] To achieve the above-mentioned objectives, the present invention provides the following technical solution:

[0009] This invention provides the use of dronedarone hydrochloride in the preparation of medicaments for treating non-alcoholic fatty liver disease and cholestatic liver disease.

[0010] Preferably, the drug comprises dronedarone hydrochloride and excipients.

[0011] Preferably, the excipients include one or more of fillers, diluents, binders, absorption enhancers, excipients, surfactants, stabilizers, flavorings, and sweeteners.

[0012] Preferably, the non-alcoholic fatty liver disease includes simple fatty liver, non-alcoholic steatohepatitis, and non-alcoholic fatty liver cirrhosis.

[0013] Preferably, the cholestatic liver disease includes obstructive cholestasis, immune-associated cholestatic liver disease, and hereditary cholestatic liver disease.

[0014] Preferably, the immune-related cholestatic liver disease includes primary biliary cholangitis, primary sclerosing cholangitis, autoimmune hepatitis, and overlap syndrome.

[0015] This invention discloses a novel pharmaceutical use of dronedarone hydrochloride, revealing its excellent efficacy in treating non-alcoholic fatty liver disease (NAFLD) and cholestatic liver disease. Dronedarone hydrochloride can effectively slow the progression of NAFLD and inhibit fat accumulation and liver fibrosis in NAFLD. It can also significantly reduce cholestatic liver damage, liver inflammation, and fibrosis. The use of dronedarone hydrochloride provides patients with NAFLD and cholestatic liver disease with a new drug treatment option, alleviating patient suffering and financial burden. This invention offers a new approach to the prevention and treatment of NAFLD and has broad application prospects in the medical field. Attached Figure Description

[0016] To more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on the provided drawings without creative effort.

[0017] Figure 1 The body weights of mice in the model group and the drug administration group after the experiment in Example 1;

[0018] Figure 2 The liver weights of mice in the model group and the drug administration group after the experiment in Example 1;

[0019] Figure 3 The ratio of liver weight to body weight in mice in the model group and the drug administration group after the experiment in Example 1;

[0020] Figure 4 The change in white adipose tissue weight in mice in the model group and the drug administration group after the experiment in Example 1;

[0021] Figure 5 The ratio of white adipose tissue weight to body weight in mice in the model group and the drug administration group after the experiment in Example 1;

[0022] Figure 6 The ALT levels in the serum of mice in the model group and the drug administration group in Example 1 are shown.

[0023] Figure 7 The AST levels in the serum of mice in the model group and the drug administration group in Example 1 are shown.

[0024] Figure 8 The TCH level in the serum of mice in the model group and the drug administration group in Example 1.

[0025] Figure 9 The TG levels in the serum of mice in the model group and the drug administration group in Example 1 are shown.

[0026] Figure 10 The results show the H&E staining and Sirius red staining of mouse livers in the model group and the drug administration group in Example 1, along with statistical results.

[0027] Figure 11 The expression of α-SMA markers for liver fibrosis in the livers of mice in the model group and the drug administration group in Example 1 is shown.

[0028] Figure 12 The expression of Col1a1, a marker of liver fibrosis, in the livers of mice in the model group and the drug administration group in Example 1.

[0029] Figure 13 The expression of Col1a2, a marker of liver fibrosis, in the livers of mice in the model group and the drug administration group in Example 1.

[0030] Figure 14 The values ​​represent the serum ALT levels of mice in the control group, model group, and drug administration group in Example 2.

[0031] Figure 15 The AST levels in the control group, model group, and drug administration group of mice in Example 2 are shown.

[0032] Figure 16 The results show the H&E staining and Sirius red staining of mouse livers in the control group, model group, and drug treatment group in Example 2, along with statistical results.

[0033] Figure 17 The expression of α-SMA, a marker of liver fibrosis, in the livers of mice in the control group, model group, and drug treatment group in Example 2.

[0034] Figure 18 The expression of Col1a1, a marker of liver fibrosis, in the livers of mice in the control group, model group, and drug treatment group in Example 2.

[0035] Figure 19 The expression of Col1a2, a liver fibrosis marker, in the livers of mice in the control group, model group, and drug treatment group in Example 2.

[0036] Figure 20 The AST levels in the model group and the drug administration group of mice in Example 3 are shown.

