Application of Alistipes putredinis in preparation of medicine for treating non-alcoholic fatty liver disease

Abstract

The invention discloses an application of Alistipes putredinis in preparation of a medicine for treating a non-alcoholic fatty liver disease. A non-alcoholic fatty liver disease animal model commonly used in the field proves that the Alistipes putredins ATCC 29800 can effectively improve the body weight, liver weight, glucose tolerance and insulin tolerance of a mouse with the non-alcoholic fatty liver disease, effectively reduce the liver lipid accumulation of the mouse with the non-alcoholic fatty liver disease, and improve the immunity of the mouse with the non-alcoholic fatty liver disease. The blood fat and the liver fat of a mouse with the non-alcoholic fatty liver disease are effectively reduced, and the liver function of the mouse with the non-alcoholic fatty liver disease is improved. Therefore, the Alistipes putredinis strain (such as Alistipes putredinis ATCC 29800) or the culture solution of the Alistipes putredinis strain has the prospect of being developed into the medicine for treating the non-alcoholic fatty liver disease.

Description

Technical Field

[0001] The present invention belongs to the field of medicine and relates to new uses of known bacterial species, and specifically relates to the use of Alistipes putredinis in preparing a drug for treating non-alcoholic fatty liver disease. Background Art

[0002] Nonalcoholic fatty liver disease (NAFLD) is a clinical and pathological syndrome characterized by excessive fat deposition in hepatocytes after the exclusion of alcohol and other clear liver-damaging factors. It is an acquired metabolic stress-induced liver injury closely related to insulin resistance and genetic susceptibility. It includes simple fatty liver (SFL), nonalcoholic steatohepatitis (NASH), and its related cirrhosis.

[0003] In addition to directly leading to decompensated cirrhosis, hepatocellular carcinoma, and recurrence of transplanted livers, nonalcoholic fatty liver disease (NAFLD) can also influence the progression of other chronic liver diseases and contribute to the pathogenesis of type 2 diabetes and atherosclerosis. Metabolic syndrome-related malignancies, arteriosclerotic cardiovascular and cerebrovascular diseases, and cirrhosis are important factors affecting the quality of life and life expectancy of patients with NAFLD. Therefore, NAFLD presents a new challenge in contemporary medicine, and its harm to human health will continue to increase.

[0004] The treatment of non-alcoholic fatty liver disease mainly focuses on improving lifestyle and drug therapy. Improving lifestyle includes controlling diet, reducing the intake of high-calorie, high-fat, and high-sugar foods, increasing dietary fiber intake, and moderately increasing exercise. Strict dietary control and regular exercise plans may impose certain restrictions on patients' daily lives, affecting the convenience and comfort of life, such as limited choices when dining out. Drug treatment has certain side effects, and different drugs have different side effects. For example, some drugs may affect gastrointestinal function, causing symptoms such as nausea, vomiting, and diarrhea.

[0005] In order to develop drugs for treating non-alcoholic fatty liver disease, the present invention is proposed. Summary of the Invention

[0006] The present invention aims to provide use of the Alistipes putredinis strain or its culture solution in preparing a medicament for treating non-alcoholic fatty liver disease.

[0007] The above-mentioned purpose of the present invention is achieved through the following technical solutions:

[0008] Application of the Alistipes putredinis strain or its culture solution in preparing a medicine for treating non-alcoholic fatty liver disease.

[0009] Preferably, the Alistipes putredinis is Alistipes putredinis ATCC 29800.

[0010] Preferably, the drug uses the strain or its culture solution as an active ingredient and is prepared into a pharmaceutically acceptable dosage form with a pharmaceutically acceptable carrier or excipient.

[0011] More preferably, the carrier or excipient is solid, liquid or semisolid.

[0012] More preferably, the dosage form is a tablet, capsule, injection or pill.

