A composition with the functions of reducing blood fat and increasing muscle and a preparation method thereof

Abstract

The present application relates to a kind of composition with the function of reducing blood fat, muscle function and its preparation method, including the following raw materials and its mass fraction: red arctic fruit 10~50 parts, human heart fruit 1~30 parts, anchor fruit 10~25 parts, chrysanthemum 1~20 parts, montmorency sour cherry 5~30 parts.Compared with prior art, human heart fruit and red arctic fruit in the composition of the present application provide active ingredients with muscle building efficacy, while anchor fruit, chrysanthemum and montmorency sour cherry provide active ingredients with blood fat reducing efficacy, and each component synergistically enhances the efficacy of reducing blood fat while building muscle, achieving day and night treatment, treating the symptoms and root cause mode of building muscle mainly during the day and reducing fat mainly at night.

Description

Technical Field

[0001] This invention relates to the field of biotechnology, and in particular to a composition having lipid-lowering and muscle-building functions and its preparation method. Background Technology

[0002] Overweight and obesity are influenced by a variety of factors, including genetics, diet, physical activity levels, lifestyle habits, and changes in the social environment. They are also significant risk factors for various cardiovascular diseases such as hypertension and hyperlipidemia, seriously affecting people's mental health and quality of life, and directly or indirectly increasing medical expenses and the socioeconomic burden.

[0003] The rising incidence of cardiovascular disease has increased the demand for lipid-lowering products. Currently, the main lipid-lowering technologies include the following: (1) Drug therapy: such as statins, which lower LDL-C by inhibiting HMG-CoA reductase, with atorvastatin as a representative drug, but with the risk of muscle toxicity (occurrence rate of about 5%); siRNA drugs, such as Inclisiran (injected twice a year), target the PCSK9 gene and have an LDL-C lowering effect of 50%-60%. (2) Gene editing: such as Verve's in vivo gene editing therapy, a single treatment can lower cholesterol for a long time. (3) Functional foods and natural extracts: such as red yeast rice containing monacolin K (similar to lovastatin), with an LDL-C lowering effect comparable to low-intensity statins, but the risk of liver toxicity should be noted; phytosterols inhibit intestinal cholesterol absorption, and 2g per day can lower LDL-C by 5%-10%, with high safety but weak effect.

[0004] While lowering blood lipids, building muscle is also a goal pursued by modern people. The common methods for building muscle currently include the following: (1) Nutritional supplementation: For example, whey protein is rapidly absorbed (reaching peak in 30 minutes) and is rich in leucine (promoting muscle synthesis), but lactose intolerant individuals need to choose isolated form; casein is slowly absorbed (lasting 7-8 hours), suitable for supplementation before bedtime to prevent muscle breakdown at night; betaine promotes muscle protein synthesis, and 1.5g per day can improve athletic performance, but excessive (>4g / d) may increase the risk of dyslipidemia; PQQ + sulforaphane synergistically protect the liver, improve the function of liver cell mitochondria, and reduce the risk of liver damage during the muscle-building period. (2) Exercise training: High-intensity interval training (HIIT): Stimulates muscle growth through short bursts of exercise, which is 30% more efficient than traditional training. (3) Cutting-edge technologies: such as: (3-1) CD47 antibody blockade: activates the AMPK pathway of skeletal muscle to achieve "fat reduction without muscle loss", animal experiments show that muscle mass increases by 18%; (3-2) Mechanical growth factor (MGF): stimulates MGF secretion through centrifugal training, promotes satellite cell proliferation and muscle nucleus increase; (3-3) Red yeast rice has a significant effect on lowering LDL-C (14.7%-48.2%), and also has anti-inflammatory effects. It contains monoclonal acid K, which may cause elevated liver enzymes and muscle symptoms. Liver function needs to be monitored.

[0005] It is evident that both the lipid-lowering and muscle-building markets are experiencing rapid growth. By fully utilizing valuable resources such as traditional food and medicine ingredients and new food raw materials, researching, utilizing, and developing products with lipid-lowering and muscle-building functions, it is expected to bring huge social and economic benefits. Summary of the Invention

[0006] The first technical problem to be solved by the present invention is to provide a composition with lipid-lowering and muscle-building functions in light of the current state of the art.

[0007] The second technical problem to be solved by the present invention is to provide a method for preparing the above-mentioned composition, in view of the current state of the prior art.

[0008] The technical solution adopted by the present invention to solve the first technical problem mentioned above is: a composition with lipid-lowering and muscle-building functions, characterized in that it includes the following raw materials and their mass fractions: 10-50 parts of red Arctic fruit, 1-30 parts of sapodilla, 10-25 parts of anchor fruit, 1-20 parts of chrysanthemum ginseng, and 5-30 parts of Montemorence sour cherry.

[0009] The above recipe uses red Arctic berries (also known as polar bear berries), which are plants of the genus Arctic berries in the family Ericaceae. They are also known as Tianlu or Dangnianku. They are deciduous shrubs of the genus Arctic berries in the family Ericaceae. The stems creep along the ground, and the berries are spherical, hairless, and glossy. When ripe, they are bright red and juicy.

[0010] The above formula uses *Ardisia crenata*, a perennial herb belonging to the genus *Ardisia* of the Boraginaceae family. In traditional Chinese medicine, *Ardisia crenata* is often used to treat skin problems caused by heat toxins, such as boils, carbuncles, and other skin conditions. Additionally, it can promote blood circulation and dispel blood stasis caused by falls and injuries, resulting in localized swelling and pain.

[0011] The chrysanthemum ginseng in the above formula is a perennial herbaceous plant belonging to the Gentianaceae family and the Gentian genus. Its roots are slightly fleshy and spindle-shaped, and its stems are clustered, purplish-red, and densely covered with papillae. The leaves are linear-lanceolate, with short white hairs (spiky lashes) along the edges, radiating outwards and spreading flat on the ground, resembling a chrysanthemum, hence the name. The root can be used medicinally; it is warm in nature and sweet in taste, possessing the effects of clearing deficiency heat, relieving cough and resolving phlegm, tonifying deficiency and benefiting the lungs, and nourishing the kidneys. It is mainly used to treat symptoms such as cough due to lung deficiency, weakness due to qi deficiency, and infantile malnutrition.

