Application of cyclic dipeptide in improvement or treatment of dyslipidemia related diseases

Abstract

The invention relates to the technical field of biological medicine, in particular to application of cyclic dipeptide to improvement or treatment of dyslipidemia related diseases. According to the present invention, the research results show that the cyclo (glycerol-casein) dipeptide has good safety and cholesterol lowering activity, and can reduce mouse serum total cholesterol, low density lipoprotein cholesterol, increase serum high density lipoprotein cholesterol, reduce the content of liver total cholesterol, reduce liver fat accumulation, and slow down the weight increase of mice. The cyclic (glycerol-casein) dipeptide can realize the effect of lowering cholesterol by regulating intestinal flora. The cyclic (glycerol-tyrosine) dipeptide is derived from probiotics, food and medicines, belongs to food-borne cyclic dipeptide, and is high in safety and small in toxic and side effects. The cyclo (glycerol-casein) dipeptide is expected to be developed into a class of innovative medicines and health-care foods with the effect of reducing cholesterol, and has a very good application prospect. The invention also provides the cyclic (sweet-tyrosine) dipeptide and a health food and a pharmaceutical composition containing the cyclic (sweet-tyrosine) dipeptide.

Description

Technical Field

[0001] The present invention relates to the field of biomedical technology, and in particular to the use of a cyclic dipeptide in improving or treating diseases related to dyslipidemia. Background Art

[0002] With the development of society and the improvement of people's living standards, the number of people with dyslipidemia has increased year by year. Hypercholesterolemia is the most common dyslipidemia. The "Guidelines for Blood Lipid Management in China (2023)" stated that in recent years, the dyslipidemia rate of Chinese adults has remained at a high level. In 2018, the number of adults with dyslipidemia reached 35.6%, among which the prevalence of hypercholesterolemia increased by nearly 1 times compared with 2015. Hypercholesterolemia can have adverse effects on blood vessels and cause cardiovascular and cerebrovascular diseases. Among them, atherosclerotic cardiovascular disease is the number one cause of death in the Chinese population. Long-term dyslipidemia can also easily lead to fatty liver and insulin resistance. At present, the first-line drugs for lowering cholesterol are statins and cholesterol absorption inhibitors, but large doses or long-term use have certain side effects. Therefore, it is necessary to develop safe and effective health products or drugs that help maintain healthy cholesterol levels.

[0003] Cyclic (sweet-tyrosine) dipeptide is a cyclic dipeptide of a diketopiperazine structure generated by dehydration cyclization of glycine and L-tyrosine. The distribution of cyclic (sweet-tyrosine) dipeptide has been found in probiotics such as Bacillus coagulans, Bacillus amyloliquefaciens, Bacillus licheniformis, and Chinese medicines such as pangolin scales and American cockroaches. The patent application with publication number CN105648002B discloses a method for extracting cyclic (sweet-tyrosine) dipeptide in Bacillus coagulans. Cyclic (sweet-tyrosine) dipeptide is naturally present in a variety of probiotics and belongs to a food-derived cyclic dipeptide with the advantages of high safety, easy absorption and good stability. At present, there are few reports on the relevant drug efficacy of cyclic (sweet-tyrosine) dipeptide, and its relevant drug efficacy for hypercholesterolemia has not been reported. Therefore, the research on the efficacy value of cyclic (sweet-tyrosine) dipeptide is worthy of further exploration. Summary of the invention

[0004] The purpose of the present invention is to overcome the shortcomings of the prior art and provide a cyclic dipeptide and / or a pharmaceutically acceptable salt thereof for use in preparing a drug or health food for improving or treating dyslipidemia-related diseases.

[0005] To achieve the above object, the technical solution adopted by the present invention is:

[0006] In a first aspect, the present invention provides a use of a cyclic dipeptide and / or a pharmaceutically acceptable salt thereof in the preparation of a drug or health food for improving or treating dyslipidemia-related diseases, wherein the cyclic dipeptide is a cyclic (glycine-tyrosine) dipeptide.

[0007] The present invention has been found through research that cyclic (glycine-tyrosine) dipeptide has good safety and cholesterol-lowering activity, can reduce serum total cholesterol and low-density lipoprotein cholesterol in mice, increase serum high-density lipoprotein cholesterol, reduce liver total cholesterol content, reduce liver fat accumulation, and slow down the weight gain of mice. Cyclic (glycine-tyrosine) dipeptide may achieve the effect of lowering cholesterol by regulating intestinal flora. Cyclic (glycine-tyrosine) dipeptide is expected to be developed into a class of innovative drugs and health foods with cholesterol-lowering effects, and has a good application prospect.

