A nutritional composition for delaying muscle attenuation and bone loss based on muscle-bone axis interaction and a method of preparing the same

Abstract

The present application belongs to the field of biomedical engineering, and relates to a nutritional composition for delaying muscle attenuation and bone mass reduction based on muscle-bone axis interaction and a preparation method thereof. Specifically, the present application relates to a nutritional composition comprising or consisting of beta-hydroxy-beta-methylbutyric acid or a salt thereof and milk-derived basic protein; wherein the mass ratio of beta-hydroxy-beta-methylbutyric acid or the salt thereof to the milk-derived basic protein is 120:1-5:1. The nutritional composition of the present application realizes a comprehensive improvement effect on muscle-bone health that cannot be achieved by a single component through the synergistic effect of beta-hydroxy-beta-methylbutyric acid or the salt thereof and the milk-derived basic protein on the regulation of muscle-bone axis.

Description

Technical Field

[0001] This invention belongs to the field of biomedical engineering and relates to a nutritional composition based on muscle-bone axis interaction for delaying muscle loss and bone loss, and its preparation method. Background Technology

[0002] Among age-related degenerative diseases, sarcopenia and osteoporosis are two closely related diseases that often occur simultaneously, seriously affecting the quality of life and mobility of older adults, and significantly increasing the risk of falls, fractures and death.

[0003] Currently, most nutritional intervention programs on the market have limitations such as single-target approach and insufficient synergy.

[0004] In muscle intervention programs, mainstream products primarily supplement protein (such as whey protein), amino acids (such as leucine), and HMB. Their mechanism of action mainly focuses on stimulating muscle protein synthesis, but their direct promotion of bone health is limited. Numerous studies have shown that high protein intake, without corresponding bone protection strategies, may even potentially burden bone metabolism by increasing urinary calcium excretion.

[0005] In bone-related intervention programs, these programs primarily focus on supplementing calcium, vitamin D, and collagen peptides. While these can provide the building blocks for bone formation or promote calcium absorption, they are ineffective in addressing age-related loss of muscle mass and function. Without strong muscle support and mechanical stimulation, simple bone nutrition supplementation often yields limited results.

[0006] In recent years, in-depth research in the life sciences has revealed that muscles and bones are not independent organs, but rather functional couples that communicate bidirectionally through the "muscle-bone axis." The mechanical signals generated by muscle contraction are key stimuli for maintaining skeletal homeostasis; at the same time, both bones and muscles are endocrine organs, influencing each other's metabolism and function by secreting various cytokines (such as osteocalcin secreted by bones and actin, such as irisin, secreted by muscles).

[0007] Therefore, existing single-target intervention strategies fundamentally neglect the important physiological interaction mechanism of the muscle-bone axis. This prevents current technological solutions from creating a virtuous cycle of synergistic muscle-bone effects, making it difficult to effectively address the comprehensive decline of the musculoskeletal system in the elderly as a whole. Summary of the Invention

[0008] Based on the shortcomings of the aforementioned technical background, this invention aims to provide a novel and systematic nutritional solution. The primary objective of this invention is to provide a nutritional composition that can simultaneously and synergistically act on muscles and bones, overcoming the limitations of existing technologies that either neglect one aspect or simply "additively combine" nutrients. The core objective of this invention is to actively utilize and regulate the "muscle-bone axis" through the scientific formulation of specific functional active ingredients, not only supplementing the nutrients needed by muscles and bones separately, but also promoting positive signal exchange between muscles and bones, forming a virtuous cycle of "muscles promoting bone growth, and bone protecting muscles."

[0009] The present invention aims to provide a formulated milk powder that is scientifically formulated, convenient to consume, and easily accepted by the elderly.

[0010] This invention provides a nutritional composition for simultaneously delaying muscle loss and bone loss. The core of the composition lies in containing an effective dose of calcium β-hydroxy-β-methylbutyrate and colostrum basic protein. β-hydroxy-β-methylbutyrate and colostrum basic protein work synergistically to regulate the muscle-bone axis, achieving a comprehensive improvement in muscle and bone health that cannot be achieved by a single component.

[0011] The first aspect of the present invention provides a nutritional composition comprising, or consisting of, β-hydroxy-β-methylbutyric acid or a salt thereof and milk-derived basic protein; wherein the mass ratio of β-hydroxy-β-methylbutyric acid or a salt thereof to milk-derived basic protein is 120:1 to 5:1.

[0012] In one or more embodiments, the β-hydroxy-β-methylbutyrate is a salt formed by β-hydroxy-β-methylbutyric acid and a metal cation.

[0013] In one or more embodiments, the metal cation includes one or more of sodium ions, potassium ions, lithium ions, calcium ions, magnesium ions, and zinc ions.

[0014] In one or more embodiments, the β-hydroxy-β-methylbutyric acid or its salts include one or more of sodium β-hydroxy-β-methylbutyrate, potassium β-hydroxy-β-methylbutyrate, lithium β-hydroxy-β-methylbutyrate, calcium β-hydroxy-β-methylbutyrate, magnesium β-hydroxy-β-methylbutyrate, and zinc β-hydroxy-β-methylbutyrate.

[0015] In one or more embodiments, the milk source includes sheep milk and / or cow milk. In one or more embodiments, the milk-derived basic protein includes colostrum basic protein and / or raw milk basic protein. In one or more embodiments, the colostrum basic protein includes bovine colostrum basic protein and / or sheep colostrum basic protein. In one or more embodiments, the raw milk basic protein includes bovine raw milk basic protein and / or sheep raw milk basic protein. In one or more embodiments, the milk-derived basic protein is colostrum basic protein, more preferably bovine colostrum basic protein.

[0016] In one or more embodiments, the nutritional composition includes β-hydroxy-β-methylbutyric acid or β-hydroxy-β-methylbutyrate, milk-derived basic protein, and optionally sodium hyaluronate.

[0017] In one or more embodiments, the mass ratio of β-hydroxy-β-methylbutyric acid or β-hydroxy-β-methylbutyrate to sodium hyaluronate in the nutritional composition is 430:1 to 22.5:1.

[0018] In one or more embodiments, the nutritional composition comprises, or consists of, calcium β-hydroxy-β-methylbutyrate and colostrum basic protein, wherein the mass ratio of calcium β-hydroxy-β-methylbutyrate to colostrum basic protein in the nutritional composition is 80:1 to 5:1.

[0019] In one or more embodiments, the mass ratio of β-hydroxy-β-methylbutyric acid or β-hydroxy-β-methylbutyrate to milk-derived basic protein in the nutritional composition is 30:1 to 6:1.

[0020] In one or more embodiments, the nutritional composition may also include, optionally, sodium hyaluronate.

[0021] In one or more embodiments, the mass ratio of the milk-derived basic protein to sodium hyaluronate is 30:1 to 3:4.

[0022] A second aspect of the present invention provides a formulation comprising a nutritional composition as described in any embodiment herein.

[0023] A third aspect of the present invention is a product comprising a nutritional composition or formulation as described in any embodiment herein.

[0024] In one or more embodiments, the product is a food or food additive. In this food or food additive, the nutritional composition does not include β-hydroxy-β-methylbutyric acid, and the β-hydroxy-β-methylbutyrate is calcium β-hydroxy-β-methylbutyrate.

[0025] In one or more embodiments, the product is a health food. In this health food, the nutritional composition does not include β-hydroxy-β-methylbutyric acid, and the β-hydroxy-β-methylbutyrate is calcium β-hydroxy-β-methylbutyrate.

[0026] In one or more embodiments, the product is a pharmaceutical composition, optionally further comprising pharmaceutically acceptable excipients.

[0027] A fourth aspect of the present invention provides the use of the nutritional composition or formulation as described in any embodiment herein in the preparation of a product;

[0028] (1) The product is a food; in the food, the nutritional composition does not include β-hydroxy-β-methylbutyric acid, and the β-hydroxy-β-methylbutyrate is calcium β-hydroxy-β-methylbutyrate;

[0029] (2) The product is a health food; in the health food, the nutritional composition does not include β-hydroxy-β-methylbutyric acid, and the β-hydroxy-β-methylbutyrate is calcium β-hydroxy-β-methylbutyrate;

[0030] (3) Preparing pharmaceutical compositions for the prevention and / or treatment of diseases; and

[0031] (4) Prepare formulations for alleviating muscle loss, muscle function decline, reducing bone loss and altering bone microstructure.

[0032] In one or more embodiments, the health product relieves physical fatigue and increases bone density by alleviating muscle loss, reducing muscle function decline, reducing bone loss, and altering bone microstructure.

[0033] In one or more embodiments, the pharmaceutical composition prevents and / or treats a disease by downregulation.

[0034] In one or more embodiments, the disease includes sarcopenia, osteoporosis, etc.

[0035] The fifth aspect of the present invention provides non-therapeutic uses of nutritional compositions, formulations or products as described in any embodiment herein, including one or more of the following: (1) reducing the content of bone resorption markers, preferably, the bone resorption markers include CTX-1; (2) increasing the content of bone formation markers, preferably, the bone formation markers include BALP; (3) regulating myokinin and bone factors, including increasing Irisin content and / or decreasing MSTN content; (4) regulating bone factors, increasing OCN content and / or decreasing SOST content; (5) activating IGF-1; (6) improving myosomal axis interaction; (7) alleviating muscle mass loss, alleviating muscle function decline, reducing bone loss and altering bone microstructure; and (8) regulating bone metabolism.

[0036] A sixth aspect of the present invention provides a method for preparing a product as described in any embodiment herein, the method comprising: mixing calcium β-hydroxy-β-methylbutyrate and an organic or inorganic acid to obtain a suspension; optionally adding a wall material to the suspension to obtain an aqueous phase for preparing the product; wherein the particle size of the calcium salt in the suspension is less than or equal to 60 μm.

[0037] In one or more embodiments, the calcium salt is an organic acid calcium salt or an inorganic acid calcium salt.

[0038] In one or more embodiments, the suspension is sheared homogenized at a shear rate of 2000-5000 r / min and / or for a shearing time of 1-5 hours, preferably 4-6 hours.

[0039] In one or more embodiments, the product is a food product, more preferably a modified milk powder. In the modified milk powder, the nutritional composition does not include β-hydroxy-β-methylbutyric acid, and the β-hydroxy-β-methylbutyrate is calcium β-hydroxy-β-methylbutyrate. In the food product, the nutritional composition does not include β-hydroxy-β-methylbutyric acid, and the β-hydroxy-β-methylbutyrate is calcium β-hydroxy-β-methylbutyrate.

[0040] In one or more embodiments, the nutritional composition comprises 0.5-30 wt% of the total weight of the modified milk powder.

[0041] In one or more embodiments, the method for preparing the product includes the following steps:

[0042] (1) Preparation of milk liquid: Sterilize and cool raw cow's milk or raw sheep's milk to obtain milk liquid;

[0043] (2) Preparation of suspension: β-hydroxy-β-methylbutyrate calcium and organic acid or inorganic acid are mixed, sheared and homogenized to obtain a suspension, wherein the particle size of the organic acid calcium or inorganic acid calcium in the suspension is less than or equal to 60 μm;

[0044] (3) Preparation of aqueous phase: Mix the raw material protein and wall material to obtain a mixture, and add the mixture to the suspension in step (2) to obtain an aqueous phase;

[0045] (4) Mix the milk and aqueous phase, homogenize, sterilize, concentrate and dry to obtain powder; and

[0046] (5) The powder is mixed with milk-derived basic protein to obtain the product.

[0047] In one or more embodiments, in step (1), the sterilization is pasteurization. In one or more embodiments, the cooling temperature is 0-4°C.

[0048] In one or more embodiments, the method further includes step (1.5) between step (1) and step (2), step (1.5) comprising: mixing phospholipids and oils to obtain an oil phase; and step (4) comprising: mixing the milk, aqueous phase and oil phase, homogenizing, sterilizing, concentrating and drying to obtain powder.

[0049] In one or more embodiments, the phospholipid comprises 5 wt% of the total mass of the oil phase.

[0050] In one or more embodiments, the oil includes one or more of vegetable oils, animal oils, and microbial oils.

