JP7837069B2 - Anti-metabolic plant peptides and their use - Google Patents

Description

This invention relates to compositions composed of low molecular weight proteins, and more particularly to anti-metabolic plant peptides and their uses.

As modern people become more health-conscious, consumers are no longer limited to purchasing basic nutritional supplements such as vitamins and minerals, but are also buying functional nutritional supplements tailored to their specific needs. With advances in biotechnology, many manufacturers are now using special manufacturing processes such as fermentation and hydrolysis to extract active ingredients with specific functions from food, and then using these as active ingredients in nutritional supplements to enhance their health-promoting effectiveness.

Statistics show that more than a quarter of the population in Taiwan suffers from metabolic diseases such as hypertension, hyperglycemia, and hyperlipidemia. The causes of these metabolic diseases are not limited to genetics; individual diet and lifestyle habits also play a role. An imbalance between work and rest, significant stress, lack of exercise, and a diet centered on fried foods are all contributing factors. While metabolic diseases may not have a significant impact on health in the short term, they can lead to serious illnesses such as stroke, heart disease, and diabetes in the long term. However, since changing diet and lifestyle habits is not easy, many people choose to control their blood pressure, blood sugar, and blood lipids with nutritional supplements. Currently, however, there are no single nutritional supplements on the market that can simultaneously improve blood pressure, blood sugar, and blood lipids.

The primary objective of this invention is to provide a group of anti-metabolic plant peptides and their use. This group of anti-metabolic plant peptides contains multiple low-molecular-weight active peptides, and these peptides, through a synergistic effect, possess the ability to simultaneously control blood glucose, blood lipids, and blood pressure. Therefore, by administering the anti-metabolic plant peptides disclosed in this invention, it is possible to simultaneously and effectively improve or treat hypertension, hyperglycemia, and hyperlipidemia-related diseases.

To achieve the above objectives, the present invention discloses a group of anti-metabolic plant peptides, which are isolated from the primary products of plant raw materials and comprise at least six low-molecular-weight peptides. These peptides can synergistically interact with each other and, by controlling pathways such as the MAPK pathway and IRS/p-IR/PI3K/Akt pathway within an organism, can simultaneously improve or treat various metabolic diseases associated with imbalances in metabolic indicators such as blood pressure, blood lipids, and blood glucose.

Here, the primary product of the plant material is obtained by sequentially performing two extraction steps on the plant material. The first extraction step is a water extraction step, and the second extraction step is a supercritical fluid extraction step.

In other words, by administering an effective amount of the anti-metabolic plant peptide group disclosed in this invention, or a composition containing said anti-metabolic plant peptide group, to an individual, it is possible to simultaneously combat or improve various metabolic diseases, and in particular, to simultaneously treat or prevent diseases related to hypertension, hyperglycemia, and hyperlipidemia. Here, the effective amount is at least 5 mg/kg/day.

In one embodiment of the present invention, the group of anti-metabolic plant peptides includes peptides with molecular sizes between 300 Da and 3000 Da, as shown in wavelength ranges (L1) to (L6) in a high-performance liquid chromatogram, where the retention time for wavelength range (L1) is 10.5 to 11.5 minutes, for wavelength range (L2) it is 12.5 to 13.5 minutes, for wavelength range (L3) it is 14.5 to 15.5 minutes, for wavelength range (L4) it is 16.5 to 17.5 minutes, for wavelength range (L5) it is 17.5 to 18.5 minutes, and for wavelength range (L6) it is 24.5 to 25.5 minutes.

Here, the high-performance liquid chromatogram was obtained by performing a pretreatment step on the primary product of the plant raw material and then analyzing it using a high-performance liquid chromatograph under predetermined analytical conditions. The analytical conditions were as follows: detection wavelength 220 nm, separation column C18 column (RP-C18), column temperature 60°C, eluent A is 5% acetonitrile containing 0.10% trichloroacetic acid, eluent B is 95% acetonitrile containing 0.085% trichloroacetic acid, and the mobile phase contains 10-20% eluent B used for 0-10 minutes, 20-30% eluent B used for 10-25 minutes, and 30-60% eluent B used for 20-60 minutes.

In another embodiment of the present invention, the pretreatment step includes dissolving the primary product of the plant raw material in 30% acetonitrile, performing ultrasonic extraction to obtain a supernatant, filtering the supernatant to obtain a filtrate, which is then used for high-performance liquid chromatography.