[0008] The amino acid composition order of glycine and tyrosine in the cyclic (glycine-tyrosine) dipeptide described in the present invention is not specific.

[0009] Preferably, the CAS number of the cyclic (glycine-tyrosine) dipeptide is: 5845-66-9.

[0010] Preferably, the dyslipidemia-related disease includes hypercholesterolemia.

[0011] Preferably, the pharmaceutically acceptable salt is a salt formed by combining a cyclic dipeptide with a pharmaceutically acceptable acid.

[0012] Preferably, the pharmaceutically acceptable acid includes at least one of an organic acid, an inorganic acid, and an amino acid.

[0013] Preferably, the medicine or health food treats dyslipidemia-related diseases by regulating cholesterol levels.

[0014] Preferably, the cholesterol levels include serum total cholesterol level, low-density lipoprotein cholesterol level, high-density lipoprotein cholesterol level, and liver total cholesterol level.

[0015] Preferably, the dosage form of the medicine or health food includes pills, tablets, capsules, and oral liquids.

[0016] In a second aspect, the present invention provides a medicine or health food for improving or treating diseases related to dyslipidemia, comprising a cyclic dipeptide and / or a pharmaceutically acceptable salt thereof; the cyclic dipeptide comprises a cyclic (glycine-tyrosine) dipeptide.

[0017] Preferably, the pharmaceutically acceptable salt is a salt formed by combining a cyclic dipeptide with a pharmaceutically acceptable acid.

[0018] Preferably, the medicine or health food further comprises pharmaceutically acceptable excipients or excipients required for food.

[0019] The beneficial effects of the present invention are:

[0020] The present invention has found in the safety study and efficacy study of cyclic (glycine-tyrosine) dipeptide that it has high safety, good cholesterol-lowering effect and certain development potential.

[0021] Cyclic (Gly-Tyr) dipeptide has shown a good cholesterol-lowering effect in a hypercholesterolemia model. The intensity of some indicators is better than or close to that of positive drugs, and it can improve pathological damage to organs. Cyclic (Gly-Tyr) dipeptide naturally exists in a variety of probiotics and animals, with high safety and low toxic side effects. Cyclic (Gly-Tyr) dipeptide is expected to be developed into a class of innovative drugs with cholesterol-lowering effects and health foods that assist in cholesterol-lowering, used to maintain cholesterol at a healthy level. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] Figure 1 The graph shows the weight changes of mice in each treatment group.

[0023] Figure 2 The graphs are the result of organ index changes of mice in each treatment group; among them, Figure A is the result of liver index change; Figure B is the result of kidney index change; Figure C is the result of spleen index change; Figure D is the result of heart index change; Figure E is the result of lung index change.

[0024] Figure 3 The following are the appearance results of the liver, kidney, spleen, heart and lung of mice in each treatment group.

[0025] Figure 4 The graphs show the pathological morphological effects (HE staining) of the liver, kidney, and spleen tissues of mice in each treatment group.

[0026] Figure 5 The graph shows the weight changes of mice in each treatment group.

[0027] Figure 6 The graph shows the changes in liver index of mice in each treatment group.

[0028] Figure 7 Figure 4 is the effect of cyclic (Gly-Tyr) dipeptide on blood lipid related indicators; Figure A is the result of serum total cholesterol (TC) level change; Figure B is the result of serum low-density lipoprotein cholesterol (LDL-c) level change; Figure C is the result of serum high-density lipoprotein cholesterol (HDL-c) level change; Figure D is the result of serum total bile acid (TBA) level change.

[0029] Figure 8 Figure 3 is the effect of cyclic (Gly-Ty) dipeptide on the total cholesterol (TC) and total bile acid (TBA) content in the liver; Figure A is the result of the change of total cholesterol (TC) content in the liver; Figure B is the result of the change of total bile acid (TBA) content in the liver.

[0030] Fig. 9 The graph shows the pathological morphology (HE staining) of the liver tissues of mice in each treatment group.

[0031] Fig.10 The graph shows the weight changes of mice in each treatment group.

[0032] Fig.11 The graph shows the changes in liver index of mice in each treatment group.

[0033] Fig.12 Figure 4 is the effect of cyclic (Gly-Tyr) dipeptide on blood lipid related indicators; Figure A is the result of serum total cholesterol (TC) level change; Figure B is the result of serum low-density lipoprotein cholesterol (LDL-c) level change; Figure C is the result of serum high-density lipoprotein cholesterol (HDL-c) level change; Figure D is the result of serum total bile acid (TBA) level change.

