Peptide for accelerating fat metabolism and use thereof

Abstract

The application aims to provide a peptide for accelerating fat metabolism and application thereof, and to prepare the peptide into a product convenient to use, and the amino acid sequence of the peptide is MVHSYSETSD. The peptide is obtained through screening of a peptide library, screening through bioinformatics analysis, and verification through fat cell decomposition experiment, and the application verifies that the active peptide screened has the effect of accelerating fat metabolism, inhibiting fat accumulation, reducing blood fat and body fat content, reducing body weight, improving insulin sensitivity and glucose tolerance, and realizing the prevention and treatment effect on obesity and its complications.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of medicine and biology, and particularly relates to a peptide for accelerating fat metabolism and application thereof. BACKGROUND

[0002] The methods for improving obesity include dietary habit adjustment, drug treatment, psychological assistance, surgical treatment, exercise intervention, etc. Commonly used drugs include appetite suppressants (such as orlistat, liraglutide), fat absorption inhibitors (such as orlistat), metabolic regulators (such as metformin), etc. Surgical treatment is suitable for patients with severe obesity or complications, such as sleeve gastrectomy, gastric bypass, etc. It is suitable for patients with BMI ≥ 40 or BMI ≥ 35 combined with severe complications. Surgical treatment requires strict indication screening, and long-term monitoring and management after surgery. The current problems include evaluation of contraindications and side effects of drug treatment, careful long-term use, and the need for more systematic, in-depth research and evaluation to prove the safety and sustainability of the treatment.

[0003] Peptide refers to a linear molecule formed by the mutual combination of peptide bond amino acid residues, which can be synthesized by the chemical synthesis method known in the art, especially the solid-phase synthesis technique.

[0004] CN117357647A 20240109Researchers of Beijing University found that VIP can promote the expression of key genes and proteins in brown adipose tissue (BAT) and increase the number of mitochondria; in normal diet mice, VIP treatment can reduce the body weight of mice, increase the body temperature of mice, and promote the expression of heat-related genes in BAT; in high-fat diet mice, VIP treatment can reduce the body weight of mice and the weight of white adipose tissue (WAT), increase glucose tolerance and insulin sensitivity, also can increase the body temperature of mice in cold stimulation environment, increase oxygen consumption, carbon dioxide production and energy expenditure, and promote the expression of heat-related genes in BAT and subcutaneous white adipose tissue (sWAT); after knocking down the VIP receptor using adeno-associated virus, the body temperature of mice in cold stimulation environment decreases, and the expression of heat-related genes in BAT decreases. CN118684758A 20240924An incretin analogue and its application. It contains a triple agonist protein that agonizes human glucose-dependent insulinotropic polypeptide (GIP) receptor, glucagon-like peptide 1(GLP 1) receptor and glucagon (GCG) receptor, which can be used for the treatment of type 2 diabetes (T2D), obesity, non-alcoholic steatohepatitis and other abnormal glucose and lipid metabolism diseases. The incretin GLP-1 receptor agonist (liraglutide) has been successfully listed as a polypeptide drug for the treatment of diabetes and obesity. These studies show that polypeptides have good application prospects in the treatment of obesity and related metabolic diseases.

[0005] The method for adjusting energy metabolism is the current research focus, in this regard, the sequence compound has higher stability (low toxicity). The inventor has researched the proteins related to the upstream and downstream signals of fat storage and fat decomposition when a human takes a high-fat diet, screened the proteins accelerating fat decomposition, and developed multiple active peptides through the above protein research and related protein research.

[0006] Therefore, the active peptide screened by the present application has the effects of accelerating fat metabolism, inhibiting fat accumulation, reducing blood lipids and body fat content, reducing body weight, improving insulin sensitivity and glucose tolerance, and achieving the prevention and treatment effects on obesity and its complications. SUMMARY

[0007] The present application aims to provide a peptide accelerating fat metabolism and its application. The active peptide has the effects of accelerating fat metabolism, inhibiting fat accumulation, reducing blood lipids and body fat content, reducing body weight, improving insulin sensitivity and glucose tolerance, and achieving the prevention and treatment effects on obesity and its complications.

