Momordica charantia polypeptide for sugar balance control by exosome regulation

Abstract

The present invention addresses the problem of providing a Momordica charantia polypeptide for sugar balance control by exosome regulation. The administration of a Momordica charantia polypeptide according to the present invention or a composition containing said Momordica charantia polypeptide to an individual has a prophylactic or therapeutic effect on patients suffering from a blood sugar level imbalance by increasing the exosome concentration in the pancreas and controlling the secretion of insulin from the pancreatic islets.

Description

Bitter melon polypeptide for carbohydrate balance control by exosome regulation

[0001] The present invention relates to uses of polypeptides, and in particular to bitter melon polypeptides for controlling carbohydrate balance by regulating exosomes.

[0002] The International Diabetes Federation predicts that the global diabetic population will exceed 600 million by 2045, becoming a national health issue in countries around the world. Diabetes is a metabolic disease related to impaired insulin secretion; in other words, when there is a lack of insulin in the body or when pancreatic islets malfunction, blood glucose cannot be absorbed into cells, causing blood sugar levels to rise, and if blood sugar levels remain unbalanced for a long period of time, it can cause diseases such as vascular disease, retinopathy, stroke, neuropathy, and nephropathy.

[0003] Exosomes are small vesicles approximately 30-150 nanometers in size secreted by cells and possess a lipid bilayer membrane structure. Exosomes are released into the extracellular space in the form of nanoparticles and are used to transmit or transport molecules such as proteins, RNA, and DNA. They also enter target cells and transmit messages between cells, thereby achieving cellular repair and regeneration. Recent studies have shown that exosomes can continuously regulate insulin function, control blood sugar levels, and improve diabetes. Therefore, the infusion of exosomes into individuals is currently being used clinically as a new treatment for diabetes and its complications. However, it faces the following challenges: First, if the exosomes are derived from unhealthy cells, unpredictable side effects may occur in the infused individual. Second, exosomes must be isolated from cells before they can be obtained. If the exosome structure is damaged during the isolation process, the exosomes lose their function and the expected improvement effects are not achieved.

[0004] Taiwan Patent No. I725326

[0005] The main object of the present invention is to provide a bitter melon polypeptide for controlling carbohydrate balance by regulating exosomes, which can exert the effect of controlling an individual's blood glucose level by directly adjusting the content and composition of autologous exosomes without supplementing exogenous exosomes.

[0006] A second object of the present invention is to provide a bitter melon polypeptide for controlling carbohydrate balance by regulating exosomes, which can provide consumers with another effective and safe method for regulating their own exosomes in preventive medicine.

[0007] Therefore, to achieve the above-mentioned object, the present invention discloses the use of a bitter melon extract containing at least one bitter melon polypeptide in the preparation of an exosome-modulating agent or a composition for preventing diabetes.

[0008] Specifically, the bitter melon polypeptide according to the present invention has the activity of increasing the exosome concentration in the pancreas and / or repairing pancreatic islets, and therefore, by administering an effective amount of the bitter melon polypeptide or a composition containing the bitter melon polypeptide to an individual, it is possible to achieve the effect of preventing or treating diseases caused by blood sugar.

[0009] The individual is a pre-diabetic patient.

[0010] In one embodiment of the present invention, the bitter melon extract is obtained from bitter melon or a Cucurbitaceae plant through an extraction technique.

[0011] In one embodiment of the present invention, the bitter melon extract is prepared as an exosome regulator and used to regulate exosome secretion in the patient's own pancreas so as to control insulin secretion.

[0012] In another embodiment of the present invention, the bitter melon extract is prepared as a composition for preventing diabetes to be administered to an individual who has abnormal blood glucose levels or glycated hemoglobin levels but has not yet reached a stage that is clinically defined as diabetes. By administering the composition for preventing diabetes to the individual, the blood glucose levels and other metabolic indices of the individual can be effectively controlled, thereby preventing the onset of diabetes or its complications.

[0013] The composition for preventing diabetes can not only control the blood sugar level of an individual, but also increase the skeletal muscle mass and decrease the body fat mass of an individual.

[0014] The effective dose of the bitter melon extract for the human body is at least 100 mg per day, and more effective is a dose of more than 300 mg per day.

