Compositions containing eicosenoic acid and its glycerides and their applications

Abstract

This invention discloses a composition containing eicosenoic acid and its glycerides, and its applications. At least one of eicosenoic acid and its glycerides is selected as the main active ingredient, enabling the obtained composition to produce a synergistic effect, significantly enhancing physiological activities such as controlling body fat, regulating intestinal flora, maintaining blood lipids / blood sugar / blood pressure, protecting the liver, controlling appetite, anti-oxidation, and anti-aging. Furthermore, all components in this composition are derived from pure natural plants, with no toxic side effects; and the preparation method is simple and easy to operate, making it suitable for preparing related health foods, pharmaceuticals, feed, and feed additives, showing promising application prospects.

Description

Technical Field

[0001] This invention relates to the fields of health food and biomedicine, and more specifically, to compositions containing eicosenoic acid and its glycerides and their applications. Background Technology

[0002] The family Taxaceae includes the genus Torreya, which comprises 6 species and 2 varieties, among which Torreya (… Torreya grandis Torreya grandis (Fort. exLindl.) is a species with high edible and utilization value. Wild, ungrafted Torreya grandis trees are called Torreya grandis or Torreya seeds, such as sesame Torreya grandis, peanut Torreya grandis, tusk Torreya grandis, and round Torreya grandis. Torreya grandis seed oil belongs to the oleic acid-linoleic acid type oil, with unsaturated fatty acids accounting for about 90% of the total fatty acids, linoleic acid accounting for 42%, oleic acid for 33%, α-linolenic acid for less than 1%, and saturated fatty acids mainly being palmitic acid (8%) and stearic acid (3%). 5,11,14-arachidonic acid, also known as pinocembryonic acid, accounts for 9%, and is a polyunsaturated fatty acid rarely found in other common oils. Torreya grandis seed oil has been shown to have the effects of inhibiting inflammatory responses, improving immune function, anti-oxidation, and delaying aging.

[0003] Patent CN202410601918.5 discloses an anti-aging composition, anti-aging cosmetic, and its preparation method and application, comprising torreya oil, Brazil walnut protein hydrolysate, Angoloside C, sarsaparilla acid, and nanovesicles; patent CN202311539075.2 discloses a hair care and growth composition containing torreya oil, its preparation method and application; CN202210168223.3 discloses a method for producing diglyceride oil by utilizing byproducts from the deep processing of special oils; and CN202111156671.3 discloses a method for preparing torreya oil. Based on currently available research, there are studies involving the preparation, extraction, structural identification, and biosynthesis of pine acid and its mono- and diglycerides, but research reports on its efficacy and the application of its analogues are relatively few.

[0004] In view of this, the present invention is proposed. Summary of the Invention

[0005] The purpose of this invention is to provide a composition containing eicosenoic acid and its glycerides and its applications, which can synergistically improve the effects of controlling body fat, regulating intestinal flora, maintaining blood lipids / blood sugar / blood pressure, protecting the liver, controlling appetite, anti-oxidation and anti-aging.

[0006] This invention is implemented as follows:

[0007] In a first aspect, the present invention provides a composition containing eicosenoic acid and its glycerides, comprising at least one of cis-5,11,14-eicosatotrienoic acid (CAS: 7019-85-4) or its monoglycerides, diglycerides, or glycerides; cis-11,14-eicosatodienoic acid (CAS: 2091-39-6) or its monoglycerides, diglycerides, or glycerides; cis-5,11-eicosatodienoic acid (CAS: 70363-48-3) or its monoglycerides, diglycerides, or glycerides; and cis-11-eicosatomonenoic acid (CAS: 5561-99-9) or its monoglycerides, diglycerides, or glycerides.

[0008] Secondly, the present invention provides the use of the above composition in the preparation of products for controlling body weight and suppressing appetite.

[0009] Thirdly, the present invention provides the use of the above composition in the preparation of products for maintaining intestinal health.

[0010] Fourthly, the present invention provides the use of the above composition in the preparation of a drug for improving insulin resistance and lowering blood sugar.

[0011] Fifthly, the present invention provides the application of the above composition in the preparation of products that lower body fat and blood lipids.

[0012] In a sixth aspect, the present invention provides the use of the above composition in the preparation of products that improve liver damage.

[0013] In a seventh aspect, the present invention provides the use of the above composition in the preparation of products for maintaining blood pressure.

[0014] Eighthly, the present invention provides the use of the above-described composition in the preparation of antioxidant and / or anti-aging products.

[0015] The present invention has the following beneficial effects:

[0016] This invention screens specific components from eicosenoic acid (EPA) and prepares a composition using at least one of them as the main active ingredient. This composition significantly enhances physiological activities such as controlling body fat, regulating intestinal flora, maintaining blood lipids / blood sugar / blood pressure, protecting the liver, controlling appetite, anti-oxidation, and anti-aging. Furthermore, all components in this composition are derived from pure natural plants, have no toxic side effects, and the preparation method is simple and easy to operate. It can be used to prepare related foods, pharmaceuticals, feeds, and feed additives, showing promising application prospects. Attached Figure Description

[0017] To more clearly illustrate the technical solutions of the embodiments of the present invention, the accompanying drawings used in the embodiments will be briefly introduced below. It should be understood that the following drawings only show some embodiments of the present invention and should not be regarded as a limitation on the scope. For those skilled in the art, other related drawings can be obtained based on these drawings without creative effort.

