Composite composition with multi-target heart protection function as well as preparation method and application thereof

Abstract

The invention discloses a compound composition with a multi-target heart protection function as well as a preparation method and application of the compound composition. The composition is composed of a basic functional oil phase, a core antioxidant / anti-inflammatory phase, a myocardial nutrition and regulation phase and a bioavailability promotion phase in a synergistic manner, and is integrated into a composite cardiac oligopeptide and microencapsulated particle key component substance through a specific enzymolysis and microencapsulation embedding process system. The invention aims to solve the technical problems of single target spot, insufficient synergistic effect of all components, low bioavailability of active components and poor stability of the existing heart protection product, and provides a stable and easily-absorbed composite composition which can realize synergistic effect from multiple channels at the same time. Technical effect verification shows that under the condition of equal-dose sample feeding, the composition can remarkably improve the bioavailability of functional components and shows excellent myocardial cell oxidative damage resistance and myocardial ischemia improvement activity in cell and animal models, the effect is remarkably superior to that of independent use or simple physical mixing of all the components, and the composition is suitable for clinical application. And the substantial progress of synergistic interaction is proved. The composition can be used for developing various medicines, health foods and functional foods beneficial to heart health.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biological medicine and special health products, and particularly relates to a composite composition with multiple cardiac protection functions such as antioxidant, anti-inflammatory and improvement of myocardial energy metabolism. BACKGROUND

[0002] Cardiac health problems are increasingly prominent, and the related products on the market at present are mostly single components or simple compounding, which have problems such as single target and limited efficacy, for example: Although fish oil and its products are beneficial to the cardiovascular system, they generally lack direct nutrition and energy metabolism support for myocardial cells; Although the functional material basis of turmeric and its extract, Dunaliella salina and its extract, and water extract of broccoli seeds (rich in glucoraphanin) and its antioxidant effect are clear, the stability and oral absorption, transformation and bioavailability are low, and the synergistic effect with other components is insufficient; As a traditional medicinal and edible material, Panax notoginseng stem and leaf extract is rich in saponins and has clear anti-myocardial ischemia, antioxidant and anti-inflammatory effects, but the synergistic effect of the existing technology with other cardiac protection components is insufficient, and the bioavailability is low; In the prior art, there is a lack of a complete technical solution for systematically integrating Omega-3 fatty acids, antioxidant network (curcumin, astaxanthin, VC, etc.), myocardial cell-specific nutrients (yak heart, horse heart, sika deer heart extract, Panax notoginseng stem and leaf extract), and broccoli seed extract through scientific proportioning and special process; In summary, it is of great significance to develop a composite cardiac protection composition with multiple components, multiple targets, multiple mechanism synergies, and stability and easy absorption. SUMMARY

[0003] PURPOSE OF THE INVENTION The purpose of the present application is to provide a composite composition which can simultaneously regulate blood lipids, inhibit oxidative stress, reduce inflammatory response, improve myocardial cell energy metabolism and other multiple pathways through scientific formula matching and dosage design, and achieve multidimensional protection of the heart. TECHNICAL SCHEME

[0004] To achieve the above-mentioned purpose, the present application adopts the following technical scheme: 1. Synergistic composition A composite composition is characterized by being made of the following raw materials by weight: Basic functional oil phase: fish oil 50-70 parts, yuanbao feng seed oil 10-20 parts; Core antioxidant / anti-inflammatory phase: Haematococcus pluvialis extract (rich in astaxanthin) 3-8 parts, turmeric extract (rich in curcumin) 5-10 parts, broccoli seed water extract (rich in glucoraphanin) 2-5 parts, purple cabbage powder 1-3 parts, Yacon fruit powder 5-15 parts, vitamin C 2-5 parts; Myocardial nutrition and regulation phase: composite heart source oligopeptide 2-5 parts obtained by enzymatic extraction of yak heart, horse heart and artificially bred sika deer heart, Panax notoginseng stem and leaf extract 5-15 parts (total saponin content not less than 50%), Chinese milk vetch seed powder 3-8 parts, red ganoderma powder 2-6 parts; Bioavailability promoting phase: black pepper powder 1-2 parts (rich in piperine); 2. Key preparation method The preparation method of the composite composition comprises the following key steps: Preparation of composite heart source oligopeptide: mix yak heart, horse heart and artificially bred sika deer heart at a mass ratio of 1:1:1, homogenize, then enzymatically digest at 50°C and pH 7.0 for 4 hours using a composite protease (trypsin and flavor protease at a ratio of 1:2), then inactivate, centrifuge, take the supernatant and spray dry to obtain oligopeptide powder with a molecular weight of less than 1000 Da; Microencapsulation: mix the water-soluble ingredients (Yacon fruit powder, vitamin C, broccoli seed water extract, etc.) in the core antioxidant / anti-inflammatory phase with the composite heart source oligopeptide and Panax notoginseng stem and leaf extract, use anthocyanins in purple cabbage powder as natural pigments and antioxidants, use meta maple seed oil and Haematococcus pluvialis extract (astaxanthin oil suspension) as part of the wall material, and use inclusion, microencapsulation, high-pressure homogenization and spray drying technology to prepare microcapsule particles; Final mixing: mix the microcapsule particles with fish oil, turmeric extract, Chinese milk vetch seed powder, red ganoderma powder, black pepper powder and other remaining components evenly, or further prepare them into soft capsules, hard capsules, tablets, granules, pills and other forms as needed.

