Composition containing silibinin

Abstract

This invention relates to the use of a silibinin-containing composition in the treatment of hepatic fibrosis. The silibinin-containing composition comprises silibinin and carvedilol as active ingredients. The silibinin composition disclosed in this invention can significantly downregulate the expression levels of Col1a1, Col1a2 (Col3a1), and Mmp2 mRNA, and can effectively improve the degree of various chronic liver diseases such as chemical liver injury, cholestatic liver disease, and non-alcoholic fatty liver disease, and the resulting hepatic fibrosis lesions.

Description

Technical Field

[0001] The present invention relates to the field of pharmaceutical technology, and specifically to the use of silybin-containing compositions in the treatment of liver fibrosis.

Background Art

[0002] The milk thistle (scientific name: Silybum marianum (L.) Gaertn.) is the dried and mature fruit of the milk thistle, a plant of the Asteraceae family, and has a bitter taste and a cool nature. It has the effects of clearing heat and detoxifying, soothing the liver and promoting bile flow. In modern clinical applications, excellent therapeutic effects on acute and chronic hepatitis, liver cirrhosis, fatty liver, etc. have been confirmed. Its active ingredient, silybin (SB), is one of the flavonoid extracts with the highest market share in the world market (Bhattacharya, S. Phytotherapeutic Properties of Milk Thistle Seeds: An Overview. J. Adv. Pharm. Educ. Res. 2011,1,69-79.).

Chemical

[0003] Silibinin reduces liver enzyme levels such as ALT and AST, preventing cell escape and loss of hepatocyte membrane functional integrity. Furthermore, silibinin exerts hepatoprotective effects by reducing CH, TG, and LDL levels. Silibinin reduces oxidative stress by activating various antioxidant enzymes and stimulating the Nrf2 pathway. From the perspective of cytokine release inhibition, silibinin has anti-inflammatory effects in NAFLD and NASH, significantly reducing levels of pro-inflammatory cytokines TNF-α, IL-6, IL-1β, and IL-12β. Chinese Patent 202111261969.0 discloses a novel manufacturing method for high-purity silibinin meglumine preparations and their application to acute and severe liver failure, while Chinese Patent 202210427370.8 discloses a manufacturing method for silibinin derivatives and their oxidative damage reduction effects. While silibinin has a definite hepatoprotective effect, its protective effect against liver fibrosis is weak. Due to its low water solubility, most current research focuses on injectable formulations for the treatment of liver fibrosis, and these studies have only resulted in a reduction of serum fibrosis indicators and proteins such as α-SMA and TGF-β1.

[0004] Hepatic fibrosis is the excessive deposition of diffusible extracellular matrix (especially collagen) within the liver, resulting from various chronic liver diseases. Its main manifestations include increased transcription of fibrosis-promoting molecules such as α-SMA protein, type I and type III collagen proteins, and substrate metalloprotease inhibitors, leading to the production of numerous fibrous and non-fibrous matrix proteins. Sustained or recurrent inflammatory death of hepatic parenchymal cells due to chronic liver disease causes excessive proliferation of fibrous connective tissue and a relative / absolute deficiency of its degradation activity, disrupting the dynamic balance of synthesis and degradation processes of various extracellular matrix components, thus leading to hepatic fibrosis. Hepatic fibrosis is a reversible stage in the progression of chronic liver disease to cirrhosis. Recent basic and clinical studies have shown that effective etiological treatment can directly suppress extracellular matrix synthesis and promote degradation, restoring the "net effect" of synthesis and degradation processes of various extracellular matrix fibers to dynamic equilibrium. Activation of hepatic stellate cells (HSCs) is a core process in hepatic fibrosis. In a normal liver, hepatocytes (HSCs) primarily store vitamin A. However, after liver injury, hepatocyte apoptosis causes apoptotic bodies to activate Kupffer cells and platelets, secreting cytokines and chemokines such as converting growth factor β (TGF-β) and platelet-derived growth factor (PDGF). Damaged hepatocytes and endothelial cells release reactive oxygen species and inflammatory factors. These factors work together to synthesize large amounts of extracellular matrix (ECM) through quiescent HSCs, converting them into activated myofibroblasts. While silibinin has a definite hepatoprotective effect, its antifibrotic effect is weak. Therefore, rationally combining silibinin with chemical agents that act on different target sites of HSC activation may provide a breakthrough in the treatment of reversing liver fibrosis caused by liver injury. [Overview of the project] [Problems that the invention aims to solve]

[0005] The present invention provides a silibinin-containing composition and the use of this composition in the treatment of hepatic fibrotic diseases.

[0006] This invention provides the following technical proposals:

[0007] A silibinin-containing composition, comprising silibinin and carvedilol as active ingredients.

[0008] a) A silibinin-containing composition comprising silibinin and carvedilol as activators; and b) a pharmaceutically acceptable additive suitable for the preparation of a solid oral dosage form; wherein the mass ratio of silibinin to carvedilol is 0.1 to 200:1.

[0009] In the silibinin-containing composition according to the present invention, the carvedilol comprises a pharmaceutically acceptable salt thereof with an organic acid or an inorganic acid. The organic acid includes, but is not limited to, oxalic acid, fumaric acid, benzoic acid, and mandelic acid, and the inorganic acid includes, but is not limited to, phosphoric acid.

[0010] In another embodiment, the present invention also discloses the following silibinin-containing compositions, the specific compositions of which are shown below.

[0011] A silibinin-containing composition comprising (a) silibinin and (b) a nonsteroidal anti-inflammatory drug as active ingredients; wherein the nonsteroidal anti-inflammatory drug is selected from the group consisting of aceclofenac and clobetasol propionic acid.

[0012] A silibinin-containing composition comprising (a) silibinin and (b) an angiotensin inhibitor as active ingredients, wherein the angiotensin inhibitor is valsartan.

[0013] A silibinin-containing composition comprising (a) silibinin and (b) a calcium channel blocker as active ingredients, wherein the calcium channel blocker is selected from the group consisting of nicardipine, manidipine, amlodipine, and cilnidipine.

[0014] A silibinin-containing composition comprising (a) silibinin and (b) a diuretic as active ingredients; wherein the diuretic is selected from the group consisting of tamsulosin and D-mannitol.

[0015] A libinin-containing composition comprising (a) silibinin and (b) a glucocorticoid as active ingredients; wherein the glucocorticoid is selected from the group consisting of desonides.

[0016] Furthermore, if silibinin and the compounds constituting the composition of the present invention (including carvedilol, nonsteroidal anti-inflammatory drugs, angiotensin inhibitors, calcium channel blockers, diuretics, and glucocorticoids) have a chiral carbon atom, then optical isomers and mixtures of isomers thereof are also included. In particular, the composition of the present invention also includes prodrugs formed from silibinin and one or more of the aforementioned compounds (carvedilol, nonsteroidal anti-inflammatory drugs, angiotensin inhibitors, calcium channel blockers, diuretics, and glucocorticoids).