[0012] The sapodilla in the above formula is a woody plant belonging to the genus *Aristolochia* of the Sapotaceae family. It is also known as Wu Feng persimmon, red iron fruit, and strange fruit, and is named for its heart-like shape. The fruit is used to treat stomach pain, the bark is used for acute gastroenteritis and tonsillitis, the seeds have a diuretic effect, and its sap is used to treat dysentery and malaria.

[0013] The Montemorence sour cherry in the above formula is one of the most popular sour cherry varieties in North America. It contains five times more anthocyanins, quercetin, and natural melatonin than ordinary cherries, and has the effects of relieving gout and improving sleep.

[0014] The technical solution adopted to further solve the second technical problem mentioned above is: a method for preparing a composition with lipid-lowering and muscle-building functions as described above, characterized by including the following steps:

[0015] (1) The Montemorence sour cherry was subjected to impurity removal, freeze drying and ultra-fine pulverization in sequence to obtain Montemorence sour cherry powder;

[0016] (2) The red Arctic fruit and sapodilla were extracted by constant pressure boiling water, filtered, enzymatically hydrolyzed and vacuum concentrated, and the Montemorence sour cherry powder obtained in step (1) was used as substrate to spray granulate to obtain composite plant oligopeptide particles.

[0017] (3) The anchor fruit and chrysanthemum ginseng are extracted by pressure boiling water, filtered, vacuum concentrated and spray granulated in sequence to obtain plant extract powder, and the plant extract powder is used as core material to make composite plant extract microcapsule particles.

[0018] (4) The composite plant extract microcapsule particles prepared in step (3) are used as the first layer, and the composite plant oligopeptide particles prepared in step (2) and excipients are used as the second layer. The tablets are then compressed to obtain the desired composition with lipid-lowering and muscle-building functions.

[0019] Furthermore, the weight of the first layer is 300mg, and the hardness is 80N~90N; the weight of the second layer is 200mg, and the hardness is 90N~110N; the disintegration time is ≤30min.

[0020] Furthermore, the Montemorence tart cherry powder in step (1) is prepared through the following steps:

[0021] (a1) Cleaning and removing impurities: Clean the fresh Montemorence cherries and remove the stems and other impurities for later use;

[0022] (b1) Vacuum freeze-drying: Slice the washed Montemorence sour cherries and then freeze-dry them under vacuum;

[0023] (c1) Low-temperature ultrafine grinding: The dried Montemorence tart cherry freeze-dried slices are ultrafinely ground at low temperature and then passed through a 200-mesh sieve to obtain Montemorence tart cherry powder.

[0024] The Montemorence sour cherry powder prepared by this invention retains the anthocyanins, polyphenols, vitamins, minerals, dietary fiber and other nutrients in fresh Montemorence sour cherries well due to the use of vacuum freeze-drying, low-temperature ultrafine grinding and other technologies, with a cell wall breakage rate of over 95%.

[0025] Furthermore, the parameters for vacuum freeze drying in step (b1) are: pre-freezing rate 0.4℃ / min, initial pre-freezing temperature -40℃, final pre-freezing temperature -70℃, drying chamber pressure 45pa, heating plate temperature 42℃, and drying time 22h.

[0026] Furthermore, the composite plant oligopeptide particles in step (2) are prepared through the following steps:

[0027] (a2) Normal pressure boiling water extraction: Mix the red Arctic fruit and sapodilla fruit evenly according to the above mass proportions, put them into the extraction tank, add 8 to 15 times the volume of the raw material of purified water, stir, heat and raise the temperature, and keep the temperature at 100±2℃, pass through a 200 mesh sieve, extract twice, and combine the filtrates.

[0028] (b2) Enzymatic hydrolysis: Heat the above-extracted filtrate to 38℃~42℃, add trypsin, adjust the pH to 9.5~10.5, and hydrolyze for 1h~3h;

[0029] (c2) Enzyme inactivation: After the enzymatic hydrolysis is completed, adjust the pH to neutral, heat to boiling, and keep warm for 10 min to 20 min;

[0030] (d2) Centrifugation, filtration and ion exchange: Centrifuge the enzyme hydrolysate after enzyme inactivation to obtain centrifuged liquid, then filter the centrifuged liquid through a membrane to collect the filtrate with a molecular weight of less than 3000 Da, and pass the obtained filtrate through an ion exchange column for later use.

[0031] (e2) Vacuum concentration: The filtrate after step (d2) is concentrated under reduced pressure to obtain a concentrated solution;

[0032] (f2) Spray drying: Using the Montemorenci tart cherry powder obtained in step (1) as a substrate, the concentrate is sterilized and then spray-dried to obtain composite plant oligopeptide particles.

[0033] Furthermore, the parameters for vacuum concentration in step (e2) are: concentration temperature 55℃~65℃, vacuum control -0.07Mpa~-0.09Mpa, and relative density 1.05~1.08;

[0034] The parameters for spray drying in step (f2) are: inlet air temperature 60℃~85℃, outlet air temperature 55℃~75℃, and feeding speed 1.0L / min~2.5L / min.

[0035] Furthermore, the composite plant extract microcapsule particles in step (3) are prepared through the following steps:

[0036] (a3) Normal pressure boiling water extraction: Mix the anchor fruit and chrysanthemum evenly according to the above mass ratio, extract twice by boiling at normal pressure, filter separately, and combine the filtrates;

[0037] (b3) Concentration under reduced pressure: The extract filtrate is concentrated under reduced pressure to obtain a concentrated solution;

[0038] (c3) Spray drying: The concentrated liquid is sterilized and then spray dried to obtain plant extract powder;

[0039] (d3) Preparation of the first wall material / first core material solution: Add an appropriate amount of water to the above plant extract powder and stir to mix evenly to prepare a first core material solution with a mass fraction of 2%~5%. Mix gum arabic, γ-cyclodextrin, sodium alginate, and mono- and diglycerides of fatty acids evenly, add an appropriate amount of water and stir to dissolve, to prepare a first wall material solution with a mass fraction of 10~40%.