[0018] Figure 1 The structural formula of cis-5,11,14-eicoseriate acid;

[0019] Figure 2 The structural formula is that of cis-5,11,14-eicosoritrienoic acid-α-monoglyceride;

[0020] Figure 3 The structural formula is that of cis-5,11,14-eicosoritrienoic acid-β-monoglyceride;

[0021] Figure 4 The structural formula of cis-5,11,14-eicosoritrienoic acid-1,2-diglyceride is given.

[0022] Figure 5 The structural formula of cis-5,11,14-eicosoritrienoic acid-1,3-diglyceride is given.

[0023] Figure 6 The structural formula of cis-5,11,14-eicosoritrienoic acid glyceride is shown.

[0024] Figure 7 The structural formula of cis-11,14-eicosadienoic acid;

[0025] Figure 8 The structural formula of cis-5,11-eicosadienoic acid;

[0026] Figure 9 The structural formula is cis-11-eicosanoic acid. Detailed Implementation

[0027] To make the objectives, technical solutions, and advantages of the embodiments of the present invention clearer, the technical solutions in the embodiments of the present invention will be clearly and completely described below. Where specific conditions are not specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all conventional products that can be purchased commercially.

[0028] The active ingredients of the compositions of the present invention are derived from eicosenoic acid and its glycerol esters. Here, eicosenoic acid and its glycerol esters refer to cis-5,11,14-eicosadienoic acid (SA, CAS: 7019-85-4), cis-11,14-eicosadienoic acid (EOA, 11E,14E-Eicosadienoic acid, CAS: 2091-39-6), cis-5,11-eicosadienoic acid (EA, 5E,11E-Eicosadienoic acid, CAS: 70363-48-3), cis-11-eicosenoic acid (ESA, 11E-Eicosenoic acid, CAS: 5561-99-9), and monoglycerides, diglycerides, and glycerol esters of the above four compounds.

[0029] Specifically, the glycerol esters of cis-5,11,14-eicosatotrienoic acid can be: cis-5,11,14-eicosatotrienoic acid-α-monoglyceride (αMSA), cis-5,11,14-eicosatotrienoic acid-β-monoglyceride (βMSA), cis-5,11,14-eicosatotrienoic acid-1,2-diglyceride (1,2-DSA), cis-5,11,14-eicosatotrienoic acid-1,3-diglyceride (1,3-DSA), and cis-5,11,14-eicosatotrienoic acid glyceride (TSA).

[0030] The glycerides of cis-11,14-eicosadienoic acid can be: cis-11,14-eicosadienoic acid-α-monoglyceride (αEOA), cis-11,14-eicosadienoic acid-β-monoglyceride (βEOA), cis-11,14-eicosadienoic acid-1,2-diglyceride (1,2-EOA), cis-11,14-eicosadienoic acid-1,3-diglyceride (1,3-EOA), and cis-11,14-eicosadienoic acid glyceride (TEOA).

[0031] Glyceryl esters of cis-5,11-eicosadienoic acid can be: cis-5,11-eicosadienoic acid-α-monoglyceride (αEA), cis-5,11-eicosadienoic acid-β-monoglyceride (βEA), cis-5,11-eicosadienoic acid-1,2-diglyceride (1,2-EA), cis-5,11-eicosadienoic acid-1,3-diglyceride (1,3-EA), and cis-5,11-eicosadienoic acid glyceride (TEA).

[0032] The glycerol esters of cis-11-eicosodeenoic acid can be: cis-11-eicosodeenoic acid-α-monoglyceride (αESA), cis-11-eicosodeenoic acid-β-monoglyceride (βESA), cis-11-eicosodeenoic acid-1,2-diglyceride (1,2-ESA), cis-11-eicosodeenoic acid-1,3-diglyceride (1,3-ESA), and cis-11-eicosodeenoic acid glyceride (TESA).

[0033] The components of the composition of the present invention are selected from at least one of the above-mentioned eicosenoic acids and their glycerol esters, wherein the structures of some compounds are as follows: Figure 1-9 As shown. For example, the composition contains at least one of cis-5,11,14-eicosatetrienoic acid glyceride, cis-11,14-eicosatedienoic acid glyceride, and cis-5,11-eicosatedienoic acid glyceride.

[0034] Preferably, the components of the composition of the present invention are selected from at least two of the above-mentioned eicosenoic acid and its glycerides. During the experiment, the inventors found that although the above-mentioned compounds have varying degrees of effect in improving the control of body fat, regulating intestinal flora, maintaining blood lipids / blood sugar / blood pressure, protecting the liver, controlling appetite, anti-oxidation, and anti-aging, only some compounds have synergistic potential when combined, such as the combination of cis-5,11,14-eicosenoic acid glyceride and cis-11,14-eicosenoic acid glyceride; or the combination of cis-5,11,14-eicosenoic acid glyceride and cis-5,11-eicosenoic acid glyceride.