[0051] In one or more embodiments, the vegetable oil includes one or more of sunflower seed oil, soybean oil, rapeseed oil, corn oil, peanut oil, rice bran oil, rapeseed oil, cottonseed oil, flaxseed oil, perilla seed oil, walnut oil, safflower seed oil, grape seed oil, pumpkin seed oil, flaxseed oil, and peony seed oil.

[0052] In one or more embodiments, the animal fat includes one or more of butter, dairy fat, lard, tallow, and fish oil.

[0053] In one or more embodiments, the microbial oil comprises DHA algal oil and / or micrococcus oil.

[0054] In one or more embodiments, the oil comprises one or more of medium- and long-chain fatty acid edible oils, phytosterol esters, conjugated linoleic acid, conjugated linoleic acid, and glycerides of Acer truncatum seed oil.

[0055] In one or more embodiments, in step (2), the organic or inorganic acid is more acidic than β-hydroxy-β-methylbutyric acid.

[0056] In one or more embodiments, the organic acid includes one or more of citric acid, gluconic acid, malic acid, tartaric acid, and lactic acid, and the inorganic acid includes hydrochloric acid.

[0057] In one or more embodiments, in step (2), the molar ratio of calcium β-hydroxy-β-methylbutyrate and the organic or inorganic acid depends on whether the organic or inorganic acid is a monobasic acid. If the organic or inorganic acid is a monobasic acid, calcium β-hydroxy-β-methylbutyrate and the organic or inorganic acid are added in a molar ratio of <1:2. If the organic or inorganic acid is a tribasic acid, calcium β-hydroxy-β-methylbutyrate and the organic or inorganic acid are added in a molar ratio of <3:2.

[0058] In one or more embodiments, in step (2), the rate of shear homogenization is 2000-5000 r / min, and / or the time of shear homogenization is 1-5 hours, preferably 4-6 hours. In one or more embodiments, the pH of the suspension is 6.6-7.2. In one or more embodiments, the suspension also contains vitamins and minerals.

[0059] In one or more embodiments, in step (3), stirring is used to assist mixing for 10-15 minutes and / or the stirring rate is 800-1000 rpm.

[0060] In one or more embodiments, in step (3), the raw material protein includes milk protein, optionally plant protein, and optionally lactoferrin or immunoglobulin. In one or more embodiments, the milk protein includes one or more of raw sheep milk protein, raw cow milk protein, whey protein powder, whey protein isolate, sodium caseinate, and calcium caseinate. In one or more embodiments, the plant protein includes one or more of soy protein isolate, pea protein isolate, walnut protein, almond protein, and rice protein. In one or more embodiments, the wall material includes starch products and / or dietary fiber. In one or more embodiments, the starch product includes one or more of corn starch, wheat starch, rice starch, barley starch, potato starch, cassava starch, sweet potato starch, mung bean starch, pea starch, banana starch, pregelatinized starch, dextrin, hydroxypropyl starch, carboxymethyl starch, hydroxypropyl distarch phosphate, acetylated distarch adipate, oxidized starch, and cationic starch. In one or more embodiments, the dextrin includes one or more of resistant dextrin, maltodextrin, and cyclodextrin. In one or more embodiments, the dietary fiber includes water-soluble dietary fiber. In one or more embodiments, the dietary fiber comprises plant-derived dietary fiber and / or microbial-derived dietary fiber. In one or more embodiments, the dietary fiber comprises one or more of pectin, guar gum, xanthan gum, sodium alginate, carrageenan, gum arabic, sodium carboxymethyl cellulose, β-glucan, inulin, fructooligosaccharides, galactooligosaccharides, polydextrose, and sodium hyaluronate. In one or more embodiments, the mass of the wall material is greater than or equal to the mass of the raw material protein. In one or more embodiments, the mass of the wall material is 1-5 times the mass of calcium β-hydroxy-β-methylbutyrate.

[0061] In one or more embodiments, in step (4), the dry matter content in the aqueous phase is 5-30 wt% based on the total weight of the mixture. In one or more embodiments, in step (4), the homogenization pressure is 140-160 bar, and / or the homogenization time is 2-3 h, and / or the homogenization flow rate is 30-40 m³ / h. In one or more embodiments, the sterilization temperature is 85-95°C, and / or the sterilization time is 30-60 s. In one or more embodiments, the concentration is a flash evaporation. In one or more embodiments, the drying includes spray drying. In one or more embodiments, the inlet air temperature of the spray drying is 160-180°C, and / or the outlet air temperature of the spray drying is 70-80°C. Attached Figure Description

[0062] Figure 1 The experiment groupings and flowcharts are shown.

[0063] Figure 2The following are the basic facts. (A) Changes in body weight of mice under intervention; (B) Body weight of mice at baseline (0 months), 3 months, and 6 months; (C) Number of mice surviving in each group after intervention; (D) Average weekly feed intake of mice under intervention; (E) Total feed efficiency of mice under intervention. N = 10-12. and These represent comparisons between the two groups. P < 0.05 and P < 0.01.

[0064] Figure 3 To investigate the effect of intervention on muscle mass in aged mice. (A) Percentage of lean body mass in mice as measured by DXA. N = 10-12; (B) Cross-sectional area of ​​mouse quadriceps femoris muscle fibers, N = 8; (C) HE staining of mouse quadriceps femoris muscle. The black arrows indicate myofibroblast nuclei. This indicates a comparison with the elderly control group (OLD). P < 0.05, # indicates comparison with product formulation group COM P < 0.05, a indicates that the COM+ monomer group is compared with the corresponding monomer group. P < 0.05.

[0065] Figure 4 To investigate the effects of intervention on muscle function in aged mice. (A) Changes in maximum limb pulling force in mice before intervention; (B) Maximum limb pulling force in mice after intervention; (C, D) Assessment of movement distance (C) and movement time (D) of mice using a small animal treadmill. N = 10-12. Among them, " "" indicates a comparison with the elderly control group (OLD). P < 0.05, "#" indicates a comparison with product formulation group COM. P < 0.05, where “a” indicates that the COM+ monomer group is compared with the corresponding monomer group. P < 0.05.

[0066] Figure 5 To investigate the effects of intervention on whole-body bone density (BMD) and femoral morphology in aged mice. (A) Whole-body BMD in mice as measured by DXA. N = 10-12; (BG) Femoral parameters detected by Micro-CT. N = 4-5: (B) Bone volume fraction; (C) Number of trabeculae; (D) Trabecular thickness; (E) Trabecular separation; (F) Trabecular connectivity; (G) Femoral bone mineral density; (H) 3D section of mouse distal femur by Micro-CT; (I) HE staining of mouse distal femur. This indicates a comparison with the elderly control group (OLD). P <0.05, # indicates a comparison with product formulation group COM. P < 0.05, a indicates that the COM+ monomer group is compared with the corresponding monomer group. P <0.05.

[0067] Figure 6 To investigate the effects of intervention on tibial morphology in aged mice. (AF) Relevant tibial parameters were detected by Micro-CT. N = 4-5: (A) Bone volume fraction %; (B) Number of trabeculae; (C) Trabeculae thickness; (D) Trabeculae separation; (E) Trabeculae connectivity; (F) Tibial bone mineral density; (G) 3D section of mouse proximal tibia using Micro-CT; (H) HE staining of mouse proximal tibia. This indicates a comparison with the elderly control group (OLD). P < 0.05, # indicates comparison with product formulation group COM P < 0.05, a indicates that the COM+ monomer group is compared with the corresponding monomer group. P < 0.05.

[0068] Figure 7 To investigate the effects of intervention on bone metabolism in aged mice. (A) Serum CTX-I levels in mice; (B) Serum BALP levels in mice; N = 6; (C) Number of osteoclasts per unit surface area of ​​femoral trabeculae; (D) Osteoclast area per unit surface area of ​​femoral trabeculae; (E) Number of osteoclasts per unit surface area of ​​tibial trabeculae; (F) Osteoclast area per unit surface area of ​​tibial trabeculae; N = 4-5; (G) TRAP staining of the distal femur of a mouse; (H) TRAP staining of the proximal tibia of a mouse. The yellow arrows indicate osteoclasts. This indicates a comparison with the elderly control group (OLD). P < 0.05, # indicates comparison with product formulation group COM P <0.05, a indicates that the COM+ monomer group is compared with the corresponding monomer group. P < 0.05.

[0069] Figure 8 To investigate the regulation of muscle-bone axis-related molecules by different interventions in aged mice. (A) Level of Irisin in mouse muscle tissue; (B) Level of IGF-1 in mouse muscle tissue; (C) Level of OCN in mouse serum; (D) Level of MSTN in mouse muscle tissue; (E) Level of SOST in mouse bone tissue. N = 6. This indicates a comparison with the elderly control group (OLD). P <0.05, # indicates a comparison with product formulation group COM. P < 0.05, a indicates that the COM+ monomer group is compared with the corresponding monomer group. P <0.05.

[0070] Figure 9 For online cutting 0h.

[0071] Figure 10 This is a comparison chart of particle size after online shearing for 1-5 hours.

[0072] Figure 11 This is a schematic diagram of the criteria for evaluating the dynamism. Detailed Implementation

[0073] To enable those skilled in the art to understand the features and effects of the present invention, the terms and expressions used in the specification and claims are explained and defined in general below. Unless otherwise specified, all technical and scientific terms used herein have the ordinary meaning understood by those skilled in the art regarding the present invention, and in case of conflict, the definitions in this specification shall prevail.

[0074] The theories or mechanisms described and disclosed herein, whether right or wrong, should not in any way limit the scope of the invention, that is, the contents of the invention can be implemented without being limited by any particular theory or mechanism.

[0075] In this document, the terms “contains,” “includes,” “containing,” and similar terms encompass the meanings of “basically composed of” and “composed of.” For example, when this document discloses “A contains B and C,” “A is basically composed of B and C” and “A is composed of B and C” should be considered as having been disclosed in this document.

[0076] In this document, all features defined in the form of numerical ranges or percentage ranges, such as numerical values, quantities, contents, and concentrations, are for the sake of brevity and convenience only. Accordingly, descriptions of numerical ranges or percentage ranges should be considered as covering and specifically disclosing all possible sub-ranges and individual numerical values ​​(including integers and fractions) within those ranges.

[0077] Unless otherwise specified, percentages refer to mass percentages and proportions refer to mass ratios in this article.

[0078] In this article, the sum of the percentages of all components in the composition is 100%.

[0079] In this document, when describing embodiments or examples, it should be understood that it is not intended to limit the invention to those embodiments or examples. Rather, all alternatives, modifications, and equivalents of the methods and materials described herein are covered within the scope defined by the claims.

[0080] For the sake of brevity, not all possible combinations of the technical features in each implementation scheme or embodiment are described herein. Therefore, as long as there is no contradiction in the combination of these technical features, the technical features in each implementation scheme or embodiment can be combined arbitrarily, and all possible combinations should be considered within the scope of this specification.

[0081] In this article, "calcium β-hydroxy-β-methylbutyrate (CaHMB)" as used herein refers to the calcium salt of β-hydroxy-β-methylbutyric acid (also known as β-hydroxy-3-methylbutyric acid, β-hydroxy-β-methylbutyric acid, β-hydroxyisovaleric acid, or HMB), usually in monohydrate form.

[0082] In this article, "raw colostrum" refers to the milk secreted within 7 days after giving birth in healthy mammals (including cows, sheep, etc.) that are normally fed and free from infectious diseases and mastitis.

[0083] In this article, "raw milk" or "fresh milk" refers to the original milk secreted by mammals without any processing. Raw milk does not include colostrum in the first seven days after calving, milk produced during the use of antibiotics and during withdrawal, or spoiled milk.

[0084] In this article, "muscle-skeleton axis" refers to the functional unit formed by the coordinated action of muscles, bones, and joints.

[0085] In this article, "Lean Body Mass Percentage" (LBM%) is the proportion of lean body mass (body weight excluding fat) to total body weight, reflecting the relative content of non-fat tissues such as muscle, bone, organs, blood, and water in the body.

[0086] In this article, "wall material" refers to a material that has the ability to form a film or wall, and can encapsulate a core material (such as protein, CaHMB or HMB) to form a microcapsule, microsphere, membrane or embedded body.

