A long-acting additive with blood sugar and blood lipid lowering effects and its preparation method

Abstract

The present invention discloses a long-acting additive with blood sugar lowering and blood lipid lowering effects and a preparation method thereof, and relates to the technical field of additives. The additive comprises the following components in mass fractions: 1-10% edestin, 1-10% edestin polypeptide, 60-90% poloxamer 188, and 5-20% fructose, which total 100%. The additive provided by the present invention can efficiently and rapidly lower the body's blood sugar and blood lipid levels within a safe range, and significantly improve bioavailability. Compared with a simple mixing method, the additive exhibits better blood sugar lowering and blood lipid lowering effects, and exceeds any other single ingredient. It is particularly suitable for the prevention of diabetes and hyperlipidemia in the daily life of the general population and for patients to assist in their treatment, and has great application potential.

Description

Technical Field

[0001] The present invention relates to the technical field of additives, and in particular to a long-acting additive with blood sugar lowering and blood lipid lowering effects and a preparation method thereof. Background Art

[0002] With economic growth, lowering blood sugar and lipid levels has become a valuable research area in areas such as drug development. The number of people with diabetes is rapidly increasing worldwide, with my country having the largest number of patients. Diabetes can easily lead to kidney damage and even renal failure, and can trigger cardiovascular and cerebrovascular diseases, while also adversely affecting nerves and metabolism. Hyperlipidemia, defined as excessive levels of lipids in the plasma, is a major cause of atherosclerosis, which in turn contributes to cardiovascular and cerebrovascular diseases such as stroke and coronary heart disease. Approximately 40% of the population in my country has dyslipidemia. Therefore, the prevention and treatment of diabetes and hyperlipidemia are imperative.

[0003] Currently, the treatment of diabetes and hyperlipidemia mainly relies on chemical drugs. While these drugs are effective, long-term or excessive use can lead to abnormal liver and kidney metabolism or damage. Regarding blood sugar reduction, the existing invention patent application CN201410374772.1 discloses a hemp seed meal polypeptide that inhibits α-glucosidase activity and its enzymatic hydrolysis preparation method. Regarding blood lipid reduction, the existing invention patent ZL202210475808.X discloses a soybean peptide and oligopeptide with lipid-lowering effects, as well as their preparation method and application. The inventors administered soybean peptides and oligopeptides to mice by gavage for 35 days and found that they significantly reduced total cholesterol and triglyceride levels in the mice's blood. Although prior art discloses techniques for using specific polypeptides to achieve blood sugar and lipid-lowering effects, the peptides are rapidly released after ingestion. While this offers the advantage of rapid onset of action, the sustained release of the peptides is poor, resulting in a lack of sustained blood sugar and lipid-lowering effects (animal experiments have shown that prolonged use is required). Summary of the Invention

[0004] In response to the shortcomings of the above-mentioned background technology, the present invention provides a long-acting additive with blood sugar and blood lipid lowering effects and a preparation method thereof, in order to solve the problems of low blood sugar and blood lipid lowering efficiency and excessively fast release rate of edestrin polypeptide.

[0005] To achieve the above objectives, the first objective of the present invention is to provide a long-acting additive with hypoglycemic and hypolipidemic effects, the additive comprising the following components by mass fraction: 1-10% edestin, 1-10% edestin polypeptide, 60-90% poloxamer 188, and 5-20% fructose, totaling 100%.

[0006] Preferably, the edestrin is prepared by a salting-out method.

[0007] Preferably, the edestrin polypeptide is a polypeptide prepared by a composite enzymatic hydrolysis method, that is, it is first enzymatically hydrolyzed with papain and then enzymatically hydrolyzed with a neutral protease to perform composite enzymatic hydrolysis.

[0008] Preferably, edestrin and edestrin polypeptide are functional factors, and poloxamer 188 and fructose are synergistic auxiliary ingredients.

[0009] The second object of the present invention is to provide a method for preparing a long-acting additive having the effects of lowering blood sugar and blood lipids, comprising the following steps:

[0010] The additive is prepared by adding edestin, edestin polypeptide and fructose into molten poloxamer 188, uniformly mixing, rapidly cooling to form a solid, and grinding the solid into powder.