[0035] The above-mentioned eicosenoic acid compounds exhibit good physiological activity when combined in any two proportions. More preferably, when the composition consists of cis-5,11,14-eicosatotrienoic acid glyceride and cis-11,14-eicosatodienoic acid glyceride, the mass ratio is 1~5:5~1; when the composition consists of cis-5,11,14-eicosatotrienoic acid glyceride and cis-5,11-eicosatodienoic acid glyceride, the mass ratio is 1~10:10~1.

[0036] The eicosenoic acid and its glycerol esters of the present invention can be derived from Torreya grandis seeds, Torreya nut seeds, and pine nuts. They can be obtained by using existing preparation methods or by directly using commercially available related products. The present invention does not limit their source or preparation method.

[0037] During the experiments, it was found that the above-mentioned eicosenoic acid and its glycerides, used alone or in combination of at least two, can enhance physiological activities such as controlling body fat, regulating intestinal flora, maintaining blood lipids / blood sugar / blood pressure, protecting the liver, controlling appetite, anti-oxidation, and anti-aging. Combinations of two of these compounds showed even better effects, exhibiting a synergistic effect. Therefore, the compositions of the present invention can have the following uses:

[0038] (1) Control weight or prepare weight control products.

[0039] (2) Suppress appetite or prepare products that suppress appetite; its specific effects include: regulating gastrointestinal hormone peptide levels and reducing food intake.

[0040] (3) Maintaining gut health or preparing products that maintain gut health; its specific effects include: reducing tumor necrosis factor-α and serum lipopolysaccharide levels.

[0041] (4) Improve insulin resistance and lower blood sugar, or prepare drugs that improve insulin resistance and lower blood sugar.

[0042] (5) Lowering lipids or preparing lipid-lowering products; its specific effects include: reducing body fat, epididymal fat, liver fat, groin fat and blood lipids; more specifically, its effects include lowering serum triglycerides, total cholesterol, low-density lipoprotein cholesterol; regulating the expression levels of leptin, adiponectin and liver peroxisome proliferator-activated receptor γ.

[0043] (6) Improve liver damage or prepare products that improve liver damage; its specific effects include: reducing serum aspartate aminotransferase activity.

[0044] (7) To maintain blood pressure or to prepare products for maintaining blood pressure.

[0045] (8) Antioxidant, anti-aging or preparation of antioxidant and anti-aging products; its antioxidant properties specifically include increasing the activity of superoxide dismutase and catalase and reducing the content of malondialdehyde.

[0046] The products mentioned in the above applications include, but are not limited to, health foods, pharmaceuticals, feed, and feed additives.

[0047] The features and performance of the present invention will be further described in detail below with reference to embodiments.

[0048] The 5,11,14-eicoseriate, cis-11,14-eicoseriate, cis-5,11-eicoseriate, and cis-11-eicoseriate used in the embodiments of this invention were all purchased from commercial chemical companies. The mono-, di-, and glycerol esters of these fatty acids were prepared in our laboratory through controlled esterification and obtained by separation and purification using molecular distillation technology.

[0049] Example 1

[0050] This embodiment provides a composition in which the active ingredient is cis-5,11,14-eicosatotrienoic acid.

[0051] Example 2

[0052] This embodiment provides a composition in which the active ingredient is cis-5,11,14-eicosoritrienoic acid-α-monoglycerate.

[0053] Example 3

[0054] This embodiment provides a composition in which the active ingredient is cis-5,11,14-eicosoritrienoic acid-1,3-diglyceride.

[0055] Example 4

[0056] This embodiment provides a composition in which the active ingredient is cis-5,11,14-eicosatotrienoic acid glyceride.

[0057] Example 5

[0058] This embodiment provides a composition in which the active ingredient is cis-11,14-eicosadienoic acid.

[0059] Example 6

[0060] This embodiment provides a composition in which the active ingredient is cis-11,14-eicosadienoic acid glyceride.

[0061] Example 7

[0062] This embodiment provides a composition in which the active ingredient is cis-5,11-eicosadienoic acid.

[0063] Example 8

[0064] This embodiment provides a composition in which the active ingredient is cis-5,11-eicosadienoic acid glyceride.

[0065] Example 9

[0066] This embodiment provides a composition in which the active ingredient is cis-11-eicosodeenoic acid.

[0067] Example 10

[0068] This embodiment provides a composition in which the active ingredient is cis-11-eicosodeenoic acid-α-monoglyceride.

[0069] Example 11

[0070] This embodiment provides a composition whose active ingredients are cis-5,11,14-eicosatetrienoic acid glyceride and cis-11,14-eicosatedienoic acid glyceride in a mass ratio of 1:5.

[0071] Example 12

[0072] This embodiment provides a composition whose active ingredients are cis-5,11,14-eicosatotrienoic acid glyceride and cis-11,14-eicosatotrienoic acid glyceride in a mass ratio of 5:1.