[0017] In some embodiments, the present invention discloses the use of silibinin-containing compositions, compositions comprising silibinin and carvedilol, and compositions comprising silibinin and nonsteroidal anti-inflammatory drugs, angiotensin inhibitors, calcium channel blockers, diuretics and glucocorticoids in the treatment of hepatic fibrosis and in the treatment of chronic liver disease.

[0018] Furthermore, the aforementioned use is in the treatment of chronic liver disease and the resulting liver fibrosis.

[0019] Furthermore, the aforementioned uses include the treatment of chronic liver diseases such as chemical liver injury, cholestatic liver disease, and non-alcoholic steatohepatitis (NASH), and the resulting liver fibrosis.

[0020] According to the present invention, the combined administration of silibinin and carvedilol shows superior efficacy compared to each drug alone, making it possible to administer them in the form of a mixed formulation. Since the administration route of the silibinin and carvedilol composition disclosed in the present invention is generally oral, both drugs can be prepared in separate unit dosage forms, or in a single unit dosage form by physically mixing them. Examples of such unit dosage forms include powders, granules, tablets, and capsules, which are prepared by mixing the active pharmaceutical ingredient with excipients, diluents, etc., and using conventional formulation techniques described later. The solid oral dosage form of the present invention contains additives commonly used in such dosage forms, and medicinal additives commonly used in tablet or capsule formulations are applied. These substances are not limited to disintegrants, binders, lubricants, stabilizers, excipients or diluents, solubilizers, etc. The formulation form of the silibinin-containing composition is a tablet or a capsule.

[0021] The following additives may be used in the formulation of the present invention in a generally known manner: excipients (e.g., sugar derivatives such as lactose, sucrose, glucose, mannitol, sorbitol; starch derivatives such as corn starch, potato starch, pregelatinized starch, dextrin; cellulose derivatives such as microcrystalline cellulose; organic excipients such as gum arabic, dextran, pullulan; silicate derivatives such as light anhydrous silicic acid, synthetic aluminum silicate, calcium silicate, magnesium aluminum silicate; phosphates such as calcium hydrogen phosphate; carbonates such as calcium carbonate; sulfates such as calcium sulfate) and other inorganic excipients. Formulators), lubricants (e.g., metal stearate salts such as stearic acid, calcium stearate, and magnesium stearate; talc; waxes such as beeswax and whale wax; sulfates such as boric acid, adipic acid, and sodium sulfate; ethylene glycol, fumaric acid, sodium benzoate, DL-leucine; lauryl sulfates such as sodium lauryl sulfate and magnesium lauryl sulfate; silicic acids such as anhydrous silicic acid and hydrated silicic acid; and the above starch derivatives), binders (e.g., hydroxypropyl cellulose, hydroxypropyl methylcellulose, polyvinylpyrrolidone, polyethylene (Cellulose and compounds similar to the above excipients), disintegrants (e.g., cellulose derivatives such as low-substituted hydroxypropylcellulose, carboxymethylcellulose, calcium carboxymethylcellulose, and internally cross-linked sodium carboxymethylcellulose; chemically modified starches and celluloses such as carboxymethyl starch, sodium carboxymethyl starch, and cross-linked polyvinylpyrrolidone; the above starch derivatives), emulsifiers (e.g., colloidal clays such as bentonite and begum; metal hydroxides such as magnesium hydroxide and aluminum hydroxide; anionic surfactants such as sodium lauryl sulfate and calcium stearate; cationic surfactants such as benzalkonium chloride; nonionic surfactants such as polyoxyethylene alkyl ethers, polyoxyethylene sorbitan fatty acid esters, and sucrose fatty acid esters; and phospholipids such as soy lecithin), stabilizers (e.g., parahydroxybenzoic acid esters such as methyl parahydroxybenzoate and propyl parahydroxybenzoate; alcohols such as chlorobutanol, benzyl alcohol, and phenethyl alcohol; benzalkonium chloride; phenols such as phenol and cresol; dehydroacetic acid;and sorbic acid), flavoring agents (e.g., commonly used sweeteners, acidulants, fragrances, etc.), diluents and other additives;

[0022] A combination with a silybin dosage range of about 0.2 - 8000 mg and a carvedilol dosage range of about 2 - 40 mg is effective for the treatment of liver fibrosis. Particularly appropriate dosage ranges are about 1.5 - 3000 mg of silybin and about 3 - 20 mg of carvedilol. More preferred unit dosages are about 3 - 1000 mg of silybin and about 3 - 10 mg of carvedilol. Even more preferred unit dosages are about 15 - 500 mg of silybin and about 3 - 10 mg of carvedilol. Even more preferably, the unit dosage is about 30 - 500 mg of silybin and about 3 - 10 mg of carvedilol. Particularly preferred unit dosages are about 90 - 500 mg of silybin and about 3 - 10 mg of carvedilol. Even more preferred unit dosages are about 120 - 500 mg of silybin and about 3 - 10 mg of carvedilol. The most preferred unit dosage is about 150 - 500 mg of silybin and about 3 - 10 mg of carvedilol.

[0023] The combined use of silybin and carvedilol disclosed in the present invention significantly down - regulates the expression levels of Col1a1 and Col1a2 (Col3a1) mRNA and effectively improves the degree of liver fibrosis lesions in various liver fibrosis models. The above drugs are preferably used in warm - blooded animals, more preferably in humans.

Brief Description of the Drawings

[0024] [Figure 1] Establishment and verification of an HSCs activation model based on Col1a1 - Luci - reporter luciferase; ***P < 0.001 compared with the empty vector group; ns > 0.05 compared with the control group, P < 0.001;

[0025] [Figure 2] Effect of silybin on HSCs activation; ***P < 0.001 compared with the empty vector group; ns > 0.05 compared with the control group;

[0026] Figure 3 shows the screening of silybin compositions for inhibiting HSCs activation. Among them,

[0027] [Figure 3-1] The effect of the combination of a compound classified as an antihypertensive drug (represented by carvedilol) and silybin on the Col1a1-Luci-reporter luciferase expression level in activated HSCs;

[0028] [Figure 3-2] The effect of the combination of silybin and other compounds on the Col1a1-Luci-reporter luciferase expression level in activated HSCs; ***P < 0.001 compared with the empty vector group; ns > 0.05, #P < 0.05, ##P < 0.01, P < 0.001 compared with the control group;

[0029] [Figure 4] Verification of the effect of the combination of silybin and carvedilol on inhibiting the activation of primary hepatic stellate cells (pHSCs). Primary mouse pHSCs were isolated and cultured, and after treatment with carvedilol alone, silybin alone or the combination of both, the mRNA expression levels of Col1a1 and Col1a2 were analyzed. ns > 0.05, **P < 0.01, ***P < 0.001;

[0030] Figure 5 shows the analysis of the administration ratio of silybin and carvedilol in the inhibitory effect on HSCs activation. Among them,

[0031] [Figure 5-1] When the concentration of carvedilol was fixed at 10 μM and the two compounds were combined at a concentration ratio of 6:1; 4:1; 2:1; 1:1; 1:2; 1:4; 1:6; 1:8, the inhibitory effect on HSCs activation;

[0032] [Figure 5-2]HSCs activation effect when the silibinin concentration was fixed at 10 μM and both compounds were combined in concentration ratios of 6:1; 4:1; 2:1; 1:2; 1:4; 1:6; 1:8. Compared to the empty vector group, ***P<0.001; compared to the control group, ns>0.05, ##P<0.01, ###P<0.001.