In the pretreatment step described above, filtration is performed using a filter medium with voids of 0.20 to 0.25 μm; the filter medium can be a filtration membrane or a mesh screen.

In another embodiment of the present invention, the molecular size of the peptides included in the group of anti-metabolic plant peptides, indicated by wavelength ranges (L1) to (L6), is between 700 Da and 2400 Da.

The chromatographic analysis results of the peptide group (1) are shown, and the peptide group is a product extracted from bitter melon 1. The results of the chromatographic analysis of the peptide group (2) are shown, and the peptide group is a product extracted from watermelon 1. The chromatographic analysis results of the peptide group (3) are shown, and the peptide group is a product extracted from watermelon 2. The chromatographic analysis results of the peptide group (4) are shown, and the peptide group is a product extracted from pumpkin 1. The chromatographic analysis results of the peptide group (5) are shown, and the peptide group is a product extracted from pumpkin 2. The chromatographic analysis results of the peptide group (6) are shown, and the peptide group is a product extracted from bitter melon 2. The chromatographic analysis results of the peptide group (7) are shown, and the peptide group is a product extracted from Trichosanthes cucumeroides 1. The chromatographic analysis results of the peptide group (8) are shown, and the peptide group is a product extracted from Trichosanthes cucumeroides 2. The chromatographic analysis results of the peptide group (9) are shown, and the peptide group is a product extracted from bitter melon 3. The chromatographic analysis results of the peptide group (10) are shown, and the peptide group is a product extracted from bitter melon 4. The chromatographic analysis results of the peptide group (11) are shown, and the peptide group is a product extracted from bitter melon 5. Figure 1 shows a comparison between the chromatogram of the chromatogram of comparative samples 1-7. This is an analysis of fasting blood glucose levels in each group of mice after the end of the 8th week of the experiment. This shows the results of the analysis of glycated hemoglobin in the blood of each mouse group after the end of the 8th week of the experiment. This is an analysis of fasting blood glucose levels in each group of mice after the end of the 8th week of the experiment. This is an analysis of the body weight of each mouse group after the end of the 8th week of the experiment. This shows the results of the body fat analysis of each mouse group after the end of the 8th week of the experiment. This shows the results of blood triglyceride analysis in each group of mice after the end of the 8th week of the experiment. This shows the results of the analysis of total blood cholesterol in each group of mice after the end of the 8th week of the experiment. This shows the results of the analysis of blood LDL cholesterol in each group of mice after the end of the 8th week of the experiment. This is an analysis of postprandial blood glucose changes in each group of subjects. This is an analysis of the area under the blood glucose curve two hours after a meal for each group of subjects.

This invention discloses a group of anti-metabolic plant peptides and their use. Specifically, the anti-metabolic plant peptides disclosed herein are obtained by separation from the primary product of plant raw materials, and the anti-metabolic plant peptides consist of peptides with molecular sizes between 300 Da and 3000 Da, indicated in the wavelength range (L1) to (L6) with retention times of 10.5 to 25.5 minutes in a high-performance liquid chromatogram.

Here, the high-performance liquid chromatogram was obtained by performing high-performance liquid chromatography under predetermined analytical conditions after pretreatment of the primary product of the plant raw material.

Specifically, the group of anti-metabolic plant peptides disclosed in this invention are obtained by separation from the primary product of a plant raw material. The primary product of the plant raw material is obtained by sequentially performing water extraction and supercritical fluid extraction on the plant raw material. Furthermore, the plant raw material possesses at least one of the following properties: inhibition of dipeptidyl peptidase 4, inhibition of α-amylase, inhibition of GLUT2, inhibition of SGLT1, inhibition of α-glucosidase, and control of insulin.

Here, the plant raw material in question is, for example, a cucurbit plant such as watermelon, pumpkin, bitter melon, or gourd.

The group of anti-metabolic plant peptides disclosed in this invention has the ability to reduce blood glucose, blood pressure, blood lipids, and lipogenesis. Therefore, by administering an effective amount of the group of anti-metabolic plant peptides or a composition containing them to an individual, it is possible to simultaneously and effectively improve indicators related to metabolic diseases. This can lead to the alleviation or treatment of diseases related to hyperglycemia, hyperlipidemia, and hypertension, such as diabetes, kidney disease, hypertension, and dyslipidemia, while also reducing the risk of complications or symptoms associated with the aforementioned diseases.

