Application of mannose in preparation of medicine for preventing and / or treating non-alcoholic fatty liver disease

Abstract

The invention discloses an application of mannose in preparation of a medicine for preventing and / or treating non-alcoholic fatty liver disease, the mannose provides a new, safe and effective medicine choice for treatment of NAFLD (non-alcoholic fatty liver disease) by regulating liver lipid metabolism and reducing liver lipid accumulation, especially in prevention and treatment of NASH (non-alcoholic fatty liver disease); besides, mannose has a remarkable effect on preventing and treating NAFLD, can control weight gain, improve liver fatty degeneration and reduce serum transaminase and blood lipid level, and has good safety. According to the application, the action mechanism of mannose is systematically researched, the application range of mannose in liver metabolic diseases is expanded, the application has important clinical significance and application prospects, a new thought and a new method are provided for treatment of NAFLD, and the blank of existing research is filled.

Description

Technical Field

[0001] The present invention relates to the technical field of new drug applications, and in particular to an application of mannose in the preparation of a drug for preventing and / or treating non-alcoholic fatty liver disease. Background Art

[0002] Non-alcoholic fatty liver disease (NAFLD) is a common chronic liver disease that is often associated with metabolic diseases such as obesity, hypertension, dyslipidemia and type 2 diabetes. Its main feature is the abnormal accumulation of lipids in the liver. This lipid accumulation not only affects liver function, but can also lead to more serious complications, including non-alcoholic steatohepatitis (NASH), liver fibrosis, and even cirrhosis and liver cancer (HCC). According to statistics, the global prevalence of NAFLD has reached 20% to 30%, and continues to grow with the prevalence of modern high-sugar and high-fat diets.

[0003] The pathogenesis of NAFLD is quite complex, involving the interaction of multiple factors such as environment, genetics and metabolism. Among them, abnormal accumulation of liver lipids is considered to be the key pathological feature of the disease. When lipid production in the liver exceeds its metabolic capacity, excessive triglycerides and fatty acids will accumulate in hepatocytes, leading to fatty degeneration. In this process, the production of toxic lipids triggers oxidative stress and inflammatory responses, which in turn accelerates the progression of NAFLD.

[0004] Although the prevalence of NAFLD is increasing, there is still a lack of effective drug treatment options. At this stage, it mainly relies on lifestyle adjustments, such as increasing exercise and improving diet, but these measures are difficult to promote in practice and have limited effects. There is a lack of effective therapeutic drugs in the clinic, and existing experimental drugs have certain side effects. Therefore, there is an urgent need to explore new low-toxic therapeutic drugs to effectively prevent and treat NAFLD.

[0005] Mannose, as a common monosaccharide, has attracted the attention of researchers in recent years. Studies have shown that mannose may have a regulatory effect on lipid metabolism and can improve the metabolic state of cells. Although the potential of mannose in other metabolic diseases has been preliminarily recognized, there is currently no research on its application in the prevention and treatment of NAFLD, especially in the prevention and treatment of NASH. Summary of the invention

[0006] The purpose of the present invention is to disclose the use of mannose in preparing a drug for preventing and / or treating non-alcoholic fatty liver disease in response to the above technical problems.

[0007] In order to achieve the above-mentioned object of the invention, the present invention provides the use of mannose in preparing a drug for preventing and / or treating non-alcoholic fatty liver disease; the drug contains mannose as the only effective active ingredient;

[0008] The non-alcoholic fatty liver disease is non-alcoholic steatohepatitis or non-alcoholic fatty liver.

[0009] Optionally, the non-alcoholic fatty liver disease is non-alcoholic fatty liver disease induced by a high-fat diet.

[0010] Optionally, the mechanism of preventing or treating nonalcoholic fatty liver disease includes one or more of the following: reducing lipid accumulation, reducing liver damage, reducing serum aspartate aminotransferase and alanine aminotransferase levels, or reducing liver triglyceride content.

[0011] Optionally, the drug is administered orally or by drinking water.

[0012] Optionally, mannose is administered in the form of an aqueous solution, and the mass concentration of the mannose aqueous solution is 5% to 40%.