[0013] Beneficial effects:

[0014] In a specific embodiment of the present invention, using a non-alcoholic fatty liver disease cell model commonly used in the art, Alistipes putredinis ATCC 29800 demonstrated a more significant therapeutic improvement in non-alcoholic fatty liver disease than Alistipes onderdonkii ATCCBAA-1178 and Alistipes finegoldii DSM17242, both members of the same genus. The former demonstrated approximately 100% higher improvement in the markers of the non-alcoholic fatty liver disease model group than the latter two strains. Those skilled in the art are aware that Alistipes is a relatively new bacterial genus, primarily isolated from the human intestinal microbiome. Bacteria of this genus not only participate in the development and progression of various diseases but also have protective effects against certain diseases. It is generally believed that Alistipes finegoldii and Alistipes onderdonkii of the genus Alistipes have better activity, but experiments in the present invention have shown that the Alistipes putredinis strain Alistipes putredinis ATCC 29800 of this genus has a significantly better therapeutic effect on non-alcoholic fatty liver disease, and the improvement rate of landmark indicators far exceeds that of Alistipes finegoldii strain Alistipes finegoldii DSM17242 and Alistipes onderdonkii strain Alistipes onderdonkii ATCC BAA-1178. This technical effect is unexpected.

[0015] In a specific embodiment of the present invention, an animal model of non-alcoholic fatty liver disease commonly used in the art was used to demonstrate that Alistipes putredinis ATCC 29800 can effectively improve the body weight, liver weight, glucose tolerance, and insulin tolerance of mice with non-alcoholic fatty liver disease, effectively reduce hepatic lipid accumulation in mice with non-alcoholic fatty liver disease, effectively reduce blood lipids and liver lipids in mice with non-alcoholic fatty liver disease, and improve liver function in mice with non-alcoholic fatty liver disease.

[0016] Therefore, the Alistipes putredinis strain (such as Alistipes putredinis ATCC 29800) or its culture fluid has the prospect of being developed into a drug for treating non-alcoholic fatty liver disease. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] Figure 1 is the TG content in HepG2 cells of each group;

[0018] Figure 2 Figure 2 shows the body weight changes (A) and liver weight (B) of mice.

[0019] Figure 3 The OGTT curve of mice (A) and the area under the curve (B); the ITT curve of mice (C) and the area under the curve (D);

[0020] Figure 4 H&E staining and Oil Red O staining of liver tissues of mice in each group;

[0021] Figure 5 The levels of TG, TC, LDL, ALT, AST in serum and TG and TC in liver of mice in each group. DETAILED DESCRIPTION

[0022] The essential contents of the present invention are described in detail below with reference to the embodiments, but the protection scope of the present invention is not limited thereto.

[0023] 1. Experimental Materials

[0024] 1. Experimental cells

[0025] Human HepG2 cell line was purchased from the Shanghai Institute of Cell Biology, Chinese Academy of Sciences and cryopreserved in our laboratory.

[0026] 2. Experimental Animals

[0027] Male C57BL / 6J mice, 6 weeks old, weighing approximately 19-21 g, source: Jiangsu Jicui Pharmaceutical Biotechnology Co., Ltd. (SCXK(Su)2023-0009).

[0028] 3. Animal feed

[0029] Ordinary feed was purchased from Jiangsu Collaborative Pharmaceutical Bioengineering Co., Ltd.; high-fat feed (US Research Diets, D12492, fat content 60%).

[0030] 4. Experimental reagents

[0031] Alistipes putredinis ATCC 29800 was purchased from the American Type Culture Collection (ATCC);

[0032] Alistipes onderdonkiiATCC BAA-1178 was purchased from the American Type Culture Collection (ATCC);

[0033] Alistipes finegoldii DSM17242 was purchased from the German DSMZ Culture Collection Management Center;

[0034] minced meat carbohydrate broth (Mingzhou Biotechnology, KDM150);

[0035] Cooked beef cubes (Shandong Top Biological, S5604);

[0036] Hemin (Shandong Top Biological, S5605);

[0037] Vitamin K1 (Shandong Top Biological, SD018);

[0038] Fetal bovine serum (GIBCO, USA, 10270-106);

[0039] 0.25% EDTA-pancreatin (GIBCO, USA, 252000072);

[0040] High-glucose DMEM medium (Keygene Biotechnology, KGMI12800-500);

[0041] Oleic acid (Sigma, O1008, USA);

[0042] Palmitic acid (Sigma, P0500, USA);

[0043] PBS powder (Nanjing Novozymes, G101);

[0044] D-Glucose (Beijing Solaibao Company, G8150);

[0045] Insulin powder (Biosharp, BS901);

[0046] Sodium chloride injection (Double Crane Pharmaceuticals);

[0047] Universal tissue fixative (Wuhan Sewell Biotechnology Co., Ltd.);