The "primary product of plant raw materials" described in this invention refers to a product obtained by performing a process combining water extraction and supercritical fluid extraction on plant raw materials, and removing residues and/or residuals. This product includes the group of anti-metabolic plant peptides disclosed in this invention.

The "plant raw materials" described in this invention include the genera Amaranthus, Phaseolus, Glycine, Avena, Cucurbitaceae, Cuminum, Chenopodium, Orus, Juglans, Hordeum, Spinacia, Vigna, Triticum, Linum, and Phalaris . These are derived from plants classified as belonging to genera such as Moringa sp., Theobroma sp., and Zea sp., including bitter melon, snake gourd, cucumber, pumpkin, gourd, watermelon, live ginger, talcum powder (Trichosanthes cucumeroides), zinnia (Zinnia elegans), meadow palm (Medicago truncatula), grapes, grapefruit, elderberry (Sambucus nigra), Arabidopsis thaliana, and rice.

The “composition” described in this invention refers to a composition containing 0.001% to 100% of the anti-metabolic plant peptide group disclosed in this invention. If the composition does not consist entirely of the anti-metabolic plant peptide group disclosed in this invention, it further includes a vehicle and/or excipients acceptable in pharmaceuticals or foods. The composition can be prepared in various dosage forms as needed, such as tablets, powders, or aqueous solutions, and may be in various doses depending on parameters such as administration needs, target population, and frequency of administration. For example, the composition may be a nutritional supplement, a food, or a pharmaceutical.

The term "effective amount" as used in this invention refers to the minimum dose at which the group of anti-metabolic plant peptides disclosed in this invention exerts activity within an individual organism.

The anti-metabolic plant peptides disclosed in this invention can be stably isolated from various types of plant materials and simultaneously have the effects of lowering blood glucose, reducing body weight, reducing body fat, lowering blood triglycerides, lowering total blood cholesterol and blood LDL cholesterol, and lowering blood pressure. In other words, by administering an effective amount of the anti-metabolic plant peptides disclosed in this invention or a composition containing such peptides to an individual, diseases caused by metabolic abnormalities of blood lipids, blood pressure, and blood glucose can be simultaneously and effectively alleviated or improved. By simultaneously preventing or improving various metabolic diseases, the discomfort consumers experience from taking various different nutritional supplements can be reduced, and the risk of interactions with nutritional supplements and/or drugs can be reduced.

The group of anti-metabolic plant peptides disclosed in the first embodiment of the present invention includes a group of peptides, which are multiple functional low-molecular-weight peptides separated from the primary product of a plant raw material. The primary product of the plant raw material is obtained by sequentially performing a water extraction step and a supercritical fluid extraction step on bitter melon. The group of anti-metabolic plant peptides are the peptides shown in the wavelength range (L1) to (L6) in Figure 1, with retention times between 10.5 and 25.5 minutes, and molecular weights between 300 Da and 3000 Da, respectively.

Here, wavelength ranges (L1) to (L6) are obtained by performing a pretreatment step on the primary product of the plant raw material and then analyzing it using high-performance liquid chromatography under predetermined analytical conditions. These analytical conditions include: detection wavelength of 220 nm, separation column of C18 column (RP-C18), column temperature of 60°C, eluent A being 5% acetonitrile containing 0.10% trichloroacetic acid, eluent B being 95% acetonitrile containing 0.085% trichloroacetic acid, and the mobile phase containing 10-20% eluent B used for 0-10 minutes, 20-30% eluent B used for 10-25 minutes, and 30-60% eluent B used for 20-60 minutes.

The holding time for wavelength range (L1) was 10.5–11.5 minutes, for wavelength range (L2) it was 12.5–13.5 minutes, for wavelength range (L3) it was 14.5–15.5 minutes, for wavelength range (L4) it was 16.5–17.5 minutes, for wavelength range (L5) it was 17.5–18.5 minutes, and for wavelength range (L6) it was 24.5–25.5 minutes.

The pre-treatment process includes the following steps:

Step a: The primary product of the plant raw material is dissolved in 30% acetonitrile, and then ultrasonic extraction is performed to obtain the supernatant.

Step b: Filter the supernatant to obtain a filtrate for high-performance liquid chromatography.