[0015] Graph showing particle size analysis of vesicles in pancreatic tissue extract of rats in the control group. Graph showing particle size analysis of vesicles in pancreatic tissue extract of rats in the experimental group. Image showing the results of microscopic observation of vesicles in pancreatic tissue extract of rats in the control group. Image showing the results of microscopic observation of vesicles in pancreatic tissue extract of rats in the experimental group. Graph showing analysis of changes in HbA1c in subjects of different age groups in each group. Graph showing analysis of changes in fasting blood glucose levels in subjects of different age groups in each group. Graph showing analysis of changes in HOMA2 secretory function index in subjects of different age groups in each group. Graph showing analysis of changes in non-HDL cholesterol content in subjects of different age groups in each group.

[0016] The present invention discloses a bitter melon polypeptide for controlling carbohydrate balance by regulating exosomes. Specifically, the bitter melon polypeptide of the present invention has the activity of increasing the number of pancreatic islet exosomes and can be used to repair pancreatic islets and / or regulate insulin secretion from pancreatic islets, thereby achieving the effect of controlling blood glucose levels in an individual. Therefore, by administering an effective amount of the bitter melon polypeptide of the present invention or a composition containing the bitter melon polypeptide to an individual, the exosome concentration in the individual's own pancreas can be increased and insulin secretion from pancreatic islets can be controlled, thereby achieving the effect of preventing or treating diseases associated with imbalances in blood glucose levels.

[0017] In one embodiment of the invention, the individual is an individual who meets the definition of pre-diabetes.

[0018] In one embodiment of the present invention, diseases associated with imbalance in blood sugar levels include metabolic syndrome, diabetes, hyperglycemia, kidney disease, cardiovascular disease, neuropathy, retinopathy, and the like.

[0019] In one embodiment of the present invention, the bitter melon polypeptide is derived from a bitter melon extract obtained by processing bitter melon with an extraction technique. Specifically, the bitter melon extract contains proteins, fats, carbohydrates, and metal ions.

[0020] Furthermore, the bitter melon polypeptide of the present invention can be isolated from extracts of other Cucurbitaceae plants such as pumpkin extract and watermelon extract, or can be directly prepared by biosynthetic methods.

[0021] In one embodiment of the present invention, when the subject is a 60 kg adult, the effective dose of the bitter melon extract in the human body is at least 100 mg per day, for example, 100 mg / day, 200 mg / day, 350 mg / day, 400 mg / day, 500 mg / day, or 600 mg / day, with at least 300 mg per day being preferred.

[0022] In another embodiment of the present invention, the effective dose of the bitter melon extract in the human body varies depending on factors such as the bitter melon polypeptide content in the bitter melon extract, the administration target, and the dosage form, and this can be achieved by a person skilled in the art through common technical means, so the adjustment of the effective dose is also within the scope of protection of the present invention.

[0023] The term "extraction technology" refers to a method for separating a desired component from a raw material, such as solvent extraction, low-temperature extraction, high-temperature extraction, supercritical extraction, ultrasonic extraction, hydrolysis separation, enzyme extraction, and the like.

[0024] The term "prediabetes" is defined as an HbA1c between 5.7% and 6.4% and / or a fasting capillary glucose between 100 and 125 mg / dL.

[0025] In order to explain the technical features and effects that can be achieved by the present invention, several experimental examples will be given below and will be described in detail with reference to the drawings.

[0026] The bitter melon peptide composition used in the following examples is a powder mixture obtained from bitter melon (Momordica charantia L.) by extraction techniques generally known to those skilled in the art, and contains a plurality of bitter melon polypeptides such as those disclosed in Patent Document 1. The bitter melon peptide composition contains protein, fat, carbohydrates, and metal ions, and the weight ratio of protein, fat, and carbohydrate was about 1.5:1:2.5-3.

[0027] Example 1: Animal experiment Type 2 diabetes model rats were established according to techniques well known in the art, and the type 2 diabetes model rats were randomly divided into two groups, each of which was kept for 8 weeks under the following conditions.

[0028] Control group: water supply.

[0029] Experimental group: fed with bitter melon peptide composition, the dosage was 31.25 mg / kg body-weight / day.