[0087] The inventors have discovered that a nutritional composition prepared by combining specific calcium β-hydroxy-β-methylbutyrate and colostrum basic protein as active components can actively target and regulate the "muscle-bone axis," achieving a systematic and synchronous improvement in musculoskeletal health. This invention is thus completed.

[0088] Nutritional composition

[0089] The present invention provides a nutritional composition comprising, or consisting of, β-hydroxy-β-methylbutyric acid or a salt thereof and a milk-derived basic protein.

[0090] In this article, β-hydroxy-β-methylbutyrate is a salt compound formed by β-hydroxy-β-methylbutyric acid and a metal cation. The metal cation can be one or more of sodium ions, potassium ions, lithium ions, calcium ions, magnesium ions, and zinc ions.

[0091] In some embodiments, the β-hydroxy-β-methylbutyrate includes one or more of sodium β-hydroxy-β-methylbutyrate, potassium β-hydroxy-β-methylbutyrate, lithium β-hydroxy-β-methylbutyrate, calcium β-hydroxy-β-methylbutyrate, magnesium β-hydroxy-β-methylbutyrate, and zinc β-hydroxy-β-methylbutyrate.

[0092] In this article, milk-derived basic protein (MBP) is a mixture of basic proteins isolated and extracted from animal milk, whey, cheese, casein byproducts, whey powder, or whey protein concentrate, with an isoelectric point greater than 7, indicating basicity. Milk-derived basic proteins typically include colostrum basic proteins, bovine milk basic proteins (including buffalo milk basic proteins), or sheep milk basic proteins, with colostrum basic proteins being preferred.

[0093] Milk-derived basic proteins can be prepared using methods commonly used in this field. Raw materials include cow's milk, whey, cheese, casein byproducts, whey powder, or whey protein concentrate. The process involves first centrifuging to defatt the milk, then treating it with rennet to remove impurities such as casein to obtain whey liquid, then enriching the basic protein components, and finally concentrating, sterilizing, and spray drying to obtain milk-derived basic proteins.

[0094] In an exemplary embodiment, the milk-derived basic protein described herein includes colostrum basic protein. Colostrum basic protein (CBP) is produced from bovine colostrum through processes such as sterilization, defatting, centrifugation, removal of casein, α-lactalbumin, and β-lactoglobulin, microfiltration, ultrafiltration, and freeze-drying. The content of colostrum basic protein in colostrum is extremely low, accounting for only about 0.1% of the total protein in colostrum.

[0095] The preparation method of the colostrum basic protein in the nutritional composition applicable to this article can be a commonly used method, or a commercially available milk-derived basic protein product can be used directly. Common methods for preparing colostrum basic proteins include defatting and casein removal, enzymatic hydrolysis, isoelectric point precipitation, ultrafiltration, purification, sterilization, and freeze-drying. The defatting and casein removal step involves centrifuging fresh bovine colostrum at 4000-5000 rpm at 4°C for 15-20 min, discarding the supernatant fat, adjusting the pH of the supernatant to 4.6 with 1 mol / L hydrochloric acid aqueous solution, allowing it to stand for 30-40 min, and then centrifuging again at 4000-5000 rpm at 4°C for 15-20 min, and filtering to obtain whey. The enzymatic hydrolysis step involves adjusting the pH of the whey to 2.0 with 2 mol / L hydrochloric acid aqueous solution and incubating in a 37°C water bath for 10-12 min. Add magnetically immobilized pepsin to whey at a mass ratio of 1:(20-25). Incubate at 37°C in a constant temperature water bath at 150-200 rpm for 1.5-2 hours. Then, separate the magnetically immobilized pepsin using an external magnetic field to obtain the whey hydrolysate. The isoelectric point precipitation step includes adjusting the pH of the whey hydrolysate to 4.8 with 1 mol / L sodium hydroxide solution, incubating at 60°C for 70-80 minutes, followed by centrifugation at 4000-5000 rpm for 25-30 minutes. Collect the supernatant to obtain the whey separation solution. The ultrafiltration step may include cross-flow ultrafiltration using an ultrafiltration membrane with a molecular weight cutoff of 30 kDa at a flow rate of 6-7 L / min and a pressure of 0.45-0.55 MPa. During ultrafiltration, collect one-third of the whey separation solution volume as filtrate and replenish with an equal volume of pure water to obtain the whey ultrafiltration filtrate. One or more filtrations can be performed. The purification steps include: pumping the whey ultrafiltration retentate into a cation exchange resin column at a rate of 1.5-2 ml / min, eluting at 40°C with 0.05 mol / L phosphate buffer (pH 7.0) at a flow rate of 2.4-2.6 ml / min until no protein is detected, obtaining the eluent. The eluent is then subjected to cross-flow ultrafiltration using an ultrafiltration membrane with a molecular weight cutoff of 1 kDa at a flow rate of 6-7 L / min and a pressure of 0.75-0.85 MPa, concentrating to 2-3% of its original volume. The retentate is then collected to obtain the whey concentrate. The sterilization and lyophilization steps may include: placing the whey concentrate in a high-voltage pulsed electric field at a field strength of 30-50 kV / cm, a pulse frequency of 200-400 Hz, and a temperature of 5-10°C for 15-20 s, followed by vacuum freeze-drying for 16-20 h to obtain colostrum basic protein.

[0096] In some embodiments, the mass ratio of β-hydroxy-β-methylbutyric acid or β-hydroxy-β-methylbutyrate to milk-derived basic protein in the nutritional composition herein can be 120:1 to 5:1, for example, 120:1, 110:1, 100:1, 90:1, 75:1, 70:1, 65:1, 60:1, 55:1, 50:1, 45:1, 40:1, 35:1, 30:1, 25:1, 20:1, 15:1, 10:1, 9:1, 8:1, 7:1, 6:1, or any two of these values.

[0097] In an exemplary embodiment, the mass ratio of calcium β-hydroxy-β-methylbutyrate to colostrum basic protein in the nutritional composition herein can be 80:1 to 5:1, for example, 75:1, 70:1, 65:1, 60:1, 55:1, 50:1, 45:1, 40:1, 35:1, 30:1, 25:1, 20:1, 15:1, 10:1, 9:1, 8:1, 7:1, 6:1, or any two of these values.

[0098] In some embodiments, the nutritional compositions described herein include β-hydroxy-β-methylbutyric acid or a salt thereof, milk-derived basic protein, and optionally sodium hyaluronate. In exemplary embodiments, the nutritional compositions described herein include calcium β-hydroxy-β-methylbutyrate (CaHMB), colostrum basic protein (CBP), and optionally sodium hyaluronate (HA), or a combination thereof.

[0099] In some embodiments, the mass ratio of β-hydroxy-β-methylbutyric acid or β-hydroxy-β-methylbutyrate to sodium hyaluronate in the nutritional composition herein can be 430:1 to 22.5:1, for example, 430:1, 400:1, 350:1, 300:1, 250:1, 200:1, 150:1, 100:1, 50:1, 40:1, 30:1, 22.5:1, or any two of these ratios. In some embodiments, the mass ratio of colostrum basic protein to sodium hyaluronate in the nutritional composition herein can be 30:1 to 3:4, for example, 25:1, 20:1, 15:1, 10:1, 8:1, 6:1, 4:1, 2:1, 1:1, 1:2, or any two of these ratios.

[0100] In some embodiments, the mass of the milk-derived basic protein in the nutritional composition herein accounts for 0.8-20% of the total mass of β-hydroxy-β-methylbutyric acid or β-hydroxy-β-methylbutyrate, milk-derived basic protein, and sodium hyaluronate, for example, 0.8%, 1.0%, 1.2%, 1.4%, 1.6%, 1.8%, 2.0%, 2.2%, 4%, 6%, 8%, 10%, 12%, 14%, 16%, 18%, 20%, or any two of these values.

[0101] In some embodiments, the β-hydroxy-β-methylbutyric acid or β-hydroxy-β-methylbutyrate in the nutritional composition comprises 85-99% of the total mass of β-hydroxy-β-methylbutyric acid or β-hydroxy-β-methylbutyrate, milk-derived basic protein, and sodium hyaluronate, for example, 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or any two of these values.

[0102] In some embodiments, the mass of the milk-derived basic protein in the nutritional composition accounts for 1-20% of the total mass of β-hydroxy-β-methylbutyric acid or β-hydroxy-β-methylbutyrate, milk-derived basic protein, and sodium hyaluronate, for example, 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, or any two of these values.

[0103] In some embodiments, the mass of sodium hyaluronate in the nutritional composition accounts for 0.1-5.0% of the total mass of β-hydroxy-β-methylbutyric acid or β-hydroxy-β-methylbutyrate, milk-derived basic protein, and sodium hyaluronate, for example, 0.2%, 0.4%, 0.6%, 0.8%, 1.0%, 1.2%, 1.4%, 1.6%, 1.8%, 2.0%, 2.2%, 2.4%, 2.6%, 2.8%, 3.0%, 3.2%, 3.4%, 3.6%, 3.8%, 4.0%, 4.2%, 4.4%, 4.6%, 4.8%, or any two of these values.

[0104] The dosage form of the compositions described herein is not particularly limited and can be any commonly used in the art, such as powder, pills, capsules, granules, tablets, liquid formulations, or gels.

[0105] food

[0106] This invention also provides a food product containing a nutritional composition as described in any embodiment herein. The food product described herein is a non-therapeutic food, such as a functional food or health supplement. In this food product, the nutritional composition does not include β-hydroxy-β-methylbutyric acid, and the β-hydroxy-β-methylbutyrate is calcium β-hydroxy-β-methylbutyrate.

[0107] In this article, food can refer to processed foods such as infant formula, baked goods or pastries, fried foods, snacks, sauces or condiments, fast food or pre-made products, dairy products, beverages, fermented foods, and candy. Baked goods or pastries include, but are not limited to, cakes, biscuits, bread, pastries, pies, mooncakes, etc.; fried foods include, but are not limited to, fried chicken, French fries, fried meatballs, fried dough sticks, spring rolls, fried fish, fried cakes, etc.; snacks include, but are not limited to, potato chips, rice crackers, puffed foods, nuts and roasted seeds, etc.; sauces or condiments include, but are not limited to, salad dressings (such as Thousand Island dressing or mayonnaise), sesame paste, peanut butter, chili oil, oyster sauce, broad bean paste, etc.; fast food or pre-made products include, but are not limited to, instant noodles, frozen dumplings, frozen buns, pre-made dishes, etc.; dairy products include, but are not limited to, milk (such as whole milk, low-fat milk, skim milk), yogurt (such as whole yogurt, low-fat yogurt, skim yogurt), cheese, whipped cream, high-fat ice cream, etc.; candies include, but are not limited to, soft candies, shortbread, milk candies, chocolate, etc.; beverages include, but are not limited to, milk-containing beverages, plant protein beverages, cream coffee, milk tea, high-fat fruit and vegetable juices, etc.; infant formula products include, but are not limited to, formula milk powder, special medical purpose formula foods, infant complementary foods or nutritional supplements, etc.; fermented foods include, but are not limited to, fermented dairy products, fermented soy products, fermented meat products, etc.

[0108] For example, the food is infant formula or its reconstituted milk, which includes the nutritional composition, protein, carbohydrates, minerals, and vitamins as described in any embodiment herein. In this infant formula, the content of the nutritional composition, based on the total weight of the formula, is typically 2%-30 wt%.

[0109] The protein can be any protein commonly added to formula milk powder, including but not limited to whey protein, casein, soy protein, and cereal protein derived from cow's milk or goat's milk, as well as partially or fully hydrolyzed whey protein, casein, and soy protein derived from cow's milk or goat's milk. Soy protein can be soy protein and / or pea protein. Cereal protein includes, but is not limited to, one or more of rice protein, rice bran protein, wheat protein, rye protein, sorghum protein, corn protein, and oat protein.

[0110] The protein can be derived from skim milk powder, whey protein powder, and cheese powder. The infant formula of this invention can be formulated using skim milk powder, whey protein powder, and cheese powder known in the art for use in infant formula. Preferred cheese powder is cow's milk cheese powder.

[0111] Carbohydrates include digestible and indigestible carbohydrates. Digestible carbohydrates are typically sugars commonly added to infant formula, including but not limited to at least one of lactose, glucose, galactose, maltose, sucrose, fructose, starch, maltodextrin, glucose syrup, and corn syrup. Indigestible carbohydrates are typically indigestible oligosaccharides, including at least one of fructooligosaccharides, galactooligosaccharides, glucose oligosaccharides, xylooligosaccharides, mannose oligosaccharides, and cyclodextrin oligosaccharides.