[0013] Optionally, the mass concentration of the mannose aqueous solution is 15% to 25%.

[0014] The present invention also provides the use of a pharmaceutical composition containing mannose in preparing a drug for preventing and / or treating non-alcoholic fatty liver disease.

[0015] Optionally, the pharmaceutical composition is prepared by using mannose as an active ingredient and adding pharmaceutically acceptable excipients.

[0016] Optionally, the drug is in the form of an oral preparation.

[0017] Optionally, the drug is in the form of tablets, capsules, granules or powders.

[0018] Compared with the prior art, the present invention has the following beneficial effects:

[0019] The present invention discovered through metabolomics analysis that mannitol is closely related to the risk of NAFLD disease, and aims to explore the application of mannose in the preparation of drugs for preventing or treating NAFLD. More importantly, the present invention also explores the application of mannose in the preparation of drugs for preventing or treating NASH, providing new ideas and methods for the treatment of NAFLD and filling the gaps in existing research. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] Figure 1A This is a graph showing the body weight changes of NAFLD mice induced by oral administration of mannose in Example 1 on a 60% HFD diet for 12 weeks;

[0021] Figure 1B This is a graph showing the ratio of liver wet weight to body weight of the NAFLD mice induced by oral administration of mannose in Example 1 for 8 and 12 weeks on a 60% HFD diet;

[0022] Figure 1C This is a graph showing the liver steatosis scores of NAFLD mice induced by oral administration of mannose in Example 1 for 8 and 12 weeks on a 60% HFD diet;

[0023] Figure 1D Figure 1 shows the effect of oral administration of mannose on the prevention of NAFLD in mice induced by 60% HFD diet for 8 and 12 weeks by HE (Hematoxylin and Eosin staining) and Oil Red staining of liver tissue;

[0024] Figure 2A This is a graph showing the prevention of the plasma ALT (Alanine Aminotransferase) level in NAFLD mice induced by 60% HFD diet for 8 and 12 weeks in Example 1 by oral administration of mannose;

[0025] Figure 2B This is a graph showing the effect of oral administration of mannose on the prevention of plasma AST (Aspartate Aminotransferase) levels in NAFLD mice induced by 60% HFD diet for 8 and 12 weeks in Example 1;

[0026] Figure 2C This is a graph showing the effect of oral administration of mannose on the prevention of plasma ALP (Alkaline Phosphatase) levels in NAFLD mice induced by 60% HFD diet for 8 and 12 weeks in Example 1;

[0027] Figure 2D The graph shows the effect of oral administration of mannose on the plasma TG (triglycerides) level in NAFLD mice induced by 60% HFD diet for 8 and 12 weeks in Example 1;

[0028] Figure 2E This is a graph showing the effect of oral administration of mannose on the prevention of plasma TC (Total Cholesterol) levels in NAFLD mice induced by 60% HFD diet for 8 and 12 weeks in Example 1;

[0029] Figure 2FThis is a graph showing the effect of oral administration of mannose on the prevention of plasma LDL-C (Low-Density Lipoprotein Cholesterol) levels in NAFLD mice induced by 60% HFD diet for 8 and 12 weeks in Example 1;

[0030] Figure 3A This is a graph showing the weight changes of NAFLD mice induced by 60% HFD diet for 12 weeks after 4 weeks of oral administration of mannose in Example 2;

[0031] Figure 3B This is a graph showing the ratio of liver wet weight to body weight of NAFLD mice treated with oral mannose for 4 weeks and induced by 60% HFD diet for 12 weeks in Example 2;

[0032] Figure 3C This is a graph showing the liver fattening score of NAFLD mice induced by 60% HFD diet for 12 weeks after oral administration of mannose for 4 weeks in Example 2;

[0033] Figure 3D HE staining and Oil Red staining of liver tissue of NAFLD mice induced by 60% HFD diet for 12 weeks after oral administration of mannose for 4 weeks in Example 2;

[0034] Figure 4A This is a graph showing the plasma ALT level of NAFLD mice induced by 60% HFD diet for 12 weeks after oral administration of mannose for 4 weeks in Example 2;