[0034] Fig.13 Figure 3 is the effect of cyclic (Gly-Ty) dipeptide on the total cholesterol (TC) and total bile acid (TBA) content in the liver; Figure A is the result of the change of total cholesterol (TC) content in the liver; Figure B is the result of the change of total bile acid (TBA) content in the liver.

[0035] Fig.14 This is a graph showing the effect of cyclic (glycine-tyrosine) dipeptide treatment on changes in liver appearance.

[0036] Fig.15 This is the result of the effect of cyclic (glycine-tyrosine) dipeptide on lipid accumulation in liver tissue (Oil Red O staining).

[0037] Fig.16 This is the result of the effect of medium-cyclic (glycine-tyrosine) dipeptide on the pathological morphology of liver tissue (HE staining). DETAILED DESCRIPTION

[0038] In order to better illustrate the purpose, technical solutions and advantages of the present invention, the present invention will be further described below in conjunction with specific embodiments.

[0039] Unless otherwise specified, the experimental methods used in the present invention are all conventional methods, and the materials, reagents, etc. used are all available from commercial channels unless otherwise specified.

[0040] The cyclo(glycine-tyrosine) dipeptide used in the present invention preferably has a purity of not less than 95%, more preferably greater than 99%. It can be obtained from commercial sources or homemade, and its molecular formula is: 11 H 12 N2O3; molecular weight: 220.22; molecular structure as shown in formula I:

[0041]

[0042] The cyclic (glycine-tyrosine) dipeptide used in the present invention is synthesized by the School of Traditional Chinese Medicine of Guangzhou University of Chinese Medicine according to the method in the literature (Brandon Cook, RR HaGE J, Efficient One-step Synthesis of Diastereoisomeric Cyclic Dipeptides from Amino Acids: Three Diastereoisomers of Cyclo-L-isoleucyl-L-isoleucine. [J]. Chemistry Department 1992, (10): 1199-1201.), and according to the literature (Mollica A, Costante R, Fiorito S, et al. Synthesis and anti-cancer activity of naturally occurring 2,5-diketopiperazines. Fitoterapia. 2014; 98: 91-7. and Li D, Gu Q, Zhu W, Liu H, Fang Y, Zhu T. Antitumor components from marine actinomycete 11014I. Cyclic dipeptides. Zhongguo Kangshengsu Zazhi.2005;30,449-452,468.), the purity of all products was greater than 95%; ezetimibe was purchased from MSD International GmbH, Singapore; poloxamer 407 was purchased from Merck, Germany; propylthiouracil was purchased from China Jinghua Pharmaceutical Group Co., Ltd.; neomycin sulfate and metronidazole were purchased from Shanghai Yuanye Company; vancomycin hydrochloride and ampicillin sodium were purchased from Shanghai McLean Company.

[0043] Example 1: Acute oral toxicity test of cyclic (glycine-tyrosine) dipeptide

[0044] 1. Experimental animals and methods

[0045] According to the maximum limit method of the National Standard of the People's Republic of China "Chemicals: Test Methods for Acute Oral Toxicity of Chemicals (GB / T21603-2008)", the median lethal dose (LD50) of cyclic (glycine-tyrosine) dipeptide (CGY) was determined. 50 ). 24 KM mice (provided by Guangdong Medical Experimental Animal Center; the same below), SPF grade, weighing 20±2g, half male and half female, were randomly divided into four groups (6 mice in each group):

[0046] Normal male control group (NM), cyclic (Gly-Tyr) dipeptide male group (CGYM), normal female control group (NF), cyclic (Gly-Tyr) dipeptide female group (CGYF). Fasted for 12 hours before the experiment, with unlimited drinking water, CGYM group and CGYF group mice were gavaged once at 0.3mL / 10g, with a dose of 6000mg / kg; NM group and NF group were given an equal volume of normal saline.

[0047] After administration, mice were given free access to food and water and were observed for 14 consecutive days. The weight changes, food intake and other daily living conditions of the mice and the death conditions were recorded every two days. In addition, within 12 hours after administration, the toxic reactions of mice in each group were observed and recorded every 3 hours, such as appearance, behavior, response to stimulation, secretions, number of deaths and time of death. After 12 hours of administration, the above indicators were observed and recorded every 6 hours. The mice were killed after the observation.

[0048] 2. Observation indicators:

[0049] (1) Mouse mortality;

[0050] (2) weight change every two days;

[0051] (3) Organ index: liver, kidney, spleen, heart, and lung;

[0052] The organ index is calculated by organ weight and body weight according to the following formula:

[0053] Organ index = organ weight (g) / body weight (g) × 100%

[0054] (4) Pathological examination: HE staining of liver, kidney, and spleen.