[0112] In this invention, the vitamins include one or more selected from vitamin A, vitamin D, vitamin E, vitamin K1, vitamin B1, vitamin B2, vitamin B6, vitamin B12, niacin, folic acid, pantothenic acid, vitamin C, and biotin. The minerals include at least one selected from sodium, potassium, copper, magnesium, iron, zinc, manganese, calcium, phosphorus, iodine, chlorine, and selenium. The complex microbial minerals may also include choline and / or inositol. Typically, in this invention, the content of the complex microbial minerals is greater than 1.0%, preferably 1.2-3%.

[0113] In this invention, the stabilizer can be a stabilizer commonly added to formula milk powder, including but not limited to one or more of carrageenan, locust bean gum, gellan gum, xanthan gum, gelatin, gum arabic, and soybean polysaccharides. In this document, the stabilizer content is typically 0.1-1%.

[0114] For example, the food product is a modified milk powder such as children's milk powder, adolescent milk powder, adult milk powder, or elderly milk powder. This adult milk powder or modified milk powder includes the nutritional composition, milk raw materials, protein, carbohydrates, minerals, and vitamins as described in any embodiment herein. The content of the nutritional composition in this adult milk powder or modified milk powder is typically 0.5%-30% by total weight of the milk powder. The components of the modified milk powder described herein comply with GB19644-2024 "Milk Powder and Modified Milk Powder" and GB 2760 "National Food Safety Standard for the Use of Food Additives".

[0115] The protein can be any protein commonly added to formula milk powder, including but not limited to whey protein, casein, soy protein, and cereal protein derived from cow's milk or goat's milk, as well as partially or fully hydrolyzed whey protein, casein, and soy protein derived from cow's milk or goat's milk. Soy protein can be soy protein and / or pea protein. Cereal protein includes, but is not limited to, one or more of rice protein, rice bran protein, wheat protein, rye protein, sorghum protein, corn protein, and oat protein.

[0116] Carbohydrates include digestible and indigestible carbohydrates. Digestible carbohydrates are typically sugars and starches added to formulated milk powder by conventional additions or processing of raw or whole milk, including but not limited to at least one of lactose, glucose, galactose, maltose, sucrose, fructose, starch, maltodextrin, glucose syrup, and corn syrup. Preferably, more than 60% of the digestible carbohydrates are lactose. Indigestible carbohydrates are typically “dietary fiber,” including at least one of fructooligosaccharides, galactooligosaccharides, xylooligosaccharides, resistant dextrin, and inulin. In this document, the total content of digestible carbohydrates is typically 30-75%, preferably 40-60%, and the total content of indigestible carbohydrates is ≤15%.

[0117] In this invention, the vitamins include one or more of vitamin A, vitamin D, vitamin E, vitamin K1, vitamin B1, vitamin B2, vitamin B6, vitamin B12, niacin, folic acid, pantothenic acid, vitamin C, and biotin, and the minerals include at least one of sodium, potassium, copper, magnesium, iron, zinc, manganese, calcium, phosphorus, iodine, chlorine, and selenium.

[0118] In this invention, the stabilizer can be a stabilizer commonly added to formulated milk powder, including but not limited to one or more of carrageenan, locust bean gum, gellan gum, xanthan gum, gelatin, gum arabic, and soybean polysaccharides. In this document, the stabilizer content is typically 0.1-1%.

[0119] The food products of this invention are generally suitable for people of any age, such as infants (including babies, older infants, toddlers), children, teenagers, adolescents, young adults, adults, middle-aged people, or the elderly. The term "infant" refers to a person aged 0-6 months. The term "older infant" refers to a person aged 6-12 months. The term "toddler" refers to a person aged 12-36 months. The term "infant" refers to a person aged 0-36 months. The term "child" refers to a person aged 3-6 years. The term "teenager" refers to a person aged 7-17 years. The term "adult" refers to a person aged 18 years and above. The term "young adult" refers to a person aged 18-40 years. The term "adolescent" refers to a person aged 7-40 years. The term "middle-aged person" refers to a person aged 41-65 years. The term "elderly person" or "senior citizen" refers to a person aged 65 years and above.

[0120] In some implementations, the food may be infant food (e.g., baby food, follow-up baby food, toddler food), children's food, adolescent food, or adult food, such as infant formula (e.g., baby formula, toddler formula), baby complementary food, nutritional or dietary supplements, children's formula, children's snacks, formula milk powder for pregnant women, or milk powder for middle-aged and elderly people.

[0121] In one embodiment, the composition may be 0.001-80% by mass, preferably 0.1-50%, for example 0.1-30% or 1-20% relative to the total mass of the food. In addition to calcium β-hydroxy-β-methylbutyrate and colostrum basic protein, the food may also contain other ingredients, such as other proteins / amino acids, carbohydrates, fats, vitamins, minerals, and other food-grade acceptable materials.

[0122] For example, when the food is modified milk powder or milk powder, in addition to the nutritional composition described in the first aspect of the present invention, the milk powder may also include proteins such as whey protein, α-lactalbumin, casein, and milk fat globule membrane protein; carbohydrates such as lactose and oligosaccharides; lipids such as medium- and long-chain fatty acids and edible oils; minerals such as calcium, iron, and phosphorus; vitamins; and other nutritional fortifiers such as whey powder, choline tartrate, L-carnitine, isomerized lactose, docosahexaenoic acid, arachidonic acid, and walnut oil.

[0123] The food described herein also includes feed, which, in addition to the nutritional composition described in any embodiment herein, may contain basic feeds commonly used in the art, such as AIN-93M, AIN-76A, or other common grain feeds. In some embodiments, in the feed described herein, the mass of CBP may be 1-15%, 3-15%, or 10-15% of the total mass of CBP, CaHMB, and HA, and / or the mass of CaHMB may be 85-99%, 90-99%, or 85-95% of the total mass of CBP, CaHMB, and HA, and / or the mass of HA may be 0.1-5.0%, 0.2-0.6%, 0.5-1.0%, or 4.0-5.0% of the total mass of CBP, CaHMB, and HA.

[0124] Preparation method

[0125] β-Hydroxy-β-methylbutyric acid (HMB) is a natural substance produced by the metabolism of the essential amino acid leucine in the human body. It generally exists in two forms: free acid and calcium salt. However, due to the highly reactive nature of the free acid, it is often converted into calcium salt (CaHMB) during the synthesis process for use as a food ingredient. β-Hydroxy-β-methylbutyric acid is widely used as a dietary supplement, primarily functioning to inhibit muscle protein breakdown and promote muscle recovery and growth. There are two conventional methods for producing HMB-containing modified milk powder: Method 1 involves directly dry-mixing CaHMB with milk raw materials; Method 2 involves simply wet-mixing CaHMB with milk raw materials followed by spray drying. However, both methods have problems: Method 1 results in flavor deterioration: Modified milk powder made by directly dry-mixing CaHMB often develops an unpleasant odor during its shelf life, especially upon first opening. This is because: firstly, CaHMB itself may degrade, producing a slight off-odor; secondly, its inherent bitterness and astringency may interact with other components during long-term storage, becoming more pronounced or producing an unpleasant aftertaste. Method 2 involves direct wet mixing followed by spray drying. Direct wet mixing results in calcium ions (from CaHMB) existing in ionic form in aqueous solution. High concentrations of calcium ions (from CaHMB) aggregate with proteins (especially casein) to form calcium bridges, leading to protein precipitation and affecting the overall reconstitution and sensory properties of the formulated milk powder.

[0126] Based on the above problems, the inventors further discovered that by using a specific method to prepare products as described in any embodiment of this document, the prepared products, such as modified milk powder, have excellent taste and stability.

[0127] Therefore, the present invention also provides a method for producing the product as described in any embodiment herein, the method comprising: mixing calcium β-hydroxy-β-methylbutyrate and an organic or inorganic acid to obtain a suspension, optionally adding a wall material to the suspension to obtain an aqueous phase, for the preparation of the product; wherein the particle size of the calcium salt in the suspension is less than or equal to 60 μm.

[0128] In some embodiments, the present invention also provides a method for preparing a modified milk powder containing β-hydroxy-β-methylbutyric acid or its salt, the method comprising adding a suspension of calcium β-hydroxy-β-methylbutyrate and an organic or inorganic acid to an aqueous phase to prepare the modified milk powder; wherein the particle size of calcium citrate in the suspension is less than or equal to 60 μm.

[0129] In this article, "suspension" refers to a macroscopically stable and uniformly dispersed system that shows no stratification or obvious sedimentation upon visual observation.

[0130] In some embodiments, the pH of the suspension of calcium β-hydroxy-β-methylbutyrate and organic or inorganic acid described herein is 6.6-7.2.

[0131] In some embodiments, the suspension described herein is sheared homogenized at a rate of 2000-5000 r / min and / or for a time of 1-5 hours, preferably 4-6 hours.

[0132] In this article, the acidity of organic or inorganic acids is greater than that of β-hydroxy-β-methylbutyric acid. Organic acids include, but are not limited to, one or more of citric acid, malic acid, gluconic acid, tartaric acid, and lactic acid. Inorganic acids include hydrochloric acid.

[0133] In the method described herein, the aqueous phase may also include other ingredients commonly used in formulating milk powder, such as proteins, dextrins, carbohydrates, minerals, vitamins, water-soluble colloids, and acidity regulators. There are no particular restrictions on the timing of adding other conventional ingredients; it can be done at any step after obtaining the homogenized mixture of milk, oil, and aqueous phases, or before spray drying, or it can be added directly to the aqueous phase and then mixed and homogenized with the milk and oil phases.

[0134] In some embodiments, the wall material comprises starch products and / or dietary fiber. In some embodiments, the starch products comprise one or more of the following: corn starch, wheat starch, rice starch, barley starch, potato starch, tapioca starch, sweet potato starch, mung bean starch, pea starch, banana starch, pregelatinized starch, dextrin, hydroxypropyl starch, carboxymethyl starch, hydroxypropyl distarch phosphate, acetylated distarch adipate, oxidized starch, and cationic starch. In some embodiments, the dextrin comprises one or more of the following: resistant dextrin, maltodextrin, and cyclodextrin. In some embodiments, the dietary fiber comprises water-soluble dietary fiber. In some embodiments, the dietary fiber comprises plant-derived dietary fiber and / or microbial-derived dietary fiber. In some embodiments, the dietary fiber comprises one or more of the following: pectin, guar gum, xanthan gum, sodium alginate, carrageenan, gum arabic, sodium carboxymethyl cellulose, β-glucan, inulin, fructooligosaccharides, galactooligosaccharides, polydextrose, and sodium hyaluronate. In the method described herein, the wall material can be a wall material commonly used in the art to encapsulate β-hydroxy-β-methylbutyric acid or protein products in the preparation of milk powders, and the mass of the wall material can be 1-5 times the mass of the calcium salt of β-hydroxy-β-methylbutyric acid or the mass of the raw protein.

[0135] The complete preparation method of modified milk powder usually involves phase separation (milk phase, oil phase and water phase), emulsification and mixing, and finally drying of the components.

[0136] In some embodiments, the method for preparing the modified milk powder of this article includes the following steps: (1) preparing milk liquid: sterilizing and cooling raw cow milk or raw sheep milk to obtain milk liquid; (2) preparing oil phase: mixing phospholipids and oils to obtain oil phase; (3) preparing suspension: mixing calcium β-hydroxy-β-methylbutyrate and organic or inorganic acids, shearing and homogenizing to obtain suspension, wherein the particle size of the organic or inorganic calcium acid in the suspension is less than or equal to 60 μm; (4) preparing aqueous phase: mixing protein, dextrin and sodium hyaluronate to obtain a mixture, adding the mixture to the suspension in step (3) to obtain aqueous phase; (5) mixing the milk liquid, oil phase and aqueous phase, homogenizing, sterilizing, concentrating and drying to obtain powder; and (6) mixing the powder with milk-derived basic protein to obtain the product.

[0137] In the method described herein, in step (1), the sterilization can be pasteurization, and the cooling temperature can be 0-4℃.