[0035] Figure 4B This is a graph showing the plasma AST level of NAFLD mice induced by 60% HFD diet for 12 weeks after oral administration of mannose for 4 weeks in Example 2;

[0036] Figure 4C This is a graph showing the plasma ALP level of NAFLD mice induced by 60% HFD diet for 12 weeks after oral administration of mannose for 4 weeks in Example 2;

[0037] Figure 4D This is a graph showing the plasma TG levels of NAFLD mice induced by 60% HFD diet for 12 weeks after oral administration of mannose for 4 weeks in Example 2;

[0038] Figure 4E This is a graph showing the plasma TC level of NAFLD mice induced by 60% HFD diet for 12 weeks after oral administration of mannose for 4 weeks in Example 2;

[0039] Figure 4FThis is a graph showing the effect of oral administration of mannose for 4 weeks on the plasma LDL-C level in NAFLD mice induced by 60% HFD diet for 12 weeks in Example 2;

[0040] Figure 5A This is a graph showing the body weight changes of NASH mice induced by oral administration of mannose in Example 3 on a CL-HFS ​​(choline-low high-fat and high-sugar diet) diet for 8 weeks;

[0041] Figure 5B This is a graph showing the ratio of liver wet weight to body weight of mannose administered orally to prevent NASH mice induced by CL-HFS ​​diet for 8 weeks in Example 3;

[0042] Figure 5C HE staining and Oil Red staining of liver tissue of NASH mice induced by CL-HFS ​​diet for 8 weeks are shown in Example 3. DETAILED DESCRIPTION

[0043] The use of mannose of the present invention in the preparation of a medicament for preventing and / or treating non-alcoholic fatty liver disease will be described in more detail below in conjunction with a schematic diagram, wherein preferred embodiments of the present invention are shown, and it should be understood that those skilled in the art may modify the present invention described herein while still achieving the beneficial effects of the present invention. Therefore, the following description should be understood as being widely known to those skilled in the art, and not as a limitation of the present invention.

[0044] For the sake of clarity, not all features of the actual embodiments are described. In the following description, well-known functions and structures are not described in detail because they would clutter the invention with unnecessary detail. It should be recognized that in the development of any actual embodiment, a large number of implementation details must be made to achieve the developer's specific goals, such as changing from one embodiment to another according to the limitations of the relevant system or the relevant business. In addition, it should be recognized that such development work may be complex and time-consuming, but it is just a routine task for those skilled in the art.

[0045] The present invention is described in more detail in the following paragraphs by way of example with reference to the accompanying drawings. The advantages and features of the present invention will become more apparent from the following description. It should be noted that the accompanying drawings are in very simplified form and are not in exact proportions, and are only used to facilitate and clearly assist in illustrating the purpose of the embodiments of the present invention.

[0046] Using a high-fat diet-induced fatty liver mouse model, this study will evaluate the effects of mannose on hepatic lipid metabolism and its potential therapeutic effects.

[0047] The molecular formula of Mannose is C6 H 12 O 6 , with a molecular weight of 180.156 g / mol. The present invention provides an application of mannose in the preparation of a drug for treating NAFLD, confirms that mannose has an anti-NAFLD efficacy, and explores its safety, in order to provide new ideas and new drugs for the clinical treatment of NAFLD.

[0048] The present invention discloses the application of mannose in preventing NAFLD. NAFLD in mice was induced by a 60% high-fat diet, and the experimental groups were supplemented with mannose through drinking water for 8 and 12 weeks respectively. Mannose can significantly reduce the severity of NAFLD, reduce liver lipid deposition, reduce the weight gain of mice caused by a high-fat diet, and reduce plasma ALT, AST, total cholesterol and LDL-C levels.

[0049] Furthermore, the present invention discloses the application of mannose in the treatment of NAFLD. The NAFLD mouse model was induced by feeding a 60% high-fat diet for 12 weeks, and the high-fat diet was continued for 4 weeks. The experimental group supplemented mannose through drinking water. Mannose can significantly improve liver pathological changes, reduce mouse body weight, reduce liver fat deposition, and significantly reduce serum ALT, AST and cholesterol levels.