[0033] Figure 6 shows the effect of concomitant use of silibinin (100 mg / kg) and carvedilol (2 mg / kg) on ​​CCl4-induced liver fibrosis.

[0034] [Figure 6-1] Analysis of liver Col1a1 and Col1a2 mRNA expression levels;

[0035] [Figure 6-2] Liver HE, Sirius Red, and Masson staining analysis;

[0036] [Figure 6-3] Comparison of volume fraction (CVF) of liver collagen in each mouse group. ns>0.05, *P<0.05, **P<0.01, ***P<0.001;

[0037] Figure 7 shows the effect of concomitant use of silibinin (100 mg / kg) and carvedilol (2 mg / kg) on ​​BDL-induced liver fibrosis.

[0038] [Figure 7-1] Analysis of liver Col1a1 and Col1a2 mRNA expression levels;

[0039] [Figure 7-2] Liver HE, Sirius Red, and Masson staining analysis;

[0040] [Figure 7-3] Comparison of volume fraction (CVF) of liver collagen in each mouse group. ns>0.05, *P<0.05, **P<0.01, ***P<0.001;

[0041] Figure 8 shows the effects of combined use of silibinin (75, 100 mg / kg) and carvedilol (2 mg / kg) on ​​MCD-induced NASH and liver fibrosis.

[0042] [Figure 8-1] Liver Col1a1, Col1a2, Col3a1 mRNA and Mmp2 expression level analysis;

[0043] [Figure 8-2] Liver HE, Sirius Red, and Masson staining analysis;

[0044] [Figure 8-3] Comparison of volume fraction (CVF) and lipid vacuolar area in the liver of each mouse group. Compared to the control group (MCS), ns > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001; compared to the MCD group, ns > 0.05, #P < 0.05, ##P < 0.01, ###P < 0.001;

[0045] Figure 9 shows the dose-dependent effects of fixed-ratio silibinin and carvedilol compositions on CCl4-induced liver fibrosis.

[0046] [Figure 9-1] Analysis of liver Col1a1 and Col1a2 mRNA expression levels;

[0047] [Figure 9-2] Liver HE, Sirius Red, and Masson staining analysis;

[0048] [Figure 9-3] Comparison of volume fraction (CVF) of liver collagen in each mouse group. Compared to the control group, ns > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001; compared to the model group, ns > 0.05, #P < 0.05, ##P < 0.01, ###P < 0.001;

[0049] Figure 10 shows the dose-dependent effects of a fixed-ratio silibinin and carvedilol composition on BDL-induced liver fibrosis.

[0050] [Figure 10-1] Analysis of liver Col1a1 and Col1a2 mRNA expression levels;

[0051] [Figure 10-2] Liver HE, Sirius Red, and Masson staining analysis;

[0052] [Figure 10-3] Comparison of volume fraction (CVF) of liver collagen in each mouse group. Compared to the control group, ns > 0.05, *P < 0.05, **P < 0.01, ***P < 0.001; compared to the model group, ns > 0.05, #P < 0.05, ##P < 0.01, ###P < 0.001. [Modes for carrying out the invention]

[0053] The present invention will be further described below through examples, but this will not limit the present invention.

[0054] The following experimental examples investigated the effectiveness of the combined use of carvedilol and silibinin against liver fibrosis.

[0055] The CAS registry number for carvedilol used in this invention is 72956-09-3, and its structural formula is as follows: [ka]

[0056] Experimental Example 1: Establishment and validation of an HSC activation model based on Col1a1-Luci-reporter luciferase.

[0057] 1. Experimental materials

[0058] The human hepatic stellate cell line LX-2 used in this invention was purchased from Wuhan Punuocai Life Technology Co., Ltd. and cultured under normal conditions at 37°C and 5% CO2. The culture medium used was DMEM (Gibco) containing 10% fetal bovine serum. Fetal bovine serum was purchased from Hyclone (Logan, Utah, USA), and trypsin from Amersco (Solon, Ohio, USA). OptiMEM culture medium was purchased from Gibco (California, USA), and Lipofectamine 3000 transfection reagent was purchased from Invitrogen (Waltham, USA). The pirfenidone used in this invention was purchased from MedChemExpress (New Jersey, USA), and the DuoLite™ Luciferase Assay System reporter assay kit was purchased from Nanjing Noweizan Biotechnology Co., Ltd. (Nanjing, China). The pEZX-FR03-hygro empty vector plasmid and the pEZX-FR03-hygro-COL1A1-WT plasmid were prepared by Cerise Co., Ltd.

[0059] 2. Experimental Method

[0060] 2.1 Culture and transfection of LX-2 cell lines

[0061] LX-2 cells were seeded in 96-well plates and cultured to an appropriate density. A transfection system was then prepared using Lipofectamine 3000 transfection reagent. 0.1 μg of plasmid was transfected into each well. The Empty vector group was transfected with the pEZX-FR03-hygro empty vector plasmid, while the Control group and other treatment groups were transfected with the pEZX-FR03-hygro-COL1A1-WT plasmid.

[0062] 2.2 Drug administration to LX-2 cells

[0063] Six hours after plasmid transfection of the LX-2 cell line, the culture media were changed to serum-free media containing 2.5 mM, 5 mM, and 10 mM of the antifibrotic agent pirfenidone, according to the group assignments. After 12 hours of incubation, the cells were analyzed using the Duo-Lite™ Luciferase Assay System kit.

[0064] 2.3 Validation using the Duo-Lite™ Luciferase Assay System kit

[0065] The cell culture plates to be tested were removed from the incubator and allowed to rise to room temperature for 30 minutes to achieve temperature equilibrium. Testing of firefly luciferase activity: The original culture medium was removed, 75 μl of PBS was added, and then an equal volume of Duo-Lite Luciferase detection reagent was added and mixed uniformly. After reacting at room temperature for 10 minutes, the luminescence of firefly luciferase was measured.