[0040] (e3) One-layer embedding: The first core material solution and the first wall material solution in step (d3) above are mixed at a mass ratio of 1~2.5:1.0~3.5, emulsified and homogenized at a speed of 1500 r / min~3000 r / min for 5 min~10 min, and allowed to stand for 2 h to obtain an emulsion for later use;

[0041] (f3) Preparation of second wall material / second core material solution: The emulsion in step (e3) above is used as the second core material solution. Maltodextrin, sucrose fatty acid ester and sodium stearoyl lactylate are mixed evenly and then water is added and stirred thoroughly to dissolve them, so as to prepare a second wall material solution with a mass fraction of 25%~40%.

[0042] (g3) Second-layer embedding: The second core material solution and the second wall material solution from step (f3) above are mixed at a mass ratio of 1:1 to 2 and stirred continuously for 20 min to 30 min to prepare a secondary embedding solution;

[0043] (h3) Spray drying: The secondary encapsulation solution in step (g3) above is spray dried to obtain composite plant extract microcapsule particles.

[0044] The composite plant extract microcapsule particles prepared by this invention have an encapsulation rate of over 95%, a particle size of 200μm~450μm, excellent emulsifying properties, and good stability to light and heat. They effectively mask the bitterness of the medicinal materials and greatly enhance the sensory quality of the product. Simultaneously, the double-layer encapsulation provides a sustained-release effect, achieving a long-lasting lipid-lowering effect.

[0045] Furthermore, in step (a3), the mass ratio of gum arabic, γ-cyclodextrin, sodium alginate, and mono- and diglycerides of fatty acids is 0.2~1.0:1.0~2.0:1.5~3.5:0.5~1.5;

[0046] In step (c3), the mass ratio of maltodextrin, sucrose fatty acid ester, and sodium stearoyl lactylate is 14~18:0.5~2:2~4.

[0047] Furthermore, the conditions for vacuum concentration in step (b3) are: concentration temperature 60℃~65℃, vacuum degree -0.07Mpa~-0.09Mpa, and relative density 1.06;

[0048] The spray drying conditions in step (c3) are: inlet air temperature 170℃~175℃, outlet air temperature 85℃~88℃, and feeding speed 2.0L / min~2.2L / min;

[0049] The parameters for spray drying in step (h3) are: inlet air temperature 160℃~185℃, outlet air temperature 80℃~90℃, and feeding speed 1.5L / min~2.0L / min.

[0050] Compared with the prior art, the advantages of the present invention are as follows:

[0051] Currently, Western medicine treatments for overweight and obesity face challenges such as side effects, adverse reactions, and safety risks, while traditional Chinese medicine treatments are known for their slow effectiveness. The composition of this invention is a natural composition, free of preservatives, artificial colors, and flavorings, ensuring its safety and quality. It relies entirely on the natural properties and flavor profiles of the raw materials. It strictly selects red Arctic fruit, sapodilla, quince, chrysanthemum ginseng, and Montemorence sour cherry as raw materials, with each ingredient formulated according to a scientific dose-response relationship. Sapodilla and red Arctic fruit provide active ingredients with muscle-building effects (represented by oligopeptides), while quince, chrysanthemum ginseng, and Montemorence sour cherry provide active ingredients with lipid-lowering effects (represented by total polysaccharides). The components work synergistically to fully realize their lipid-lowering and muscle-building effects.

[0052] Generally, higher activity levels during the day are more conducive to muscle growth, while lower activity levels at night result in less energy expenditure, potentially leading to energy surplus and elevated blood lipids. This invention utilizes a dual-layer encapsulation and dual-layer tableting technology to synergistically enhance the sustained-release effect, ensuring that daytime muscle building and nighttime fat reduction do not interfere with each other, but rather complement each other. This achieves a new model for improving body shape: a day-night approach that prioritizes muscle building while focusing on fat reduction, addressing both the symptoms and the root cause.

[0053] Furthermore, the preparation method of the composition of the present invention can retain the main active ingredients of each raw material to the greatest extent, thereby improving the bioavailability of each raw material and effectively reducing the dosage. Attached Figure Description

[0054] Figure 1 The results of the simulated gastric juice dissolution experiment of the sample obtained in Example 1 of Experiment 4 of this invention;

[0055] Figure 2 The results of the simulated gastric juice dissolution experiment of the sample obtained in Example 2 of Experiment 4 of this invention;

[0056] Figure 3 The results of the simulated gastric juice dissolution experiment of the sample in Example 3 of Test Example 4 of this invention;

[0057] Figure 4 This is the result of the simulated gastric juice dissolution experiment of the sample in Comparative Example 1 in Test Example 4 of the present invention. Detailed Implementation

[0058] The present invention will be further described in detail below with reference to the accompanying drawings and embodiments.

[0059] The composition of the present invention having the functions of lowering blood lipids and increasing muscle mass includes the following raw materials and their mass fractions: 10-50 parts of red Arctic fruit, 1-30 parts of sapodilla, 10-25 parts of anchor fruit, 1-20 parts of chrysanthemum ginseng, and 5-30 parts of Montemorence sour cherry.

[0060] Example 1: Preparation of a composition with lipid-lowering and muscle-building functions

[0061] Example 1.1: Preparation of Montemoren Simontemorensi Sour Cherry Powder

[0062] The specific preparation process of Montemoren Simontemorensi tart cherry powder is as follows:

[0063] (a1) Cleaning and removing impurities: Wash 5 portions of fresh Montemoranse tart cherries thoroughly with purified water to remove stems and other impurities and set aside.