[0055] 3. Experimental results

[0056] After the administration, the above indices were observed and measured, and the experimental data were expressed as X±SEM.

[0057] Table 1 Results of acute toxicity test in mice (unit: mice)

[0058]

[0059]

[0060] Table 2 Changes in body weight of mice in acute toxicity experiment (unit: g)

[0061]

[0062] Table 3 Changes in organ indexes of mice in acute toxicity experiment (unit: %)

[0063]

[0064] The results are shown in Table 1-3. Figure 1-4 The results showed that within 12 hours of administration, mice did not show any toxic reaction after administration of cyclic (glycine-tyrosine) dipeptide. No obvious abnormalities or deaths were observed in the cyclic (glycine-tyrosine) dipeptide group within 14 days. Cyclic (glycine-tyrosine) dipeptide LD 50 The value is greater than 6000mg / kg, which is a low-toxic chemical.

[0065] There was no significant change in the body weight and organ index of mice in the cyclic (Gly-Ty) dipeptide group ( Figure 1-2 ); After treatment with cyclic (glycine-tyrosine) dipeptide, the appearance of the heart, liver, spleen, lung, and kidney of the mice showed no significant damage ( Figure 3 ).

[0066] HE staining was used to perform pathological sections on the liver, kidney, and spleen tissues of mice. The results of pathological sections ( Figure 4 ) showed that after treatment with cyclic (Gly-Tyr) dipeptide, there was no significant change in the pathological morphology of the liver, kidney, and spleen of mice, indicating that cyclic (Gly-Tyr) dipeptide had no obvious liver, kidney, and spleen toxicity.

[0067] Example 2: Treatment of poloxamer 407-induced hypercholesterolemia with cyclic (Gly-Ty) dipeptide 1. Experimental animals and methods

[0068] In this experiment, poloxamer 407 (P407) was used to induce hypercholesterolemia in mice to explore the effect of cyclic (Gly-Ty) dipeptide on hypercholesterolemia mice.

[0069] Forty-eight SPF male C57BL / 6J mice, weighing 20±2g, were randomly divided into a normal control group (N), a model group (M), a positive drug control group (ezetimibe group, EZ, 10mg / kg), and low-, medium-, and high-dose groups of cyclic (glycine-tyrosine) dipeptide (CGY, corresponding to doses of 25mg / kg, 50mg / kg, and 100mg / kg, respectively), with 8 mice in each group. Except for the N group, the other groups were intraperitoneally injected with P407 at a dose of 400mg / kg daily, and the N group was intraperitoneally injected with an equal volume of normal saline to establish a hypercholesterolemia model. The mice in the EZ group and the low-, medium-, and high-dose groups of CGY were gavaged with the corresponding drugs and doses daily, and the N and M groups were gavaged with an equal volume of normal saline. The body weight of all mice was measured daily. After 7 consecutive days of intervention in each group, the samples were collected for analysis.

[0070] 2. Observation indicators:

[0071] (1) Daily weight changes;

[0072] (2) Lipid-related biochemical indicators: TC, HDL-c, LDL-c, TBA;

[0073] TC was measured using a total cholesterol test kit; LDL-c was measured using a low-density lipoprotein cholesterol test kit; HDL-c was measured using a high-density lipoprotein cholesterol test kit; the above kits were purchased from Nanjing Jiancheng Bioengineering Institute, and the specific operating steps were carried out according to the kit instructions.

[0074] (3) liver index;

[0075] (4) Liver biochemical indicators: TC, TBA;

[0076] (5) Pathological examination: HE staining of liver.

[0077] 3. Experimental results

[0078] After the administration, the above indices were observed and measured, and the experimental data were expressed as X±SEM.

[0079] Table 4 Body weight changes of mice in each group (unit: g)

[0080]

[0081]

[0082] Table 5 Changes of serum biochemical indexes in each group of mice

[0083]

[0084] Note: #p<0.05, ##p<0.01, compared with N group; *p<0.05, **p<0.01, compared with M group.

[0085] Table 6 Changes of liver index in each group of mice

[0086]

[0087] Note: #p<0.05, ##p<0.01, compared with N group; *p<0.05, **p<0.01, compared with M group.

[0088] Table 7 Changes of liver biochemical indexes in each group of mice

[0089]

[0090]

[0091] Note: #p<0.05, ##p<0.01, compared with N group; *p<0.05, **p<0.01, compared with M group.

[0092] The results are shown in Table 4-7. Figure 5-9The results showed that compared with the model group, cyclic (glycine-tyrosine) dipeptide could significantly reduce the levels of total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-c) in serum; reduce the content of total cholesterol (TC) in the liver; and significantly reduce the liver index.

