Application of N-acetyl-L-aspartic acid in the preparation of drugs or foods for preventing or treating obesity

Abstract

The present invention provides a new use of N-acetyl-L-aspartic acid — its application in the preparation of drugs or foods for preventing or treating obesity, belonging to the technical field of biomedicine and health care. Research shows that N-acetyl-L-aspartic acid can inhibit adipocyte differentiation and reduce lipid accumulation, thereby inhibiting fat formation, and thus has the effect of inhibiting lipid accumulation. Therefore, N-acetyl-L-aspartic acid can be used as an active substance for inhibiting fat accumulation in cells and used in the preparation of drugs or health care preparations for preventing or treating weight loss. That is, N-acetyl-L-aspartic acid is used as an active component to prepare weight loss drugs or weight loss foods. Or it is compounded with a compatible weight loss active ingredient to prepare a compound weight loss drug or health care preparation.

Description

Technical Field

[0001] The present invention relates to a new use of N-acetyl-L-aspartic acid - its application in inhibiting fat accumulation, belonging to the technical fields of biomedicine and health care. Technical Background

[0002] Obesity is a chronic metabolic disease caused by multiple factors such as genetics, biological behavior, and environment, and it is one of the most easily overlooked diseases currently faced by humans. Obesity is a state caused by excessive accumulation of fat in the body, especially triglycerides, and excessive subcutaneous fat accumulation. When humans accumulate too much fat due to excessive food intake or changes in body metabolism, it will cause excessive weight gain and lead to pathophysiological changes in the human body, gradually evolving into obesity. With the development of technology and the improvement of people's living standards, combined with an unhealthy lifestyle, obesity has become an epidemic globally. According to the statistics of the World Health Organization, as of 2022, more than 1 billion people worldwide suffer from obesity, including 650 million adults, 340 million adolescents, and 39 million children. In China, obesity is an even more notable public health issue. According to the data of "Report on Nutrition and Chronic Diseases of Chinese Residents (2020)", the overweight rate of Chinese adults (≥18 years old) is 34.3%, and the obesity rate is 16.4%.

[0003] Obesity is mainly divided into two types: simple obesity and pathological obesity. Simple obesity usually has no obvious cause, and those with obvious causes are called pathological obesity. Obesity can cause a series of changes in human physiology, pathology, nerves, and humoral regulation, reducing the working ability of the human body. Severely obese people have a decreased resistance to diseases and even a significantly shortened lifespan. Modern medical research reveals that obese people have a lifespan 5 - 20 years shorter than those with normal body weight. In addition, obesity can cause a series of complications, including diabetes, coronary heart disease, hypertension, hyperlipidemia, atherosclerosis and plaques, fatty liver, hyperuricemia, etc., which not only affect the patients themselves but also impose high medical costs on families.

[0004] At present, the pathogenic mechanism of obesity has not been fully clarified, so there is a lack of effective treatment methods. The commonly used treatment methods include lifestyle intervention, drug treatment and surgical treatment, among which drug treatment is the main clinical treatment method. Commonly used weight loss drugs on the market at present include orlistat, liraglutide, etc. The content of lipase in obese people is relatively high. Orlistat can selectively inhibit the activity of pancreatic lipase and reduce the absorption of about 30% of the fat in the diet, but it will cause intestinal adverse reactions and inhibit the absorption of fat-soluble vitamins. Long-term use may even have potential cardiovascular risks; Liraglutide is a glucagon-like peptide-1 (GLP-1) receptor agonist, which can stimulate the production of insulin, inhibit the secretion of glucagon, reduce appetite and food intake. However, after injecting liraglutide, it is easy to cause gastrointestinal side effects such as abdominal distension, diarrhea, nausea, and vomiting. The above-mentioned weight loss drugs have low efficacy and obvious side effects, so there is an urgent need to develop weight loss drugs and health products with good curative effects and few side effects.

[0005] N-acetyl-L-aspartic acid (NAA) is an important fine organic chemical intermediate, which is widely used in the fields of medicine, pesticides, chemical industry, etc. NAA is a biochemical index of the severity of neuronal damage. The decrease of NAA in ischemic brain tissue after stroke indicates neuronal metabolic disorder. It has been confirmed that NAA and lactate are of great significance for the determination of the ischemic penumbra after stroke, the treatment of acute stroke and the judgment of prognosis. The structural formula of N-acetyl-L-aspartic acid is as follows:

[0006]

[0007] We unexpectedly found in the research that N-acetyl-L-aspartic acid can significantly inhibit the accumulation of fat in cells and can be used for the treatment or prevention of obesity and its complications. Summary of the Invention

[0008] The purpose of the present invention is to provide a new use of N-acetyl-L-aspartic acid - its application in the preparation of drugs or foods for preventing or treating obesity.