[0066] 3. Experimental Results

[0067] As shown in Figure 1, in in vitro cultured LX-2 cells, control cells transfected with the Col1a1 reporter gene consistently expressed high levels of Col1a1-Luci-reporter luciferase based on the characteristics of the astrocytic cell activation state. In the experimental group, the antifibrotic agent Pirfenidone was administered at concentrations of 2.5 mM, 5 mM, and 10 mM. Of these, 5 mM and 10 mM Pirfenidone significantly downregulated luciferase expression levels, showing a clear dose-dependent relationship. This established an HSC activation model based on Col1a1-Luci-reporter luciferase and validated it with the positive agent Pirfenidone.

[0068] Experimental Example 2: Evaluation of the effect of silibinin on HSC activation.

[0069] 1. Experimental materials

[0070] The silybin used in this invention was purchased from Nanjing Zelang Biotechnology Co., Ltd.

[0071] The other experimental materials were the same as in Experimental Example 1.

[0072] 2. Experimental Method

[0073] 2.1 Culture and transfection of LX-2 cell lines

[0074] The specific method was the same as in section 2.1 of Experimental Example 1.

[0075] 2.2 Drug administration to LX-2 cells

[0076] Six hours after plasmid transfection of the LX-2 cell line, the culture medium was replaced with serum-free medium containing 10 μM, 20 μM, and 50 μM silybin. After 12 hours of incubation, the cells were analyzed using the Duo-Lite™ Luciferase Assay System kit.

[0077] 2.3 Validation using the Duo-Lite™ Luciferase Assay System kit

[0078] The specific method was the same as in section 2.3 of Experimental Example 1.

[0079] 3. Experimental Results

[0080] As shown in Figure 2, the efficacy of silibinin at concentrations of 10 μM, 20 μM, and 50 μM was evaluated in an HSC activation model based on Col1a1-Luci-reporter luciferase using LX-2 cells. The results showed that in vitro experiments, silibinin at concentrations of 50 μM and lower did not show a significant inhibitory effect on stellate cell activation.

[0081] Experimental Example 3: Screening of silibinin compositions for inhibiting HSC activation

[0082] 1. Experimental materials

[0083] All drugs screened (rofecoxib, pranoprofen, meloxicam, hydrocortisone acetate, sulindac, fluocinolone acetonide, loxoprofen sodium, desonide, etc.) were prepared using the FDA-approved drug library (MCE-FDA-approved drug library-HY-L1022) and the PerkinElmer high-throughput drug discovery screening workstation at China Pharmaceutical University.

[0084] The other experimental materials were the same as in Experimental Examples 1 and 2.

[0085] 2. Experimental Method

[0086] 2.1 Culture and transfection of LX-2 cell lines

[0087] The specific method was the same as in section 2.1 of Experimental Example 1.

[0088] 2.2 Drug administration to LX-2 cells

[0089] Six hours after plasmid transfection of the LX-2 cell line, cells were treated in three groups: 10 μM silibinin alone, 10 μM screening agent alone, and a combination of 20 μM silibinin and 20 μM screening agent in a 1:1 ratio. After incubation for 12 hours in serum-free culture medium, assays were performed using the Duo-Lite™ Luciferase Assay System kit.

[0090] 2.3 Validation using the Duo-Lite™ Luciferase Assay System kit

[0091] The specific method was the same as in section 2.3 of Experimental Example 1.

[0092] 2.4 Analysis of compound action in combination

[0093] Cell activation inhibition rate % = (Firefly luminescence value of control group - Firefly luminescence value of drug-administered group) / Firefly luminescence value of control group × 100%. The CI value was calculated using CompuSyn software. 0.90 < CI < 1.10: additive effect, CI < 0.90: synergistic effect (the smaller the CI value, the stronger the synergistic effect).

[0094] 3 Experimental results

[0095] As shown in Figures 3-1 and 3-2, silibinin showed a synergistic effect with the following drugs: carvedilol (CI = 0.11), nifedipine (CI = 0.31), valsartan (CI = 0.47), tamsulosin (CI = 0.50), manidipine (CI = 0.54), amlodipine (CI = 0.62), D-mannitol (CI = 0.66), cilnidipine (CI = 0.79), aceclofenac (CI = 0.46), clobetasol propionate (CI = 0.86). It showed an additive effect with desonide (CI = 0.91). In particular, the combination of silibinin and carvedilol showed the strongest synergistic effect.

[0096] Note: Screening of silibinin compositions for inhibition of HSCs activation