[0073] Example 13

[0074] This embodiment provides a composition whose active ingredients are cis-5,11,14-eicosatetrienoic acid glyceride and cis-5,11-eicosatedienoic acid glyceride in a mass ratio of 10:1.

[0075] Example 14

[0076] This embodiment provides a composition whose active ingredients are cis-5,11,14-eicosatetrienoic acid glyceride and cis-5,11-eicosatedienoic acid glyceride in a mass ratio of 1:10.

[0077] Comparative Example 1

[0078] This comparative example provides a composition whose active ingredients are cis-5,11,14-eicosatetrienoic acid glyceride and cis-11-eicosatemonenoic acid in a mass ratio of 1:5.

[0079] Comparative Example 2

[0080] This comparative example provides a composition whose active ingredients are cis-5,11,14-eicosatetrienoic acid glyceride and cis-5,11-eicosatedienoic acid in a mass ratio of 10:1.

[0081] The functional evaluation protocols based on laboratory mice were all implemented in accordance with the "Methods for Functional Testing and Evaluation of Health Foods (2023 Edition)" issued by the National Health Commission.

[0082] Experimental Example 1

[0083] The effects of the compositions of Examples 1-14 and Comparative Examples 1-2 on weight loss, appetite control, and improved gut health were verified. The specific methods are as follows:

[0084] High-fat diet mouse model: 4-week-old healthy male C57BL / 6 mice (SPF grade) were selected, with 12 mice in 3 cages in each treatment group. They were given free access to food and water for 8 weeks.

[0085] The experimental groups are as follows:

[0086] The control group (CHOW) was fed a standard maintenance diet;

[0087] The high-fat diet model group (HFD) was fed a commercially available high-fat diet (purified high-fat diet with a fat energy ratio of 45%).

[0088] The experimental intervention group was fed a commercially available high-fat diet with equal energy, and the diet was supplemented with the compositions of Examples 1-14 and Comparative Examples 1-2 at a dose of 500 mg / kg.

[0089] Mouse model of compound antibiotic treatment: 4-week-old healthy male C57BL / 6 mice (SPF grade) were selected, with 12 mice in 3 cages in each group. They were given free access to food and water for 8 weeks.

[0090] The experimental groups were as follows: The model control group (NCD) was fed a commercially available high-fat diet (purified high-fat diet with a fat energy ratio of 45%) and was provided with normal drinking water. Except for this group, all other groups had drinking water supplemented with compound antibiotics (Abx, penicillin 1 g / L, neomycin 1 g / L, metronidazole 1 g / L, vancomycin 0.5 g / L). The high-fat diet model group (NCD+Abx) was fed a commercially available high-fat diet, and the intervention group was fed a commercially available high-fat diet with the same energy content. The diet was supplemented with the compositions of Examples 1-14 and Comparative Examples 1-2 at a dose of 500 mg / kg, and the above-mentioned compound antibiotics were added to the normal drinking water.

[0091] Biochemical indicators were measured: gastrointestinal hormone peptide (PYY), tumor necrosis factor-α (TNF-α), and serum lipopolysaccharide (LPS) levels were determined using an enzyme-linked immunosorbent assay (ELISA) kit.

[0092] The experimental results are shown in Table 1-2:

[0093] Table 1. Effects of different experimental groups on body weight, appetite, and gut health in a high-fat diet model.

[0094]

[0095] Note: Content of gastrointestinal hormone peptide (PYY), tumor necrosis factor-α (TNF-α), and lipopolysaccharide (LPS). *Indicates compared to NCD or CHOW.* p <0.05,** p <0.01, *** p <0.001. $ indicates that compared to the HFD group, $ p <0.05, $$ p <0.01, $$$ p <0.001.

[0096] Table 2. Effects of each experimental group on body weight and appetite in the combined antibiotic treatment experiment.

[0097]

[0098] Note: * indicates comparison with NCD or CHOW. p <0.05,** p <0.01, *** p <0.001. $ indicates that compared to the HFD group or the NCD+Abx group, $ p <0.05, $$ p <0.01, $$$ p <0.001.

[0099] Regarding weight control: The relative weight of the HFD group was 2.51 times that of the CHOW group (p<0.05), indicating that the high-fat diet-induced obesity model was successfully established. The relative weight of the treatment groups in Examples 1-10 was significantly lower than that of the HFD group, indicating that eicosenoic acid and its mono-, di-, and glycerol esters can significantly reduce mouse weight and have the potential to control weight. The relative weight of the same dose of eicosenoic acid and its mono-, di-, and glycerol ester combinations (Examples 11-14) was reduced by 47.28%, 47.65%, 52.35%, and 55.72% respectively compared to the HFD group, and the relative weight was close to that of the CHOW group. The weight control efficacy was significantly better than that of eicosenoic acid and its mono-, di-, and glycerol esters used alone. The combinations of the two showed a significant synergistic effect. In contrast, the weight of the control groups 1-2 did not change significantly compared to the HFD group, and no significant synergistic effect was observed when these eicosenoic acids and their mono-, di-, and glycerol esters were used alone.