[0037] Figure 21 The values ​​represent the serum ALT levels of mice in the model group and the drug administration group in Example 3.

[0038] Figure 22 The serum ALP levels of mice in the model group and the drug administration group in Example 3 are shown.

[0039] Figure 23 The TBA levels in the model group and the drug administration group of mice in Example 3 are shown.

[0040] Figure 24 The results show the H&E staining and Sirius red staining of mice in the model group and the drug administration group in Example 3, as well as the statistical results.

[0041] Figure 25 The expression of α-SMA, a marker of liver fibrosis, in the livers of mice in the model group and the drug administration group in Example 3.

[0042] Figure 26 The expression of Col1a1, a marker of liver fibrosis, in the livers of mice in the model group and the drug administration group in Example 3.

[0043] Figure 27 The expression of Col1a2, a marker of liver fibrosis, in the livers of mice in the model group and the drug administration group in Example 3. Detailed Implementation

[0044] The technical solutions provided by the present invention will be described in detail below with reference to the embodiments, but they should not be construed as limiting the scope of protection of the present invention.

[0045] The experimental mice, agents, and drugs used in the following examples, and their sources:

[0046] dronedarone hydrochloride was purchased from McLean, lot number 2016031601, CAS number 141625-93-6.

[0047] Tween 80 was purchased from McLean, part number T6336, CAS number 9005-65-6.

[0048] High-fat, high-fructose, and high-cholesterol diet (ResearchDiets, D09100310), high-fat diet control diet (ResearchDiets, D09100304), 3,5-Diethoxycarbonyl-1,4-dihydrocollidine (DDC) (Sigma, 137030).

[0049] H&E staining reagent (Solarbio, G1120) and Sirius Red staining reagent (Solarbio, G1471).

[0050] Reverse transcription reagent (PrimeScript RT reagent Kit with gDNA, Takara, RR047A), Real-time qPCR reagent (Premix Ex Taq, Takara, RR390A; TB GreenPremix Ex Taq II, Takara, RR820A).

[0051] C57BL / 6 mice were purchased from Jiangsu Jicui Pharmaceutical Biotechnology Co., Ltd.

[0052] Example 1

[0053] dronedarone hydrochloride treatment of mice on a high-fat, high-fructose, high-cholesterol (HFFC) diet

[0054] Thirteen 6-8 week old C57BL / 6 mice were randomly divided into an HFFC model group (n=6) and a dronedarone hydrochloride treatment group (n=7). Both groups were fed a high-fat, high-fructose, and high-cholesterol diet. After eight weeks of feeding, the dronedarone hydrochloride treatment group received an injection every other day for eight consecutive weeks of a solvent (100 μL 0.1% Tween 80) containing 50 mg / kg dronedarone hydrochloride, while the HFFC model group received an injection daily for eight consecutive weeks of the same solvent (100 μL 0.1% Tween 80). The mice were sacrificed after eight weeks.

[0055] 1. Serum, liver, and white fat were collected from mice in the model group and the drug treatment group, respectively. The body weight of the mice and the collected liver and white fat were weighed. The liver was fixed with 4% paraformaldehyde. The results are as follows: Figures 1-5 As shown, by Figures 1-5 It is known that dronedarone hydrochloride can significantly inhibit the body weight and liver weight of mice and reduce the deposition of white fat.

[0056] 2. The levels of serum liver injury markers (ALT, AST) and lipid markers (TCH, TG) in the mouse serum collected in step 1 were detected. The results are as follows: Figures 6-9 As shown, mice given dronedarone hydrochloride can significantly inhibit the levels of serum liver injury markers (ALT, AST) and lipid markers (TCH, TG).

[0057] 3. The liver sections collected in step 1 were processed into sections, and the sections were stained with H&E and Sirius red for histopathological analysis. The results are as follows: Figure 10 As shown, treatment with dronedarone hydrochloride significantly reduced triglyceride deposition in the liver of mice.

[0058] 4. Total RNA was extracted from the liver tissue obtained in step 1 and reverse transcribed. Real-time qPCR was used to detect changes in liver fibrosis markers. Results are as follows: Figures 11-13 As shown, mice given dronedarone hydrochloride can inhibit the expression of liver fibrosis markers α-SMA, Col1a1, and Col1a2 in the liver.