[0093] The mouse liver tissue was pathologically sectioned by HE staining, and the results of pathological sections showed that ( Fig. 9 ), the liver lobule structure of the normal group was clear, the cytoplasm was evenly stained, and there were no vacuoles in the cells; the liver cells in the model group showed enlarged and damaged liver cells, transparent cytoplasm, and balloon-like changes. After the administration of cyclic (glycine-tyrosine) dipeptide, the liver damage of mice in each group was improved and the cell swelling was reduced.

[0094] Example 3: Treatment of hypercholesterolemia induced by high-fat emulsion with cyclic (glycine-tyrosine) dipeptide

[0095] 1. Experimental animals and methods

[0096] In this experiment, a hypercholesterolemia model was induced by intragastric administration of high-fat emulsion to explore the effect of cyclic (Gly-Ty) dipeptide on hypercholesterolemia mice.

[0097] 64 SPF male KM mice, weighing 22±2g, were randomly divided into normal control group (N), model group (M), positive drug control group (ezetimibe group, EZ, 10mg / kg), low, medium and high dose groups of cyclic (glycine-tyrosine) dipeptide (CGY, corresponding to doses of 25mg / kg, 50mg / kg, 100mg / kg, respectively), antibiotic group (ABX), antibiotic combined with cyclic (glycine-tyrosine) dipeptide group (ABX+CGY), 8 mice in each group. Except for group N, the other groups were given high-fat emulsion by gavage every day at a dose of 0.2mL / 10g, and group N was gavaged with an equal volume of normal saline to establish a hypercholesterolemia model. The antibiotic group and the antibiotic combined with cyclic (glycine-tyrosine) dipeptide group were given antibiotic solution 0.2mL / mouse every day, and the antibiotic combined with cyclic (glycine-tyrosine) dipeptide group was given 100mg / kg of cyclic (glycine-tyrosine) dipeptide. The mice in the EZ group, CGY low-, medium-, and high-dose groups were given corresponding drugs and doses by gavage every day, and the mice in the N and M groups were given an equal volume of normal saline by gavage. The body weights of all mice were measured daily. Each group was intervened for 8 consecutive days.

[0098] 2. Preparation method of high fat emulsion

[0099] Take 1g of propylthiouracil and grind it in a mortar. Take it out for later use. Take 20g of lard and heat it in a 40℃ water bath to melt it. Put it in a mortar, add 10g of cholesterol and 1g of propylthiouracil, stir it thoroughly and dissolve it. Then add 20mL each of Tween-80 and propylene glycol, grind and mix well, then slowly add 20mL of 10% sodium deoxycholate aqueous solution, grind and emulsify it. Then add distilled water to 100mL. Put it in a sealed container and refrigerate it. When using it, melt it in a 37℃ water bath first.

[0100] 3. Preparation method of antibiotic solution

[0101] Take 0.5 g of neomycin sulfate, 0.5 g of vancomycin hydrochloride, 1 g of ampicillin sodium, and 1 g of metronidazole, mix them, add 1 L of normal saline, dissolve and mix thoroughly, store at 4°C, and return to room temperature before use.

[0102] 4. Observation indicators:

[0103] (1) Daily weight changes;

[0104] (2) liver index;

[0105] (3) Lipid-related biochemical indicators: TC, HDL-c, LDL-c, TBA;

[0106] (4) Liver-related biochemical indicators: TC, TBA;

[0107] (5) Pathological examination: liver HE staining and Oil Red O staining.

[0108] 5. Experimental results

[0109] After the administration, the above indices were observed and measured, and the experimental data were expressed as X±SEM.

[0110] Table 8 Body weight changes of mice in each group (unit: g)

[0111]

[0112]

[0113] Note: #p<0.05, ##p<0.01, compared with N group; *p<0.05, **p<0.01, compared with M group; &p<0.05, &&p<0.01, compared with ABX group.

[0114] Table 9 Changes of liver index in each group of mice

[0115]

[0116] Note: #p<0.05, ##p<0.01, compared with N group; *p<0.05, **p<0.01, compared with M group; &p<0.05, &&p<0.01, compared with ABX group.

[0117] Table 10 Changes of serum biochemical indexes in each group of mice

[0118]

[0119] Note: #p<0.05, ##p<0.01, compared with N group; *p<0.05, **p<0.01, compared with M group; &p<0.05, &&p<0.01, compared with ABX group.

[0120] Table 11 Changes of liver biochemical indexes in each group of mice

[0121]

[0122] Note: #p<0.05, ##p<0.01, compared with N group; *p<0.05, **p<0.01, compared with M group; &p<0.05, &&p<0.01, compared with ABX group.