[0011] Preferably, during the solidification process, the molten mixture of edestrin, edestrin polypeptide, poloxamer 188 and fructose is poured into an aluminum or copper container and quickly immersed in a CaCl2 aqueous solution at a temperature of -20°C. The molten mixture will quickly solidify into a solid.

[0012] The third object of the present invention is to provide a long-acting additive with hypoglycemic and hypolipidemic effects for use in the preparation of hypoglycemic and hypolipidemic medicines.

[0013] Preferably, during application, it is prepared into powder, tablet or capsule.

[0014] Compared with the prior art, the present invention has the following beneficial effects:

[0015] The present invention provides a long-acting additive with hypoglycemic and hypolipidemic effects and a preparation method thereof. The combination formula of the present invention includes edestin, edestin polypeptide, poloxamer 188, and fructose. To further optimize the hypoglycemic and hypolipidemic properties of the additive, edestin is prepared by a sodium chloride precipitation method, and edestin polypeptide is prepared by a composite enzymatic hydrolysis method. edestin and edestin polypeptide are used as functional factors, poloxamer 188 and fructose are used as synergistic auxiliary ingredients, and the additive is prepared by a special process of melt mixing, rapid cooling, and grinding. As verified by the examples and comparative examples, this composition has the effect of significantly and persistently reducing the body's blood glucose concentration, total cholesterol concentration, triglyceride concentration, and low-density lipoprotein concentration, and can exert its effects efficiently and quickly within a safe range. It is particularly suitable for the prevention of diabetes and hyperlipidemia in the daily life of the general population, and as an auxiliary measure for the treatment of patients, and has broad application prospects.

[0016] This invention utilizes an innovative "protein + polypeptide" combination, allowing the polypeptide to be rapidly absorbed and effective upon entry into the body. Furthermore, once the protein enters the gastrointestinal tract, it is continuously degraded by the digestive system, forming new polypeptides and amino acids that are continuously absorbed by the body, extending the release time of the active ingredients.

[0017] The present invention adopts a combination of poloxamer 188 and fructose, which can help improve the solubility of functional factor components (protein + polypeptide) in a water-based environment and increase the bioavailability of functional factor components. Therefore, it not only has a fast onset of effect and good lasting effect, but also has high bioavailability.

[0018] The present invention adopts a special melt-state quick-freezing process to achieve the homogenization of the internal structure of the solid mixture product, so that the components of the final additive product are more evenly distributed, and the various components can more effectively play a synergistic role, and the blood sugar and blood lipid lowering effects are better.

[0019] In summary, the novel additive disclosed in this invention utilizes a novel formula and unique preparation process, resulting in not only rapid onset of action but also sustained blood sugar and lipid-lowering effects. Compared to simple blends of individual components, this additive exhibits superior blood sugar and lipid-lowering effects, and its duration of action also exceeds that of any other single ingredient. DETAILED DESCRIPTION

[0020] In order to enable those skilled in the art to better understand and implement the technical solution of the present invention, the present invention is further described below with reference to specific embodiments, but the embodiments are not intended to limit the present invention.

[0021] This invention addresses the current issues of edestin polypeptides, such as low blood sugar and lipid lowering efficiency and excessively rapid release rates. This invention utilizes edestin and edestin polypeptides as functional factors, with poloxamer 188 and fructose as synergistic auxiliary ingredients. The additive is prepared through a special process of melt mixing, rapid cooling, and grinding.

[0022] A first aspect of the present invention provides a long-acting additive having the effects of lowering blood sugar and blood lipids, characterized in that the additive comprises the following components by mass fraction: 1-10% edestin, 1-10% edestin polypeptide, 60-90% poloxamer 188, and 5-20% fructose, totaling 100%.

[0023] The edestin is prepared by salting out method, specifically using hemp seed meal as raw material and adopting salting out method to prepare edestin.

[0024] The edestin polypeptide is prepared by a combined enzymatic hydrolysis method, i.e., first hydrolyzing the edestin with papain and then hydrolyzing it with a neutral protease. Specifically, the edestin is initially hydrolyzed with papain and then hydrolyzed with a neutral protease to obtain an edestin polypeptide with excellent blood sugar and lipid-lowering activity.

[0025] Edethin and edethin polypeptide are functional factors, and poloxamer 188 and fructose are synergistic auxiliary ingredients.