[0064] (b1) Vacuum freeze drying: The Montemoranse tart cherry slices after the above cleaning were sliced ​​with an average thickness of 4 mm. The slices were then put into a vacuum freeze drying equipment for drying. The parameters were set as follows: pre-freezing rate 0.4℃ / min, initial pre-freezing temperature -40℃, final pre-freezing temperature -70℃, drying chamber pressure 45pa, heating plate temperature 42℃; drying time 22h.

[0065] (c1) Low-temperature ultra-fine grinding: After drying, the freeze-dried Montemorence cherry slices are put into a low-temperature ultra-fine grinder for low-temperature ultra-fine grinding to obtain Montemorence cherry powder. The obtained Montemorence cherry powder is required to pass through a 200-mesh sieve 100%.

[0066] Example 1.2: Preparation of composite plant oligopeptide particles

[0067] (a2) Normal pressure boiling water extraction: Mix 10 parts of red Arctic fruit and 1 part of sapodilla evenly, put them into the extraction tank and extract twice under normal pressure boiling. For the first extraction, add 10 times the amount of water and extract for 2 hours. For the second extraction, add 8 times the amount of water and extract for 1 hour. Filter and combine the filtrates.

[0068] (b2) Enzymatic hydrolysis: Heat the above extract filtrate to 38°C, add trypsin, adjust the pH to 9.6, and hydrolyze for 2 hours;

[0069] (c2) Enzyme inactivation: After the enzymatic hydrolysis is completed, adjust the pH to neutral, heat to boiling, and keep warm for 15 minutes;

[0070] (d2) Centrifugation: Centrifugation using a disc or tubular centrifuge;

[0071] (e2) Membrane filtration: The centrifuged liquid is subjected to membrane filtration, and the filtrate with a molecular weight of less than 3000 Da is collected;

[0072] (f2) Ion exchange: Desalination via an ion exchange column;

[0073] (g2) Vacuum concentration: The extract filtrate is concentrated under reduced pressure. The concentration temperature is controlled at 59℃~63℃, the vacuum degree is controlled at -0.07Mpa~-0.09Mpa, and the relative density is controlled at 1.07 to obtain the concentrate.

[0074] (h2) Spray drying: Using the Montemoren Simonte Morence tart cherry powder from Example 1 as the substrate, the concentrate was sterilized by UHT and then spray-dried into granules. The process parameters were: inlet air temperature 60℃~85℃, outlet air temperature 55℃~75℃, and feeding speed 1.0L / min~2.5 L / min, to obtain composite plant oligopeptide particles.

[0075] Example 1.3: Preparation of plant extracts

[0076] (a3) Normal pressure boiling water extraction: Mix 10 parts of anchor fruit and 1 part of chrysanthemum ginseng evenly, put them into the extraction tank and extract twice under normal pressure boiling. For the first extraction, add 8 times the amount of water and extract for 2 hours. For the second extraction, add 6 times the amount of water and extract for 1 hour. Filter and combine the filtrates.

[0077] (b3) Vacuum concentration: The extract filtrate is concentrated under reduced pressure. The concentration temperature is controlled at 60℃~65℃, the vacuum degree is controlled at -0.079Mpa~-0.09Mpa, and the relative density is controlled at 1.06 to obtain the concentrate.

[0078] (c3) Spray drying: The concentrate is sterilized by UHT and then spray dried. The process parameters are: inlet air temperature 170℃~175℃, outlet air temperature 85℃~88℃, and feeding speed 2.0L / min~2.2L / min to obtain plant extract powder.

[0079] Example 1.4: Preparation of microcapsule particles of compound plant extract

[0080] (a4) Preparation of the first wall material / core material solution: Add an appropriate amount of water to the plant extracts in the above mass ratio and stir to mix evenly to obtain a core material solution with a mass fraction of 2%~5%. Mix gum arabic, γ-cyclodextrin, sodium alginate, and mono- and diglycerides of fatty acids evenly, add an appropriate amount of water and stir to dissolve, to prepare a first wall material solution with a mass fraction of 20% (m / m). The mass ratio of the above gum arabic, γ-cyclodextrin, sodium alginate, and mono- and diglycerides of fatty acids is 0.5:1.2:1.8:1.0.

[0081] (b4) One-layer embedding: The first core material solution and the first wall material solution in step (a4) above are mixed at a mass ratio of 1.5:1.3, and emulsified and homogenized at a speed of 2500 r / min for 10 min using an emulsifying homogenizer, and then left to stand for 2 h for later use.

[0082] (c4) Preparation of the second wall material / core material solution: The emulsion in step (b4) above is used as the second core material solution. Maltodextrin, sucrose fatty acid ester and sodium stearoyl lactylate are mixed evenly and then an appropriate amount of water is added and stirred thoroughly to dissolve them, so as to prepare a second wall material solution with a mass fraction (m / m) of 25~40%. The mass ratio of the above maltodextrin, sucrose fatty acid ester and sodium stearoyl lactylate raw materials is 15:0.8:2.

[0083] (d4) Second-layer embedding: The second core material solution and the second wall material solution from step (c4) above are mixed at a mass ratio of 1:1.5 and stirred continuously for 20 min to 30 min to prepare a secondary embedding solution;

[0084] (e) Spray drying: The secondary encapsulation solution in step (d) above is spray dried with the following process parameters: inlet air temperature 160℃~165℃, outlet air temperature 80℃~185℃, and feeding speed 1.5L / min~2.0L / min to obtain composite plant extract microcapsule particles.

[0085] Example 1.5: Preparation of double-layer tablets

[0086] The first layer consists of microcapsules of the composite plant extract prepared in Example 1.4, while the second layer consists of a mixture of composite plant oligopeptide particles prepared in Example 1.2, 1% silica by weight, and 1% magnesium stearate by weight. The first layer weighs 300 mg and has a hardness of 80N-90N, while the second layer weighs 200 mg and has a hardness of 90N-110N. The resulting bilayer tablet has a disintegration time ≤30 min.