[0123] The results are shown in Tables 8-11. Figure 10-16 As shown in the figure, after the mice were given high-fat emulsion by gavage, they became irritable and their fur was messy and dull. After antibiotics were given, the mice's food intake was significantly reduced. After 5 days of intervention with ezetimibe and cyclo(glycine-tyrosine) dipeptide, the weight gain of the mice was suppressed and the above symptoms were significantly improved.

[0124] After intervention with cyclic (glycine-tyrosine) dipeptide, the liver index decreased significantly; lipid-related indicators showed that cyclic (glycine-tyrosine) dipeptide could significantly reduce the levels of total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-C) in serum, and significantly increase the level of high-density lipoprotein cholesterol (HDL-c); significantly reduce the content of total cholesterol (TC) in the liver. Compared with the model group mice, the weight gain of mice was inhibited after antibiotic administration; serum total cholesterol (TC) and low-density lipoprotein cholesterol (LDL-c) were significantly increased, while the liver index, high-density lipoprotein cholesterol (HDL-c), total bile acid (TBA), liver TC and TBA did not change significantly. Compared with the antibiotic group, the weight, liver index, serum TC, LDL-c, HDL-c, TBA, liver TC and TBA of mice in the antibiotic combined with 100 mg / kg cyclic (glycine-tyrosine) dipeptide treatment group did not change significantly.

[0125] The above results show that cyclic (glycine-tyrosine) dipeptide can inhibit the weight gain of mice, improve the increase of liver index, improve dyslipidemia and liver cholesterol accumulation. Antibiotics can block the effects of cyclic (glycine-tyrosine) dipeptide on liver index, dyslipidemia and liver cholesterol accumulation. The administration of antibiotics can cause intestinal flora disorders in mice, so it is speculated that the cholesterol-lowering effect of cyclic (glycine-tyrosine) dipeptide may be achieved by regulating intestinal flora.

[0126] The results of observation of mouse liver appearance are as follows Fig.14As shown in the figure, the liver of mice in the normal group was dark red in color, soft in texture, smooth in surface and thin in edge; the liver of mice in the model group and antibiotic group was yellow in appearance, with increased volume, brittle texture, easy to break, blunt edges and rough surface, indicating pathological changes in the liver. After intervention with cyclic (glycine-tyrosine) dipeptide, the above symptoms could be alleviated, while the antibiotic combined with cyclic (glycine-tyrosine) dipeptide group could not improve liver damage.

[0127] The mouse liver tissue was stained using the Oil Red O fat staining method. The results of Oil Red O staining are shown in Fig.15 As shown in the figure, compared with the normal group, the fat accumulation in the liver of the mice in the model group and the antibiotic group increased significantly, the boundaries of the liver cells were unclear, the fat droplets were irregular in shape and large in area, and the fat accumulation in the liver was significantly reduced after the intervention of cyclic (glycine-tyrosine) dipeptide, and the area of ​​red fat droplets in the liver cells decreased. The fat accumulation in the liver of the antibiotic combined with cyclic (glycine-tyrosine) dipeptide group could not be reduced, and the fat accumulation was severe.

[0128] The mouse liver tissue was pathologically sectioned by HE staining. The results of pathological sections were as follows: Fig.16 As shown in the figure, the liver lobule structure of the normal group was clear, the cytoplasm was evenly stained, and there were no vacuoles in the cells; the liver cells in the model group and the antibiotic group showed swelling of the liver cells, with many round vacuoles of different sizes in the cytoplasm, a large number of fatty changes, and some cells were squeezed and deformed. After the administration of cyclic (glycine-tyrosine) dipeptide, the liver vacuoles of mice in each group decreased and the cell swelling was alleviated. The liver fat vacuoles in the antibiotic combined with cyclic (glycine-tyrosine) dipeptide group did not decrease significantly, and the fatty lesions in the liver could not be alleviated. The administration of antibiotics can cause intestinal flora disorders in mice. It is speculated that the effect of cyclic (glycine-tyrosine) dipeptide on improving liver fat accumulation may be achieved by regulating intestinal flora.

[0129] Example 4: Preparation of medicines and health foods

[0130] 1. Preparation of drugs

[0131] Example 1: (Drop pills) Take 500g of cyclo(glycine-tyrosine) dipeptide, add 1500g of polyethylene glycol 6000 as a matrix, heat to melt and mix, then drop into an appropriate amount of liquid paraffin to make a drop pill. Each pill contains 45mg of cyclo(glycine-tyrosine) dipeptide, and the net weight of each pill is 135mg. Oral administration, 2 times a day, 8 pills each time, for the prevention and treatment of hypercholesterolemia.