[0048] Total cholesterol (T-CHO) detection kit (Nanjing Jiancheng Biological Co., Ltd., A111-1-1);

[0049] Triglyceride (TG) detection kit (Nanjing Jiancheng Biological Co., Ltd., A110-1-1);

[0050] Low-density lipoprotein cholesterol (LDL-C) test kit (Nanjing Jiancheng Biological Co., Ltd., A113-1-1);

[0051] Alanine aminotransferase (ALT) test kit (Nanjing Jiancheng Biological Co., Ltd., C009-2-1);

[0052] Aspartate aminotransferase (AST) test kit (Nanjing Jiancheng Biological Co., Ltd., C010-2-1);

[0053] Omron blood glucose meter (HGM-114) and matching test strips (AS1).

[0054] 2. Experimental Methods

[0055] 1. Preparation of solution

[0056] Preparation of minced meat carbohydrate liquid culture medium: Weigh 57.5 g minced meat carbohydrate broth powder into 1000 mL of purified water, heat to boil, and dispense into test tubes (7 mL / tube). Add 3-4 pieces of cooked beef cubes to each tube and add 0.5 mg of hemin chloride and 5 mg of vitamin K1 per 100 mL of each tube. Sterilize at 121°C for 15 min, cool quickly, and place in an anaerobic incubator until used.

[0057] Preparation of sodium oleate (OA) solution: weigh 117.4 mg of oleic acid and add 20 mL of 0.1 mM NaOH solution. React at 70°C until clear. Add to 20 mL of 4% BSA solution to prepare 10 mM OA stock solution and store at 4°C.

[0058] Preparation of sodium palmitate (PA) solution: weigh 53.3 mg of palmitic acid and add 20 mL of 0.1 mM NaOH solution. React at 70°C until clear. Add to 20 mL of 4% BSA solution to prepare 5 mM PA stock solution and store at 4°C.

[0059] Glucose solution: Weigh glucose powder and add purified water to dissolve it to a final concentration of 0.2 g / mL glucose gavage solution for OGTT test.

[0060] Insulin solution: Accurately weigh 1.00 mg of insulin powder and add 1.00 mL of normal saline. Vortex for 1 min, place in an ice-water bath and sonicate for 15 min. Pipette 146 μL of the solution and add normal saline to make up to 50 mL to obtain the insulin injection solution for ITT test.

[0061] 2. Bacterial culture and sample preparation

[0062] (1) Bacterial culture

[0063] Alistipes putredinis ATCC 29800, Alistipes onderdonkii ATCC BAA-1178, and Alistipes finegoldii DSM17242 were cultured in minced meat carbohydrate liquid medium in an anaerobic incubator containing 10% CO2, 10% H2, and 80% N2.

[0064] (2) Sample preparation

[0065] Preparation of cell experiment samples: The growth curves of each bacteria were determined by preliminary experiments, and the mid-exponential harvest point was determined. Then, the cells were harvested at the same initial bacterial density (1×10 8 CFU / mL) was inoculated and cultured. The strain was collected when the target OD value was ±10%, and centrifuged at 13,000 rpm and 4°C for 10 min. The total colony count corresponding to each supernatant was determined after centrifugation. The supernatant was diluted / concentrated according to the CFU ratio to an equivalent bacterial secretion amount. The supernatant with an equivalent bacterial secretion amount was then filtered through a 0.22 μm sterile filter membrane to remove residual bacteria and particulate matter before use for cell administration.

[0066] Animal experiment sample preparation: Bacteria were collected in the logarithmic phase, centrifuged at 13000 rpm and 20°C for 10 min, and diluted to 2×10 8 CFU / mL was used for gavage.

[0067] 3. Effects on non-alcoholic fatty liver disease cell models

[0068] (1) Cell culture

[0069] HepG2 cells were cultured in DEME high-glucose medium (penicillin 100 U / mL, streptomycin 100 μg / mL) containing dual-antibody antibodies, followed by addition of inactivated fetal bovine serum (FBS) to a complete medium containing 15% FBS. The cells were incubated at 37°C in a 5% CO2 incubator and passaged using 0.25% trypsin when they reached 70% to 80% confluency.