[0008] To achieve the above-mentioned purpose, the present application is implemented by the following technical solutions:

[0009] In one aspect, the present application provides a peptide accelerating fat metabolism, wherein the peptide has the amino acid sequence of MVHSYSETSD (SEQ ID NO: 1).

[0010] In one aspect, the present application provides the use of the peptide in the manufacture of a product for preventing or treating obesity or improving metabolism or reducing blood lipids or reducing blood sugar, wherein the peptide accelerates fat metabolism, the peptide reduces blood lipids and body fat content, the peptide inhibits fat accumulation, the peptide can reduce body weight, and the peptide improves insulin sensitivity and glucose tolerance.

[0011] In one aspect, the present application provides a composition containing the peptide accelerating fat metabolism, wherein the composition comprises the peptide, and the composition is pharmaceutically acceptable.

[0012] The present application has the following beneficial effects:

[0013] The application is obtained through screening of a peptide library, screening through bioinformatics analysis, and verification through fat cell decomposition experiment. The application also verifies that the active peptide obtained through screening has the effect of accelerating fat metabolism, improves the excessive increase of body weight of obese mice, and also reflects the improvement in weight gain and weight growth rate, inhibits fat accumulation, and the effect is similar to that of the positive control; significantly reduces the proportion of inguinal white adipose tissue and epididymal white adipose tissue. After the intervention of the active peptide, the mice reduce food intake, decompose accumulated fat, reduce blood lipids and body fat content, the active peptide significantly improves insulin sensitivity and glucose tolerance, reduces fasting blood glucose, and the effect is comparable to that of the positive drug. The active peptide group shows superior effects of reducing triglycerides, total cholesterol, low-density lipoprotein cholesterol, and increasing high-density lipoprotein cholesterol. BRIEF DESCRIPTION OF DRAWINGS

[0014] Fig. 1 is the body weight change of each group of mice, the rhombus represents the blank control group, the square represents the HFD group, the triangle represents the liraglutide group, and the cross represents the active peptide group.

[0015] Fig. 2 is the change of body fat rate of each group of mice, the horizontal coordinate represents the group, and the vertical coordinate represents the body fat rate, unit: %.

[0016] Fig. 3 is the change of white adipose tissue rate of each group, the horizontal coordinate represents the group, and the vertical coordinate represents the white adipose tissue rate. DETAILED DESCRIPTION

[0017] The embodiments of the application will be further described below in combination with examples.

[0018] The concentration of the active peptide is 1 uM (this concentration has been screened, the minimum effective concentration, and the actual industrial application can be increased), the synthesis of the peptide is completed by Hangzhou Taijia Biotechnology Co., Ltd., and other compounds or kits are purchased from commercial brands.

[0019] Fat tissue decomposition rate

[0020] Fat tissue acquisition: after the C57BL / 6J mice (n=3) were sacrificed, the inguinal fat tissue was quickly separated and rinsed in pre-cooled physiological saline to remove impurities. The fat tissue was cut into small pieces (about 50-100 mg), washed with Krebs-Ringer buffer solution (containing 3% BSA) for 3 times to remove free fatty acids.

[0021] The fat tissue block is placed in a Krebs-Ringer buffer containing 0.5% BSA, and incubated at 37°C for 1-2 hours. Add blank saline, active peptide or adrenaline hydrochloride to stimulate lipolysis, and incubate at 37°C for 72 hours. Collect the supernatant and detect the glycerol release (uM / g / hr) (colorimetric method), and calculate the glycerol release rate, i.e. the fat decomposition rate. Compared with the blank control (100%), the active peptide fat decomposition rate reaches 183.54±12.30%, and the adrenaline hydrochloride fat decomposition rate reaches 210.60±15.80%. It can be seen that the active peptide can decompose the accumulated fat.

[0022] High-fat diet (HFD) model construction process and evaluation method

[0023] Forty male C57BL / 6J mice (15-20 g, 4 weeks old, specific pathogen-free level, SPF level) with uniform development and consistent growth performance were purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd. and were allowed to freely eat and drink in an IVC system (room temperature 23±2°C, light-dark cycle 12 h). After 1 week of adaptation, the mice with similar average body weight were randomly divided into 4 groups according to a random number table: a normal diet group (normal breeding feed HD8015, blank control, n=10), a high-fat diet group (HFD (D12451, 45 kcal% fat), n=10), a liraglutide group (n=10), and an active peptide group (n=10). The blank control group was fed with normal breeding feed, and the other groups were fed with high-fat feed (the proportion of high-fat feed in daily feeding was gradually increased until it was completely replaced).