[0138] In the method described herein, in step (2), the phospholipids account for 5 wt% of the total mass of the oil phase. In the method described herein, the oils include one or more of vegetable oils, animal oils, and microbial oils. Preferably, the vegetable oils include one or more of sunflower seed oil, soybean oil, rapeseed oil, corn oil, peanut oil, rice bran oil, rapeseed oil, cottonseed oil, flaxseed oil, perilla seed oil, walnut oil, safflower seed oil, grape seed oil, pumpkin seed oil, flaxseed oil, and peony seed oil. The animal oils include one or more of butter, milk fat, lard, tallow, and fish oil. The microbial oils include DHA algal oil and / or *Micrococcus pluvialis* oil. Other functional oils may also be included, such as medium- and long-chain fatty acid edible oils, phytosterol esters, etc. In some embodiments, the oils include one or more of medium- and long-chain fatty acid edible oils, phytosterol esters, conjugated linoleic acid, conjugated linoleic acid, glycerides, and *Acer truncatum* seed oil.

[0139] In the method described herein, in step (3), the organic acid includes citric acid and gluconic acid, and the inorganic acid includes hydrochloric acid. The shear homogenization rate can be 2000-5000 r / min, and / or the shear homogenization time can be 1-5 hours, preferably 4-6 hours. In the method described herein, the pH value of the suspension is 6.6-7.2, and the suspension may also contain vitamins and minerals;

[0140] In the method described herein, in step (3), the concentration of the aqueous solution of calcium β-hydroxy-β-methylbutyrate is 20-25 wt%. The molar ratio of calcium β-hydroxy-β-methylbutyrate to organic or inorganic acid depends on whether the organic or inorganic acid is a monoprotic acid. If the organic or inorganic acid is a monoprotic acid, calcium β-hydroxy-β-methylbutyrate and organic or inorganic acid are added in a molar ratio of <1:2. If the organic or inorganic acid is a triprotic acid, calcium β-hydroxy-β-methylbutyrate and organic or inorganic acid are added in a molar ratio of <3:2.

[0141] In the method described herein, in step (4), stirring is used to assist mixing, the stirring time is 10-15 min, and / or the stirring rate is 800-1000 rpm.

[0142] In the method described herein, in step (4), the raw material protein includes milk protein, optional plant protein, and optional lactoferrin or immunoglobulin. In the method described herein, the milk protein includes one or more of raw sheep milk protein, raw cow milk protein, whey protein powder, whey protein isolate, sodium caseinate, and calcium caseinate; preferably, the plant protein includes one or more of soy protein isolate, pea protein isolate, walnut protein, almond protein, and rice protein.

[0143] In the method described herein, in step (5), the dry matter content in the aqueous phase is 5-30 wt%, preferably 5-29.8 wt%, based on the total weight of the mixture. Preferably, in the method described herein, in step (5), the dry matter content of the milk is 70-90 wt%, and the oil phase content is 0-5 wt%, based on the total weight of the mixture.

[0144] In the method described herein, in step (5), the homogenization pressure is 140-160 bar, and / or the homogenization time is 2-3 hours, and / or the homogenization flow rate is 30-40 m³ / h. 3 / h; preferably, the sterilization temperature is 85-95℃, and / or the sterilization time is 30-60s; preferably, the concentration is flash evaporation; preferably, the drying includes spray drying; preferably, the inlet air temperature of the spray drying is 160-180℃, and / or the exhaust air temperature of the spray drying is 70-80℃.

[0145] In the method described herein, the final product contains β-hydroxy-β-methylbutyric acid or its salt in a mass ratio of β-hydroxy-β-methylbutyric acid to milk-derived basic protein in a mass ratio that meets the requirements of any embodiment described herein.

[0146] This article does not impose any particular restrictions on the conditions for homogenization, sterilization, concentration, and drying; these conditions can be those conventional in the field. Homogenization is typically performed using a high-pressure homogenizer, with a pressure of 140-160 bar and a flow rate of 30-40 m³ / s. 3 / h. Homogenization time can be adjusted according to the homogenization rate, for example, 1-6 h or 10-15 min. Sterilization can be high-pressure instantaneous sterilization, such as direct steam injection (DSI). Concentration can be a combination of flash evaporation and filtration. Drying can be spray drying, which is a key step in converting liquid materials into powder; the spray drying temperature can be 160-180℃.

[0147] This document also provides a method for improving the stability, reconstitution properties, and / or sensory properties of a modified milk powder containing HMB, the method comprising the steps of preparing the modified milk powder as described in any embodiment herein.

[0148] In this article, "stability" refers to the ability of milk powder or formulated milk powder to resist deterioration in quality during storage.

[0149] In this article, "reconstitution" refers to the solubility, dispersion, or suspension properties of milk powder or modified milk powder in water.

[0150] In this article, the sensory indicators include color, taste, odor, and state as specified in GB19644 National Food Safety Standard for Milk Powder and Modified Milk Powder.

[0151] Pharmaceutical Composition

[0152] The present invention also provides a pharmaceutical composition comprising a nutritional composition as described in any embodiment herein and a pharmaceutically acceptable carrier.

[0153] The present invention does not impose any particular limitation on the dosage form of the pharmaceutical composition, and can be any oral dosage form commonly used in the art, including but not limited to plasters, ointments, lotions, tinctures, slurries, powders, tablets, granules, pills, capsules, drop pills, granules, etc., preferably powders, granules, etc.

[0154] In some embodiments, the applied mass of colostrum basic protein in the pharmaceutical composition accounts for 3-50% of the total applied mass of colostrum basic protein and sodium hyaluronate, for example, 4%, 6%, 8%, 10%, 12%, 14%, 16%, 18%, 20%, 22%, 24%, 26%, 28%, 30%, 32%, 34%, 36%, 38%, 40%, 42%, 44%, 46%, 48%, or any two of these values.

[0155] In some embodiments, the applied mass of β-hydroxy-β-methylbutyric acid or a salt thereof in the pharmaceutical composition accounts for 85-99% of the total applied mass of β-hydroxy-β-methylbutyric acid or a salt thereof, colostrum basic protein and sodium hyaluronate, for example 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or any two of these values.

[0156] In some embodiments, the applied mass of colostrum basic protein in the pharmaceutical composition is 1-20% of the total applied mass of β-hydroxy-β-methylbutyric acid or its salt, colostrum basic protein and sodium hyaluronate, for example 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19% or any two of these values.

[0157] In some embodiments, the applied mass of sodium hyaluronate in the pharmaceutical composition is 0.1-5.0% of the total applied mass of β-hydroxy-β-methylbutyric acid or its salt, colostrum basic protein, and sodium hyaluronate, for example, 0.2%, 0.4%, 0.6%, 0.8%, 1.0%, 1.2%, 1.4%, 1.6%, 1.8%, 2.0%, 2.2%, 2.4%, 2.6%, 2.8%, 3.0%, 3.2%, 3.4%, 3.6%, 3.8%, 4.0%, 4.2%, 4.4%, 4.6%, 4.8%, or any two of these values.

[0158] β-hydroxy-β-methylbutyric acid (BMA) or its salts and colostrum basic protein can be formulated into a composition or formulation for administration to a subject. Alternatively, BMA and colostrum basic protein can be administered separately, for example, either concurrently or separately, without being formulated into the same composition. When administered separately, BMA and colostrum basic protein can be taken at intervals throughout the day. BMA and colostrum basic protein can also be taken in several divided doses throughout the day. The interval between administrations of the two components or the number of separate administrations is readily determined by those skilled in the art.

[0159] The pharmaceutical composition described herein can be administered to animals, such as mammals, rodents, etc., preferably humans (including adolescents or the elderly). In this document, animals can be livestock, poultry, aquatic animals, or rodents or mammals, such as pigs, cattle, sheep, chickens, ducks, geese, fish, shrimp, crabs, mice, rabbits, etc. In an exemplary embodiment, the animal is a rodent, preferably a mouse or rat. In a specific embodiment of the invention, mice are used as experimental animals, and a dosing regimen for improving myosomal axis interaction using a composition of calcium β-hydroxy-β-methylbutyrate and colostrum basic protein is proposed. It should be understood that converting the dosage from mouse to human dosage is easily done by those skilled in the art; for example, the theoretical human dosage can be calculated using the formula: Human dosage (mg / kg·BW) = W (conversion factor, usually taken as 0.11). The mouse dosage (mg / kg BW) is further calculated based on the theoretical human dosage being 0.1-4 times the actual human dosage. When the compositions of the present invention are used in humans (e.g., orally), the effective intake of CAHMB is 24.5-100 mg / kg BW, and / or the effective intake of CBP is 0.8-4.2 mg / kg BW, and / or the effective intake of HA is 0.15-1.1 mg / kg BW.

[0160] The pharmaceutical compositions applicable to this invention may be in the form of supplements or pharmaceutical preparations, and may further include a pharmaceutically available carrier, such as a solid, liquid, or gas. Examples of solid carriers include lactose, kaolin, sucrose, talc, gelatin, agar, pectin, gum arabic, magnesium stearate, and stearic acid. Examples of liquid carriers include syrup, peanut oil, olive oil, and water. Examples of gaseous carriers include carbon dioxide and nitrogen.

[0161] In preparing compositions for oral dosage forms, any convenient pharmaceutical medium can be used. For example, water, ethanol, oils, alcohols, flavoring agents, preservatives, coloring agents, etc., can be used to form oral liquid dosage forms, such as suspensions and solutions; while carriers, such as starch, sugars, microcrystalline cellulose, diluents, granulators, emulsifiers, lubricants, binders, and disintegrants, can be used to form oral solid dosage forms, such as powders, capsules, and tablets. Tablets and capsules are preferred oral dosage units using solid pharmaceutical carriers due to their ease of administration. Tablets can be coated using standard aqueous or non-aqueous techniques.

[0162] In this document, tablets of pharmaceutical compositions may be prepared by compression or molding, with the option of using one or more excipients or adjuvants. Tableting can be performed by compression of the active ingredient in a free-flowing form (e.g., powder or granules) in a suitable machine, optionally mixed with binders, lubricants, inert diluents, surfactants, or dispersants. Molded tablets can be formed in a suitable machine from a mixture of powdered compounds moistened with an inert liquid diluent. Each tablet preferably contains about 0.05 mg to about 5 g of active ingredient, and each sachet or capsule preferably contains about 0.05 mg to about 5 g of active ingredient. For example, formulations intended for oral administration to humans may contain about 0.5 mg to about 5 g of active pharmaceutical ingredient, mixed with a suitable and convenient carrier material, which may comprise about 5% to 95% of the total composition. Unit dosage forms typically contain about 1 mg to about 2 g of active ingredient, typically in doses of 25 mg, 50 mg, 100 mg, 200 mg, 300 mg, 400 mg, 500 mg, 600 mg, 800 mg, or 1000 mg.

[0163] The present invention also provides a method of administering a pharmaceutical composition as described in any embodiment herein, the method comprising administering the pharmaceutical composition as described in any embodiment herein to a subject to be treated and / or prevented.

[0164] In this document, the object can be an animal, such as a mammal or rodent, preferably a human (including infants, toddlers, adolescents, or the elderly). The animal can also be livestock, poultry, aquatic animals, or rodents or mammals, such as pigs, cattle, sheep, chickens, ducks, geese, fish, shrimp, crabs, mice, rabbits, etc. In an exemplary embodiment, the animal is a rodent, preferably a mouse or rat.

[0165] Applications and methods

[0166] The present invention also provides the use of the nutritional composition or formulation as described in any embodiment herein in the preparation of one or more of the following products: (1) the product is a food; (2) the product is a health product; (3) the preparation of a pharmaceutical composition for the prevention and / or treatment of diseases; and (4) the preparation of a formulation for the relief of muscle loss, the relief of muscle function decline, the reduction of bone loss and the alteration of bone microstructure.

[0167] In one or more embodiments, the disease includes sarcopenia, osteoporosis, etc.