[0050] Furthermore, the present invention discloses the application of mannose in preventing severe NASH of NAFLD. NASH in mice was induced by CL-HFS ​​diet, and the experimental group was supplemented with mannose through drinking water for 8 weeks. Mannose can significantly reduce liver lipid deposition.

[0051] Furthermore, the present invention discloses and provides a safety evaluation of in vivo supplementation of mannose.

[0052] Furthermore, the present invention observed the behavior, weight changes and physiological indicators of important organs of mice during the feeding process of 20% mannose water, and the results showed that mannose had no obvious toxic effects within the dosage range used, and no obvious side effects were found. These results provide reliable safety support for the clinical application of mannose.

[0053] The above-mentioned non-alcoholic fatty liver disease is non-alcoholic fatty liver disease induced by a high-fat diet.

[0054] Furthermore, the mechanism of the present invention for preventing or treating non-alcoholic fatty liver disease includes one or more of the following: reducing lipid accumulation, reducing liver damage, reducing serum AST and ALT levels, or reducing liver triglyceride content.

[0055] Furthermore, mannose is administered in the form of an aqueous solution, and the mass concentration of the mannose aqueous solution is 5% to 40%.

[0056] Preferably, the mass concentration of the mannose aqueous solution is 15% to 25%.

[0057] Preferably, the mass concentration of the mannose aqueous solution is 20%.

[0058] The present invention also provides the use of a pharmaceutical composition containing mannose in preparing a drug for preventing and / or treating non-alcoholic fatty liver disease. The pharmaceutical composition is prepared by using mannose as an active ingredient and adding pharmaceutically acceptable excipients.

[0059] The pharmaceutically acceptable excipients described in the present invention refer to various conventional excipients required when preparing different dosage forms, such as diluents, binders, disintegrants, glidants, lubricants, flavoring agents, inclusion materials, adsorbents, etc., which are prepared into any commonly used oral preparations by conventional preparation methods, such as tablets, capsules, granules or powders.

[0060] Example 1

[0061] Experimental purpose: To verify the effect of mannose in preventing NAFLD induced by high-fat diet.

[0062] Experimental design and grouping: C57BL / 6J male mice, 8 weeks old, were used and randomly divided into normal diet + normal drinking water group, normal diet + 20% mannose drinking water group, high-fat diet + normal drinking water group, high-fat diet + 20% mannose drinking water group, 4 mice in each group. Normal diet used ordinary feed, and high-fat diet (HFD diet) was fed with 60% high-fat feed (purchased from Research Diet: D12492) for 8 weeks or 12 weeks.

[0063] It should be noted that the 20% mannose drinking water means that the 20% mannose replaces the normal drinking water of mice. The 60% high-fat feed means that 60% of the total calories provided in the feed come from fat.

[0064] Experimental steps:

[0065] 1. After the model is established, weigh the mouse, remove one eyeball and collect blood after anesthesia. After the blood sample is left at room temperature for 2 hours, centrifuge at 3000g for 15 minutes at 4°C to separate the serum for later use.

[0066] 2. After blood collection, the mice were killed by cervical dislocation, the intact liver was removed and weighed, and the ratio of liver weight to mouse body weight was calculated.

[0067] 3. After formalin fixation, the liver tissue is embedded in paraffin, routinely or frozen sectioned, and stained.

[0068] 4. The level of hepatic lipid deposition was assessed by HE staining and Oil red staining, and the fat content of liver sections was scored.

[0069] 5. The serum samples were used to detect ALT, AST, ALP, TG, TC and LDL-C indicators of each group of mice using a fully automatic biochemical analyzer.

[0070] Experimental results:

[0071] 1. Mannose significantly slowed down the weight gain induced by high-fat diet: The weight of mice in the high-fat diet + normal drinking water group increased significantly, while the weight of mice in the high-fat diet + 20% mannose drinking water group did not increase significantly (see Figure 1A ).

[0072] 2. Mannose delayed the severity of NAFLD and lipid deposition induced by high-fat diet: The ratio of liver weight to body weight of mice in the high-fat diet + normal drinking water group increased significantly, and obvious liver fat was observed; in contrast, the increase in liver weight and lipid deposition of mice in the high-fat diet + mannose drinking water group were not obvious (see Figure 1B , Figure 1C as well as Figure 1D ).