[0087] Example 2:

[0088] In this embodiment, the ingredients are: 20 parts Montemorence sour cherry, 30 parts red Arctic fruit, 15 parts sapodilla, 10 parts chrysanthemum ginseng, and 17 parts anchor thorn fruit. All other preparation steps are the same as in Example 1.

[0089] Example 3:

[0090] In this embodiment, the ingredients are: Montemorence sour cherry (30 parts), red Arctic fruit (50 parts), sapodilla fruit (30 parts), chrysanthemum ginseng (20 parts), and anchor thorn fruit (25 parts). All other preparation steps are the same as in Example 1.

[0091] Comparative Example 1:

[0092] The composite plant extract microcapsule particles prepared in Example 1.4, the composite plant oligopeptide particles prepared in Example 1.2, 1% silica by weight, and 1% magnesium stearate by weight were mixed evenly and then compressed into tablets to obtain the tablets of Comparative Example 1, with a weight of 500 mg, 80 N~110 N, and a disintegration time ≤10 min.

[0093] Comparative Example 2:

[0094] The composite plant extract microcapsule particles prepared in Example 1.4 were mixed with microcrystalline cellulose (equal in weight to the composite plant oligopeptide particles in Example 1), silicon dioxide accounting for 1% of the tablet weight, and magnesium stearate accounting for 1% of the tablet weight, followed by tableting. The tablet weight is 500mg, the hardness is 80N~110N, and the disintegration time is ≤10min.

[0095] Comparative Example 3:

[0096] The composite plant oligopeptide particles prepared in Example 1.2, microcrystalline cellulose (equal in weight to the composite plant extract microcapsule particles in Example 1), silicon dioxide accounting for 1% of the tablet weight, and magnesium stearate accounting for 1% of the tablet weight were mixed uniformly and then tableted. The tablet weight is 500mg, the hardness is 80N~110N, and the disintegration time is ≤10min.

[0097] Test Example 1: Muscle gain test in a zebrafish model of sarcopenia

[0098] 1.2.1 Formulation preparation

[0099] VD, HMBCa (calcium β-hydroxy-β-methylbutyrate) and MLCT (medium- and long-chain fatty acid oil) are prepared at an actual content ratio of 1:350000:200000. VD is prepared into a 2.00mg / mL stock solution with standard dilution water, and HMBCa and MLCT are respectively prepared into 20.0mg / mL stock solutions with standard dilution water. The formulation containing 1.96μg / mL VD, 1250μg / mL HMBCa and 1000μg / mL MLCT is prepared with standard dilution water.

[0100] 1.2.2 Zebrafish feeding and embryo collection

[0101] Adult zebrafish are reared in fish culture water at 28℃ (water quality: 200mg of instant sea salt is added per 1L of reverse osmosis water, with a conductivity of 450~550μS / cm; pH of 6.8; hardness of 50~100mg / L as CaCO₃), under a light cycle of 14 / 10h (light / dark), and are fed with artemia once every morning and evening. One day before spawning, female and male fish (at a ratio of 1:2) are mated, the fish eggs are collected and cleaned in time, and then cultured in a incubator at 28.5±1℃. The license number for the use of experimental animals is: SYXK (Zhejiang) 2022-0004, and the rearing management meets the requirements of international AAALAC accreditation (accreditation number: 001458).

[0102] Wild-type AB strain zebrafish (3dpf) were randomly selected and placed in 6-well plates, with 30 zebrafish per well. Standard dilution water was brought to a final volume of 3 mL. Each example, comparative example, and positive control (125 μg / mL whey protein powder) were administered in water. A normal control group and a model control group were also established. The normal control group was fed a standard diet without special treatment. All other experimental groups received 2.00 μmol / L dexamethasone in water concurrently with the drug administration. The treatment was carried out at 28°C for 96 h (with medium changes every 48 h) to establish a zebrafish sarcopenia model. The model control group was fed only a standard diet.

[0103] 1.2.3 Detection Indicators

[0104] HE staining: After exposure, zebrafish in each group were fixed overnight at 4°C with 4% PFA. After fixation, the zebrafish samples were washed with 70% ethanol, and then dehydrated by 70% ethanol / 0.5h, 85% ethanol / 0.5h, 95% ethanol / 0.5h, and 100% ethanol / 0.5h, respectively, before being embedded in paraffin. The paraffin-embedded tissues were serially sectioned at a thickness of 5μm and stained with hematoxylin and eosin (H&E). The muscle regions stained with H&E were observed blinded, and the intersegmental muscle fiber area and interstitial ratio of the zebrafish were analyzed using ImageProplus 6.0 software.

[0105] Statistical analysis of the average area of ​​intersegmental muscle fibers and the proportion of interfibrillary stroma in the normal control group showed that the average area of ​​intersegmental muscle fibers in the normal control group was 183,691 pixels, and the proportion of interfibrillary stroma was 10.52%. In contrast, the average area of ​​a single intersegmental muscle fiber in the model control group was significantly reduced (P<0.05) to 121,357 pixels, and the proportion of interfibrillary stroma reached 34.6%. Compared with the model control group, the average area of ​​intersegmental muscle fibers in the positive control group increased to 169,206 pixels, and the proportion of interfibrillary stroma decreased to 18.9%. After treatment in the examples, the average area of ​​a single muscle fiber between sarcomeres in Example 1 was 173,651 pixels, with a stroma ratio of 9.03%; in Example 2, the average area of ​​a single muscle fiber between sarcomeres was 180,679 pixels, with a stroma ratio of 9.54%; and in Example 3, the average area of ​​a single muscle fiber between sarcomeres was 179,305 pixels, with a stroma ratio of 9.32%. After treatment in the comparative examples, the average area of ​​a single muscle fiber between sarcomeres in Comparative Example 1 was 130,938 pixels, with a stroma ratio of 12.23%; and in Comparative Example 3, the average area of ​​a single muscle fiber between sarcomeres was 120,317 pixels, with a stroma ratio of 13.47%, slightly higher than the normal group, and the muscle-building effect was significantly better than whey protein powder (P<0.05). In Comparative Example 2, the average area of ​​a single muscle fiber between sarcomeres was 126,729 pixels, with a stroma ratio of 28.09%.