[0168] The present invention also provides the use of the nutritional compositions or preparations described in any embodiment herein for non-therapeutic purposes, the use of which includes one or more of the following: (1) reducing the content of bone resorption markers, preferably, the bone resorption markers include CTX-1; (2) increasing the content of bone formation markers, preferably, the bone formation markers include BALP; (3) regulating myokinin and bone factors, including increasing Irisin content and / or decreasing MSTN content; (4) regulating bone factors, increasing OCN content and / or decreasing SOST content; (5) activating IGF-1; (6) improving myosomal axis interaction; (7) alleviating muscle mass loss, alleviating muscle function decline, reducing bone loss and altering bone microstructure; and (8) regulating bone metabolism.

[0169] The proteins or factors mentioned in this article have their conventional meanings in the field. For example, MSTN is a secretory glycoprotein encoded by the Myostatin gene, SOST is a secretory glycoprotein encoded by the Sclerostin gene, CTX-1 is a C-terminal cross-linked telopeptide of type I collagen, BALP is bone alkaline phosphatase, Irisin is a muscle factor secreted by muscles, IGF-1 (Insulin-like Growth Factor 1) is a protein hormone, and OCN (Osteocalcin) is a non-collagenous bone matrix protein specifically synthesized and secreted by osteoblasts.

[0170] In this document, "bone" or "skeleton" includes the periosteum, bone tissue, and bone marrow, and according to the origin, structure, function, and location of the skeleton in the body, it includes exoskeletons and endoskeletons, cartilage and bone, axial skeletons (including the skull, spine, sternum, and ribs) and appendage bones, long bones, short bones, flat bones (parietal bones, zygomatic bones, maxilla), and irregular bones. In an exemplary embodiment, the skeleton is the femur or tibia.

[0171] The present invention has the following beneficial effects:

[0172] 1. Theoretical Innovation: A Paradigm Shift from "Single Target" to "System Regulation"

[0173] Breaking through the limitations of existing technologies: Existing solutions often target muscle or bone nutrition in isolation, neglecting the intrinsic connection between muscle and bone as a functional couple. This invention, for the first time in the field of nutrition, actively targets and regulates the "muscle-bone axis" through a specific combination of active ingredients, achieving a systematic and synchronous improvement in musculoskeletal health.

[0174] Establishing a two-way virtuous cycle: This invention aims to form a two-way positive cycle of "muscle-driven bone growth" (providing mechanical stimulation by enhancing muscle function) and "bone-driven muscle protection" (improving muscle metabolism by optimizing skeletal endocrine function feedback), fundamentally solving the problem of limited effectiveness of single intervention measures.

[0175] 2. Composition Innovation: "Dose-Reduced Synergistic Effect" of Calcium β-Hydroxy-β-Methylbutyrate and Colostrum Basic Protein

[0176] Equivalent low dose: The effect of a very low dose of calcium β-hydroxy-β-methylbutyrate and colostrum basic protein combination is equivalent to or even better than several times the dose of calcium β-hydroxy-β-methylbutyrate monomer or colostrum basic protein monomer.

[0177] Multiple benefits: This synergy directly translates into higher safety (avoiding the potential risks of high doses), better economics (reduced raw material costs), and better product applicability.

[0178] 3. Innovation in the chain of evidence: A complete explanation of the mechanism from macro to micro levels.

[0179] This invention, through robust animal experiments, constructs a complete chain of evidence from macroscopic function to microscopic molecules, confirming its effects and mechanisms:

[0180] Macroscopic structure and function: Based on relevant indicators of muscle and bone, the nutritional composition of the present invention can significantly reduce muscle mass loss and muscle function decline, reduce bone loss and bone microstructure changes.

[0181] Metabolic dynamic balance: Biochemical markers confirm that the nutritional composition of the present invention can bidirectionally regulate bone metabolism (reduce the bone resorption marker CTX-I and increase the bone formation marker BALP).

[0182] Muscle-bone axis molecular dialogue: Through the detection of key factors, we have revealed the deep mechanism by which it regulates muscle-bone cross-communication by modulating myogenic factors (such as increasing Irisin and decreasing MSTN) and bone factors (such as increasing OCN and decreasing SOST) and activating common hub factors (such as IGF-1).

[0183] 4. Process Innovation: Dedicated preparation process designed to achieve synergistic effects.

[0184] HMB itself is a directly absorbable molecular form, with higher absorption and utilization compared to CaHMB, but it has the disadvantages of strong hygroscopicity and a pungent taste. A solution containing HMB and calcium citrate is dispersed in a wall material solution (such as resistant dextrin, concentrated whey protein powder, or sodium hyaluronate). A homogeneous system is formed through emulsification and homogenization. Then, raw milk, carbohydrates such as syrup, and oils are mixed in, and the mixture is emulsified and homogenized again to form a homogeneous system. This mixture is then dried using an optimized low-temperature spray drying process. During drying, the wall material forms a dense protective film, encapsulating HMB and calcium citrate within the particles. This method improves the flavor degradation of dry-process CaHMB-added modified milk powder products and avoids the protein denaturation and precipitation problems caused by the reaction between proteins and CaHMB in the wet process. It produces modified milk powder with excellent taste and stability, significantly reducing heat contact, masking undesirable flavors, reducing hygroscopicity, and improving dispersion uniformity.

[0185] The present invention will be further described below by way of specific embodiments. It should be understood that these embodiments are merely illustrative and are not intended to limit the scope of the invention. Unless otherwise stated, the methods and reagents used in the embodiments are conventional methods and reagents in the art.

[0186] Observation indicators:

[0187] 1. Muscle Indicators:

[0188] 1) Muscle mass: (A) Percentage of lean body mass in mice as measured by dual-energy X-ray absorptiometry (DXA); (B) Cross-sectional area of ​​quadriceps femoris muscle fibers in mice; (C) HE staining of quadriceps femoris muscle in mice;

[0189] 2) Muscle function: (A) Maximum limb pulling force; (B) Assessment of movement distance and movement time in mice using a small animal treadmill;

[0190] 2. Skeletal parameters:

[0191] 1) Whole-body bone mineral density in mice;

[0192] 2) Bone morphology: (A) Relevant indicators of femur and tibia detected by Micro-CT, including bone volume fraction, number of trabeculae, trabeculae thickness, trabeculae separation, trabeculae connectivity, and bone mineral density; (B) 3D sections of distal femur and proximal tibia of mouse obtained by Micro-CT; (C) HE staining of distal femur and proximal tibia of mouse.

[0193] 3) Bone metabolism: (A) CTX-I level in mouse serum; (B) BALP level in mouse serum; (C) Number of osteoclasts per unit surface area of ​​trabecular bone in femur and tibia; (D) Osteoclast area per unit surface area of ​​trabecular bone in femur and tibia; (E) TRAP staining in distal femur and proximal tibia of mouse.

[0194] 3. Molecular levels related to the muscle-skeleton axis:

[0195] 1) The level of irisin in mouse muscle tissue; 2) The level of insulin-like growth factor-1 (IGF-1) in mouse muscle tissue; 3) The level of osteocalcin (OCN) in mouse serum; 4) The level of myostatin (MSTN) in mouse muscle tissue; 5) The level of osteostatin (SOST) in mouse bone tissue.

[0196] 4. Knee joint parameters:

[0197] 1) The degree of swelling in the knee joint;

[0198] 2) Cartilage thickness of the medial tibial and femoral joints in mice;

[0199] 3) Morphology of mouse knee joint: (A) HE staining of mouse knee joint; (B) Safranin-Fix-Green staining of mouse knee joint; (C) Masson staining of mouse knee joint;

[0200] 4) Percentage of glycosaminoglycans in the medial tibial cartilage of mice (%)

[0201] 5) Percentage of collagen area in the medial tibial cartilage of mice.

[0202] 5. Feed efficiency: Total weight gain / Total feed intake 100%

[0203] Experimental Design:

[0204] The intervention animals were 2- or 12-month-old male C57BL / 6J mice (purchased from Beijing Vital River Laboratory Animal Technology Co., Ltd.), housed in an SPF-barrier environment at the Animal Laboratory Department of the North Campus of Sun Yat-sen University, with a 12-hour diurnal cycle, a temperature of 25°C, and a relative humidity of 40%-60%. All mice had free access to water and food during the experiment. The intervention method was a special diet intervention, lasting for 6 months. Housing conditions: Mice were housed individually in an SPF-barrier environment with a 12-hour diurnal cycle, a temperature of 25°C, and a relative humidity of 40%-60%. All mice had free access to water and food during the experiment. The diet was changed weekly, with an addition of 35g / mice (5g / mice / day). Mice had free access to food, and the remaining feed was weighed. During the acclimatization period, all mice were fed AIN-93G diet. After one week of acclimatization, 12 2-month-old mice were used as the young control group, and the remaining 108 12-month-old mice were randomly divided into 9 groups of 12 mice each.

[0205] 1) Young control group (YOU, 2 months old): fed AIN-93G feed (purchased from Jiangsu Medison Biomedical Co., Ltd.);

[0206] 2) Elderly control group (OLD, 12 months old): fed AIN-93G diet;

[0207] 3) CaHMB group (HMB, 12 months old): CaHMB at a dose of 2400 mg / kg of feed was added to the AIN-93G diet;

[0208] 4) CBP group (CBP, 12 months old): CBP was fed AIN-93G diet supplemented with 150mg / kg diet.

[0209] 5) HA group (HA, 12 months old): HA was added at a rate of 40 mg / kg of feed to the AIN-93G diet.

[0210] 6) Core Formula Group (COM, 12 months): Feed the animal an AIN-93G diet supplemented with 900 mg / kg, 30 mg / kg, and 5.6 mg / kg of CaHMB, CBP, and HA respectively; (composition)

[0211] 7) Core formula + CaHMB group (COM + HMB, 12 months): In addition to AIN-93G feed, 2400mg / kg, 30mg / kg and 5.6mg / kg of feed CaHMB, CBP and HA were added respectively;

[0212] 8) Core Formula + CBP Group (COM + CBP, 12 months old): On the basis of feeding AIN-93G feed, 900mg / kg, 150mg / kg and 5.6mg / kg of feed CaHMB, CBP and HA were added respectively;

[0213] 9) Core Formula + HA Group (COM+HA, 12 months): On the basis of feeding AIN-93G feed, add 900mg / kg, 30mg / kg and 40mg / kg of feed respectively to CaHMB, CBP and HA;

[0214] 10) Whey protein + core formula group (W-COM, 12 months): On the basis of feeding AIN-93G feed, replace 10% of the casein by weight with whey protein, and add the same amount of CaHMB, CBP and HA as the core product formula group.

[0215] Among them, groups 3-10 were the intervention group, groups 1-2 were the control group, groups 3-5 were the monomer group, and groups 7-9 were the COM+ monomer group.

[0216] Mice were weighed weekly using an electronic balance, measuring their body weight (g) and feed intake (g). After a 6-month intervention, mice were fasted for 10-12 hours, and blood was collected from their eyeballs under anesthesia. They were then euthanized by cervical dislocation. Blood samples were left at room temperature for at least 30 minutes before being processed at 1000 mL / min. The serum supernatant was obtained by centrifugation at 4°C for 15 min and aliquoted and stored at -80°C for subsequent analysis. The femur, tibia, knee joint, lower limb muscles (including quadriceps, tibialis anterior, and gastrocnemius), and vital organs (liver, kidney, and heart) of mice were isolated. Some bones and quadriceps muscles were fixed with 4% (w / v) paraformaldehyde, and the remaining samples were stored at -80°C for later use. The animal experimental protocol involved in this study was approved by the Laboratory Animal Ethics Committee of the School of Public Health, Sun Yat-sen University (No. 2024-066) before commencement.

[0217] The manufacturer of colostrum basic protein is Seperex Nutritionals Ltd.; the supplier is Shanghai Prochin International Trading Co., Ltd.

[0218] Example 1: Basic Situation

[0219] Basic information on the OLD, YOU group, and each intervention group (including survival status, body weight, feed intake, and feed efficiency) is as follows: Figure 2 As shown. Because the experiment used aged mice, spontaneous disease-related deaths occurred. Therefore, after 6 months of intervention, one mouse died in the OLD, HA, COM+HMB, COM+CBP, and COM+HA groups, two mice died in the COM group, and no mice died in the other groups (e.g., ...). Figure 2 .C).