[0009] The following is a specific study on the effect of N-acetyl-L-aspartic acid in inhibiting the accumulation of fat in cells through experiments.

[0010] 1. Experimental materials and reagents

[0011] 3T3-L1 mouse embryonic fibroblasts (Procell, CL0006); Dulbecco’s Modified Eagle Medium (DMEM, Biological Industries, 01-052-1A); Fetal Bovine Serum (FBS, Biological Industries, 04-001-1A); Penicillin-streptomycin (PS, Biological Industries, 03-031-1B); 3-isobytyl-1-methylxanthine (IBMX, Sigma Aldrich, I7018); Dexamethasone (Aladdin, D392303); Insulin (Ronn, R008974); N-acetyl-L-aspartic acid (Aladdin, N305071).

[0012] 2. Experimental grouping

[0013] The cultured cells were divided into three groups: blank control group MDI (-) + N-Acetyl-L-aspartic acid (-), model control group MDI (+) + N-Acetyl-L-aspartic acid (-), and experimental group MDI (+) + N-Acetyl-L-aspartic acid (+).

[0014] 3. Experimental methods

[0015] 3.1 Cell differentiation

[0016] (1) Seed 3T3-L1 cells in a 6-well plate and incubate them with 10% FBS / 1% PS / DMEM at 5% CO2 and 37 °C until cell contact inhibition;

[0017] (2) Incubate 3T3-L1 cells with 1% PS / DMEM medium containing 10% bovine calf serum (BCS) at 5% CO2 and 37 °C for 2 days;

[0018] (3) Two groups of MDI(+)+N-Acetyl-L-aspartic acid(-) and MDI(+)+N-Acetyl-L-aspartic acid(+) were added with differentiation inducers (1 μM dexamethasone, 0.5 mM IBMX, 10 μg / mL insulin, MDI), and incubated in 10% FBS / 1% PS / DMEM, 5% CO₂ at 37 °C for 2 days. N-Acetyl-L-aspartic acid was added to the MDI(+)+N-Acetyl-L-aspartic acid(+) group, and N-Acetyl-L-aspartic acid should be added every time the medium was changed for this group. The concentration of N-Acetyl-L-aspartic acid was 100 nM;

[0019] (4) The culture medium was changed to 10% FBS / 1% PS / DMEM containing 10 μg / mL insulin, and incubated again at 5% CO₂, 37 °C for 2 days;

[0020] (5) The culture medium was changed to 10% FBS / 1% PS / DMEM, and incubated continuously at 5% CO₂, 37 °C, and the culture medium was changed every two days. Differentiation was successful around 10 - 14 days of the experiment, lipid droplets accumulated, presenting an adipocyte phenotype, and subsequent experiments were carried out.

[0021] 3.2 Oil Red staining

[0022] The cells were stained with Oil Red as follows:

[0023] (1) Carefully pour out the culture medium, and wash the cells 2 - 3 times with 1 mL PBS;

[0024] (2) Add 1 mL of 4% paraformaldehyde solution and fix for 30 min;

[0025] (3) Aspirate the paraformaldehyde solution and wash 2 - 3 times with 1 mL PBS;

[0026] (4) Add 1 mL of 60% isopropanol and soak for about 1 minute to make the lipid particles stain better; add 1 mL of 0.3% Oil Red, and place it in an oven at 37 °C for 30 minutes for staining;

[0027] (5) After staining, wash once with distilled water, then quickly wash once with 1 mL of 60% isopropanol, and then wash 2 - 3 times with distilled water. Add 1 mL of PBS and observe the results under a microscope.

[0028] 4. Experimental results

[0029] Figure 1Oil red staining results of cells in each group of the present invention: Among them, MDI is a differentiation inducer, and N-Acetyl-L-aspartic acid is N-acetyl-L-aspartic acid. "MDI (-) + N-Acetyl-L-aspartic acid (-)" is the blank control group without adding the differentiation inducer and N-acetyl-L-aspartic acid, "MDI (+) + N-Acetyl-L-aspartic acid (-)" is the model control group with the addition of the differentiation inducer and without the addition of N-acetyl-L-aspartic acid, and "MDI (+) + N-Acetyl-L-aspartic acid (+)" is the experimental group with the addition of the differentiation inducer and N-acetyl-L-aspartic acid. As Figure 1 shown, under the action of the differentiation inducer, intracellular lipids in 3T3-L1 cells continuously accumulate. The lipid accumulation in the cells of the model control group "MDI (+) + N-Acetyl-L-aspartic acid (-)" is significantly higher than that in the blank control group "MDI (-) + N-Acetyl-L-aspartic acid (-)". Compared with the model control group "MDI (+) + N-Acetyl-L-aspartic acid (-)", the lipid accumulation in the experimental group "MDI (+) + N-Acetyl-L-aspartic acid (+)" with the addition of N-acetyl-L-aspartic acid is significantly reduced and approaches the level of the blank control group "MDI (-) + N-Acetyl-L-aspartic acid (-)". Thus, it can be seen that N-acetyl-L-aspartic acid can inhibit adipocyte differentiation, thereby inhibiting fat formation, and therefore has the effect of inhibiting lipid accumulation.