[0005] The creativity, novelty and significant progress of the present application 1. Novelty different from prior art Process and form novelty: the present application innovatively uses animal heart sources (yak heart, horse heart and artificially bred sika deer heart at a mass ratio of 1:1:1) of specific sources and proportions, and uses a specific preparation technology to prepare composite heart source oligopeptide with a molecular weight of less than 1000 Da, which provides specific and easily absorbed nutrients (amino acids and small molecule peptides) for myocardial cells and supports energy metabolism of the heart; the present application innovatively microencapsulates the composite heart source oligopeptide and antioxidant ingredients, and uses meta maple seed oil and Haematococcus pluvialis (rich in astaxanthin) oil as part of the wall material, which solves the problem of stable coexistence of water-soluble ingredients and oil phase, and the problem of easy inactivation of various active ingredients during storage and digestion; Ingredient combination novelty: The prior art does not disclose the combination of complex heart-derived oligopeptides (yak heart, horse heart, sika deer heart) of specific sources, fish oil (rich in Omega-3), water extract of broccoli seeds (rich in glucoraphanin), ginkgo biloba seed oil (rich in nervonic acid), Haematococcus pluvialis extract (rich in astaxanthin), turmeric extract (rich in curcuminoids), and Panax notoginseng stem and leaf extract (rich in total ginsenosides), supplemented with black pepper powder (to improve bioavailability), as a systematic solution; 2. Creative outstanding features Multi-target synergistic mechanism: The present application is not a simple stacking of constituent substances or ingredients, but is based on the complex pathophysiological mechanism of heart protection, and a clear synergistic pathway is designed, in which: The first pathway (comprehensive antioxidant and anti-inflammatory) synergistically removes multiple free radicals through an antioxidant network composed of Haematococcus pluvialis extract, turmeric extract, vitamin C, roxburgh rose fruit powder, and red cabbage powder; and inhibits key inflammatory factors through the combined action of turmeric extract and water extract of broccoli seeds, thereby reducing myocardial inflammatory damage; The second pathway (myocardial cell-specific energy support and nutrition): Through the synergistic action of complex heart-derived oligopeptides and Panax notoginseng stem and leaf extract, small molecule peptides and saponins are provided to myocardial cells, thereby specifically improving myocardial energy metabolism and anti-ischemic injury; The third pathway (cardiovascular function regulation and cell membrane repair): The function of the heart autonomic nervous system and the body's stress state are regulated by Zizyphus jujuba powder and Red Ganoderma lucidum powder; the damaged myocardial cell membrane structure is repaired and maintained by nervonic acid in Ginkgo biloba seed oil; The fourth pathway (systemic bioavailability improvement): Piperine in black pepper powder non-selectively inhibits intestinal metabolic enzymes, thereby improving the oral bioavailability of the above-mentioned multiple active ingredients as a whole; 3. Significant progress achieved Synergistic enhancement of efficacy (creative core): Through the systematic design of the above-mentioned multi-target synergistic mechanism, the protective effect of the composition on the heart is significantly better than the simple addition of any single ingredient or conventional composition (see Technical Effect Verification Examples 1-3 for details); Improved product stability and compliance: The microencapsulation technology effectively protects the activity of sensitive ingredients and masks the undesirable flavor, making the product more stable and the user experience better; Widely applicable: The composition can be flexibly applied to various dosage forms, such as soft capsules, solid beverages, and functional foods, to meet different market demands. DETAILED DESCRIPTION