[0220] During the feeding period, the weight of all groups showed a gradual upward trend, with the YOU group showing the largest increase (e.g., ...). Figure 2 A). For example Figure 2 As shown in (B), at baseline (0 months), the body weight of the YOU group was significantly lower than that of the animals in each of the older groups. P < 0.05), this weight difference still existed at 3 months of intervention, but after the intervention, there was no significant difference in weight between the YOU group and the other older groups; while at baseline, at 3 months of intervention, and after the intervention, there were no significant differences in weight between the OLD group and the other intervention groups. During the feeding period, there were no significant differences in average weekly feed intake between the OLD group and other groups (e.g., Figure 2 .D). The overall feed efficiency of the YOU group was significantly higher than that of the OLD group ( P < 0.05), and there was no statistically significant difference in total feed efficiency between the OLD group and other groups (e.g., Figure 2 .E).

[0221] Example 2: Effects of different interventions on muscle mass and function in aged mice

[0222] 2.1 Effects of different interventions on muscle mass in aged mice

[0223] Firstly, regarding overall muscle mass, the results are as follows: Figure 3 As shown. After 6 months of intervention, DXA analysis showed that the lean body percentage of mice in the OLD group was significantly reduced ( P < 0.05 indicates muscle loss during aging. The COM group maintained the same level of overall muscle mass as the high-dose monomeric HMB group and was significantly better than the high-dose monomeric CBP group. P < 0.05), the W-COM group showed a trend of better performance than the COM group, but the difference was not statistically significant. There were no significant differences between the three COM+ monomer groups and the COM group (e.g., Figure 3 A).

[0224] Secondly, through HE staining of the quadriceps femoris muscle (e.g. Figure 3 In the OLD group, the quadriceps muscle fibers were observed to be significantly atrophied and thinner, while the muscle fibers in the HMB, COM, COM+HMB, and W-COM groups were relatively fuller and thicker. Further quantitative assessment of muscle fiber CSA revealed a significant decrease in CSA in the OLD group mice. P < 0.05%, further demonstrating the muscle loss during aging. The COM group achieved the same effect on maintaining muscle fibers as the high-dose monomeric HMB group, and was significantly superior to the high-dose monomeric CBP group. P <0.05, the W-COM group showed a trend of better maintenance effect than the COM group, but the difference was not statistically significant. Meanwhile, the COM+monomer group was significantly better than its corresponding monomer group ( P < 0.05), there were no significant differences between the three COM+ monomer groups and the COM group (e.g., Figure 3 .B).

[0225] 2.2 Effects of different interventions on muscle function in aged mice

[0226] First, the maximum pulling force of the mice's limbs was measured using a dynamometer. The results are as follows: Figure 4 As shown in the figure, during the 6-month intervention period, the pulling force of aged mice showed a gradual decreasing trend, indicating that muscle function also weakens during aging. Among them, the OLD group showed the most significant decreasing trend, while the YOU group showed an overall gradual increasing trend (as shown in the figure). Figure 4 A). After the intervention, the pull strength in the OLD group was significantly reduced ( ). P < 0.05), the pull of the COM group was significantly higher than that of the high-dose monomeric HMB and monomeric CBP groups ( P< 0.05), the tensile strength of the W-COM group was trending higher than that of the COM group, but the difference was not statistically significant. Meanwhile, the tensile strength of the COM+HMB group was significantly higher than that of the HMB group, and there were no significant differences between the three COM+monomer groups and the COM group (e.g., ...). Figure 4 .B).

[0227] Secondly, after the intervention, the treadmill test was used to assess the mice's motor ability, which can indirectly reflect muscle function. It was observed that the distance and time of movement in the OLD group mice were significantly reduced. P < 0.05), while the movement distance and time of mice in the COM group were significantly higher than those in the high-dose monomeric HMB and CBP groups ( P < 0.05), the W-COM group showed higher movement distance and time trends than the COM group, but the difference was not statistically significant. Meanwhile, the movement distance and time of the COM+ monosodium group were significantly higher than their corresponding monosodium groups ( P < 0.05), there were no significant differences between the three COM+ monomer groups and the COM group (e.g., Figure 4 (C, D).

[0228] In summary, the COM group showed significantly better muscle mass maintenance than the high-dose monomeric CBP group, and trended higher than the high-dose monomeric HMB group, but the difference was not statistically significant. Its effect on promoting muscle function was significantly better than both high-dose monomeric groups. The W-COM group showed better results than the COM group in both maintaining muscle mass and promoting muscle function. Meanwhile, the COM+HMB, COM+CBP, and COM+HA groups showed significantly better beneficial effects on muscle health than their corresponding monomeric groups, but no significant improvement over the COM group was observed. In other words, the core formulation of this invention (CaHMB+CBP+HA) is significantly more effective in maintaining muscle mass than the group that only added a high dose of CBP.

[0229] Example 3: Effects of different interventions on bone mass, bone morphology and bone metabolism in aged mice

[0230] The effects of different interventions on bone mass and bone morphology in aged mice are as follows: Figure 5 and 6 As shown. After 6 months of intervention, whole-body BMD was measured by DXA, and it was observed that the whole-body BMD of mice in the OLD group was significantly reduced ( P < 0.05 indicates bone loss during aging. The COM group maintained systemic BMD at a level comparable to the high-dose monomeric CBP group, while there was no significant difference in effect between the W-COM group and the COM group. Furthermore, the COM + monomeric group was significantly more effective than its corresponding monomeric group, and the COM + CBP group was significantly more effective than the COM group. P < 0.05) (e.g. Figure 5 A).

[0231] Morphological characteristics of the femur and tibia in mice were assessed using micro-CT scanning and HE staining. For the femur (FE), the OLD group mice showed significantly decreased BV / TV%, trabecular bone number, trabecular bone thickness, trabecular bone connectivity, and femoral BMD, while trabecular bone separation was significantly increased. P < 0.05 indicates that aging is accompanied not only by bone loss but also by changes in bone microstructure, resulting in thinning and weakening of the bone. The COM group showed an improvement in these indicators comparable to the high-dose monomeric CBP group ( P < 0.05), the W-COM group showed a better trend in improving the above indicators than the COM group, but the difference was not statistically significant. The COM+CBP group was significantly better than the COM and CBP groups (except for trabecular bone thickness and femoral BMD). P < 0.05) (e.g. Figure 5 BG). Observation of 3D sections of the distal femur obtained from Micro-CT and HE staining shows (e.g.) Figure 5 In the H and I groups, the cortical bone of the distal femur in the OLD group was relatively thin, and the trabeculae in the cancellous bone were sparse, scattered, fewer in number, and thinner; while in the CBP, COM, COM+CBP and W-COM groups, the cortical bone was relatively thick, and the trabeculae in the cancellous bone were more dense, more numerous, and thicker.

[0232] For the tibia (TI), the BV / TV%, trabecular bone number, trabecular bone thickness, trabecular bone connectivity, and tibial BMD were significantly decreased in the OLD group mice, while trabecular bone separation was significantly increased. P < 0.05%, the COM group achieved the same improvement in the above indicators as the high-dose monomeric CBP group (except for trabecular bone thickness and trabecular bone connectivity). The W-COM group was significantly better than the COM group in improving BV / TV%, trabecular bone thickness, and trabecular bone connectivity. Meanwhile, the COM+CBP group showed a better effect in improving trabecular bone connectivity than the COM group. P < 0.05) (e.g. Figure 6 ,AF). Observation of 3D sections of the proximal tibia obtained from Micro-CT and HE staining shows (e.g. Figure 6 In the G and H groups, the cortical bone of the proximal tibia in the OLD group was relatively thin, and the trabeculae in the cancellous bone were sparse, separated, fewer in number, and thinner; while in the CBP, COM, COM+CBP and W-COM groups, the cortical bone was relatively thick, and the trabeculae in the cancellous bone were more dense, more numerous, and thicker.

[0233] To further elucidate the dynamic regulatory effect of the composition of this invention on bone metabolism, the inventors detected bone metabolism markers in mouse serum. The results of the effects of different interventions on bone metabolism in aged mice are as follows: Figure 7As shown. After 6 months of intervention, serum bone metabolism markers were first measured. It was observed that the OLD group showed elevated levels of type I collagen C-terminal cross-linked telopeptide (CTX-I), a marker of bone resorption, and decreased levels of bone-specific alkaline phosphatase (BALP), a marker of bone formation. P < 0.05), the COM group could inhibit the changes in these two indicators, and the effect was comparable to that of the high-dose monomeric CBP group and significantly better than that of the HMB group ( P < 0.05, the W-COM group was generally better than the COM group, but there was no statistically significant difference. Meanwhile, for CTX-I levels, the COM+HMB group was significantly better than both the COM and HMB groups; for BALP levels, the COM+CBP group was significantly better than the CBP group. P < 0.05) (e.g. Figure 7 (A and B). This indicates that the present invention can effectively inhibit excessive bone resorption, slow down bone destruction, and simultaneously activate osteoblasts to promote the formation of new bone.

[0234] Next, TRAP staining was performed on the distal femur and proximal tibia of the mouse (e.g., Figure 7 The activity of osteoclasts was assessed using G and H, reflecting the level of bone resorption. It was observed that, in both the femur and tibia, the number of osteoclasts on the trabecular surface was significantly increased in the OLD group, while the number of osteoclasts in the CBP, COM, COM+CBP, and W-COM groups was relatively lower. Further quantitative analysis clarified that, firstly, in the femur, the number and area of ​​osteoclasts per unit trabecular surface area were significantly increased in the OLD group (G and H), reflecting the level of bone resorption. P <0.05%, the COM group can reduce the number of osteoclasts, and the effect is comparable to that of the high-dose CBP group and significantly better than that of the HMB group. P <0.05), the W-COM group showed a trend of better reduction in osteoclast number and area than the COM group, but this was not statistically significant. Meanwhile, the COM+HMB group was significantly better than the HMB group in reducing osteoclast number (…). P < 0.05), but there was no significant difference compared to the COM group (e.g., Figure 7 (C and D). Secondly, in the tibia, the number and area of ​​osteoclasts per unit trabecular surface area were significantly increased in the OLD group ( P (< 0.05), the COM group can reduce the number and area of ​​osteoclasts, with the effect reaching that of the high-dose monomeric CBP group. The W-COM group showed a trend of better effect in reducing the number and area of ​​osteoclasts than the COM group, but the difference was not statistically significant.

[0235] Synergistic changes in bone metabolism indicators suggest that this composition can bidirectionally regulate bone metabolism, redirecting the imbalanced state of bone metabolism (high absorption, low formation) to a healthy balance, namely "low absorption, high formation." This explains, from a biochemical and kinetic perspective, why this invention can effectively reduce bone loss and improve bone microstructure.

[0236] In summary, the composition can maintain bone mass and bone microstructure and improve bone metabolism in aged mice, and the effect is significantly better than that of the high-dose monomeric HMB group; the W-COM group is significantly better than the COM group in maintaining tibial bone mass, and also shows a better trend in improving femoral bone mass and bone metabolism; at the same time, the beneficial effects of COM+HMB, COM+CBP and COM+HA groups on bone health are significantly better than their corresponding monomeric groups.

[0237] Example 4: The product formulation may exert its promoting effect by regulating molecules related to the "muscle-bone axis".

[0238] To further elucidate the possible molecular mechanisms underlying the aforementioned promoting effects, this embodiment examined the levels of "muscle-bone axis"-related molecules in mice. The effects of different interventions on the regulation of "muscle-bone axis"-related molecules in aged mice were as follows: Figure 8 As shown. First, regarding the positive regulators of the "muscle-bone axis," the levels of Irisin and IGF-1 in the quadriceps femoris tissue of the OLD group were found to be significantly decreased ( P < 0.05), except for the CBP and HA groups, all other interventions increased Irisin and IGF-1 levels. The COM group achieved the same effect as the high-dose monomeric HMB group and was significantly better than the CBP group ( P < 0.05), there was no significant difference in efficacy between the W-COM group and the COM group. Meanwhile, the COM+CBP and COM+HA groups were significantly better than their respective monomeric groups ( P < 0.05), but there was no significant difference compared to the COM group ( Figure 8 (A, B). Furthermore, a significant decrease in serum OCN levels was found in the OLD group ( P < 0.05), except for the HMB and HA groups, all other interventions increased OCN levels. The COM group achieved the same effect as the high-dose monomeric CBP group and was significantly better than the HMB group ( P < 0.05), there was no significant difference in efficacy between the W-COM group and the COM group. Meanwhile, the COM+HMB and COM+HA groups were significantly better than their respective monomeric groups ( P < 0.05), but there was no significant difference compared to the COM group ( Figure 8 C).