[0059] Example 2

[0060] Dronedarone hydrochloride treatment of mice with cholestatic liver disease in 0.1% DDC

[0061] Six- to eight-week-old C57BL / 6 mice were randomly divided into a control group, a model group, and a drug treatment group. The control group (n=6) was fed a normal diet, while the model group (n=6) and the drug treatment group (n=7) were fed a diet containing 0.1% DDC.

[0062] Starting with a diet containing 0.1% DDC, mice in the treatment group were intraperitoneally injected once daily for two weeks with a solvent containing 100 μL of dronedarone hydrochloride at a dose of 100 mg / kg (100 μL 0.1% Tween 80). Mice in the model group and control group were also intraperitoneally injected once daily for two weeks with the same solvent (100 μL 0.1% Tween 80). Two weeks after administration, the mice were sacrificed, and serum and liver samples were collected from all three groups. The mice were weighed, and the livers were fixed with 4% paraformaldehyde.

[0063] 1. The levels of ALT and AST in the collected mouse serum were detected, and the results are as follows: Figures 14-15 As shown, administration of dronedarone hydrochloride to mice significantly inhibited serum ALT and AST levels in mice with 0.1% DDC cholestasis.

[0064] 2. Liver sections collected from the three groups of mice were processed and stained with H&E and Sirius red. The results are as follows: Figure 16 As shown, mice treated with dronedarone hydrochloride showed a significant reduction in liver inflammation and liver fibrosis in mice with 0.1% DDC cholestasis.

[0065] 3. Total RNA was extracted from liver tissue collected from the three groups of mice, and reverse transcription was performed. Real-time qPCR was used to detect changes in liver fibrosis markers. The results are as follows: Figures 17-19 As shown, administration of dronedarone hydrochloride to mice can inhibit the expression of liver fibrosis markers α-SMA, Col1a1, and Col1a2 in the liver of cholestatic mice.

[0066] Example 3

[0067] Dronedarone hydrochloride treatment of Mdr2 gene knockout mice (a model of primary sclerosing cholangitis).

[0068] Mdr2 gene knockout mice aged 6-8 weeks were randomly divided into a model group (n=6) and a treatment group (n=7). The treatment group received intraperitoneal injections once daily for four weeks of a solvent containing 100 mg / kg dronedarone hydrochloride (100 μL 0.1% Tween 80). The control group received intraperitoneal injections once daily for four weeks of the same solvent (100 μL 0.1% Tween 80). Four weeks after treatment, the mice were sacrificed, and their serum and livers were collected. The mice were weighed, and the livers were fixed using 4% paraformaldehyde.

[0069] 1. The levels of ALT, AST, ALP, and TBA in the collected mouse serum were measured, and the results are as follows: Figures 20-23 As shown, administration of dronedarone hydrochloride to mice can significantly inhibit serum AST, ALT, TBA, and ALP levels in Mdr2 gene knockout mice.

[0070] 2. The collected liver sections were processed and stained with H&E and Sirius red before being subjected to liver pathological analysis. The results are as follows: Figure 24 As shown, treatment with dronedarone hydrochloride significantly reduced liver inflammation and liver fibrosis in Mdr2 gene knockout mice.

[0071] 3. Total RNA was extracted from the livers of the collected mice, reverse transcribed, and real-time qPCR was used to detect changes in liver fibrosis markers. The results are as follows: Figures 25-27 As shown, administration of dronedarone hydrochloride can inhibit the expression of liver fibrosis markers α-SMA, Col1a1, and Col1a12 in the liver of Mdr2 gene knockout mice.

[0072] In summary, the embodiments of this invention demonstrate that dronedarone hydrochloride can effectively alleviate liver damage, liver inflammation, and liver fibrosis during the process of cholestasis, thus playing a role in the treatment of cholestatic liver disease.

[0073] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and modifications can be made without departing from the principle of the present invention, and these improvements and modifications should also be considered within the scope of protection of the present invention.

Claims

1. Application of dronedarone hydrochloride as the sole active ingredient in the preparation of drugs for the treatment of primary sclerosing cholangitis.

2. The application as described in claim 1, characterized in that, The drug includes dronedarone hydrochloride and excipients.

3. The application as described in claim 2, characterized in that, The excipients include one or more of the following: fillers, binders, absorption enhancers, surfactants, stabilizers, flavorings, and sweeteners.

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