[0106] In summary, Examples 1, 2, 3, Comparative Example 1, and Comparative Example 3 all improved muscle fiber quality and muscle tissue structure based on the treatment of the sarcopenia model.

[0107] Experimental Example 2: Zebrafish Lipid-Lowering Experiment

[0108] 2.1 Sample preparation

[0109] Atorvastatin calcium stock solution with a mass concentration of 24 mg / mL was prepared using dimethyl sulfoxide (DMSO) and stored at 4°C for later use. During the experiment, the mass concentration of the atorvastatin calcium solution was adjusted to 2.4 μg / mL. 0.5 g of Oil Red O was weighed and added to 100 mL of 1,2-propanediol solution. The solution was stirred with a magnetic stirrer in the dark for 8 hours at room temperature until fully dissolved. The solution was then filtered through qualitative filter paper to prepare a fresh Oil Red O staining solution. A 1% glucose solution was prepared in the culture water. Solutions with concentrations of 20 μg / mL, 60 μg / mL, and 120 μg / mL were prepared for subsequent experiments in each experimental case.

[0110] 2.2 Model Construction and Intervention

[0111] Thirty zebrafish, 5 hours post-fertilization, were placed in each group in a 6-well plate. The control group received 4 mL of E3 culture water and was fed normal zebrafish fry feed. The other groups received 4 mL of glucose solution and were fed a high-fat zebrafish diet. Incubation was carried out at 28°C for 5 days, with the solution changed daily. After model construction, the solutions were discarded. The control and model groups received 4 mL of E3 culture water, while the positive control group received 4 mL of atorvastatin calcium solution. The high, medium, and low concentrations in this example were 20 μg / mL, 60 μg / mL, and 120 μg / mL, respectively. Each group was incubated at 28°C for 72 hours. Each well was fed 5 mg of feed daily, with the solution changed daily. Except for the control group, which was fed normal zebrafish feed, the other groups continued to be fed a high-fat diet (15% fat, Shanghai Feixi Biotechnology Co., Ltd.). The success of the high-fat model was determined by Oil Red O staining and the results of TG (triglycerides) and TC (total cholesterol) tests.

[0112] 2.3 Oil Red O staining incubation

[0113] After the treatment, the juvenile fish were rinsed with E3 culture water and collected in 1.5 mL centrifuge tubes. 1 mL of 4% paraformaldehyde solution was added, and the fish were fixed at 4°C for 1 day. The fixed juvenile fish were then transferred to 24-well plates, rinsed twice with E3 culture water, and subjected to permeation with different volume fractions of 1,2-propanediol solution in the following order: 25% 1,2-propanediol solution for 25 min → 50% 1,2-propanediol solution for 25 min → 75% 1,2-propanediol solution for 25 min → 100% 1,2-propanediol solution for 25 min. 2 mL of 0.5% Oil Red O stain (completely immersing the juvenile fish) was added, and the fish were protected from light with aluminum foil and stained overnight at room temperature. The stain was washed off, and 100% 1,2-propanediol solution was added to remove the background stain for 20 min (the effect of background removal was observed under a microscope). The staining of the zebrafish juveniles was observed under a microscope (adipocytes and hepatic steatosis stained red). ImageJ 6.0 software was used for image analysis and data acquisition. The total optical density (S) of blood lipids in the zebrafish tail vessels was analyzed and statistically analyzed. Based on the statistical results of the total optical density, the relative fat level was calculated according to formula (2) to evaluate the lipid-lowering effect of each embodiment and each comparative example. Relative fat level / % = (S) 模型组 -S 干预组 ) / S 模型组 ×100.

[0114] 2.4 Determination of TG and TC content in zebrafish

[0115] A zebrafish model of hyperlipidemia was constructed, with the following groups: a blank group (150 mL 0.01% DMSO + normal diet), a model group (150 mL 0.01% DMSO + high-fat diet), a positive group (150 mL 2.5 μg / mL atorvastatin calcium solution + high-fat zebrafish diet), and experimental groups (150 mL 20 μg / mL + high-fat zebrafish diet; 150 mL 60 μg / mL + high-fat zebrafish diet; 150 mL 120 μg / mL + high-fat zebrafish diet). Each group was incubated at 28℃ for 14 days. After the intervention, zebrafish were collected in 2mL EP tubes (30 fish / tube per group). The liquid in the centrifuge tubes was aspirated, and 200μL of PBS was added to each tube. The collected zebrafish were homogenized using a tissue homogenizer until no obvious tissue fragments were found. After centrifugation at 4℃ and 10000r / min for 10min, the supernatant was collected, and the TG and TC contents were determined according to the instructions of the kits (TC assay kit and TG assay kit, Shenzhen Raydu Life Sciences Co., Ltd.). The results are shown in Table 1.

[0116] Table 1 shows the effects of each example and comparative example on the TG, TC, and fat content of zebrafish. ±s, n=30)

[0117]

[0118] Note: # indicates P < 0.05 compared to the control group; * indicates P < 0.05 compared to the model group; ** indicates P < 0.01 compared to the model group.

[0119] As shown in Table 1, compared with the model group, the dose and / or high dose in each embodiment significantly reduced TC, TG and relative fat. Comparative Examples 1 and 2 showed only significant decreases in some indicators, while Comparative Example 3 had no significant effect.

[0120] Experiment Example 3: Human Food Trial

[0121] A fat loss survey was conducted to recruit 140 volunteers (BMI ≥ 24, age 50-65, 70 men and 70 women) to try the sample. They were randomly divided into 7 groups: 1 placebo group and 6 experimental groups, with 10 men and 10 women in each group. Each group received a placebo (starch tablets, 500mg / tablet), and the experimental and comparative groups received 4g daily before breakfast for 60 days. After the treatment, body fat percentage and muscle mass were measured using a BCA-1C body fat analyzer manufactured by Tsinghua Tongfang Co., Ltd.