[0100] Regarding appetite control: The food intake of the treatment groups in Examples 1-10 was significantly lower than that of the HFD group, and the serum level of the appetite-suppressing hormone PYY was significantly increased, indicating that eicosenoic acid and its mono-, di-, and glycerol esters can effectively control appetite. The same dose of eicosenoic acid and its mono-, di-, and glycerol ester combinations (Examples 11-14) significantly reduced food intake in mice, while significantly increasing PYY, which was significantly better than using eicosenoic acid and its mono-, di-, and glycerol esters alone. The combinations of these two types of eicosenoic acid showed a significant synergistic effect. In contrast, the food intake of the control groups 1-2 did not change significantly compared to the HFD group, and no significant synergistic effect was observed when these eicosenoic acids and their mono-, di-, and glycerol esters were used alone.

[0101] Regarding the improvement of gut health: The relative body weight and food intake of mice in the treatment groups of Examples 1-10 showed no significant changes compared to the NCD+Abx group, indicating that eicosenoic acid (EPA) and its mono-, di-, and glycerol esters affect mouse body weight and food intake through the gut microbiota. LPS is an extracellular toxin secreted by Gram-negative intestinal bacteria. Due to changes in intestinal pathways, it infiltrates into the host's bloodstream, triggering systemic low-grade inflammation. It is a host endotoxin and an important biomarker of gut health. Serum LPS and TNF-α levels in the treatment groups of Examples 1-10 were significantly lower than those in the HFD group, indicating that EPA and its mono-, di-, and glycerol esters can significantly improve gut microbiota dysbiosis and systemic inflammation caused by a high-fat diet, thus improving gut health. Furthermore, mice with the same dose of EPA and its mono-, di-, and glycerol ester combinations (Examples 11-14) showed even lower LPS and TNF-α levels, demonstrating a significant synergistic effect. Furthermore, the Bifidobacterium content in the treatment groups of Examples 1-10 was significantly increased compared to the HFD group, indicating that eicosenoic acid and its mono-, di-, and glycerol esters can significantly improve gut health. The same dosage of eicosenoic acid and its mono-, di-, and glycerol ester compositions (Examples 11-14) also showed a synergistic effect in improving gut health. These indicators of the compositions in Comparative Examples 1-2 showed no significant changes compared to the HFD group, and no significant synergistic effect was observed compared to the use of these eicosenoic acids and their mono-, di-, and glycerol esters alone.

[0102] Experimental Example 2

[0103] The effects of the compositions of Examples 1-14 and Comparative Examples 1-2 on maintaining blood glucose levels and improving insulin resistance were verified. The specific methods are as follows:

[0104] High-fat diet mouse model: 4-week-old healthy male C57BL / 6 mice (SPF grade) were selected, with 12 mice in 3 cages in each treatment group. They were given free access to food and water for 16 weeks.

[0105] The experimental groups are as follows:

[0106] The control group (NCD) was fed a standard maintenance diet;

[0107] The high-fat diet model group (HFD) was fed a commercially available high-fat diet;

[0108] The intervention group was fed an isoenergetic commercial high-fat diet at 75 mg / kg body weight daily via gavage with the combination of Examples 1-14 and Comparative Examples 1 and 2 for 10 weeks.

[0109] db / db mouse model: Four-week-old healthy male C57BLKS / JGpt mice were used as the normal control group (CON), and were administered the experimental solvent by gavage at a rate of 5 mL / kg body weight daily; other groups used C57BLKS / JGpt mice. dbdb diabetic mice: The diabetic mouse control group (DBDB) was administered the experimental solvent by gavage at 5 mL / kg body weight daily; the metformin positive control group (Met) was administered metformin by gavage at 200 mg / kg body weight daily (200 mg metformin dissolved in 2.5 mL of PEG400 + 2.5 mL of physiological saline); the intervention group was administered the combination of Examples 1-14 and Comparative Examples 1 and 2 by gavage at 75 mg / kg body weight daily for 10 weeks.

[0110] Glucose tolerance test: At the end of week 15, mice were fasted for 12 hours, followed by intraperitoneal injection of glucose solution (2 g / kg body weight). Blood glucose levels were measured in each mouse at 0, 30, 60, 90, and 120 minutes, and blood glucose change curves were plotted and the area under the blood glucose change curve (AUC) was calculated.

[0111] Biochemical assays: Serum glucose (GLU) and insulin levels were measured using commercially available standard kits or enzyme-linked immunosorbent assay (ELISA) kits. Insulin resistance index: Insulin level × blood glucose concentration / 22.5.

[0112] The results are shown in Table 3:

[0113] Table 3 Effects of each group on blood glucose and insulin resistance

[0114]

[0115] AUC: Area under the glucose tolerance test curve; GLU: Serum glucose level (mM); IRI: Insulin resistance index. *Indicates comparison with NCD or CON.* p <0.05,** p <0.01, *** p <0.001. $ indicates comparison with the HFD or DBDB group. p <0.05, $$ p <0.01, $$$ p <0.001. # indicates comparison with Met, # p <0.05, ## p <0.01, ### p <0.001.