[0097] The drugs screened from the FDA-approved drug library are as follows: lofecoxib, pranoprofen, meloxicam, hydrocortisone acetate, ramipril, irbesartan, rasibipine, sulindac, fluocinonide, loxoprofen sodium, desonide, idebenone, amodiaquine hydrochloride, azelaic acid, clobetasol propionate, halosinonide, benfotiamine, flumethasone, mannitol, flufenamic acid, 5-aminosalicylic acid, ketoprofen, bromf Enac sodium hydrate, isopropylantipyrine, valdecoxib, troglitazone, L-ascorbic acid, imatinib mesylate, regorafenib hydrochloride, ponatinib, cabozantinib malate, quinapril hydrochloride, benazepril hydrochloride, perindopril tert-butylamine, cilazapril hydrate, zofenpril, imidapril hydrochloride, moekipril, temocapril hydrochloride, captopril, enalapril maleate, losartan, candesartan, valsartan, eprosartan, olmesartan, medoxomil, fimasartan, telmisartan, olmesartan, azilsartan, nisoldipine, benidipine hydrochloride, efodipine hydrochloride Monoethanolamine salt, nitrendipine, manidipine hydrochloride, crebidipine, nifedipine, cilnidipine, nimodipine, amlodipine, nicardipine hydrochloride, nivadipine, felodipine, verapamil hydrochloride, cinnarizine, aliskiren, carvedilol, spironolactone, hosinopril sodium, enalaprilate, triamcinolone, zamifibrate, troxipide, tiotropium bromide, flurbiprofen, roflumilast, tyrolidine hydrochloride, budesonide, etoricoxib, sunitinib, homohalintonin, zofer Nopril calcium, amlexanox, salmeterol xinafoate, lenvatinib, aceclofenac, nintedanib, apatinib, ruxolitinib, gefitinib, benzdamine hydrochloride, regorafenib, firocoxib, dimethyl fumarate, beclomethasone dipropionate, mometasone furoate, ganciclovir, articaine hydrochloride, sapropterin hydrochloride, dextromethorphan (hydrobromide), riluzole, avanafil, clenbuterol hydrochloride, dapoxetine hydrochloride, naltrexone hydrochloride, amisulpride,Candesartan cilexetil, vardenafil hydrochloride, indacaterol maleate, hexamethylenetetramine, desmopressin acetate, fosfomycin calcium, tyrofiban (hydrochloride monohydrate), betamethasone dipropionate, dapiprazole hydrochloride, parecoxib, isoniazid, telaprevir, sulfaphylazole, pyrazinamide, methylprednisolone, sodium cromoglycate, sodium resinoxad, metoprolol succinate, atenolol, acebutolol hydrochloride, betaxolol, labetalol hydrochloride, arotinolol, propranolol hydrochloride, (S)-timolol maleate, phentolamine mesylate, trazoline hydrochloride, prazosin hydrochloride, tamsulosin, bisoprolol Hemi fumarate, esmolol hydrochloride, sotalol hydrochloride, carteolol hydrochloride, alprenolol hydrochloride, phenoxybenzamine hydrochloride, yohimbine, rauwalscin hydrochloride, icotinib, fidaxomicin, barnemurin, lamivudine, prelixafoll, azafen, cefoperazone, tegafur, diflorazone butyrate, sulbactam, citrate, trametinib, estrompiperazine sulfate, alprostadil, moclobemide, capecitabine, phentolamine, sulindac, sodium gluconate, acetylglutamine, bromhexine, sevelamer, dimemorphan, L-adrenaline, mangafodipyr, nitrogen mustard, hydroxyphasudil, akabastine, irinotecan, sulfaguanidine, cobimetinib, taltirelin, jiyo Dohydroxyquinoline, ropinirole, deoxycholic acid, terconazole, prostaglandin E2, clinofibrate, loxoprofen, brigatinib, pentostatin, medroxyprogesterone, albendazole, vitamin B1, fosphenytoin sodium, fusidic acid, natamycin, dacarbazine, doxate sodium, balofloxacin, suramin, thiamazole, epirubicin, mitoxantrone, bosentan, resinoxad, azatadine, trazoline, doripenem, vonoprazan fumarate, milnacipran, venlafaxine, dantrolene, tepillonic acid, palonosetron, fenspiride, isotretinoin, fenofibrate, nedocromil sodium, lafutidine, fluphenazine, fluticasone, cabergoline,Carmofur, Strophanthidin G, Tenofovir Alafenamide, Drospirenone, Lidocaine, Mianserin, Pyridostigmine, Sodium Salicylate, Cefotaxime, Sulfamethoxydine, Vincristine, Daptomycin, Doxycycline, Phyloquinone, Lapatinib, Irinotecan, Pravastatin, L-Arginine, Nimorazole, Fluconazole, Perphenazine, Diprophylline, Pimavanserin, Dapsone, Tyamulin, Risedronic Acid, Olanzapine, Levobupivacaine, Lovastatin, Fluvastatin, Teicoplani Ferbamate, Pivmecilinum, Iguratimod, Rifabutin, Berberine, Tauroursodeoxycholic acid, Progesterone, Altoretamine, Levodopa, Duloxetine, Galantamine, Tinidazole, Tolvaptan, Guanidine thiocyanate, Altemeter, Ivabradine, Mosapride, Tadalafil, Ramelteon, Bosentan, Trifluridine, Decroxydine, Tamibarotene, Gemcitabine, Phenoxybenzamine, 3,4-Diaminopyridine, Primaquine, Olopatadine, Diethylcarbamazine citrate, Pancuronium bromide Cefometasol, carbamazepine, tedizolid, ipratropium bromide, brexpiprazole, plucarprid, diclofenac sodium, aminoglutethimide, clotrimazole, tranexamic acid, hydrochlorothiazide, ibuprofen, rifampicin, moxisilyl, piroxicam, fludarabine, delamanid, levodropropidine, rucaparib, granisetron, metacholine, ribavirin, novobiocin, levonorgestrel, clozapine, febuxostat, tranilast, solifenacin, valganciclovir, ketothi Fen, Milnacipran, Dalifenacin, Lenalidomide, Ondansetron, Procainamide, Fexofenadine, Fenofibrate, Prulifloxacin, Sulfadimethoxine, Bezafibrate, Sumatriptan, Levetiracetam, Dorzolamide, Letapamlin, Diphenidol, Glibenclamide, Acetazolamide, Hexylamine Hydrochloride, Irbesartan, 9-Hydroxyrisperidone, Ciprofibrate, Glimepiride, Cefdinir, Decitabine, Vecuronium Bromide, Ticlopidine, Aspirin, Haloperidol, Gestodine,Trimethadione, fluorouridine, estradiol, rasagiline, nitrofurantoin, ezetimibe, acarbose, 5-fluorouracil, mexiletine, treosulfan, voglibose, leflunomide, rivastigmine, acitretin, promethazine, hydroxyzine, glucosamine, iproniazid, safinamide, prazosin, histamine, ethambutol, cephamandol, diltiazem, lincomycin, bazedoxifen, cidofovir, tranexamic acid, tropisetron, tizanidine, scopolamine, etoposide , folinic acid, nelfinavir, chlorthalidone, carbidopa, diphenhydramine citrate, salicylic acid, ubenimex, probenecid, enoxacin, anagrelide, enacidonib, levosulpiride, quinine, nicotine, heparin, penicillamine, neomycin, itraconazole, prelixafo, dipyridamole, methoxsalen, sodium nitroprusside, sodium ertapenem, ornidazole, peramivir trihydrate, zanamivir, titanium dioxide, sodium glutamate, L-ornithine, a total of 397 types.

[0098] Of these, the following showed synergistic or additive effects in inhibiting HSC activation when used in combination with silibinin:

[0099] Synergistic effects: Carvedilol (CI=0.11), nicardipine (CI=0.31), valsartan (CI=0.47), tamsulosin (CI=0.50), manidipine (CI=0.54), amlodipine (CI=0.62), D-mannitol (CI=0.66), cilnidipine (CI=0.79), aceclofenac (CI=0.46), clobetasol propionate (CI=0.86)

[0100] Additive effect: desonide (CI=0.91)

[0101] [Table 1]

[0102] Experimental Example 4: Verification of the effect of combined use of carvedilol and silibinin on the inhibition of primary hepatic stellate cell (p-HSC) activation.

[0103] 1. Experimental materials

[0104] Male C57BL / 6J mice (8 weeks old, 20-24g body weight) were purchased from Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd. Heparin sodium, EGTA, Type IV collagenase, Pronase E, and Dnase I were purchased from Sigma-Aldrich, and Nycodenz was purchased from Axis-Shield / Alere Technologies AS. The RNA extraction kit (RNAiso Plus reagent, Cat# 9109) was purchased from Takara (Japan), and HiScript Q RT SuperMix for qPCR and ChamQ SYBR qPCR Master Mix were purchased from Vazyme (Cat# Q311-02 / 03, Nanjing, China). Carvedilol was purchased from MedChemExpress (New Jersey, USA).