[0132] Example 2: (Drop pills) Take 500g of cyclo(glycine-tyrosine) dipeptide, add 1500g of poloxamer as a matrix, heat and melt and mix, then drop into an appropriate amount of liquid paraffin to make drop pills. Each pill contains 45mg of cyclo(glycine-tyrosine) dipeptide, and the net weight of each pill is 135mg. Oral administration, 2 times a day, 8 pills each time, for the prevention and treatment of hypercholesterolemia.

[0133] Example 3: (Tablets) Take 900g of cyclo(glycine-tyrosine) dipeptide, add 910g of starch, 500g of microcrystalline cellulose, and 50g of cross-linked polyvinylpyrrolidone, mix well, use 125g of 10% starch slurry as a binder, wet granulate, dry, use 15g of magnesium stearate as a lubricant, mix well, and press into 10,000 tablets containing 90mg of cyclo(glycine-tyrosine) dipeptide per tablet, with a net weight of 0.25g per tablet. Oral administration, 2 times a day, 4 tablets each time, for the prevention and treatment of hypercholesterolemia.

[0134] Example 4: (Tablets) Take 900g of cyclo(glycine-tyrosine) dipeptide, add 710g of starch, 500g of lactose, and 250g of dry starch, mix well, use 125g of 10% starch slurry as a binder, wet granulate, dry, use 15g of magnesium stearate as a lubricant, mix well, and press into 10,000 tablets containing 90mg of cyclo(glycine-tyrosine) dipeptide per tablet, each tablet has a net weight of 0.25g. Oral, twice a day, 4 tablets each time, for the prevention and treatment of hypercholesterolemia.

[0135] Example 5: (Tablets) Take 900g of cyclo(glycine-tyrosine) dipeptide, add 885g of lactose, 500g of hydroxypropylmethylcellulose (20%), 200g of stearyl alcohol (8%), mix well, wet granulate, dry, add 15g of magnesium stearate (0.6%) as a lubricant, mix well, and press into 10,000 sustained-release tablets containing 90mg of cyclo(glycine-tyrosine) dipeptide per tablet, with a net weight of 0.25g per tablet. Oral administration, 2 times a day, 4 tablets each time, for the prevention and treatment of hypercholesterolemia.

[0136] Example 6: (Tablets) Take 900g of cyclo(glycine-tyrosine) dipeptide, add 710g of lactose, 625g of high-substituted hydroxypropylcellulose (25%), 250g of povidone (10%), mix well, wet granulate, dry, add 15g of magnesium stearate (0.6%) as a lubricant, mix well, and press into 10,000 sustained-release tablets containing 90mg of cyclo(glycine-tyrosine) dipeptide per tablet, with a net weight of 0.25g per tablet. Oral administration, 2 times a day, 4 tablets each time, for the prevention and treatment of hypercholesterolemia.

[0137] Example 7: (Capsules) Take 900g of cyclo(glycine-tyrosine) dipeptide, add 960g of starch and 500g of microcrystalline cellulose, mix well, use 125g of 10% starch slurry as a binder, wet granulate, dry, add 15g of magnesium stearate, mix well, fill into No. 1 capsules to make 10,000 capsules containing 90mg of cyclo(glycine-tyrosine) dipeptide per capsule, and each capsule has a net weight of 0.25g. Oral administration, 2 times a day, 4 capsules each time, for the prevention and treatment of hypercholesterolemia.

[0138] Example 8: (Capsules) Take 900g of cyclo(glycine-tyrosine) dipeptide, add 960g of starch and 500g of lactose, mix well, use 125g of 10% starch slurry as a binder, wet granulate, dry, add 15g of magnesium stearate, mix well, fill into No. 1 capsules to make 10,000 capsules containing 90mg of cyclo(glycine-tyrosine) dipeptide per capsule, and the net weight of each capsule is 0.25g. Oral administration, 2 times a day, 4 capsules each time, for the prevention and treatment of hypercholesterolemia.

[0139] Example 9: (Oral solution) Take 39g of cyclic (glycine-tyrosine) dipeptide, add 100mL of polyethylene glycol 400 and 400mL of 50% ethanol, fully dissolve, add 2g of aspartame, 0.5g of stevioside, 0.5g of o-phenylphenol, add pure water to 1000mL, mix well, filter, put the contents into a glass bottle of oral solution, and sterilize. The specification is that each bottle contains 390mg of cyclic (glycine-tyrosine) dipeptide, and the net content of each bottle is 10mL. Oral, twice a day, 1 bottle each time, for the prevention and treatment of hypercholesterolemia.