[0030] After the experiment, pancreatic tissue from each of the two groups of rats was collected and subjected to extraction to obtain pancreatic tissue extracts. The particle size of the vesicles (exosomes) in the pancreatic tissue extracts from each group of rats was analyzed, and the results are shown in Table 1 and Figures 1 to 4. The mode diameter in Table 1 refers to the particle size with the highest frequency of occurrence in the particle size distribution, also known as the mode value. D50 refers to the particle size corresponding to a cumulative frequency of 50% in the particle size distribution for each group, also known as the median diameter.

[0031]

[0032] The results of this example show that compared with the control group, the rats in the experimental group had significantly higher vesicle concentrations in the pancreatic tissue fluid, and the composition of the pancreatic secretions was also different. These results indicate that administering the bitter melon peptide composition of the present invention to diabetic individuals can effectively regulate exosomes in the individual's pancreatic islets, repair or regulate the insulin secretion function of the pancreatic islets, and thereby improve or treat diabetes or its complications. In other words, the bitter melon polypeptide or bitter melon polypeptide-containing composition of the present invention has the activity of regulating autologous exosomes, and by regulating the secretion of autologous exosomes, it exerts the effect of controlling blood glucose levels.

[0033] (Example 2: Human Experiment) Seventy-five pre-diabetic patients who met the conditions were selected as subjects for this example, and were randomly divided into three groups, and a 12-week experiment was carried out under the following conditions.

[0034] Group 1, Placebo Group: Each subject received 600 mg of placebo (sugar-free, non-viscous soluble fiber) per day.

[0035] Second group, low dose group: Each subject was given 300 mg of bitter melon peptide composition and placebo (sugar-free, non-viscous soluble fiber) per day.

[0036] Group 3, high dose group: Each subject was given 600 mg of bitter melon peptide composition per day.

[0037] During the experimental period, the subjects in each group did not consume bitter melon or any food made from bitter melon, except for the bitter melon peptide extract.

[0038] The selection criteria for subjects were as follows: age range: 18 to 74 years, BMI: 25.0 to 39.9 kg / m ２Those who fall into any of the following categories will not be able to participate in the study: those who have been taking antihypertensive or lipid-altering medications (either just started taking them or had their dose changed) within the four weeks prior to the start of the study, those who have been taking hypoglycemic drugs, systemic corticosteroids, or dietary supplements that regulate lipid or carbohydrate metabolism or body weight (e.g., chromium, omega-3 fatty acid supplements, plant sterols, or stanols) within the four weeks prior to the start of the study, and those who have been using PCSK9 (proprotein convertase subtilisin kexin type 9) within the 12 weeks prior to the start of the study. 9) Taking an inhibitor, having a history of bariatric surgery, currently taking diet drugs, having a weight change of 4.5 kg within 12 weeks prior to the start of the study, extreme diets (vegan, Atkins diet, very low carbohydrate diet, etc.), intention to gain or lose weight, taking antibiotics for infection or clinically significant symptoms within 5 days prior to the start of the study, but within the washout period clinically significant laboratory test results: positive urine drug screen; clinically significant medical diagnoses: uncontrolled hypertension (systolic blood pressure ≥ 160 mmHg and / or diastolic blood pressure ≥ 100 mmHg); allergy, hypersensitivity, and / or intolerance to the study product; history of primary cancer within 2 years (excluding non-melanoma skin cancer or primary cervical cancer); severe drug or alcohol abuse within 12 months of the end of the study; contact with unregistered drugs within 30 days of the end of the study; pregnant, planning pregnancy, or breastfeeding during the study period, or capable of becoming pregnant but not using medically approved contraception.

[0039] Experimental Procedure One week before the experiment: HbA1c and fasting blood glucose level of each subject were measured, and height, weight, heart rate and blood pressure were measured. Urine was collected and analyzed.

[0040] Experimental period (weeks 0 to 12): At weeks 0, 6, and 12, blood was collected from each subject and HbA1c, fasting blood glucose, insulin, and lipoprotein lipids were measured. At week 12, urine was collected from each subject and analyzed, as well as blood biochemistry analysis and complete blood count analysis.

[0041] The analysis and measurement results for each group during the experimental period are shown in Tables 2 to 4 below. The changes in HbA1c, HOMA2%S, and HOMA2%B at week 6 or week 12 in each table are numerical values ​​rather than percentages, while the changes in other items are expressed as percentages, and the values ​​in each column in the tables are expressed as averages or medians.