[0105] The other experimental materials were the same as in Experimental Example 2.

[0106] 2. Experimental Method

[0107] 2.1 Isolation of primary mouse hepatic stellate cells (p-HSCs)

[0108] After weighing the mice, they were anesthetized by intraperitoneal administration of 5% chloral hydrate at a dose of 0.1 ml / 10 g. The entire body was disinfected with ethanol spray, and after fixation, the abdomen was disinfected with alcohol swab, the abdominal cavity was opened, the intestines were pushed aside, and the portal vein and inferior vena cava were exposed. The portal vein was dissected, and after pre-inserting sutures beneath the vessel, a cannula was inserted into the portal vein and ligated with sutures. The terminal end of the inferior vena cava was cut open, and the thoracic portion of the inferior vena cava was closed. Then, pre-warmed EGTA solution, Pronase E solution, and Collagenase IV solution were perfused for approximately 3 minutes, 5 minutes, and 7 minutes, respectively, at a flow rate setting of 6. After perfusion, the livers were removed and transferred to EP tubes containing 10 ml of enzyme buffer solution, and stored at 4°C. After all livers were collected, they were digested together. The Pronase E / Collagenase IV solution was pre-warmed. All livers were transferred to a clean bench for primary culture, then to a sterile petri dish. 10 ml of an equal volume of Pronase E / Collagenase IV solution was added, and the livers were sheared to thoroughly disperse them. This was then transferred to a 50 ml EP tube, and the residue in the petri dish was washed with enzyme buffer solution and Pronase E / Collagenase IV solution (1:1) to adjust the total volume so as not to exceed 40 ml. DNase I solution equivalent to 1% of the total volume was added, and digestion was carried out at 37°C and 180 rpm for 17 minutes. The centrifuge was pre-cooled. The digested suspension was filtered through a 70 μm filter and collected in a 50 ml EP tube. All suspensions were dispensed into appropriate volumes, filled to 50 ml with cooled GBSS B solution, and centrifuged at 4°C and 580 g for 10 minutes. Remove the supernatant, leaving 10 ml, add 120 μl of Dnase I solution and suspend, fill with cooled GBSS B solution to 50 ml, and centrifuge at 4°C and 580 g for 10 minutes. Remove the supernatant, leaving 5-10 ml, add 120 μl of Dnase I solution and suspend, fill with cooled GBSS B solution to 15 ml, and mix. Add 30 ml of 18% Nycodenz solution to the above cell suspension, mix, and dispense equal volumes into 15 ml EP tubes. Slowly overlay 2 ml of cooled GBSS B solution into each tube (first moisten the tube wall, then gently add using a syringe). Centrifuge at 4°C and 1380 g for 17 minutes (acceleration 1, deceleration 0).After centrifugation, the white cell layer formed at the interface was collected by aspirating with a Pasteur pipette, washed 1-2 times with cooled GBSS B solution, and then centrifuged at 4°C and 580g for 10 minutes. The resulting cell precipitate was suspended in 3 ml of 20% FBS-containing DMEM medium, and the cell count and viability were measured by trypan blue staining. The cells were seeded at a density of 5 × 10⁵ cells / well in a 12-well plate, uniformly dispersed after microscopic examination, and cultured in an incubator for 24 hours before being used in experiments.

[0109] 2.2 Mouse hepatic astrocytocyte spontaneous activation model and drug administration

[0110] Isolated primary mouse hepatic stellate cells were adherently cultured and spontaneously activated for 3 days. The serum-free medium was then replaced with the following drug-treated media, and the cells were incubated for 12 hours: 25 μM silibinin (Sil), 10 μM carvedilol (Car), or a combination of 50 μM silibinin and 20 μM carvedilol in a 1:1 ratio. After treatment, cells were harvested, and the mRNA expression levels of fibrosis-related genes Col1a1 and Col1a2 were measured by qRT-PCR.

[0111] 2.3 qRT-PCR (Reverse Transcription Polymerase Chain Reaction)

[0112] The cell culture plate was collected, and 1 mL of RNAiso Plus was added. The mixture was thoroughly mixed using a 1 mL micropipette until the liquid was clear and no cell clumps remained, then transferred to a 1.5 mL EP tube. Total RNA was extracted according to the protocol of the RNAiso Plus reagent total RNA extraction kit, and RNA quantification was performed. mRNA was reverse transcribed to cDNA according to the procedure of the RT-PCR kit. The reaction mixture and cDNA were mixed to a total volume of 15 μL according to the instructions of the qRT-PCR kit, and the PCR reaction was performed using the specified program. The reaction conditions for quantitative PCR were as follows:

[0113] Step 1: 95°C, 1 minute (preliminary denaturation)

[0114] Step 2: PCR reaction: 95°C for 15 seconds; 60°C for 15 seconds; 72°C for 30 seconds, 40 cycles

[0115] Step 3: Melting curve analysis: 65℃~95℃

[0116] 2.4 Analysis of Compound Combination Action Index (CI)

[0117] The analysis method was the same as in section 2.4 of Experimental Example 3.

[0118] 3. Experimental Results

[0119] As shown in Figure 4, in a mouse primary hepatic stellate cell activation model, the group receiving carvedilol and silibinin showed significant downregulation of mRNA expression levels of Col1a1 and Col1a2, genes specifically expressed in activated hepatic stellate cells, demonstrating a superior inhibitory effect compared to the groups receiving each drug alone. Analysis of the combined effect of the two drugs using CompuSyn software yielded CI values ​​of Col1a1 (CI=0.54) and Col1a2 (CI=0.36), confirming a strong synergistic effect.

[0120] Experimental Example 5: Examination of the dosage ratio when carvedilol and silibinin are used in combination.

[0121] 1. Experimental materials

[0122] The experimental materials were the same as those used in Experimental Examples 1 and 2.

[0123] 2. Experimental Method

[0124] 2.1 Culture and transfection of LX-2 cell lines

[0125] The specific method was the same as in section 2.1 of Experimental Example 1.

[0126] 2.2 Drug administration to LX-2 cells

[0127] The carvedilol dosage concentration was fixed at 10 μM, and it was added in combination with silibinin in dosage ratios of 6:1, 4:1, 2:1, 1:1, 1:2, 1:4, 1:6, and 1:8. Similarly, the silibinin dosage concentration was fixed at 10 μM, and it was added in combination with carvedilol in dosage ratios of 6:1, 4:1, 2:1, 1:2, 1:4, 1:6, and 1:8.

[0128] The specific method was the same as in section 2.2 of Experimental Example 3.