[0006] The present application is described in detail below through examples, but the scope of protection of the present application is not limited thereto;

[0007] Example 1: Preparation of composite heart-derived oligopeptide 1. Fresh or frozen yak heart, horse heart, artificially bred sika deer heart were taken, and fat and connective tissue were removed, and then washed with clean water; 2. The three kinds of heart sources were weighed according to a mass ratio of 1:1:1, mixed after cutting, and 2 times by weight of water was added for high-speed homogenization; 3. The homogenate was adjusted to pH 7.0 and the temperature was raised to 50℃. 1.5% of complex protease (trypsin: flavor protease = 1:2) of the total mass of the heart was added, and the enzyme was hydrolyzed for 4 hours under the conditions of incubation and slow stirring; 4. The enzyme solution was heated to 90℃ for 10 minutes to inactivate the enzyme; 5. The inactivated enzyme solution was centrifuged at 8000 r / min for 15 minutes at 4℃, and the supernatant was collected; 6. The supernatant was spray dried (import temperature 180℃, export temperature 80℃), and a light yellow composite heart-derived oligopeptide powder with a special meat aroma was obtained (referred to as sample No. 1).

[0008] Example 2: Preparation of microcapsule particles 1. Water phase preparation: 10 parts of Rosa roxburghii fruit powder, 3 parts of vitamin C, 3 parts of the composite heart-derived oligopeptide prepared in Example 1, 3 parts of water extract of broccoli seeds, 2 parts of purple cabbage powder, and 10 parts of Panax notoginseng stem and leaf extract were added to an appropriate amount of water, and then stirred, dissolved and homogenized; 2. Oil phase preparation: 15 parts of Acronychia fruit oil and 5 parts of Dunaliella salina extract (astaxanthin content 5%) were heated to 60℃ and stirred uniformly; 3. Emulsification and homogenization: the water phase was slowly added to the oil phase under high-speed shearing to preliminarily emulsify. Then, the stable O / W emulsion was formed by homogenizing 3 times through a high-pressure homogenizer under a pressure of 40 MPa; 4. Spray drying: the emulsion was spray dried (import temperature 160℃, export temperature 70℃) to obtain microencapsulated particles (referred to as sample No. 2);

[0009] Example 3: Preparation of the composite composition of the application 60 parts of fish oil, 30 parts of the microcapsule particles prepared in Example 2, 8 parts of turmeric extract (which needs to be dispersed with a small amount of fish oil in advance), 5 parts of Semen Ziziphi Spinosae powder, 4 parts of black fungus powder, and 1.5 parts of black pepper powder were taken. All ingredients were put into a vacuum tank with stirring, and stirred and mixed uniformly under vacuum conditions to ensure that there were no dry powder particles, and the composite composition of the application was obtained (referred to as sample No. 3).

[0010] Example 4: Preparation of a simple superposition composition (comparative example) To provide a comparison, a simple physical mixture sample containing all the component substances in claim 1 but not using the core preparation process of the application (i.e. not preparing the composite cardiogenic oligopeptide, not performing microencapsulation embedding) is prepared in this embodiment: 1. Take the same raw materials and weight parts as in Example 3: fish oil 60 parts, yuanbao maple seed oil 15 parts, Haematococcus pluvialis extract 5 parts, turmeric extract 8 parts, broccoli seed water extract 3 parts, purple cabbage powder 2 parts, roxburgh rose fruit powder 10 parts, vitamin C 3 parts, jujube kernel powder 5 parts, red ganoderma powder 4 parts, black pepper powder 1.5 parts, and freeze-dried mixed powder of non-enzymatic yak heart, horse heart, and sika deer heart in a mass ratio of 1:1:1 (instead of composite cardiogenic oligopeptide), notoginseng stem and leaf extract 10 parts; 2. Simply mechanically mix all the solid powder raw materials (freeze-dried mixed heart powder, jujube kernel powder, red ganoderma powder, turmeric extract, purple cabbage powder, roxburgh rose fruit powder, vitamin C, notoginseng stem and leaf extract) to obtain a mixed powder; 3. Directly stir and mix the mixed powder in item 2 with the liquid oil phase raw materials (fish oil, yuanbao maple seed oil, Haematococcus pluvialis extract) to obtain a mixture with the same composition but different process, i.e. a simple additive composition (referred to as sample No. 4).