[0070] (2) Establishment of non-alcoholic fatty liver disease cell model, grouped drug administration, and determination of markers

[0071] HepG2 cells were plated in 6-well plates (about 8 × 10 cells per well). 5 cells), cultured in 15% FBS DMEM high-glucose medium containing double antibodies, and administered the next day. A control group (replaced with normal medium containing 15% FBS), a model group (replaced with 15% FBS medium containing 400 μM OA, 200 μM PA, and 20% minced meat carbohydrate culture solution), an Ao-administered group (replaced with 15% FBS medium containing 400 μM OA, 200 μM PA, and 20% Alistipes onderdonkii supernatant), an Ap-administered group (replaced with 15% FBS medium containing 400 μM OA, 200 μM PA, and 20% Alistipes putredinis supernatant), and an Af-administered group (replaced with 15% FBS medium containing 400 μM OA, 200 μM PA, and 20% Alistipes finegoldii supernatant) were set up. 24 hours after administration, discard the culture medium and rinse twice with PBS buffer. Trypsinize for 2 minutes, then pipette cells with PBS and transfer to a 1.5 ml centrifuge tube. Centrifuge at 1000 rpm for 5 minutes, discard the supernatant, and retain the cell pellet. Homogenize the cell pellet with 0.2 ml of PBS and disrupt with ultrasound in an ice-water bath. The homogenate was then assayed for TG content according to the TG kit instructions without centrifugation.

[0072] 4. Effects on animal models of non-alcoholic fatty liver disease

[0073] (1) Establishment of animal model of nonalcoholic fatty liver disease and group-based drug administration

[0074] After one week of adaptive feeding, mice were randomly divided into two groups: a control group (Chow, n=8) was fed a normal diet, while a model group (HFD, n=8) and an Ap administration group (HFD+Ap, n=8) were fed a high-fat diet for 8 weeks. Mice in the HFD+Ap group were gavaged daily with Alistipes putredinis ATCC 29800 dissolved in sterile PBS, while mice in the Chow and HFD groups were gavaged daily with an equal volume of PBS. During the daily dosing period, mice had unlimited access to water and food intake, and their body weights were recorded weekly.

[0075] (2) Oral glucose tolerance test (OGTT) and insulin tolerance test (ITT)

[0076] An OGTT test was performed after 8 weeks of dosing. After fasting for 12 hours, the mice in each group had their blood glucose levels measured at 0 minutes. Subsequently, they were gavaged with a 2g / kg glucose solution, and their blood glucose levels were measured at 15, 30, 60, 90, and 120 minutes. A line graph was plotted with time on the X-axis and blood glucose on the Y-axis, and the area under the curve (AUC) was calculated.

[0077] An ITT study was performed 8 weeks after administration. After fasting for 4 hours, blood glucose levels were measured at 0 minutes. Each mouse was then intraperitoneally injected with insulin solution (0.75 U / kg). Blood glucose levels were measured at 15, 30, 60, 90, and 120 minutes. A line graph was plotted with time as the X-axis and blood glucose as the Y-axis, and the AUC was calculated.

[0078] (3) Collection and processing of samples at the end of animal experiments

[0079] After 8 weeks of dosing, various indicators were measured. After the mice stabilized for 3 days, they were fasted for 12 hours. Blood was collected by removing the eyeballs and allowed to stand at room temperature for 4 hours. The cells were then centrifuged at 5000 rpm for 10 minutes at 4°C. The supernatant was collected and aliquoted for later use (stored at -80°C). The liver tissue was removed and weighed. After washing with saline, tissue from three mice in each group was fixed with a universal tissue fixative. The remaining tissue was quickly frozen in liquid nitrogen and then transferred to -80°C for storage.

[0080] (4) Observation of liver tissue pathology

[0081] Liver samples were fixed in 4% PFA and embedded in paraffin. The embedded sections were stained with hematoxylin-eosin (H&E). Some frozen liver tissue was stained with Oil Red O to observe lipid accumulation in the liver.

[0082] (5) Detection of biochemical indicators in mouse serum and liver

[0083] The kits were used to determine total cholesterol (TC), triglycerides (TG), low-density lipoprotein cholesterol (LDL-C), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in serum as well as total cholesterol (TC) and triglycerides (TG) in liver according to the instructions.

[0084] (6) Data analysis methods

[0085] Statistical analysis was performed using GraphPad Prism 9.5.0 software, and all experimental data are expressed as mean ± SEM. One-way ANOVA was used to compare differences between multiple groups, with P < 0.05 indicating statistical significance (P < 0.05 is marked with "*"; P < 0.01 is marked with "**"; and P < 0.001 is marked with "***").