[0116] In a high-fat diet mouse model (Table 3), the AUC, GLU, and IRI of the treatment groups in Examples 1-10 were significantly lower than those in the HFD group, indicating that eicosenoic acid and its mono-, di-, and glycerol esters can effectively reduce the increase in blood glucose levels induced by HFD, maintain normal blood glucose levels, and improve insulin resistance. Furthermore, the AUC, GLU, and IRI of mice treated with the same dose of eicosenoic acid and its mono-, di-, and glycerol ester combinations (Examples 11-14) were significantly lower, showing a significant synergistic effect compared to the use of the same dose of eicosenoic acid and its mono-, di-, and glycerol esters alone. These indicators in Comparative Examples 1-2 showed no significant changes compared to the HFD group, and no significant synergistic effect was observed compared to the use of these eicosenoic acids and their mono-, di-, and glycerol esters alone. Based on these findings, the ability of eicosenoic acid and its mono-, di-, and glycerol esters and their combinations to maintain blood glucose levels was specifically verified using a diabetic mouse model.

[0117] As shown in Table 3, the AUC of the treatment groups in Examples 1-10 was significantly lower than that of the DBDB group, confirming that eicosenoic acid and its mono-, di-, and glycerol esters can effectively reduce the elevation of blood glucose levels in DBDB mice and maintain normal blood glucose levels. The AUC of mice treated with the same dose of eicosenoic acid and its mono-, di-, and glycerol ester compositions (Examples 11-DB, 12-DB, 13-DB, and 14-DB) was also significantly reduced, demonstrating a significant synergistic effect in maintaining blood glucose levels compared to the use of these eicosenoic acids and their mono-, di-, and glycerol esters alone. The AUC of the Comparative Example 1-DB and Comparative Example 2-DB treatment groups showed no significant change compared to the HFD group, and no significant synergistic effect was observed compared to the use of these eicosenoic acids and their mono-, di-, and glycerol esters alone.

[0118] Experimental Example 3

[0119] The effects of the compositions of Examples 1-14 and Comparative Examples 1-2 on lipid-lowering and liver-damage-improving effects were verified. The specific methods are as follows:

[0120] High-fat diet mice: 4-week-old healthy male C57BL / 6 mice (SPF grade) were selected, with 12 mice in 3 cages in each treatment group. They were given free access to food and water, and the experiment lasted for 10 weeks.

[0121] The experimental groups are as follows:

[0122] The control group (CHOW) was fed a standard maintenance diet;

[0123] The high-fat diet model group (HFD) was fed a commercial high-fat diet (a high-fat diet with a fat energy ratio of 45%).

[0124] The experimental intervention group was fed a commercially available high-fat diet with equal energy, supplemented with the compositions of Examples 1-14 and Comparative Examples 1 and 2 at a dose of 750 mg / kg of feed.

[0125] Measurement of body fat and other indicators: After anesthesia, body fat content was measured using a digital dual-energy X-ray scanner. After dissection, epididymal fat and inguinal fat were quickly dissected and weighed, and their ratios to fasting body weight were recorded as epididymal fat and inguinal fat, respectively.

[0126] Biochemical index determination: Lipids were extracted from the liver using the Folch method, dried under nitrogen, and the resulting liver lipid samples were weighed and divided by fasting body weight to determine the liver fat content. The lipids were reconstituted in ethanol, and triglyceride (TG) and total cholesterol (TC) levels were determined according to the procedures of commercially available kits. Serum triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDLC), high-density lipoprotein cholesterol (HDLC), serum aspartate aminotransferase (AST) activity, leptin (Lep), and adiponectin (Adi) were measured using commercially available standard kits or enzyme-linked immunosorbent assay (ELISA). Peroxisome proliferator-activated receptor gamma (PPARγ), a ligand-dependent transcription factor, plays an important role in maintaining triglyceride homeostasis, improving fatty liver degeneration, and visceral fat deposition. Its gene expression in the liver and extraction analysis were performed using standard real-time quantitative PCR methods.

[0127] The experimental results are shown in Table 4-5:

[0128] Table 4. Effects of each experimental group on reducing body fat

[0129]

[0130] Note: * indicates comparison with CHOW, * p <0.05,** p <0.01, *** p <0.001. $ indicates comparison with the HFD group. p <0.05, $$ p <0.01, $$$ p <0.001.

[0131] Table 5. Effects of each experimental group on lipid-lowering and liver damage.

[0132]

[0133] Note: Triglycerides (TG), Total Cholesterol (TC), Low-Density Lipoprotein Cholesterol (LDLC), High-Density Lipoprotein Cholesterol (HDLC, mM), Serum Aspartate Aminotransferase Activity (AST), Leptin (Lep), Adiponectin (Adi). *Indicates comparison with CHOW,* p <0.05,** p <0.01, *** p <0.001. $ indicates comparison with the HFD group. p <0.05, $$ p <0.01, $$$ p <0.001.