[0026] Preparation of edema protein: Add defatted hemp meal to a 5% sodium chloride solution, stir and heat for 1 hour, then control the heating temperature to 60±2°C. Maintain a constant temperature of 60°C and let it stand for another 1 hour. Remove the supernatant, wait for the temperature to cool to room temperature, and place it in a 4°C refrigerator to allow the protein to naturally settle. Remove the precipitate, add 20 times the volume of distilled water, and stir thoroughly to completely disperse the protein in the distilled water. Place it in the refrigerator again and let it stand. Remove the precipitate. Repeat this process 5 times and then dialyze to obtain desalted hemp protein. After freeze-drying, edema protein is obtained.

[0027] At the same time, edestrin was used as raw material and a composite enzymatic hydrolysis method was adopted to prepare polypeptides, that is, papain was first used for enzymatic hydrolysis, and then neutral protease was used for enzymatic hydrolysis. The enzymatic hydrolysis conditions were pH 7.0, enzymatic hydrolysis temperature 50°C, enzyme activity 10000U / g, and enzymatic hydrolysis time 5 hours to obtain edestrin polypeptide.

[0028] The second aspect of the present invention provides a method for preparing a long-acting additive having the effects of lowering blood sugar and blood lipids, comprising the following steps:

[0029] The additive is prepared by adding edestin, edestin polypeptide and fructose into molten poloxamer 188, uniformly mixing, rapidly cooling to form a solid, and grinding the solid into powder.

[0030] In the process of forming the solid, a molten mixture of edestin, edestin polypeptide, poloxamer 188 and fructose is poured into an aluminum or copper container and quickly immersed in a CaCl2 aqueous solution at a temperature of -20°C. The molten mixture will quickly solidify into a solid.

[0031] Specifically, the molten mixture is prepared by heating Poloxamer 188 to 60° C., and adding the edestrin, edestrin polypeptide and fructose to the Poloxamer 188 to obtain a molten mixture.

[0032] Preparation of solid mixture: Pour the molten mixture into an aluminum or copper container and quickly immerse it in a CaCl2 aqueous solution at a temperature of -20°C. The molten mixture will quickly solidify to form a solid mixture.

[0033] The preparation of the additive comprises grinding the solid mixture and sieving it through an 80-mesh sieve to obtain a powdery substance, which is the additive of the present invention.

[0034] In one embodiment, a method for preparing a long-acting additive having the effects of lowering blood sugar and blood lipids comprises the following steps:

[0035] Poloxamer 188 is heated to 60°C, and the edestin, edestin polypeptide, and fructose are added to the poloxamer 188 in appropriate proportions to obtain a molten mixture. The molten mixture is poured into an aluminum or copper container and quickly immersed in a -20°C CaCl2 aqueous solution. The molten mixture will rapidly solidify to form a solid mixture, which is then ground and sieved through an 80-mesh sieve to obtain the additive.

[0036] The third aspect of the present invention provides a use of a long-acting additive with hypoglycemic and hypolipidemic effects in the preparation of hypoglycemic and hypolipidemic medicines.

[0037] During application, it is prepared into powder, tablets or capsules.

[0038] Statistical analysis: All experimental data were analyzed using GraphPad Prism 8.0 software (GraphPad-software Inc., CA, USA).

[0039] The additive provided by this invention can efficiently and rapidly lower blood sugar and lipid levels within a safe range, significantly improving bioavailability. Compared with simple mixing methods, this additive exhibits superior blood sugar and lipid-lowering effects, surpassing any other single ingredient. It is particularly suitable for preventing diabetes and hyperlipidemia in the general population and assisting patients in their treatment, demonstrating significant potential for application.

[0040] It should be noted that the experimental methods used in the present invention are all conventional methods unless otherwise specified; the reagents and materials used are all commercially available unless otherwise specified.

[0041] The complex enzymatic hydrolyzed polypeptide used in the following examples refers to edestrin polypeptide.

[0042] Example 1

[0043] A long-acting additive with the effects of lowering blood sugar and blood lipids is prepared according to the following formula:

[0044] The mass fraction of edestrin is 2%; the mass fraction of complex enzymatic hydrolyzed polypeptide is 10%; the mass fraction of poloxamer 188 is 70%; and the mass fraction of fructose is 18%.