Each wavelength range (L1) to (L6) in Figure 1 was analyzed by gel permeation chromatography (GPC), and the molecular weight for each wavelength range (L1) to (L6) was estimated by comparing it with the GPC chromatogram of a standard sample, confirming that it was between 300 and 3000 Da. The standards used were cytidine (molecular weight 243 Da), vitamin B12 (molecular weight 1350 Da), aprotinin (molecular weight 6500 Da), cytochrome (molecular weight 12000 Da), and myoglobin (molecular weight 17600 Da). The order of molecular weights of these substances was as follows: myoglobin (10.226 min), cytochrome (10.658 min), aprotinin (11.864 min), vitamin B12 (13.787 min), and cytidine (14.835 min).

Furthermore, simulated digestion tests were conducted on a group of anti-metabolic disorder plant peptides, and the difference in area percentages in each wavelength range before and after the test was analyzed using high-performance liquid chromatography. The results are shown in Table 1.

Table 1: Analysis results for each wavelength range (L1) to (L6) in Figure 1.

To demonstrate the stable isolation of the anti-metabolic plant peptide group disclosed in this invention, in the second embodiment disclosed in this invention, various cucurbitaceous plants from different origins, including pumpkin, watermelon, bitter melon, and snake gourd, were prepared, and a water extraction step and a supercritical fluid extraction step were sequentially performed on each to obtain primary products of the plant raw materials. After treating the primary products of each plant raw material according to the above-described pretreatment steps, analysis was performed using high-performance liquid chromatography under the above-described analytical conditions to obtain the chromatograms shown in Figures 2 to 11. Here, in the explanation of each figure, the numbers after the cucurbitaceous plants are used to distinguish between batches or sources of the same cucurbitaceous plant. Subsequently, the content of each wavelength range in each chromatogram was analyzed as shown in Table 2 below.

As can be seen from the chromatograms in Figures 2 to 11, wavelength ranges (L1) to (L6) appeared with retention times of 10.5 to 25.5 minutes, respectively. Of these, the retention time for wavelength range (L1) was 10.5 to 11.5 minutes, for wavelength range (L2) 12.5 to 13.5 minutes, for wavelength range (L3) 14.5 to 15.5 minutes, for wavelength range (L4) 16.5 to 17.5 minutes, for wavelength range (L5) 17.5 to 18.5 minutes, and for wavelength range (L6) 24.5 to 25.5 minutes.

Table 2: Content in wavelength ranges (L1) to (L6) in Figures 2 to 11

As can be seen from the chromatograms disclosed in Figures 1 to 11, the group of anti-metabolic plant peptides disclosed in this invention stably appears in the primary products of the plant raw materials in each batch, and stably appears in all different types of plant raw materials. Therefore, it is clear that the group of anti-metabolic plant peptides disclosed in this invention possesses both availability and reproducibility.

The group of anti-metabolic plant peptides disclosed in the third embodiment of the present invention is the same as in the above-described embodiments, but the difference lies in the pretreatment step performed on the primary product of the plant raw material. The specific step is as follows:

After dissolving the primary product of the plant raw material in 30% acetonitrile, a first ultrasonic extraction is performed for 20 minutes. The water bath temperature is lower than room temperature, and the mixture is separated by centrifugation at 9000 rpm for 5 minutes to obtain the first supernatant and first precipitate. After dissolving the first precipitate in 30% acetonitrile, a first ultrasonic extraction is performed for 20 minutes. The water bath temperature is lower than room temperature, and the mixture is separated by centrifugation at 9000 rpm for 5 minutes to obtain the second supernatant. The first and second supernatants are collected and filtered through a filtration membrane with a void size of 0.20–0.25 μm. The resulting filtrate can be used for subsequent high-performance liquid chromatography.

In addition, to demonstrate the novelty and uniqueness of the anti-metabolic plant peptide group disclosed in this invention, in this example, commercially available bitter melon extracts (comparative samples 1 to 7) were each subjected to the pretreatment method disclosed in the first example of this invention, analyzed by high-performance liquid chromatography under the same analytical conditions, and the chromatograms obtained from comparative samples 1 to 7 were combined with the chromatogram shown in Figure 1. The results are shown in Figure 12 .

As can be seen from the results in Figure 1 and 2 , the group of anti-metabolic plant peptides disclosed by the present invention includes peptides in the wavelength range (L1) to (L6), while comparative sample 1 does not have wavelength ranges (L1), (L2), and (L6); comparative sample 2 does not have wavelength ranges (L1), (L4), and (L5); comparative sample 3 does not have wavelength ranges (L1) to (L6); comparative sample 4 does not have wavelength ranges (L1) to (L6); comparative sample 5 does not have wavelength ranges (L1) to (L6); comparative sample 6 does not have wavelength ranges (L1), (L3) to (L6); and comparative sample 7 does not have wavelength ranges (L4) to (L6). These results demonstrate that the group of anti-metabolic plant peptides disclosed by the present invention is indeed novel and unique, and is not obtained by isolation from other plant extracts of the same origin.