[0129] 2.3 Validation using the Duo-Lite™ Luciferase Assay System kit

[0130] The specific method was the same as in section 2.3 of Experimental Example 1.

[0131] 2.4 Analysis of Compound Combination Action Index (CI)

[0132] The specific method was the same as in section 2.4 of Experimental Example 3.

[0133] 3. Experimental Results

[0134] As shown in Figures 5-1 and 5-2, synergistic effects were observed in activated LX-2 cells when carvedilol 10 μM was combined with silibinin in concentration ratios of 2:1, 1:1, 1:2, 1:4, 1:6, and 1:8, and when silibinin 10 μM was combined with carvedilol in concentration ratios of 1:2, 1:4, 1:6, and 1:8. The details of each combination were as follows: Fixed Car: 10 μM, 2:1 (Sil: 5 μM; CI = 0.39); 1:1 (Sil: 10 μM; CI = 0.08); 1:2 (Sil: 20 μM; CI = 0.04); 1:4 (Sil: 40 μM; CI = 0.02); 1:6 (Sil: 60 μM; CI = 0.04); 1:8 (Sil: 80 μM; CI = 0.02); Fixed Sil: 10 μM, 1:2 (Car: 20 μM; CI = 0.86); 1:4 (Car: 40 μM; CI = 0.21); 1:6(Car:60μM;CI=0.01);1:8(Car:80μM;CI=0.30).

[0135] Experimental Example 6: Verification of the effects of combined use of carvedilol and silibinin on liver fibrosis in mice.

[0136] 1. Experimental materials

[0137] Male C57BL / 6J mice (8 weeks old, 20-24g body weight) were purchased from Beijing Weitong Lihua Laboratory Animal Technology Co., Ltd. Carbon tetrachloride (CCl4) was purchased from Shanghai Lingfeng Chemical Reagents Co., Ltd., and mineral oil was purchased from Sigma-Aldrich Corporation (St. Louis, MO, USA).

[0138] The other experimental materials were the same as in Experimental Example 4.

[0139] 2. Experimental Method

[0140] 2.1 Effects of concomitant use of silibinin (100 mg / kg) and carvedilol (2 mg / kg) on ​​CCl4-induced liver fibrosis

[0141] After acclimatizing the animals for one week, the mice were randomly divided into five groups (6 mice per group, 30 mice in total): a control group, a model group, a silibinin monotherapy group, a carvedilol monotherapy group, and a Sil+Car combination therapy group. The model group and the treatment groups received 5 ml / kg of carbon tetrachloride (CCl4, dissolved in mineral oil at a 1:8 ratio) intraperitoneally twice a week for 6 weeks. The control group received the same amount of solvent (mineral oil). Two weeks after the start of model creation, 100 mg / kg of silibinin and 2 mg / kg of carvedilol were orally administered once daily for 4 weeks. After the end of administration, the mice were euthanized, and their livers were collected and preserved.

[0142] 2.2 Effects of concomitant use of silibinin (100 mg / kg) and carvedilol (2 mg / kg) on ​​BDL-induced liver fibrosis

[0143] After acclimatizing the animals for one week, the mice were randomly divided into five groups (6 mice per group, 30 mice in total): a sham surgery group, a BDL surgery model group, a silibinin monotherapy group, a carvedilol monotherapy group, and a Sil+Car combination therapy group. The model and treatment groups were prepared as a cholestatic liver fibrosis model by bile duct ligation (BDL), while the sham surgery group underwent a sham surgery. From postoperative day 1, silibinin 100 mg / kg and carvedilol 2 mg / kg were orally administered once daily for 7 days. After the completion of administration, the mice were euthanized, and their livers were collected and preserved.

[0144] 2.3 Effects of concomitant use of silibinin (75, 100 mg / kg) and carvedilol (2 mg / kg) on ​​MCD-induced diet-induced liver fibrosis (NASH)

[0145] After acclimatizing the animals for one week, the mice were randomly divided into five groups (6 mice per group, 30 mice in total): an MCS control group, an MCD model group, a silibinin monotherapy group, a carvedilol monotherapy group, and a Sil+Car combination therapy group. The MCD model group and the therapy groups were fed a high-lipid MCD model diet for 6 weeks, while the control group was fed an MCS control diet to induce a non-alcoholic steatohepatic fibrosis (NASH) model. Starting 2 weeks after the start of model diet feeding, silibinin 75 or 100 mg / kg and carvedilol 2 mg / kg were orally administered once daily for 4 weeks. After the end of administration, the mice were euthanized, and their livers were collected and preserved.

[0146] 2.4 qRT-PCR (Reverse Transcription Polymerase Chain Reaction)

[0147] Target genes for measurement: Col1a1, Col1a2 (Col3a1)

[0148] The specific method was the same as in section 2.3 of Experimental Example 4.

[0149] 2.5 Liver histopathological analysis

[0150] After fixing a portion of liver tissue with 4% paraformaldehyde, it was sent to Wuhan Guge Biotechnology Co., Ltd. (Wuhan, China) for double-blind analysis using HE staining, Masson staining, and Sirius Red staining. For Masson stained images, the volume fraction (CVF) of collagen was calculated using ImageJ software.

[0151] 3. Experimental Results

[0152] As shown in Figures 6, 7, and 8, qRT-PCR expression analysis of liver tissue fibrosis-related genes (Col1a1, Col1a2 (Col3a1), Mmp2) and liver tissue pathological analysis revealed that in all liver fibrosis models—CCl4-induced, BDL-induced, and MCD-induced—the combination of silibinin and carvedilol (dose ratios: 50:1, 100:2, 75:2) significantly downregulated the mRNA expression levels of Col1a1, Col1a2 (Col3a1), and Mmp2, effectively improving the degree of fibrotic lesions in each liver fibrosis model. Furthermore, the combined effect of both drugs was superior to that of each drug administered alone, and synergistic effects were confirmed in analyses based on the CI values ​​of each indicator.

[0153] Experimental Example 7: Study on the dose-dependent pharmacological effects of a fixed-ratio silibinin and carvedilol composition on liver fibrosis caused by various factors.

[0154] 1. Experimental materials

[0155] The experimental materials were the same as those used in Experimental Example 6.

[0156] 2. Experimental Method

[0157] 2.1 Study on the dose-dependent pharmacological effects of a fixed-ratio silibinin and carvedilol composition on CCl4-induced chemotoxic liver fibrosis.