[0045] Preparation method:

[0046] Poloxamer 188 is heated to 60°C, and edestin, edestin polypeptide, and fructose are added to the poloxamer 188 in appropriate proportions to obtain a molten mixture. The molten mixture is poured into an aluminum or copper container and quickly immersed in a -20°C CaCl2 aqueous solution. The molten mixture will rapidly solidify to form a solid mixture, which is then ground and sieved through an 80-mesh sieve to obtain the additive.

[0047] Example 2

[0048] The difference from Example 1 is that the formula of this embodiment is:

[0049] The mass fraction of edestrin is 8%; the mass fraction of complex enzymatic hydrolyzed polypeptide is 8%; the mass fraction of poloxamer 188 is 69%; and the mass fraction of fructose is 15%.

[0050] Example 3

[0051] The difference from Example 1 is that the formula of this embodiment is:

[0052] The mass fraction of edestrin is 4%; the mass fraction of complex enzymatic polypeptide is 7%; the mass fraction of poloxamer 188 is 74%; and the mass fraction of fructose is 15%.

[0053] Example 4

[0054] The difference from Example 1 is that the formula of this embodiment is:

[0055] The mass fraction of edestrin is 1%; the mass fraction of complex enzymatic hydrolyzed polypeptide is 1%; the mass fraction of poloxamer 188 is 85%; and the mass fraction of fructose is 13%.

[0056] Example 5

[0057] The difference from Example 1 is that the formula of this embodiment is:

[0058] The mass fraction of edestrin is 3%; the mass fraction of complex enzymatic polypeptide is 6%; the mass fraction of poloxamer 188 is 78%; and the mass fraction of fructose is 13%.

[0059] Example 6

[0060] The difference from Example 1 is that the formula of this embodiment is:

[0061] The mass fraction of edestrin is 5%; the mass fraction of complex enzymatic hydrolyzed polypeptide is 5%; the mass fraction of poloxamer 188 is 80%; and the mass fraction of fructose is 10%.

[0062] Comparative Example 1

[0063] The difference from Example 6 is that the formula of this comparative example is:

[0064] The mass fraction of edestrin is 20%; the mass fraction of complex enzymatic polypeptide is 30%; the mass fraction of poloxamer 188 is 30%; and the mass fraction of fructose is 20%.

[0065] Comparative Example 2

[0066] The difference from Example 6 is that the formula of this comparative example is:

[0067] The mass fraction of edestrin was 0.5%; the mass fraction of complex enzymatic polypeptide was 0.8%; the mass fraction of poloxamer 188 was 68.2%; and the mass fraction of fructose was 30.5%.

[0068] Comparative Example 3

[0069] The difference from Example 6 is that the formula of this comparative example is:

[0070] The mass fraction of edestrin is 15%; the mass fraction of complex enzymatic hydrolyzed polypeptide is 25%; the mass fraction of poloxamer 188 is 33%; and the mass fraction of fructose is 27%.

[0071] Comparative Example 4

[0072] The difference from Example 6 is that the formula of this comparative example is:

[0073] The mass fraction of edestrin is 12%; the mass fraction of complex enzymatic hydrolyzed polypeptide is 12%; and the mass fraction of poloxamer 188 is 76%.

[0074] Comparative Example 5

[0075] The difference from Example 6 is that the formula of this comparative example is:

[0076] The mass fraction of edestrin is 35%, the mass fraction of complex enzymatic hydrolyzed polypeptide is 35%, and the mass fraction of fructose is 30%.

[0077] Comparative Example 6

[0078] The difference from Example 6 is that the preparation process of this comparative example is:

[0079] The mass fraction of edestrin is 5%, the mass fraction of complex enzymatic hydrolyzed polypeptide is 5%, the mass fraction of poloxamer 188 is 80%, and the mass fraction of fructose is 10%. The components are simply mixed.

[0080] In order to illustrate the relevant properties of the additive provided by the present invention, the additives obtained in Examples 1 to 6 and Comparative Examples 1 to 6 were tested for relevant properties.