[0140] Example 10: (Oral solution) Take 39g of cyclic (glycine-tyrosine) dipeptide, add 150mL of propylene glycol (15%) and 400mL of 50% ethanol, fully dissolve, add 2g of aspartame, 0.5g of stevioside, 1mL of ethyl parahydroxybenzoate, 1mL of propyl parahydroxybenzoate, add pure water to 1000mL, mix well, filter, put the contents into a glass bottle of oral solution, and sterilize. The specification is that each bottle contains 390mg of cyclic (glycine-tyrosine) dipeptide, and the net content of each bottle is 10mL. Oral, twice a day, 1 bottle each time, for the prevention and treatment of hypercholesterolemia.

[0141] 2. Preparation of health food

[0142] Example 11: (Tablets) Take 900g of cyclo(glycine-tyrosine) dipeptide, add appropriate excipients, and prepare 10,000 tablets (each tablet contains 90mg of cyclo(glycine-tyrosine) dipeptide) according to conventional methods. Take orally, twice a day, 4 tablets each time, for dietary cholesterol-lowering in patients with obesity and hypercholesterolemia.

[0143] Example 12: (Capsules) Take 900g of cyclo(glycine-tyrosine) dipeptide, add appropriate excipients, and prepare 10,000 capsules (each containing 90mg of cyclo(glycine-tyrosine) dipeptide) according to conventional methods. Oral administration, 2 times a day, 4 capsules each time, for dietary cholesterol-lowering in patients with obesity and hypercholesterolemia.

[0144] Example 13: (Gel) Take 10000g of cyclo(glycine-tyrosine) dipeptide, add appropriate gel, water-soluble dietary fiber and other food raw materials, and prepare into gel according to conventional methods, each 100g contains 200mg of cyclo(glycine-tyrosine) dipeptide. It is used as food to assist in lowering cholesterol in patients with obesity and hypercholesterolemia.

[0145] Example 14: (Tea) Take 2000g of cyclo(glycine-tyrosine) dipeptide and 20000g of tea leaves, prepare 10000 bags of teabags (each bag contains 200mg of cyclo(glycine-tyrosine) dipeptide) according to conventional methods, and drink with boiling water. It can be used as food to assist in lowering cholesterol in patients with obesity and hypercholesterolemia.

[0146] Example 15: (Instant) Take 2000g of cyclo(glycine-tyrosine) dipeptide, add appropriate auxiliary materials, prepare 10,000 packets of instant instant according to conventional methods (each packet contains 200mg of cyclo(glycine-tyrosine) dipeptide), and brew with boiling water. It can be used as a food supplement to lower cholesterol in patients with obesity and hypercholesterolemia.

[0147] Finally, it should be noted that the above embodiments are only used to illustrate the technical solution of the present invention rather than to limit the scope of protection of the present invention. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solution of the present invention can be modified or replaced by equivalents without departing from the essence and scope of the technical solution of the present invention.

Claims

1. Use of a cyclic dipeptide and / or a pharmaceutically acceptable salt thereof in the preparation of a drug or health food for improving or treating a disease related to dyslipidemia, characterized in that: The cyclic dipeptide is a cyclic (glycine-tyrosine) dipeptide.

2. The use according to claim 1, characterized in that The CAS number of the cyclic (glycine-tyrosine) dipeptide is: 5845-66-9.

3. The use according to claim 1, characterized in that The dyslipidemia-related diseases include hypercholesterolemia.

4. The use according to claim 1, characterized in that The pharmaceutically acceptable salt is a salt formed by combining the cyclic dipeptide with a pharmaceutically acceptable acid.

5. The use according to claim 4, characterized in that The pharmaceutically acceptable acid includes at least one of an organic acid, an inorganic acid, and an amino acid.

6. The use according to claim 1, characterized in that The medicine or health food treats diseases related to dyslipidemia by regulating cholesterol levels.

7. The use according to claim 6, characterized in that The cholesterol levels include serum total cholesterol level, low-density lipoprotein cholesterol level, high-density lipoprotein cholesterol level, and liver total cholesterol level.

8. The use according to claim 1, characterized in that The dosage forms of the medicine or health food include pills, tablets, capsules and oral liquids.

9. A medicine or health food for improving or treating diseases related to dyslipidemia, characterized in that: Including cyclic dipeptides and / or pharmaceutically acceptable salts thereof; the cyclic dipeptide includes cyclic (Gly-Tyr) dipeptide.

10. The medicine or health food according to claim 9, characterized in that: The medicine or health food also includes pharmaceutically acceptable excipients or excipients required for food.

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