[0158] After acclimatizing the animals for one week, the mice were randomly divided into eight groups (6 mice per group, 48 mice in total). The groups were: control group, model group, 1.5 mg / kg obeticholic acid (OCA) administration group, 5 mg / kg OCA administration group, 50 mg / kg Sil+1 mg Car composition administration group, 100 mg / kg Sil+2 mg Car composition administration group, 150 mg / kg Sil+3 mg Car composition administration group, and 200 mg / kg Sil+4 mg Car composition administration group. The model group and administration groups were intraperitoneally administered 5 ml / kg of carbon tetrachloride (CCl4, dissolved in mineral oil at a 1:8 ratio) twice a week for 6 weeks. The control group was administered the same amount of solvent (mineral oil). Starting two weeks after the start of model creation, each drug was orally administered once daily for 4 weeks. After the end of administration, the mice were euthanized, and their livers were collected and preserved.

[0159] 2.2 Study on the dose-dependent pharmacological effects of a fixed-ratio silibinin and carvedilol composition on BDL-induced cholestatic hepatic fibrosis.

[0160] After acclimatizing the animals for one week, the mice were randomly divided into nine groups (6 mice per group, 54 mice in total). The groups were: sham surgery group, BDL surgery model group, 120 mg / kg ursodeoxycholic acid (UDCA) administration group, 1.5 mg / kg obeticholic acid (OCA) administration group, 5 mg / kg OCA administration group, 50 mg / kg Sil+1 mg Car composition administration group, 100 mg / kg Sil+2 mg Car composition administration group, 150 mg / kg Sil+3 mg Car composition administration group, and 200 mg / kg Sil+4 mg Car composition administration group. The model group and administration groups were set up as a cholestatic liver fibrosis model by bile duct ligation (BDL), and the sham surgery group underwent sham surgery. Each drug was administered orally once daily for 7 days starting 1 day post-surgery. After administration, the mice were euthanized, and their livers were collected and preserved.

[0161] 2.3 qRT-PCR (Reverse Transcription Polymerase Chain Reaction)

[0162] Target genes for measurement: Col1a1, Col1a2

[0163] The specific method was the same as in section 2.3 of Experimental Example 4.

[0164] 2.4 Liver histopathological analysis

[0165] The specific method was the same as in section 2.5 of Experimental Example 6.

[0166] 3. Experimental Results

[0167] As shown in Figures 9 and 10, expression analysis of fibrosis-related genes (Col1a1, Col1a2) in liver tissue by qRT-PCR experiments and liver tissue pathology analysis revealed that in both CCl4-induced and BDL-induced liver fibrosis models, the fixed-ratio silibinin and carvedilol composition clearly downregulated the mRNA expression levels of Col1a1 and Col1a2, effectively improving the degree of fibrotic lesions in both liver fibrosis models and showing a clear dose-dependent effect. In the CCl4-induced chemotoxic liver fibrosis model, the antifibrotic effect of the fixed-ratio composition consisting of 200 mg / kg silibinin and 4 mg / kg carvedilol was superior to that of 1.5 mg / kg OCA (oveticolic acid) and equivalent to that of 5 mg / kg OCA. In a BDL-induced cholestatic hepatic fibrosis model, the antifibrotic effect of a fixed-ratio composition consisting of 200 mg / kg silibinin and 4 mg / kg carvedilol was superior to that of the positive agents 120 mg / kg UDCA and 1.5 mg / kg OCA, and equivalent to that of 5 mg / kg OCA.

[0168] Example 1

[0169] The composition of the silibinin carvedilol composition is as follows: [Table 2]

[0170] Preparation method: Silibinin, carvedilol, and soybean phospholipid were dissolved in anhydrous ethanol, and this was mixed with lactose and carboxymethyl starch sodium as a binder. After granulation, drying, and particle sizing, it was uniformly mixed with talc and filled into capsules to form the formulation.

[0171] Example 2

[0172] The composition of the silibinin carvedilol composition is as follows: [Table 3]

[0173] Preparation method: Silibinin, carvedilol, and soybean phospholipid were dissolved in anhydrous ethanol, and this was mixed with lactose and carboxymethyl starch sodium as a binder. After granulation, drying, and particle sizing, it was uniformly mixed with talc and filled into capsules to form the formulation.

Claims

1. A silibinin-containing composition characterized by containing silibinin and carvedilol.

2. a) A silibinin-containing composition according to claim 1, characterized in that it contains silibinin and carvedilol as active ingredients, and b) contains pharmaceutically acceptable additives suitable for the preparation of a solid oral dosage form, wherein the mass ratio of silibinin to carvedilol is 0.1 to 200:

1.

3. The silibinin-containing composition according to claim 2, characterized in that the mass ratio of silibinin to carvedilol among the aforementioned active ingredients is 1 to 50:

1.

4. The silibinin-containing composition according to claim 2, characterized in that the mass ratio of silibinin to carvedilol among the aforementioned active ingredients is 2 to 50:

1.

5. The silibinin-containing composition according to claim 2, characterized in that the mass ratio of silibinin to carvedilol among the aforementioned active ingredients is 1 to 8:

1.

6. The silibinin-containing composition according to claim 2, characterized in that the mass ratio of silibinin to carvedilol among the aforementioned active ingredients is 2 to 8:

1.

7. The silibinin-containing composition according to claim 2, characterized in that the mass ratio of silibinin to carvedilol among the above active ingredients is 37.5 to 50:

1.

8. The silibinin-containing composition according to claim 2, characterized in that the mass ratio of silibinin to carvedilol among the aforementioned active ingredients is 50:

1.

9. The silibinin-containing composition according to claim 1, characterized in that the carvedilol comprises a pharmaceutically acceptable salt thereof made of an organic acid or an inorganic acid.

10. The silibinin-containing composition according to claim 9, characterized in that the organic acid includes oxalic acid, fumaric acid, benzoic acid, and mandelic acid, and the inorganic acid includes phosphoric acid.

11. A silibinin-containing composition characterized by comprising (a) silibinin and (b) a nonsteroidal anti-inflammatory drug, an angiotensin inhibitor, a calcium channel blocker, a glucocorticoid, or a diuretic.

12. The silibinin-containing composition according to claim 11, characterized in that the nonsteroidal anti-inflammatory drug is selected from the group consisting of aceclofenac and clobetasol propionic acid, the angiotensin inhibitor is valsartan, the calcium channel blocker is selected from the group consisting of nicardipine, manidipine, amlodipine and cilnidipine, the glucocorticoid is desonide, and the diuretic is selected from the group consisting of tamsulosin and D-mannitol.

13. Use of the silibinin-containing composition according to claims 1 to 12 in the preparation of a therapeutic agent for liver fibrosis.

14. Use of the silibinin-containing composition according to claims 1 to 12 in the preparation of a therapeutic drug for chronic liver disease.

15. The use according to claim 13, characterized in that the use is used in the preparation of a therapeutic drug for hepatic fibrosis caused by chronic liver disease.

16. The use according to claim 13, characterized in that it is used in the preparation of therapeutic drugs for chemical liver injury, cholestatic liver disease, and non-alcoholic fatty liver disease and resulting liver fibrosis.

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