[0122] The experimental results are shown in Table 2. Statistical analysis showed that, compared with before the trial, the body fat percentage and muscle mass of the placebo group did not change significantly. The body fat percentage of each example group decreased significantly, and the muscle mass increased significantly. In Comparative Example 2, only the male group showed a significant decrease in body fat percentage. In Comparative Example 1 and Comparative Example 3, only some gender groups showed a significant increase in body fat percentage or muscle mass.

[0123] Table 2 shows the statistics of the trial results for the target population.

[0124]

[0125] Note: # indicates P < 0.05 compared to before the food trial; ## indicates P < 0.01 compared to before the food trial.

[0126] Experiment Example 4: Simulated Gastric Juice Dissolution Experiment of Lipid-Lowering and Muscle-Building Bilayer Tablets

[0127] Referring to the General Chapter 0931 Dissolution and Release Determination Method of the 2020 Edition of the Chinese Pharmacopoeia, six tablets of each example and Comparative Example 1 were placed in 0.1 mol / L hydrochloric acid solution. The content of oligopeptides and taurine in the solution was tested at different time periods. The total polysaccharide (Wang Guangya. Detection Method of Efficacy Components of Health Food [M] Beijing: China Light Industry Press, 2002: 19-23) and oligopeptide (GB / T22492-2008 Soybean Peptide Powder) content and in vitro release amount in the double-layer tablets and acid solution were measured.

[0128] A graph is plotted with time on the x-axis and release rate on the y-axis. The results of the in vitro release rate test are as follows: Figure 1 , Figure 2 , Figure 3 as well as Figure 4 As shown. By Figure 1 , Figure 2 as well as Figure 3 It can be seen that the oligopeptides in the sample concentration solutions of the three examples were released at approximately 90% within 4 hours, followed by a sustained-release plateau. Release accelerated again after 7 hours, and was essentially complete after 8 hours, indicating that the oligopeptides in the bilayer tablets have an immediate-release effect. The total polysaccharides were released at approximately 40% within 3 hours, followed by a sustained-release plateau. Release accelerated again after 5 hours, reaching nearly 80% after 8 hours, after which the release rate slowed down again, and was essentially complete after 12 hours. Therefore, the samples in each example all exhibited good sustained-release effects.

[0129] And by Figure 4 It can be seen that approximately 80% of the oligopeptides and total polysaccharides in the Comparative Example 1 sample solution were released within 0.5 hours, and almost all of them were released within 2 hours. This demonstrates that the composition of the present invention exhibits good sustained-release properties, while the sustained-release effect of the Comparative Example 1 sample is poor, indicating that the bilayer compression technology plays a synergistic role in the sustained-release effect of the composition of the present invention.

[0130] The composition of the present invention contains both active ingredients with muscle-building effects (represented by oligopeptides) and active ingredients with lipid-lowering effects (represented by total polysaccharides). The dissolution test of Example 4 shows that the composition of the present invention can achieve sustained release of lipid-lowering components such as polysaccharides. Therefore, when the composition of the present invention is taken before breakfast, each component can exert its effect at different times, thereby achieving the effect of mainly building muscle during the day and lowering blood lipids at night.

[0131] Generally, people are more active during the day, which is more conducive to muscle growth. Using muscle-building ingredients at this time can achieve better muscle-building results. At night, people are less active and consume less energy, which can easily lead to excess energy and elevated blood lipids. At this time, the lipid-lowering effect is better, and at the same time, it will not consume muscle due to excessive increase in metabolic capacity, thus shaping the body more healthily.

[0132] The time-phased dissolution and release of the lipid-lowering and muscle-building components in the composition of Experiment 4 above indicates that the active ingredients in the composition of this invention intervene in zebrafish in a time-phased manner. At night, the lipid-lowering components of the composition primarily exert their effect, while during the day, the muscle-building components primarily exert their effect. Most zebrafish exhibit typical diurnal rhythms. In the experimental example, the drug was administered during the day, and after 8 hours, most of the total polysaccharides were released into the water. During the release of polysaccharides, the muscle-building components first exerted their muscle-building effect, primarily during the 8 hours of daylight. After 8 hours, with the dissolution of the lipid-lowering components, the lipid-lowering effect became dominant; the same principle applies to humans.

Claims

1. A composition having lipid-lowering and muscle-building functions, characterized in that... The ingredients include the following raw materials and their weight proportions: 10-50 parts of red Arctic fruit, 1-30 parts of sapodilla, 10-25 parts of anchor fruit, 1-20 parts of chrysanthemum ginseng, and 5-30 parts of Montemorence sour cherry.

2. A method for preparing the composition with lipid-lowering and muscle-building functions as described in claim 1, characterized in that... Includes the following steps: (1) The Montemorence sour cherry was subjected to impurity removal, freeze drying and ultra-fine pulverization in sequence to obtain Montemorence sour cherry powder; (2) The red Arctic fruit and sapodilla were extracted by constant pressure boiling water, filtered, enzymatically hydrolyzed and vacuum concentrated, and the Montemorence sour cherry powder obtained in step (1) was used as substrate to spray granulate to obtain composite plant oligopeptide particles. (3) The anchor fruit and chrysanthemum ginseng are extracted by pressure boiling water, filtered, vacuum concentrated and spray granulated in sequence to obtain plant extract powder, and the plant extract powder is used as core material to make composite plant extract microcapsule particles. (4) The composite plant extract microcapsule particles prepared in step (3) are used as the first layer, and the composite plant oligopeptide particles prepared in step (2) and excipients are used as the second layer. The tablets are then compressed to obtain the desired composition with lipid-lowering and muscle-building functions.

3. The method for preparing the composition with lipid-lowering and muscle-building functions as described in claim 2, characterized in that, The first layer weighs 300mg and has a hardness of 80N~90N; the second layer weighs 200mg and has a hardness of 90N~110N; the disintegration time is ≤30min.