[0073] 3. Mannose reduces liver enzymes and blood lipid indexes: ALT (Alanine Aminotransferase, alanine aminotransferase), AST (Aspartate Aminotransferase, aspartate aminotransferase), TC (Total Cholesterol, total cholesterol) and LDL-C (Low-Density Lipoprotein Cholesterol, low-density lipoprotein cholesterol) of mice in the high-fat diet + normal drinking water group increased significantly, while the liver enzymes and blood lipid indexes of mice in the high-fat diet + mannose drinking water group did not increase significantly (please refer to Figure 2A , Figure 2B , Figure 2C , Figure 2D , Figure 2E as well as Figure 2F ).

[0074] 4. Safety assessment of mannose: During the administration of 20% mannose water to mice, their behaviors, weight changes and physiological indices were observed, and no obvious toxic effects or side effects were found, indicating that the dose used was safe and non-toxic.

[0075] Example 2

[0076] Experimental purpose: To verify the effect of mannose in treating NAFLD induced by high-fat diet.

[0077] Experimental design and grouping: C57BL / 6J male mice, 8 weeks old, were fed with 60% high-fat diet + normal drinking water for 12 weeks to induce NAFLD, and then randomly divided into: high-fat diet + normal drinking water group, high-fat diet + 20% mannose drinking water group, with 4 mice in each group, and continued feeding for 4 weeks.

[0078] Experimental steps:

[0079] 1. After the model is established, weigh the mouse, remove one eyeball and collect blood after anesthesia. After the blood sample is left at room temperature for 2 hours, centrifuge at 3000g for 15 minutes at 4°C to separate the serum for later use.

[0080] 2. After blood collection, the mice were killed by cervical dislocation, the intact liver was removed and weighed, and the ratio of liver weight to mouse body weight was calculated.

[0081] 3. After formalin fixation, the liver tissue is embedded in paraffin, routinely or frozen sectioned, and stained.

[0082] 4. The level of hepatic lipid deposition was assessed by HE staining and Oil red staining, and the fat content of liver sections was scored.

[0083] 5. The serum samples were used to detect ALT, AST, ALP, TG, TC and LDL-C indicators of each group of mice using a fully automatic biochemical analyzer.

[0084] Experimental results:

[0085] 1. Mannose significantly reduces the body weight of obese mice induced by a high-fat diet: Supplementing mannose with drinking water on a high-fat diet can significantly reduce the body weight of mice (please refer to Figure 3A ).

[0086] 2. Mannose reduces the severity of NAFLD and lipid deposition: After 12 weeks of high-fat diet modeling, supplementing the drinking water with mannose for 4 weeks can significantly reduce the ratio of liver weight to body weight in mice and significantly reduce liver fat (see Figure 3B , Figure 3C as well as Figure 3D ).

[0087] 3. Mannose reduces liver enzymes and blood lipid indexes: After 12 weeks of high-fat diet, supplementing drinking water with mannose for 4 weeks can significantly reduce the levels of serum ALT, AST, TC and LDL-C in mice (please refer to Figure 4A , Figure 4B , Figure 4C , Figure 4D , Figure 4E as well as Figure 4F ).

[0088] Example 3

[0089] Experimental purpose: To verify the effect of mannose in preventing NASH induced by CL-HFS ​​diet.

[0090] Experimental design and grouping: C57BL / 6J male mice, 8 weeks old, were used and randomly divided into the following groups after 48 hours of adaptive feeding: normal diet + normal drinking water group, normal diet + 20% mannose drinking water group, low choline high-fat and high-sugar diet (choline-low high-fat and high-sugar diet, CL-HFS ​​diet) + normal drinking water group, CL-HFS ​​diet + 20% mannose drinking water group, 4 mice in each group. The normal diet used ordinary feed, and the CL-HFS ​​diet was a customized diet (Synergy Bio Co., Ltd.), with a caloric composition of 40% calories from fat, 20% calories from fructose, 2% cholesterol and 0.05% choline, which was used to induce the mouse NASH model. Feeding for 8 weeks.