[0134] As shown in Table 4, the body fat and epididymal fat content of the treatment groups in Examples 1-10 were significantly lower than those in the HFD group, and the liver fat and groin fat were also significantly lower than those in the HFD group, indicating that eicosenoic acid and its mono-, di-, and glycerol esters can significantly reduce body fat content and have the potential to control body fat. However, the same dose of eicosenoic acid and its mono-, di-, and glycerol ester combinations (Examples 11-14) reduced body fat by 58%, 56%, 53%, and 54% compared to the HFD group, liver fat by 38%, 40%, 36%, and 42%, epididymal fat by 63%, 65%, 71%, and 70%, and groin fat by 60%, 69%, 78%, and 79%, respectively. This indicates that eicosenoic acid and its mono-, di-, and glycerol ester combinations can significantly reduce body fat content, control body fat, and have a significant synergistic effect in reducing body fat. The body fat index of Comparative Examples 1-2 was slightly lower than that of the HFD group, but no significant synergistic effect was observed compared to the use of these eicosenoic acids and their mono-, di-, and glycerol esters alone.

[0135] As shown in Table 5, the levels of TG, TC, LDLC, Lep, and Adil in the treatment groups of Examples 1-10 were significantly lower than those in the HFD group, while the relative expression levels of PPARγ in each group were significantly higher. This indicates that these eicosenoic acids and their mono-, di-, and glycerol esters can significantly reduce serum TG, TC, and LDLC levels, and control blood lipid levels through key lipid metabolism hormones (Lep and Adil) and PPARγ, clearly demonstrating the lipid-lowering pathway and mechanism. In the treatment groups of Examples 11-14, TG, TC, and LDLC were significantly lower than those in the HFD group, showing a significant synergistic effect compared to the use of the same dose of eicosenoic acids and their mono-, di-, and glycerol esters alone. In Comparative Examples 1-2, these indicators were slightly lower than those in the HFD group, and no significant synergistic effect was observed compared to the use of these eicosenoic acids and their mono-, di-, and glycerol esters alone.

[0136] Experiment Example 4

[0137] The effects of the compositions of Examples 1-14 and Comparative Examples 1-2 in maintaining blood pressure were verified. The specific methods are as follows:

[0138] Experimental setup:

[0139] Normal control group (NCD): Wistar Kyoto rats of the same population but without hypertension and with stable blood pressure. Hypertension model group (CON), positive control group, and experimental group: Spontaneously hypertensive rats (SHRs) were selected, with 10 rats in each group. They were fed a normal diet and had free access to food and water. The experimental period was 4 weeks. The hypertension model group was administered physiological saline by gavage at 5 mL / kg body weight daily (CON), and the positive control group was administered captopril at gavage at 10 mg / kg body weight / day (CAP). The dosage of eicosapentaenoic acid and its mono-, di-, and glycerol esters in the experimental groups was set as follows: 75 mg / kg body weight / day, respectively, by gavage of the compositions of Examples 1-14 and Comparative Examples 1 and 2.

[0140] Blood pressure measurement: Blood pressure was measured weekly in the tail artery using the BP-2000 non-invasive rat blood pressure monitoring system. The results are shown in Table 6.

[0141] Table 6. Effects of each experimental group on blood pressure in spontaneously hypertensive rats

[0142]

[0143] Note: * indicates comparison with CON, * p <0.05,** p <0.01, *** p <0.001. $ indicates comparison with CAP, $ p <0.05, $$ p <0.01, $$$ p <0.001.

[0144] As shown in Table 6, the systolic blood pressure of rats in the CAP (positive control group) and the treatment groups of Examples 1-10 decreased by 28%, 20%, 20%, 21%, 25%, 25%, 23%, 17%, 22%, 25%, and 17% respectively compared with the CON group, while the diastolic blood pressure decreased by 31%, 17%, 16%, 14%, 22%, 21%, 17%, 10%, 16%, 16%, and 10% respectively compared with the CON group. This indicates that eicosenoic acid and its mono-, di-, and glycerol esters all have significant blood pressure-lowering effects. In rats using the same dose of eicosenoic acid and its mono-, di-, and glycerol ester combinations (Examples 11-14), systolic blood pressure decreased by 27%, 29%, 33%, and 33% respectively compared to the CON group, and was only 17%, 13%, 6%, and 7% higher than the NCD group, respectively. Diastolic blood pressure decreased by 26%, 27%, 30%, and 29% respectively compared to the CON group, and was only 48%, 46%, 40%, and 42% higher than the NCD group, respectively. The effects were far superior to the blood pressure-lowering capacity of these eicosenoic acids and their mono-, di-, and glycerol esters used alone, and remained at levels similar to those of normal rats, indicating that the combination of eicosenoic acid and its mono-, di-, and glycerol esters has a significant synergistic blood pressure-lowering effect. The blood pressure indicators in Comparative Examples 1-2 showed no significant changes compared to the CON group, and no significant synergistic effect was observed compared to the use of these eicosenoic acids and their mono-, di-, and glycerol esters alone.