[0024] According to the body weight conversion, each group was given medication or water, and each group of mice was intraperitoneally injected once every other day. The liraglutide group (n=10, 50 μg / kg·bw physiological saline, injection volume 120 μL) and the active peptide group (n=10, 50 μg / kg·bw (this concentration has been screened for the lowest effective concentration) physiological saline, injection volume 120 μL) mice. The blank control group and the HFD group were injected with 120 μL of physiological saline. During the experiment, the tail tip blood of the mice was collected every week for fasting blood glucose detection. After 12 weeks of intervention, glucose tolerance test (GTT) and insulin tolerance test (ITT) were performed. Before the GTT experiment, the mice were fasted for 10 hours and then intraperitoneally injected with a glucose solution (2 g / kg body weight). Before the ITT experiment, the mice were allowed to freely eat, and in the morning, they were intraperitoneally injected with insulin (1 U / kg body weight). The GTT experiment and the ITT experiment were performed before injection (0 min), 15, 30, 60, 90 and 120 minutes after injection, respectively. Mouse tail blood was collected for measurement, and the blood glucose level was measured using a blood glucose meter. The blood glucose change curve was drawn and the area under the glucose curve was calculated.

[0025] Twelve weeks after the intervention, mice in each group were anesthetized and euthanized. Serum and white adipose tissue were collected and stored at -80°C for further analysis. During the experiment, the mice's food intake was recorded daily. They were weighed after the first 10 days, and then every two weeks thereafter. The animal experimental methods were in accordance with standard procedures and were approved by the laboratory animal ethics review committee.

[0026] Weight gain (g) = Weight at the end of the experiment - Weight at the beginning of the experiment

[0027] Weight gain rate (%) = Weight gain / Initial weight (at the start of the experiment) × 100%

[0028] Weigh the white adipose tissue of the mouse epididymis and the white adipose tissue of the groin.

[0029] White fat percentage (%) = white fat weight / body weight at the end of the experiment × 100%, results are shown in [link to results]. Figs. 1-3 .

[0030] Frozen mouse serum samples were thawed at 4°C, and serum triglycerides (TG), total cholesterol (TC), high-density lipoprotein cholesterol (HDL-C), and low-density lipoprotein cholesterol (LDL-C) were measured according to the kit instructions. The results are shown in Table 1 below.

[0031] Table 1 shows the measurement results of TC, TG, HDL-C, and LDL-C in each group of mice.

[0032]

[0033] The results show that the HFD group had the fastest weight gain. Compared with the blank control group and the HFD group, the active peptide improved the excessive weight gain in obese mice, showing improvement in both weight gain and weight growth rate, and inhibiting fat accumulation, with effects similar to the positive control. The HFD group significantly increased the proportion of white adipose tissue in the groin and epididymis of mice, while the active peptide group significantly improved the proportion of white adipose tissue in the groin and epididymis. Monitoring the daily food intake of mice revealed that the active peptide intervention reduced food intake, broke down accumulated fat, and reduced blood lipids and body fat content. Combined with the results of insulin sensitivity and glucose tolerance, the active peptide significantly improved insulin tolerance (AUC) and glucose tolerance (AUC) (396, 1257), and reduced fasting blood glucose to 6.2 mM (compared with the blank control group, the HFD group significantly increased fasting blood glucose by 11 mM, and insulin tolerance (AUC) and glucose tolerance (AUC) by 396, 1257), roughly equivalent to the effect of the positive control. Compared with the blank control group, the HFD group showed significantly increased TC and LDL-C (p<0.05), and TG showed an increasing trend but not significantly. The active peptide group showed superior effects in reducing triglycerides, total cholesterol, and low-density lipoprotein cholesterol, and increasing high-density lipoprotein cholesterol, and has development potential.

[0034] The above embodiments are only used to illustrate the technical solutions of the present invention, and are not intended to limit it. Although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features. Such modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A peptide that accelerates fat metabolism, characterized in that, The amino acid sequence of the peptide is MVHSYSETSD.

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