[0011] Example 5: Preparation of other formulations of the composite composition of the application Drop pills: use the composite composition prepared in Example 3 as the raw material, add a matrix (such as polyethylene glycol 6000) to melt and mix, drop into a condensate under heat preservation conditions, and shrink to form drop pills; Soft capsules: use the composite composition prepared in Example 3 as the raw material, add an appropriate amount of soybean oil to mix uniformly as the content, use gelatin, glycerol, and water as the capsule skin material, and press the mixed content into soft capsules with a net content of 1000 mg per capsule; Tablets: use the composite composition prepared in Example 3 as the raw material, add an appropriate amount of filler (such as microcrystalline cellulose), disintegrant (such as sodium carboxymethyl starch), and lubricant (such as magnesium stearate) to mix uniformly, granulate, dry, and press into tablets; Granules: use the composite composition prepared in Example 3 as the raw material, add an appropriate amount of sucrose powder and dextrin to mix, use ethanol as a wetting agent to prepare a soft material, sieve and granulate, dry the granules, and pack them into granules.

[0012] Technical effect verification example To prove the significant progress and substantial significance of the composition of the present application (sample No. 3), the following series of tests were designed. All tests followed the principle of equal dosage, i.e. the dosage of the core functional components (such as astaxanthin, nervonic acid, glucoraphanin and xinyuan oligopeptide) of each sample group was kept consistent to ensure the comparability of the results. The experimental data were expressed as mean ± standard deviation (Mean ± SD), and one-way ANOVA was used for inter-group comparison, with p<0.05 and p<0.01 as the standard for statistically significant differences.

[0013] Verification Example 1: Absorption and bioavailability test Test samples: sample No. 2 (microcapsule particles), sample No. 3 (composition of the present application), sample No. 4 (simple superposition composition); Test method: healthy SD rats were selected and randomly divided into 3 groups, 12 rats in each group. After fasting for 12 hours, the corresponding samples were administered by gavage, and the dosage was converted to astaxanthin 5 mg / kg, nervonic acid 50 mg / kg, glucoraphanin 10 mg / kg, and total saponins 50 mg / kg. Blood was collected from the tail vein before administration and at 0.5, 1, 2, 4, 8, and 12 hours after administration, and plasma was separated. The plasma concentrations of nervonic acid, astaxanthin, glucoraphanin, and total saponins were detected by HPLC-MS / MS method. The area under the concentration-time curve (AUC0-12h) from 0 to 12 hours was calculated for each experimental group and subjected to statistical analysis and comparison; Test results: The pharmacokinetic parameters (AUC0-12h) of the four functional components in SD rats are shown in Table 1; Table 1. Pharmacokinetic parameters (AUC0-12h, Mean ± SD, n=12) of the four functional components in rats

[0014] Statistical note: "**" indicates p<0.01 compared with sample No. 4; "##" indicates p<0.01 compared with sample No. 2; Test conclusion: Table 1 shows that the AUC of each component in sample No. 3 is significantly higher than that in the other two groups, indicating that the bioavailability of the four functional components in sample No. 3 (composition of the present application) is significantly higher than that in sample No. 2 and sample No. 4. This fully proves that the complete preparation process (microencapsulation + black pepper powder + oil phase system) of the present application produces a synergistic absorption-promoting effect.

[0015] Verification Example 2: Myocardial cell oxidative damage test Test samples: sample No. 1 (xinyuan oligopeptide), sample No. 2 (microcapsule particles), sample No. 3 (composition of the present application), sample No. 4 (simple superposition composition); Test method: SD rat H9c2 myocardial cells were cultured in vitro, and normal control group and model group were set up. Except for the normal control group, the rest of the groups were treated with 200 μm hydrogen peroxide (H2O2) for 6 hours to induce oxidative damage. Each sample group was pretreated with the culture medium containing the corresponding sample (the final concentration was equivalent to 10 μg / mL of isodose heart source oligopeptide) for 24 hours before modeling. The cell survival rate was detected by CCK-8 method, and the SOD activity and MDA content in the cells were detected by kit; Test results: The cell survival rate, SOD activity and MDA content test statistical results of each group are shown in Table 2; Table 2. Protective effect of each sample on H9c2 myocardial cell oxidative damage (Mean ± SD, n = 6)

[0016] Statistical note: "*" indicates p < 0.05 compared with the model group; "**" indicates p < 0.01 compared with the model group; "##" indicates p < 0.01 compared with sample groups 1, 2 and 4; Test conclusion: Table 2 shows that sample No. 3 has the most significant protective effect on hydrogen peroxide (H2O2) induced oxidative damage. It is shown that the anti-myocardial cell oxidative damage activity of sample No. 3 (the composition of the present application) which is based on the same dose of heart source oligopeptide and fused with components such as notoginseng stem and leaf extract is obviously better than that of other samples, which proves the synergistic effect of the complete prescription and process.