4. The method for preparing the composition with lipid-lowering and muscle-building functions as described in claim 2, characterized in that, The Montemorence tart cherry powder in step (1) is prepared through the following steps: (a1) Cleaning and removing impurities: Clean the fresh Montemorence cherries and remove the stems and other impurities for later use; (b1) Vacuum freeze-drying: Slice the washed Montemorence sour cherries and then freeze-dry them under vacuum; (c1) Low-temperature ultrafine grinding: The dried Montemorence tart cherry freeze-dried slices are ultrafinely ground at low temperature and then passed through a 200-mesh sieve to obtain Montemorence tart cherry powder.

5. The method for preparing the composition with lipid-lowering and muscle-building functions as described in claim 4, characterized in that, The parameters for vacuum freeze drying in step (b1) are: pre-freezing rate 0.4℃ / min, initial pre-freezing temperature -40℃, final pre-freezing temperature -70℃, drying chamber pressure 45pa, heating plate temperature 42℃, and drying time 22h.

6. The method for preparing the composition with lipid-lowering and muscle-building functions as described in claim 2, characterized in that, The composite plant oligopeptide particles in step (2) are prepared through the following steps: (a2) Normal pressure boiling water extraction: Mix the red Arctic fruit and sapodilla fruit evenly according to the above mass proportions, put them into the extraction tank, add 8 to 15 times the volume of the raw material of purified water, stir, heat and raise the temperature, and keep the temperature at 100±2℃, pass through a 200 mesh sieve, extract twice, and combine the filtrates. (b2) Enzymatic hydrolysis: Heat the above-extracted filtrate to 38℃~42℃, add trypsin, adjust the pH to 9.5~10.5, and hydrolyze for 1h~3h; (c2) Enzyme inactivation: After the enzymatic hydrolysis is completed, adjust the pH to neutral, heat to boiling, and keep warm for 10 min to 20 min; (d2) Centrifugation, filtration and ion exchange: Centrifuge the enzyme hydrolysate after enzyme inactivation to obtain centrifuged liquid, then filter the centrifuged liquid through a membrane to collect the filtrate with a molecular weight of less than 3000 Da, and pass the obtained filtrate through an ion exchange column for later use. (e2) Vacuum concentration: The filtrate after step (d2) is concentrated under reduced pressure to obtain a concentrated solution; (f2) Spray drying: Using the Montemorenci tart cherry powder obtained in step (1) as a substrate, the concentrate is sterilized and then spray-dried to obtain composite plant oligopeptide particles.

7. The method for preparing the composition with lipid-lowering and muscle-building functions as described in claim 6, characterized in that, The parameters for vacuum concentration in step (e2) are: concentration temperature 55℃~65℃, vacuum degree -0.07Mpa~-0.09Mpa, and relative density 1.05~1.

08. The parameters for spray drying in step (f2) are: inlet air temperature 60℃~85℃, outlet air temperature 55℃~75℃, and feeding speed 1.0L / min~2.5L / min.

8. The method for preparing the composition with lipid-lowering and muscle-building functions as described in claim 2, characterized in that, The composite plant extract microcapsule particles in step (3) are prepared through the following steps: (a3) Normal pressure boiling water extraction: Mix the anchor fruit and chrysanthemum evenly according to the above mass ratio, extract twice by boiling at normal pressure, filter separately, and combine the filtrates; (b3) Concentration under reduced pressure: The extract filtrate is concentrated under reduced pressure to obtain a concentrated solution; (c3) Spray drying: The concentrated liquid is sterilized and then spray dried to obtain plant extract powder; (d3) Preparation of the first wall material / first core material solution: Add an appropriate amount of water to the above plant extract powder and stir to mix evenly to prepare a first core material solution with a mass fraction of 2%~5%. Mix gum arabic, γ-cyclodextrin, sodium alginate, and mono- and diglycerides of fatty acids evenly and add an appropriate amount of water to stir and dissolve to prepare a first wall material solution with a mass fraction of 10%~40%. (e3) One-layer embedding: The first core material solution and the first wall material solution in step (d3) above are mixed at a mass ratio of 1~2.5:1.0~3.5, emulsified and homogenized at a speed of 1500r / min~3000r / min for 5min~10min, and allowed to stand for 2h to obtain an emulsion for later use; (f3) Preparation of second wall material / second core material solution: The emulsion in step (e3) above is used as the second core material solution. Maltodextrin, sucrose fatty acid ester and sodium stearoyl lactylate are mixed evenly and then water is added and stirred thoroughly to dissolve them, so as to prepare a second wall material solution with a mass fraction of 25%~40%. (g3) Second-layer embedding: The second core material solution and the second wall material solution from step (f3) above are mixed at a mass ratio of 1:1 to 2 and stirred continuously for 20 min to 30 min to prepare a secondary embedding solution; (h3) Spray drying: The secondary encapsulation solution in step (g3) above is spray dried to obtain composite plant extract microcapsule particles.

9. The method for preparing the composition with lipid-lowering and muscle-building functions as described in claim 8, characterized in that, In step (a3), the mass ratio of gum arabic, γ-cyclodextrin, sodium alginate, and mono- and diglycerides of fatty acids is 0.2~1.0:1.0~2.0:1.5~3.5:0.5~1.

5. In step (c3), the mass ratio of maltodextrin, sucrose fatty acid ester, and sodium stearoyl lactylate is 14~18:0.5~2:2~4.

10. The method for preparing the composition with lipid-lowering and muscle-building functions as described in claim 8, characterized in that, The conditions for vacuum concentration in step (b3) are: concentration temperature 60℃~65℃, vacuum degree -0.07Mpa~-0.09Mpa, and relative density 1.06; The spray drying conditions in step (c3) are: inlet air temperature 170℃~175℃, outlet air temperature 85℃~88℃, and feeding speed 2.0L / min~2.2L / min; The parameters for spray drying in step (h3) are: inlet air temperature 160℃~185℃, outlet air temperature 80℃~90℃, and feeding speed 1.5L / min~2.0L / min.

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