[0145] Experimental Example 5

[0146] The effects of the compositions of Examples 1-14 and Comparative Examples 1-2 on antioxidation and anti-aging were verified. The specific methods are as follows:

[0147] Experimental Setup: Lifespan is a key indicator for evaluating the anti-aging properties of interventions. The significantly longer lifespan of nematodes compared to naturally aging organisms indicates effective anti-aging. During the lifespan experiment, synchronized nematodes were randomly selected and placed into the drug-treated group. Each group had three parallel plates, with 30 nematodes per plate, marked as day 0. The nematodes were cultured at 20 °C, and their survival was recorded daily. Nematode death and removal criteria: No reaction within 10 seconds of being lightly touched with a platinum wire; nematodes that disappeared or died after crawling out of the culture dish, or whose eggs hatched into bagworms within the nematode, were not included in the data.

[0148] The concentrations of each drug addition group are as follows: the nematode food without any treatment is recorded as the NCD group; the food containing 5 mg / mL nicotinamide mononucleotide is recorded as the NMN group; and the addition amount of the compositions of Examples 1-14 and Comparative Examples 1-2 is 1 mM.

[0149] Antioxidant index determination: Superoxide dismutase (SOD) and catalase (CAT) activities, and malondialdehyde (MDA) content were determined strictly in accordance with the kit instructions from Nanjing Jiancheng Biotechnology Co., Ltd.

[0150] The results are shown in Table 7:

[0151] Table 7. Effects of each experimental group on the lifespan and in vivo antioxidant levels of wild-type Caenorhabditis elegans.

[0152]

[0153] Note: Superoxide dismutase (SOD), catalase (MDA) activity, and malondialdehyde (MDA) content. *Indicates comparison with NCD. p <0.05,** p <0.01, *** p <0.001. $ indicates comparison with NMN. p <0.05, $$ p <0.01, $$$ p <0.001.

[0154] As shown in Table 7, the average lifespan of nematodes in the NMN (positive control group) and the treatment groups of Examples 1-10 were 1.35, 1.06, 1.11, 1.19, 1.22, 1.26, 1.26, 1.08, 1.25, 1.19, and 1.18 times that of NCD, respectively. The lifespan extension rate was 34.71%, 6.33%, 11.25%, 19.48%, 22.28%, 25.87%, 25.81%, 8.29%, 25.14%, 19.43%, and 17.53% higher than that of NCD, respectively. This indicates that eicosenoic acid and its mono-, di-, and glycerol esters all have significant lifespan extension and anti-aging effects. The average lifespan of nematodes fed the same dose of eicosenoic acid and its mono-, di-, and glycerol esters (Examples 11-14) was 1.32, 1.35, 1.34, and 1.36 times that of NCD, respectively, with lifespan extension rates of 32.25%, 35.27%, 34.15%, and 35.78%, respectively. These extensions were significantly higher than the anti-aging effects of eicosenoic acid and its mono-, di-, and glycerol esters used alone, indicating that the combination of eicosenoic acid and its mono-, di-, and glycerol esters has a significant synergistic anti-aging effect. In Comparative Examples 1-2, these anti-aging indicators showed no significant changes compared to the NCD group, and no significant synergistic effect was observed compared to the use of eicosenoic acid and its mono-, di-, and glycerol esters alone.

[0155] Correspondingly, the SOD and CAT enzyme activities in nematodes treated with NMN (positive control group) and Examples 1-10 were significantly enhanced compared to NCD, while the content of the peroxidation product MDA decreased, indicating that eicosenoic acid and its mono-, di-, and glycerol esters significantly improved the antioxidant capacity of nematodes. However, the same dose of the combination of eicosenoic acid and its mono-, di-, and glycerol esters (Examples 11-14) showed significantly higher SOD and CAT enzyme activities and significantly lower MDA content compared to NCD, indicating that the combination also significantly improved the antioxidant capacity of nematodes. Furthermore, the improvement in antioxidant capacity of the combination was significantly greater than that of eicosenoic acid and its mono-, di-, and glycerol esters used alone, indicating that the combination also has a significant synergistic antioxidant effect. These antioxidant indicators in Comparative Examples 1-2 showed no significant changes compared to the NCD group, and no significant synergistic effect was observed compared to the use of these eicosenoic acid and its mono-, di-, and glycerol esters alone.

[0156] The above description is merely a preferred embodiment of the present invention and is not intended to limit the invention. Various modifications and variations can be made to the present invention by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of the present invention should be included within the scope of protection of the present invention.

Claims

1. Use of a composition for the manufacture of a product according to any one of (1) to (6), characterized in that The composition comprises cis-5,11,14-eicosatetrienoic acid glyceride and cis-11,14-eicosatedienoic acid glyceride in a mass ratio of 1~5: 5~1; or The composition comprises cis-5,11,14-eicosatotrienoic acid glyceride and cis-5,11-eicosatodienoic acid glyceride in a mass ratio of 1~10: 10~1; The product is: (1) Health foods that help maintain healthy blood sugar levels; (2) Health foods that help control body fat; (3) Health foods that help maintain healthy blood lipid levels; (4) Health foods that help maintain healthy blood pressure levels; (5) Antioxidant health foods; (6) Anti-aging drugs.

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