[0017] Verification example 3: myocardial ischemia model test Test sample: sample No. 1 (heart source oligopeptide), sample No. 2 (microcapsule granules), sample No. 3 (the composition of the present application), sample No. 4 (simple superposition composition); Test method: SD rats were used to establish myocardial ischemia model by ligating the left anterior descending branch of the coronary artery. Sham operation group, model group and four sample groups (n = 12) were set up. After the operation, each drug group was given corresponding sample (the dose was equivalent to 50 mg / kg / d of isodose heart source oligopeptide) by gavage every day, for 28 consecutive days. At the end of the experiment, left ventricular ejection fraction (LVEF) was detected by echocardiography, and infarct size was calculated by TTC staining; Test results: The left ventricular ejection fraction and myocardial infarct size test data of each group are shown in Table 3; Table 3. Effect of each group on heart function and infarct size of myocardial ischemia rats (Mean ± SD, n = 12)

[0018] Statistical notes: "*" means p < 0.05 compared with the model group; "**" means p < 0.01 compared with the model group; "##" means p < 0.01 compared with the 1st, 2nd and 4th sample groups; Test conclusion: Table 3 shows that the 3rd sample has the best effect on improving heart function and reducing infarct area, indicating that the 3rd sample (the composition of the application) has extremely significant advantages over other groups in improving myocardial ischemia at the whole animal level, fully verifying that the heart protection effect of the application through multi-target point synergy has substantial progress.

Claims

1. A composite composition with multi-target cardioprotective function, characterized in that... Made from the following raw materials in parts by weight: 50-70 parts fish oil, 10-20 parts Acer truncatum seed oil, 3-8 parts Haematococcus pluvialis extract, 5-10 parts turmeric extract, 2-5 parts broccoli seed water extract, 1-3 parts purple cabbage powder, 5-15 parts prickly pear fruit powder, 2-5 parts vitamin C, 2-5 parts compound heart-derived oligopeptides, 3-8 parts jujube seed powder, 2-6 parts Ganoderma lucidum powder, 1-2 parts black pepper powder, and 5-15 parts Panax notoginseng stem and leaf extract; the compound heart-derived oligopeptides are obtained by enzymatic hydrolysis, extraction, and drying of yak heart, horse heart, and artificially raised sika deer heart in a mass ratio of 1:1:1, resulting in oligopeptide powder with a molecular weight of less than 1000 Da; the total saponin content of the Panax notoginseng stem and leaf extract is not less than 50%; the preparation method of the compound composition includes the following steps: a) Preparation of compound heart-derived oligopeptides: Yak heart, horse heart and artificially bred sika deer heart are mixed and homogenized in proportion, and then oligopeptide powder is obtained after enzymatic hydrolysis, enzyme inactivation, centrifugation and drying. b) Preparation of microcapsule particles: Prickly pear fruit powder, vitamin C, complex heart-derived oligopeptide, broccoli seed water extract, purple cabbage powder, Panax notoginseng stem and leaf extract are mixed with water to form an aqueous phase, which is then mixed with an oil phase composed of Acer truncatum seed oil and Haematococcus pluvialis extract. After emulsification, homogenization and spray drying, the microcapsule particles are obtained. c) The microcapsule particles obtained in step b) are mixed with fish oil, turmeric extract, jujube seed powder, Ganoderma lucidum powder, and black pepper powder to prepare the final composite composition.

2. The composite composition according to claim 1, characterized in that, The enzymatic hydrolysis in step a) uses a complex protease that is a mixture of trypsin and flavor protease in a mass ratio of 1:

2.

3. The composite composition according to claim 1, characterized in that, The enzymatic hydrolysis conditions in step a) are: temperature 50℃, pH 7.0, and time 4 hours.

4. A formulation, characterized in that, The composite composition comprising any one of claims 1 to 3 and pharmaceutically or food-grade excipients.

5. The formulation according to claim 4, characterized in that, The preparation is a soft capsule, pellet, tablet or granule.

6. The use of the composite composition of any one of claims 1 to 3 or the formulation of claim 4 or 5 in the preparation of pharmaceuticals, health foods, or functional foods for protecting heart health, assisting in improving myocardial energy metabolism, and for antioxidation or anti-inflammation.

Images