[0081] Test 1: In vitro α-amylase inhibition rate and α-glucosidase inhibition rate determination

[0082] The additives obtained in Examples 1 to 6 and Comparative Examples 1 to 6 were subjected to in vitro α-amylase inhibition rate and α-glucosidase inhibition rate determination to determine their ability to lower blood glucose levels in vitro.

[0083] The experimental results are shown in Table 1. The in vitro hypoglycemic effect of Example 6 is the most significant, which is significantly higher than that of the comparative example group. Its in vitro α-amylase inhibition rate is 98.89%±0.57%, and its α-glucosidase inhibition rate is 94.23%±3.23%.

[0084] Table 1 Determination of in vitro α-amylase inhibition rate and α-glucosidase inhibition rate of each group

[0085]

[0086] Test 2: Determination of in vitro bile salt adsorption activity

[0087] The additives obtained in Examples 1 to 6 and Comparative Examples 1 to 6 were subjected to in vitro bile salt adsorption activity assay to determine their ability to reduce blood lipid levels in vitro.

[0088] The experimental results are shown in Table 2. Example 6 had the most significant in vitro lipid-lowering effect, which was significantly higher than that of the control group. Its in vitro bile salt adsorption rate was the highest, among which the adsorption rate of sodium taurocholate was 98.13% ± 0.012%, the adsorption rate of sodium glycocholate was 99.51% ± 0.014%, and the adsorption rate of sodium cholate was 98.50% ± 0.011%.

[0089] Table 2 Determination of bile salt adsorption activity in vitro in each group

[0090]

[0091] Test 3: In vivo hypoglycemic test verification in mice

[0092] Eighty-four Kunming mice were divided equally according to body weight into a blank group, a model group, Example 1-6 groups, and Comparative Example 1-6 groups, with six mice in each group. The mice in the blank group received no treatment. The remaining mice were intraperitoneally injected with 0.2 g / kg·BW of alloxan to establish a diabetic model. Five hours after the injection, the mice were gavaged again with a 50% glucose solution to alleviate the stress response caused by the hypoglycemic process. Seventy-two hours after the injection, blood was drawn from the tail vein to measure fasting blood glucose levels. A successful diabetic model was considered established in mice when the blood glucose level was greater than 11.1 mmol / L.

[0093] After the model was successfully established, mice were gavage-treated daily for seven consecutive days. During the experiment, the mice's mental state, fur changes, and urination and defecation patterns were observed daily. In addition, food and water intake were recorded daily, and fasting blood glucose levels were measured before the first and last gavages.

[0094] Table 3 Effects of different additives in each group on blood glucose levels in alloxan-induced diabetic mice

[0095]

[0096] As shown in Table 3, on day 0, the blood glucose levels of all mice (except the blank group) were greater than 11.1 mmol / L, indicating that the diabetic model was successfully established. Subsequently, the additive was administered orally. The results showed that after 7 days of oral administration, the blood glucose levels of the mice in the Example groups all decreased significantly. Specifically, the blood glucose levels of the mice in Example 6 group decreased by 40.14% compared to the model group.

[0097] Test 4: Lipid-lowering experimental verification in mice

[0098] One hundred and twelve SPF healthy male Kunming mice were adapted to a normal diet for 5 days and then divided equally according to body weight into a blank control group (8 mice) and a model group (104 mice). The model group was fed a high-fat diet, while the blank control group was fed a normal diet. After 14 days of feeding, eye blood was collected, and serum was rapidly separated. Serum levels of high-density lipoprotein cholesterol (HDL-C), low-density lipoprotein cholesterol (LDL-C), total cholesterol (TC), and triglyceride (TG) were measured. A hyperlipidemia mouse model was established if serum triglycerides, total cholesterol, or low-density lipoprotein cholesterol (LDL-C) were significantly elevated in the model group compared to the blank group.

[0099] After successful model establishment, mice were gavage-treated once daily for seven consecutive days. During the gavage period, the blank control group continued to be fed a standard diet, while the model control group and the various Example and Comparative Example groups were fed a high-fat diet. Animal body weights were recorded daily. After the final gavage, blood was collected from the mice's eyeballs, and serum was rapidly isolated and measured for HDL-C, LDL-C, TC, and TG levels.