[0042]

[0043]

[0044]

[0045] Comparing the results in Tables 2 to 4, it can be seen that the triglycerides in the low-dose group (Group 2) and the high-dose group (Group 3) decreased by 5.5% and 10.5%, respectively, at week 12 compared to week 0, and the skeletal muscle mass increased by 1.7% and 1.6%, respectively, at week 12 compared to week 0, whereas the triglycerides in the placebo group (Group 1) increased by 7.5% at week 12 compared to week 0, and the skeletal muscle mass decreased by 0.2% at week 12 compared to week 0.

[0046] Furthermore, the mean change in fasting blood glucose levels at week 12 in the low-dose group (Group 2) and the high-dose group (Group 3) was smaller than the increase in the placebo group; specifically, the mean change in fasting blood glucose levels at week 12 in the high-dose group (Group 3) (1.33 ± 2.77%) was significantly smaller than the change rate in Group 1 (9.08 ± 2.66%).

[0047] Furthermore, the subjects in each group were classified by median age (55.6 years), and the effects of the placebo or bitter melon peptide composition on changes in HbA1c and blood glucose levels in subjects of different age groups were analyzed, with the results shown in Figures 5 and 6. The subjects in each group were also classified by median blood glucose level (108 mg / dL), and the effects of the placebo or bitter melon peptide composition on the HOMA2 secretory function index and non-HDL cholesterol content in subjects with hyperglycemia or hypoglycemia were analyzed, with the results shown in Figures 7 and 8.

[0048] The results in Figures 5 and 6 show that in older subjects (55.6 years or older), high-dose bitter melon peptide compositions were able to reduce HbA1c and fasting blood glucose levels more than the placebo group, while in younger subjects (under 55.6 years old), both high- and low-dose bitter melon peptide compositions were able to reduce fasting blood glucose levels more than the placebo group. The results in Figures 7 and 8 show that both high- and low-dose bitter melon peptide compositions were able to improve HOMA2 secretory function and reduce non-HDL cholesterol levels more than the placebo group, with the high-dose bitter melon peptide composition showing better improvement in subjects with hyperglycemia (≥108 mg / dL).

[0049] The results of this example demonstrate that the bitter melon peptide composition of the present invention can reduce the risk of developing diabetes and prevent diabetes and its complications, thereby helping prediabetic individuals maintain healthy blood glucose levels. In other words, if prediabetic individuals can continue to take the bitter melon peptide composition of the present invention, they can normally control their blood glucose levels, which helps reduce the risk of developing blood glucose-related metabolic diseases, and can prevent diabetes and maintain metabolic health.

Claims

1. Use of a bitter melon extract containing at least one bitter melon polypeptide in the preparation of an exosome modulating agent.

2. Use of bitter melon extract in the preparation of the exosome modulating agent according to claim 1, wherein the bitter melon polypeptide can increase the exosome concentration in the pancreas.

3. Use of bitter melon extract in the preparation of the exosome modulating agent according to claim 2, wherein the exosome modulating agent can be used to restore pancreatic islet function.

4. Use of bitter melon extract in the preparation of the exosome modulating agent according to claim 2, wherein the exosome modulating agent can be used to regulate insulin secretion from pancreatic islets.

5. Use of a bitter melon extract containing at least one bitter melon polypeptide in the preparation of a composition for preventing diabetes.

6. Use of bitter melon extract in the preparation of a composition for preventing diabetes according to claim 5, wherein said composition for preventing diabetes is used for individuals with prediabetes.

7. Use of bitter melon extract in the preparation of a composition for preventing diabetes according to claim 6, wherein the age of the individual is over 50 years.

8. Use of bitter melon extract in the preparation of a composition for preventing diabetes according to claim 5, wherein the composition for preventing diabetes can be used to increase skeletal muscle mass.

9. Use of bitter melon extract in the preparation of a composition for preventing diabetes according to claim 5, wherein the composition for preventing diabetes can be used to reduce body fat mass.

10. The use of bitter melon extract in the preparation of a composition for preventing diabetes according to claim 5, wherein the effective dose of said bitter melon extract in the human body is at least 100 mg per day.

11. The use of bitter melon extract in the preparation of a composition for preventing diabetes according to claim 6, wherein the effective dose of said bitter melon extract in the human body is at least 300 mg per day.