The following is an efficacy study conducted to demonstrate that the anti-metabolic plant peptides disclosed in this invention have therapeutic or ameliorative effects on diseases related to abnormalities in blood pressure, blood glucose, and blood lipids.

First, the human dose was based on 300 mg/60 kg adult body weight/day. This was converted to a rat dose according to common knowledge known to those skilled in the art, and the following tests were conducted according to various animal model designs. The test period for all tests was 8 weeks.

Animal study (1): SD rats were randomly divided into three groups. Group 1 consisted of normal rats, while Groups 2 and 3 were rats that were type 2 diabetic models induced by simultaneous administration of the drug STZ (25 mg/kg body weight) and a high-fat diet. Group 3 received continuous administration of the anti-metabolic plant peptides disclosed in this invention throughout the study period. After the 8th week of the study, fasting blood glucose levels and glycated hemoglobin levels were examined in each group of mice. The results are shown in Figures 13 and 14.

Animal study (2): SD rats were randomly divided into three groups. Group 1 consisted of normal rats, while Groups 2 and 3 were rats that were type 1 diabetic models induced by the drug STZ (75 mg/kg body weight). Group 3 received continuous administration of the anti-metabolic plant peptides disclosed in this invention throughout the study period. After the 8th week of the study, fasting blood glucose levels were measured in each group of mice. The results are shown in Figure 15.

Animal study (3): SD rats were randomly divided into two groups. Both groups were given a high-fat diet, increasing the body weight of both groups by 10%. An 8-week study was then conducted. The high-fat diet was maintained throughout the study period, except for the second group of rats, which were given the anti-metabolic plant peptide group disclosed in this invention. After the study, body weight, body fat, blood triglycerides, blood total cholesterol, and blood LDL cholesterol were measured for each group of rats. The results are shown in Figures 16 to 20.

Furthermore, multiple subjects were gathered and randomly divided into two groups. Group 1 did not receive the anti-metabolic plant peptides disclosed in this invention, while Group 2 received the anti-metabolic plant peptides disclosed in this invention (300 mg/60 kg adult body weight/day). Each subject in each group was given 75 g of glucose on an empty stomach. Blood glucose levels were measured at 0, 30, 60, 90, and 120 minutes after ingestion, and the area under the curve (OGTT AUC) of the oral glucose tolerance test was analyzed 2 hours later. The results are shown in Figures 21 and 22.

The results of the animal and human studies described above are summarized in Table 3 below. As can be seen from the results in Table 3 and Figures 12 to 21, the group of anti-metabolic plant peptides disclosed in this invention has an improving effect on both congenital and acquired blood glucose metabolic disorders. Furthermore, even under conditions of continuous high-fat diets, it was possible to effectively reduce the effects of high-fat diets on blood lipids and cholesterol. From this, it can be seen that the group of anti-metabolic plant peptides disclosed in this invention can indeed simultaneously and effectively improve abnormal metabolic indicators, simultaneously improve symptoms related to hyperglycemia, hyperlipidemia, and obesity, and achieve a reduction in the risk of developing diseases caused by the aforementioned metabolic disorders.

Table 3: Analysis results of efficacy

Furthermore, to explain the difference in efficacy when comparing the anti-metabolic plant peptide group disclosed in the present invention with compositions that do not contain the peptide group indicated in other wavelength ranges (L1) to (L6), the anti-metabolic plant peptide group of the present invention, comparative sample 1, and comparative sample 4 were each administered to rats of a type 2 diabetes model, and the changes in body weight, FBG , and HbA1c of the rats in each experimental group were analyzed. The results are shown in Table 4. For details of the experimental procedure, please refer to the explanation of animal test (1) above. Also, the data in Table 4 represent the percentage change for each experimental group relative to the diabetic model rats.

Furthermore, after administering the anti-metabolic plant peptide group of the present invention and comparative sample 4 to rats of a high-fat diet model, changes in body weight, body fat, triglycerides, and total cholesterol in each experimental group of rats were analyzed. The results are shown in Table 5. Please refer to the explanation of the animal test (3) above for the experimental procedure. Also, the data in Table 5 represent the percentage change for each experimental group compared to diabetic model rats.