[0091] Experimental steps:

[0092] 1. After the model is established, weigh the mice, kill them by cervical dislocation, remove the intact liver and weigh it, and calculate the ratio of liver weight to mouse body weight.

[0093] 2. After formalin fixation, the liver tissue was paraffin-embedded, routinely or frozen-sectioned, and stained.

[0094] 3. The level of hepatic lipid deposition was assessed by HE staining and Oil red staining, and the fat content of liver sections was scored.

[0095] Experimental results:

[0096] 1. Mannose significantly slowed down the weight gain induced by CL-HFS ​​diet: The weight of mice in the CL-HFS ​​diet + normal drinking water group increased significantly, while the weight of mice in the CL-HFS ​​diet + 20% mannose drinking water group did not increase significantly (see Figure 5A ).

[0097] 2. Mannose delays CL-HFS ​​diet-induced liver lipid deposition in mice: Compared with the CL-HFS ​​diet + normal drinking water group, the increase in liver weight and lipid deposition in the CL-HFS ​​diet + mannose drinking water group were significantly reduced (see Figure 5B and Figure 5C ).

[0098] It should be noted that all data in the figures of the specification are expressed as mean ± SEM, *P < 0.05, **P < 0.01, ***P < 0.001.

[0099] The results of the above three groups of experiments all confirmed that mannose has significant effects in preventing and treating NAFLD, and in particular, it also verified the role of mannose in preventing NASH induced by CL-HFS ​​diet; mannose can control weight gain, improve liver fatty degeneration, reduce serum transaminase and blood lipid levels, and has good safety. These experimental results provide sufficient experimental basis for the use of mannose in the prevention and treatment of NAFLD.

[0100] In summary, the use of mannose provided by the present invention in the preparation of a drug for preventing and / or treating non-alcoholic fatty liver disease reveals that mannose regulates liver lipid metabolism and reduces liver lipid accumulation, thereby providing a new, safe and effective drug option for the treatment of NAFLD. In addition, the present invention expands the scope of application of mannose in liver metabolic diseases by systematically studying the mechanism of action of mannose, which has important clinical significance and application prospects.

[0101] The above is only a preferred embodiment of the present invention and does not limit the present invention in any way. Any technician in the relevant technical field, without departing from the scope of the technical solution of the present invention, makes any form of equivalent replacement or modification to the technical solution and technical content disclosed in the present invention, which does not depart from the content of the technical solution of the present invention and still falls within the protection scope of the present invention.

Claims

1. Use of mannose in the preparation of a drug for preventing and / or treating non-alcoholic fatty liver disease; the drug contains mannose as the only effective active ingredient; The non-alcoholic fatty liver disease is non-alcoholic steatohepatitis or non-alcoholic fatty liver.

2. The use according to claim 1, characterized in that: The non-alcoholic fatty liver disease is non-alcoholic fatty liver disease induced by a high-fat diet.

3. The use according to claim 1, characterized in that: The mechanisms of prevention or treatment of nonalcoholic fatty liver disease include one or more of the following: reduced lipid accumulation, reduced liver damage, lower serum aspartate aminotransferase and alanine aminotransferase levels, or lower hepatic triglyceride content.

4. The use according to claim 1, characterized in that: Administer via oral administration or drinking water.

5. The use according to claim 1, characterized in that: Mannose is administered in the form of an aqueous solution, and the mass concentration of the mannose aqueous solution is 5% to 40%.

6. The use according to claim 5, characterized in that: The mass concentration of the mannose aqueous solution is 15% to 25%.

7. Use of a pharmaceutical composition containing mannose in the preparation of a drug for preventing and / or treating non-alcoholic fatty liver disease.

8. The use according to claim 7, characterized in that: The pharmaceutical composition is prepared by taking mannose as an active ingredient and adding pharmaceutically acceptable auxiliary materials.

9. The use according to claim 7, characterized in that: The dosage form of the drug is an oral preparation.

10. The use according to claim 7, characterized in that: The dosage form of the medicine is tablet, capsule, granule or powder.

Images