[0030] Figure 2 Bar graph of the area ratio occupied by lipid-accumulating adipocytes after oil red staining in each group of the present invention. Among them, MDI is a differentiation inducer, and N-Acetyl-L-aspartic acid is N-acetyl-L-aspartic acid. "MDI (-) + N-Acetyl-L-aspartic acid (-)" is the blank control group without adding the differentiation inducer and N-acetyl-L-aspartic acid, "MDI (+) + N-Acetyl-L-aspartic acid (-)" is the model control group with the addition of the differentiation inducer and without the addition of N-acetyl-L-aspartic acid, and "MDI (+) + N-Acetyl-L-aspartic acid (+)" is the experimental group with the addition of the differentiation inducer and N-acetyl-L-aspartic acid. As Figure 2As shown, compared with the blank control group "MDI(-)+N-Acetyl-L-aspartic acid(-)", the model control group "MDI(+)+N-Acetyl-L-aspartic acid(-)" ### showed p<0.001, indicating successful differentiation of 3T3-L1 adipocytes; compared with the model control group "MDI(+)+N-Acetyl-L-aspartic acid(-)", the experimental group "MDI(+)+N-Acetyl-L-aspartic acid(+)" *** showed p<0.001, indicating that N-acetyl-L-aspartic acid has a good effect of inhibiting adipocyte differentiation and reducing lipid accumulation.

[0031] Based on the above research, the present invention can use N-acetyl-L-aspartic acid as an active substance for inhibiting fat accumulation in cells, and is used for preparing weight loss drugs or health care preparations for prevention or treatment. That is, taking N-acetyl-L-aspartic acid as an active component to prepare weight loss drugs or weight loss foods. Or compounding with a compatible weight loss active ingredient to prepare compound weight loss drugs or health care preparations. The dosage forms of the preparations include powders, injections, tablets, capsules, granules, pills, oral liquids, etc. Description of the Drawings

[0032] Figure 1 It is a graph showing the results of oil red staining of cells in each group of the present invention.

[0033] Figure 2 It is a bar graph showing the area ratio of oil red-stained accumulated adipocytes in each group of the present invention. Detailed Embodiments

[0034] Example 1: Using N-acetyl-L-aspartic acid as an active component, a weight loss drug preparation is prepared with pharmaceutically or physiologically acceptable excipients and conventional drug preparation processes. The drug preparation is powder, granule, capsule, soft capsule, powder, pill, tablet, oral liquid.

[0035] Example 2: Using a compound of N-acetyl-L-aspartic acid and a compatible weight loss reagent as an active component, a compound weight loss drug preparation is prepared. The oral drug preparation is powder, granule, capsule, soft capsule, powder, pill, tablet, oral liquid.

[0036] Example 3: Using N-acetyl-L-aspartic acid as an active component, a weight loss food is prepared.

[0037] Example 4: Using a compound of N-acetyl-L-aspartic acid and a compatible weight loss active ingredient as an active component, a weight loss food is prepared.

Claims

1. Use of N-acetyl-L-aspartic acid in the preparation of a drug for preventing or treating obesity, characterized in that: The structural formula of the N-acetyl-L-aspartic acid is as follows: 。 2. Use of N-acetyl-L-aspartic acid in the preparation of a drug for preventing or treating obesity according to claim 1, characterized in that: Using N-acetyl-L-aspartic acid as the active ingredient, a weight loss pharmaceutical preparation is made with pharmaceutically or physiologically acceptable excipients and the preparation process of conventional pharmaceutical preparations.

3. Use of N-acetyl-L-aspartic acid in the preparation of a drug for preventing or treating obesity according to claim 1, characterized in that: Using a complex of N-acetyl-L-aspartic acid and a compatible weight loss active ingredient as the active ingredient, a weight loss pharmaceutical preparation is made with pharmaceutically or physiologically acceptable excipients and the preparation process of conventional pharmaceutical preparations.

Images