The results in Tables 4 and 5 demonstrate that the anti-metabolic plant peptide group disclosed in this invention has a significant effect in improving blood glucose and blood lipid control compared to compositions that do not contain the peptide group shown in other wavelength ranges (L1) to (L6).

4: Percentage change in each experimental group compared to diabetic model rats

Table 5: Percentage change in each experimental group relative to diabetic model rats

Furthermore, the blood pressure-lowering activity of the anti-metabolic plant peptides disclosed in this invention was analyzed through activity analysis of LC angiotensin-converting enzyme (ACE) using liquid chromatography. Specifically, although the peptides use hippryl-L-histidyl-L-leucine (HHL, Hip-His-Leu) as a substrate, ACE action generates hippuric acid (HA, hippuric acid) and histidylleucine (His-Leu). Therefore, if ACE activity is inhibited after the addition of the test sample and the amount of hippuric acid and histidylleucine produced decreases, it means that the test sample can effectively treat or improve hypertension. In this example, the blood pressure-lowering ability of the anti-metabolic plant peptides disclosed in this invention was analyzed by comparing the amount of hippuric acid produced with and without the addition of the anti-metabolic plant peptides disclosed in this invention. Here, the inhibition rate of ACE activity was calculated by measuring the amount of hippuric acid produced (integrated area) using HPLC analysis. The test results showed that the group treated with the anti-metabolic plant peptides disclosed in this invention (10 mg/mL) had a blood pressure reduction of 30.63% compared to the control group without the anti-metabolic plant peptides. This demonstrates that the anti-metabolic plant peptides disclosed in this invention do indeed have the ability to lower blood pressure and can be used in the preparation of compositions for treating or improving hypertension and related diseases.

Claims (6)

The isolated group of anti-metabolic plant peptides is isolated from the primary product of a cucurbitaceous plant raw material, and the primary product of the cucurbitaceous plant raw material is obtained by sequentially performing water extraction and supercritical fluid extraction on the cucurbitaceous plant raw material.
The aforementioned group of antimetabolic plant peptides includes peptides with molecular sizes between 700 Da and 2400 Da, as shown in wavelength ranges (L1) to (L6) in a high-performance liquid chromatogram, with retention times of 10.5 to 11.5 minutes for wavelength range (L1), 12.5 to 13.5 minutes for wavelength range (L2), 14.5 to 15.5 minutes for wavelength range (L3), 16.5 to 17.5 minutes for wavelength range (L4), 17.5 to 18.5 minutes for wavelength range (L5), and 24.5 to 25.5 minutes for wavelength range (L6).
The high-performance liquid chromatogram was obtained by performing a pretreatment step on the primary product of the cucurbitaceous plant raw material, and then analyzing it using a high-performance liquid chromatograph under predetermined analytical conditions.
The aforementioned analytical conditions are as follows: detection wavelength is 220 nm, separation column is C18 column (diameter 4.6 mm x length 250 mm, flow rate approximately 1.0 ml/min) , column temperature is 60°C, eluent A is 5% acetonitrile containing 0.10% trichloroacetic acid, eluent B is 95% acetonitrile containing 0.085% trichloroacetic acid, and the mobile phase includes 10-20% eluent B used for 0-10 minutes, 20-30% eluent B used for 10-25 minutes, and 30-60% eluent B used for 20-60 minutes .
The aforementioned pretreatment step is
Step a: After dissolving the primary product of the cucurbitaceous plant raw material in 30% acetonitrile, ultrasonic extraction is performed to obtain the supernatant.
A group of separated anti-metabolic plant peptides , comprising step b, which involves filtering the supernatant to obtain a filtrate for high-performance liquid chromatography . The separated group of anti-metabolic plant peptides according to claim 1 , wherein in step b, filtration is performed using a filter material with voids of 0.20 to 0.25 μm. The use of the isolated group of anti-metabolic plant peptides described in claim 1 or claim 2 in the preparation of a composition for preventing or treating hyperlipidemia, hypertension, and hyperglycemia. The use according to claim 3 , wherein the human dose of the isolated group of anti-metabolic plant peptides contained in the composition is 5 mg/kg/day. Use of the isolated group of anti-metabolic plant peptides described in claim 1 or claim 2 in the preparation of a diet composition. The use according to claim 5 , wherein the human dose of the isolated group of anti-metabolic plant peptides contained in the composition is 5 mg/kg/day.