[0086] 3. Experimental Results

[0087] 1. Improvement of non-alcoholic fatty liver disease cell model

[0088] The experimental results are shown in Figure 1 and Table 1. Compared with the control group, the TG content in the model group cells was significantly increased, indicating successful model establishment. Compared with the model group, the TG content in the groups treated with supernatants from Alistipes putredinis ATCC 29800, Alistipes onderdonkii ATCC BAA-1178, and Alistipes finegoldii DSM 17242 bacteria was significantly decreased. The supernatant from Alistipes putredinis ATCC 29800 showed the greatest improvement, with the improvement rate of the marker relative to the model group approximately 100% higher than that of the other two strains. Therefore, Alistipes putredinis ATCC 29800 was selected for subsequent experiments.

[0089] Table 1 TG content in HepG2 cells in each group and improvement rate of the drug-treated group

[0090] Group TG content (mmol / gprot) Improvement rate of landmark indicators relative to the model group Blank group 0.079±0.006*** / Model Group 0.367±0.013 / Drug administration group (Ao bacteria supernatant) 0.300±0.026** 18.3％ Drug administration group (Ap bacteria supernatant) 0.206±0.008*** 43.9％ Drug administration group (Af bacteria supernatant) 0.281±0.034** 23.4％

[0091] 2. Improvement of body weight and liver weight in mice with non-alcoholic fatty liver disease

[0092] The experimental results are shown in Figure 2 and Table 2. Compared with the control group, the body weight and liver weight of the mice in the model group were significantly increased, indicating that the model was successfully established; compared with the model group, the body weight and liver weight of the mice in the treatment group were significantly decreased, indicating that Alistipes putredinis ATCC29800 can significantly improve the body weight and liver weight of mice with non-alcoholic fatty liver disease.

[0093] Table 2 Body weight and liver weight of mice in each group after 8 weeks of drug administration

[0094] Group Weight (g) Liver weight (g) Control group (Chow) 25.68±0.38*** 0.83±0.03*** Model group (HFD) 35.54±1.98 0.90±0.07 Drug group (HFD+Ap) 31.04±2.51*** 0.77±0.03***

[0095] 3. Improvement of oral glucose tolerance and insulin tolerance in mice with non-alcoholic fatty liver disease

[0096] The results are shown in Tables 3, 4 and Figure 3 As shown in the figure, compared with the control group, the areas under the curves of oral glucose tolerance and insulin tolerance of the mice in the model group were significantly increased, indicating that the model was successfully established; compared with the model group, the areas under the curves of oral glucose tolerance and insulin tolerance of the mice in the drug-treated group were significantly decreased, indicating that Alistipes putredinis ATCC 29800 can significantly improve the glucose tolerance and insulin tolerance of mice with non-alcoholic fatty liver disease.

[0097] Table 3 OGTT curves and area under the curves of mice in each group

[0098]

[0099]

[0100] Table 4 Statistics of ITT curves and area under the curves of mice in each group

[0101]

[0102] 4. Protective effect on liver tissue in mice with non-alcoholic fatty liver disease

[0103] The results of H&E staining of liver tissue are shown in Figure 4 Compared with the control group (Chow), the model group (HFD) mice had a large number of vacuoles and obvious liver tissue damage, indicating that the model was successfully established; compared with the model group (HFD), the proportion of liver vacuoles in the drug-treated group (HFD+Ap) mice was significantly reduced, and liver damage was improved, indicating that Alistipes putredinis ATCC 29800 can significantly improve liver damage in mice with non-alcoholic fatty liver disease. The results of Oil Red O staining of liver tissue are shown in Figure 4 Compared with the control group (Chow), a large amount of fat accumulation appeared in the liver cells of the model group (HFD) mice, indicating that the model was successful; compared with the model group (HFD), the fat accumulation in the liver cells of the drug-treated group (HFD+Ap) mice was significantly reduced, indicating that Alistipes putredinis ATCC 29800 can significantly reduce the liver lipid accumulation in mice with non-alcoholic fatty liver disease.