[0100] As shown in Table 4, after 14 days of high-fat diet feeding, the mice in each group did not experience any significant discomfort. Except for the blank group, the blood lipid data of all other groups increased to significant levels, indicating that the hyperlipidemia model was successfully established. The mice were then gavaged with the additive.

[0101] Table 4 Effects of high-fat diet for 14 days on blood lipids in each group of mice

[0102]

[0103] The results of the weight changes of mice are shown in Table 5. Among them, the weight of mice fed with a high-fat diet was significantly higher than that of mice in the blank control group. After oral administration, the weight of mice in each group decreased significantly, indicating that each example group can effectively improve the obesity of mice and reduce their weight. Among them, Example 6 had the most significant effect.

[0104] Table 5 Effects of each group of additives on the body weight of mice

[0105]

[0106] The test results of the effects of each group of additives on the blood lipids of mice are shown in Table 6. As can be seen from Table 6, compared with the model group, each example group can significantly reduce the levels of total cholesterol, triglycerides and low-density lipoprotein, while increasing the level of high-density lipoprotein, with a significant lipid-lowering effect, among which Example 6 has the most significant lipid-lowering effect.

[0107] Table 6 Effects of each group of additives on blood lipids in mice

[0108]

[0109] After ingestion, the additive provided by the present invention is digested and absorbed first, rapidly onset of action. Once in the gastrointestinal tract, edestin is continuously degraded by the digestive system, forming new polypeptides and amino acids that are continuously absorbed by the body, extending the release time of the active ingredient. Poloxamer 188 and fructose are used as carriers to solubilize edestin, aiding digestion and increasing the bioavailability of edestin and its polypeptides. This results in a rapid onset of action, long-lasting effects, and high bioavailability. Compared to a simple blend of the components (Comparative Example 6), the composition prepared by the melt method exhibits a greater blood sugar and lipid-lowering effect.

[0110] Based on the above embodiments and comparative examples, it is found that the additive can efficiently and rapidly reduce the body's blood sugar and blood lipid levels within a safe range, significantly improve bioavailability, and is particularly suitable for the prevention of diabetes and hyperlipidemia in the daily life of the general population and for assisting patients in their treatment, and has great application potential.

Claims

1. A long-acting additive with the effects of lowering blood sugar and blood lipids, characterized in that: The additive comprises the following components by mass fraction: edestin is 1-10%, edestin polypeptide is 1-10%, poloxamer 188 is 60-90%, and fructose is 5-20%, and the total mass fraction of each component is 100%. The long-acting additive is prepared according to the following steps: The additive is obtained by adding edestin, edestin polypeptide and fructose to molten poloxamer 188, uniformly mixing, rapidly cooling to form a solid, and grinding the solid into powder. The edestrin polypeptide is a polypeptide prepared by a composite enzymatic hydrolysis method, that is, it is first enzymatically hydrolyzed with papain and then enzymatically hydrolyzed with a neutral protease to perform composite enzymatic hydrolysis.

2. The long-acting additive with hypoglycemic and hypolipidemic effects according to claim 1, characterized in that: The edestrin is prepared by a salting-out method.

3. The long-acting additive with hypoglycemic and hypolipidemic effects according to claim 1, characterized in that: Edethin and edethin polypeptide are functional factors, and poloxamer 188 and fructose are synergistic auxiliary ingredients.

4. A method for preparing the long-acting additive with hypoglycemic and hypolipidemic effects according to any one of claims 1 to 3, characterized in that: The following steps are involved: The additive is prepared by adding edestin, edestin polypeptide and fructose into molten poloxamer 188, uniformly mixing, rapidly cooling to form a solid, and grinding the solid into powder.

5. The method for preparing the long-acting additive with hypoglycemic and hypolipidemic effects according to claim 4, characterized in that: In the process of forming the solid, a molten mixture of edestin, edestin polypeptide, poloxamer 188 and fructose is poured into an aluminum or copper container and quickly immersed in a CaCl2 aqueous solution at a temperature of -20°C. The molten mixture will quickly solidify into a solid.

6. Use of the long-acting additive having hypoglycemic and hypolipidemic effects according to any one of claims 1 to 3 in the preparation of a drug for lowering blood sugar and blood lipids.

7. The use according to claim 6, characterized in that During application, it is prepared into powder, tablets or capsules.

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