[0104] 5. Improvement of blood lipids, liver lipids and liver function in mice with non-alcoholic fatty liver disease

[0105] The results of blood lipid level test in mouse serum samples are shown in Table 5, Table 6, Table 7 and Figure 5As shown in the figure, compared with the control group (Chow), the levels of TG, TC, LDL, ALT, and AST in the serum of the mice in the model group (HFD) were significantly increased, indicating that the model was successfully established; compared with the model group (HFD), the levels of TG, TC, LDL, ALT, and AST in the serum of the mice in the drug administration group (HFD+Ap) were significantly decreased, indicating that Alistipes putredinis ATCC 29800 can significantly reduce blood lipids and liver lipids in mice with non-alcoholic fatty liver disease and improve liver function in mice with non-alcoholic fatty liver disease.

[0106] Table 5 Blood lipid levels in serum of mice in each group

[0107]

[0108] Table 6 ALT and AST levels in the serum of mice in each group

[0109] Group Alanine aminotransferase (U / L) Aspartate aminotransferase (U / L) Control group (Chow) 7.20±2.12*** 15.74±2.92*** Model group (HFD) 11.62±2.90 21.11±2.97 Drug group (HFD+Ap) 6.30±2.47*** 15.17±2.15***

[0110] Table 7 TG and TC levels in the liver of mice in each group

[0111] Group Total triglycerides (mmol / gprot) Total cholesterol (mmol / gprot) Control group (Chow) 11.84±1.17*** 3.63±0.40*** Model group (HFD) 17.13±1.84 5.30±0.56 Drug group (HFD+Ap) 12.03±1.63*** 4.32±0.36***

[0112] In summary:

[0113] The above experiments, using a non-alcoholic fatty liver disease cell model commonly used in the art, demonstrated that Alistipes putredinis ATCC 29800 exhibited a more significant therapeutic improvement in non-alcoholic fatty liver disease than Alistipes onderdonkii ATCC BAA-1178 and Alistipes finegoldii DSM17242, both of the same genus Alistipes. The former demonstrated approximately 100% higher improvement in the markers of the non-alcoholic fatty liver disease model group than the latter two strains. Those skilled in the art are aware that Alistipes is a relatively new bacterial genus, primarily isolated from the human intestinal microbiome. Bacteria of this genus not only participate in the development and progression of various diseases but also have protective effects against certain diseases. It is generally believed that Alistipes finegoldii and Alistipes onderdonkii of the genus Alistipes have better activity, but the above experiments have proved that the Alistipes putredinis strain Alistipes putredinis ATCC 29800 of this genus has a significantly better therapeutic effect on non-alcoholic fatty liver disease, and the improvement rate of landmark indicators far exceeds that of Alistipes finegoldii strain Alistipes finegoldii DSM17242 and Alistipes onderdonkii strain Alistipes onderdonkii ATCC BAA-1178. This technical effect is unexpected.

[0114] The above experiments demonstrated, using an animal model of non-alcoholic fatty liver disease commonly used in the art, that Alistipesputredinis ATCC 29800 can effectively improve the body weight, liver weight, glucose tolerance, and insulin tolerance of mice with non-alcoholic fatty liver disease, effectively reduce hepatic lipid accumulation in mice with non-alcoholic fatty liver disease, effectively reduce blood lipids and liver lipids in mice with non-alcoholic fatty liver disease, and improve liver function in mice with non-alcoholic fatty liver disease.

[0115] Therefore, the Alistipes putredinis strain (such as Alistipes putredinis ATCC 29800) or its culture fluid has the prospect of being developed into a drug for treating non-alcoholic fatty liver disease.

[0116] The purpose of the above embodiments is to specifically introduce the essential content of the present invention, but those skilled in the art should know that the protection scope of the present invention should not be limited to this specific embodiment.

Claims

1. Use of the Alistipes putredinis strain or its culture medium in the preparation of a drug for treating non-alcoholic fatty liver disease.

2. The use according to claim 1, wherein Alistipes putredinis is Alistipes putredinis ATCC 29800.

3. The use according to claim 1 or 2, wherein the drug uses the strain or its culture solution as an active ingredient and is prepared into a pharmaceutically acceptable dosage form with a pharmaceutically acceptable carrier or excipient.

4. The use according to claim 3, characterized in that: The carrier or auxiliary material is solid, liquid or semi-solid.

5. The use according to claim 3, characterized in that: The dosage form is tablet, capsule, injection or pill.

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