[0239] Secondly, regarding the negative regulators in the "muscle-bone axis," the levels of MSTN in the quadriceps femoris tissue and SOST in the femur tissue were found to be significantly increased in the OLD group. P < 0.05), all interventions reduced MSTN and SOST levels. For MSTN, the COM group achieved the same effect as the high-dose monomeric HMB group and was significantly superior to the CBP group ( P < 0.05), there was no significant difference in efficacy between the W-COM group and the COM group. Meanwhile, the COM+CBP and COM+HA groups were significantly better than their respective monomeric groups ( P < 0.05), but there was no significant difference compared to the COM group ( Figure 8 For SOST, the COM group achieved the same effect as the high-dose monomeric CBP group and was significantly superior to the HMB group (D); P < 0.05), there was no significant difference in efficacy between the W-COM group and the COM group. Meanwhile, the COM+HMB and COM+HA groups were significantly better than their respective monomeric groups ( P < 0.05), but there was no significant difference compared to the COM group ( Figure 8 E).

[0240] In summary, this composition establishes a virtuous cycle of muscle-bone interaction by regulating key factors of the "muscle-bone axis," specifically manifested as follows:

[0241] (1) On the muscle side: significantly increased the level of beneficial myotrophic factor Irisin and decreased the level of inhibitory myotrophic factor MSTN, which not only directly promoted the increase of muscle mass, but also enabled Irisin to act as a messenger to transmit positive mechanical and metabolic signals to the skeleton.

[0242] (2) On the skeletal side: it significantly increased the level of the bone formation marker OCN and decreased the level of the bone formation inhibitor SOST, which created an excellent microenvironment for bone formation.

[0243] (3) At the core hub: IGF-1 levels were increased simultaneously, providing a common driving force for the anabolism of the two major tissues, musculoskeletal tissues.

[0244] Furthermore, elevated levels of bone-derived OCN may feed back to muscles, improving their function and thus completing a full communication loop from 'muscle→bone' to 'bone→muscle'.

[0245] In summary, compared with the elderly control group, the high-dose HMB monomer group showed significant improvements in muscle mass and muscle function (p<0.05), but no significant improvements in bone microstructure (especially the tibia) and bone metabolism parameters; the high-dose CBP monomer group showed significant improvements in bone mass, bone microstructure, and bone metabolism (p<0.05), but limited improvements in overall lean body mass. The composition of this application showed significant improvements in all observed muscle and bone parameters, and was superior to any single high-dose component group in terms of muscle mass and muscle function (p<0.05).

[0246] The inventors further investigated the effect of calcium salt particle size on the suspension obtained by mixing HMB and calcium citrate. The steps for preparing the suspension included: weighing 10 moles of CaHMB, adding 11 kg of deionized water to prepare a 20% CaHMB aqueous solution, adding 6.7 moles of citric acid to the CaHMB aqueous solution, reacting for 15 min, then homogenizing by shearing at 3000 r / min for 8 min, adding 0.8 kg of potassium hydroxide to adjust the pH to 6.6-7.2, controlling the temperature at 45-55℃, and continuing shearing (for 1 h, 2 h, 3 h, 4 h, and 5 h respectively). The calcium salt particle size and suspension condition were observed, and the calcium salt particle size was measured using a Bettersize 2600E laser particle size analyzer. The results are shown below. Figure 9-10 As shown, the particle size of insoluble calcium gradually decreased with the optimization of the formulation reaction ratio and the increase of online cyclic shear reaction time. The particle size of the suspension with a shear time of more than 4 hours was all <60μm.

[0247] Example 1 of preparing modified milk powder:

[0248] The preparation method of the whey protein-rich modified milk powder containing calcium β-hydroxy-β-methylbutyrate (CaHMB), colostrum basic protein (CBP), and sodium hyaluronate (HA) includes the following steps:

[0249] 1) Preparation of milk: Raw milk is preheated to 45°C and then put into a sterilization separator for cleaning and sterilization. It is then pasteurized and cooled to 4°C for storage to obtain milk A1.

[0250] 2) Preparation of oil solution: Weigh medium- and long-chain fatty acid edible oil and soybean phospholipids (phospholipids account for 2wt% of the oil mass) according to the formula, mix them and heat to 65℃ to melt the oil, stir at 500r / min for 10min until the phospholipids are completely dissolved, filter through a 200-mesh filter, transfer to a constant temperature temporary storage tank, keep warm at 40℃ for later use, and obtain oil solution A2.

[0251] 3) Preparation of HMB and calcium citrate suspension: Weigh 10 moles of CaHMB and add 11 kg of deionized water to prepare a CaHMB aqueous solution with a mass concentration of 20%. Add about 6.7 moles of citric acid to the CaHMB aqueous solution and react for 15 min. After homogenization by shearing at 3000 r / min for 8 min, add 0.8 kg of potassium hydroxide to adjust the pH to 6.8 and continue shearing. Observe the calcium salt particle size and the suspension of the solution. Use a laser particle size analyzer (Bettersize2600E) to detect the calcium salt particle size. When the calcium salt particle size is less than or equal to 60 μm, stop shearing. Add 0.2 kg of vitamins and 2 kg of minerals and mix evenly to obtain suspension B1 containing HMB and calcium citrate.

[0252] 4) Preparation of wet mixture for wall material: 8 kg of desalted whey powder, 3 kg of concentrated whey protein powder, 4 kg of resistant dextrin, 0.02 kg of sodium hyaluronate, etc. are put into the powder hopper and mixed in the powder hopper. The mixed powder is added to suspension B1 to obtain suspension B2. Stir at 1000 rpm for 15 min until HMB and vitamin minerals are evenly dispersed in the wall material. Then, it is temporarily stored in a vacuum mixing tank through a vacuum conveying system.

[0253] 5) Preparation of total mixture: Mix milk A1, oil solution A2 and suspension B2. After all the mixing is completed, the total mixture C is obtained. Based on the total weight of the total mixture C, the dry matter content of milk A1 is 75.7 wt%, the content of oil solution A2 is 0.2 wt%, and the dry matter content of suspension B2 is 24.1 wt%.

[0254] 6) The total mixture C is subjected to the following steps: homogenization at a pressure of 140-160 bar for 3 hours, with a homogenization flow rate of 30-40 m³ / h. 3 / h; Sterilize in DSI sterilizer at 85-95℃ for 30-60s; Flash evaporation in flash evaporator, followed by filtration through dual filter and into falling film evaporator; After filtration, spray-dry in drying tower by high pressure pump at 160℃ and 80℃ to obtain material D1;

[0255] 7) Mixing heat-sensitive materials and packaging: Mix approximately 92 kg of material D1 with 0.092 kg of colostrum basic protein evenly, pass metallization and other tests, and then package and seal it in a suitable container to obtain the modified milk powder described in this invention.

[0256] Example 2 of preparing modified milk powder:

[0257] The preparation method of the whey protein-rich modified milk powder containing calcium β-hydroxy-β-methylbutyrate (CaHMB), colostrum basic protein (CBP), and sodium hyaluronate (HA) includes the following steps:

[0258] 1) Preparation of milk: Raw milk is preheated to 45°C and then put into a sterilization separator for cleaning and sterilization. It is then pasteurized and cooled to 4°C for storage to obtain milk A1.

[0259] 2) Preparation of oil solution: Weigh medium- and long-chain fatty acid edible oil and soybean phospholipids (phospholipids account for 50 wt% of the oil mass) according to the formula, mix them and heat to 65℃ to melt the oil, stir at 500 r / min for 10 min until the phospholipids are completely dissolved, filter through a 200 mesh filter, transfer to a constant temperature temporary storage tank, keep warm at 40℃ for later use, and obtain oil solution A2.

[0260] 3) Preparation of HMB and calcium chloride suspension: Weigh 10 moles of CaHMB and add 11 kg of deionized water to prepare a CaHMB aqueous solution with a mass concentration of 20%. Add 20 moles of hydrochloric acid to the CaHMB aqueous solution and react for 15 min. Then, shear homogenize at 3000 r / min for 8 min. Add 0.8 potassium hydroxide to adjust the pH to 6.6-7.2. Control the temperature at 45℃-55℃ and continue shearing for 1 h. Observe the calcium salt particle size and the suspension of the solution. Use a Bettersize2600E laser particle size analyzer to detect the calcium salt particle size. When the calcium salt particle size is less than or equal to 60 μm, stop shearing. Add 0.2 kg of vitamins and 2 kg of minerals and mix evenly to obtain suspension B1 containing HMB and calcium chloride.

[0261] 4) Preparation of wet mixture for wall material: 8 kg of desalted whey powder, 3 kg of concentrated whey protein powder, 4 kg of resistant dextrin and 0.02 kg of sodium hyaluronate are put into the powder hopper and mixed in the powder hopper. The mixed powder is added to suspension B1 to obtain suspension B2. Stir at 1000 rpm for 15 min until HMB and vitamin minerals are evenly dispersed in the wall material. Then, it is temporarily stored in a vacuum mixing tank through a vacuum conveying system.

[0262] 5) Preparation of total mixture: Mix milk A1, oil solution A2 and suspension B2. After all the mixtures are mixed, the total mixture C is obtained. Based on the total weight of the total mixture C, the dry matter content of milk A1 is 76.6 wt%, the content of oil solution A2 is 0.2 wt%, and the dry matter content of suspension B2 is 23.2 wt%.

[0263] 6) The total mixed liquor C is subjected to the following steps: homogenization at a pressure of 140-160 bar for 3 hours, with a homogenization flow rate of 30-40 m3 / h; sterilization in a DSI sterilizer at a temperature of 85-95℃ for 30-60 seconds; flash evaporation in a flash evaporator, followed by filtration through a dual filter and entry into a falling film evaporator; filtration through a filter and spray drying in a drying tower by a high-pressure pump at an inlet air temperature of 160℃ and an outlet air temperature of 80℃ to obtain material D1;

[0264] 7) Mixing heat-sensitive materials and packaging: Mix approximately 92 kg of material D1 with 0.092 kg of colostrum basic protein evenly, pass metallization and other tests, and then package and seal it in a suitable container to obtain the modified milk powder described in this invention.

[0265] Example 3 of modified milk powder preparation:

[0266] The preparation method of the whey protein-rich modified milk powder containing calcium β-hydroxy-β-methylbutyrate (CaHMB), colostrum basic protein (CBP), and sodium hyaluronate (HA) includes the following steps:

[0267] 1) Preparation of milk: Raw milk is preheated to 45°C and then put into a sterilization separator for cleaning and sterilization. It is then pasteurized and cooled to 4°C for storage to obtain milk A1.

[0268] 2) Preparation of oil solution: Weigh medium- and long-chain fatty acid edible oil and soybean lecithin according to the formula (oil mass ratio 0-10wt%), mix and heat to 65℃ to melt the oil, stir at 500r / min for 10min until the lecithin is completely dissolved, filter through a 200-mesh filter, transfer to a constant temperature temporary storage tank, keep warm at 40℃ for later use, and obtain oil solution A2.

[0269] 3) Preparation of HMB and calcium lactate suspension: Weigh 10 moles of CaHMB and add 11 kg of deionized water to prepare a CaHMB aqueous solution with a mass concentration of 20%. Add 20 moles of lactic acid to the CaHMB aqueous solution and react for 15 min. Then, shear homogenize at 3000 r / min for 8 min. Add 0.8 potassium hydroxide to adjust the pH to 6.6-7.2. Control the temperature at 45℃-55℃ and continue shearing. Use a Bettersize2600E laser particle size analyzer to detect the calcium salt particle size. Stop shearing when the calcium salt particle size is less than or equal to 60 μm. Add 0.2 kg of vitamins and 2 kg of minerals and mix evenly to obtain suspension B1 containing HMB and calcium lactate.

[0270] 4) Preparation of wet mixture for wall material: 8 kg of desalted whey powder, 3 kg of concentrated whey protein powder, 4 kg of resistant dextrin and 0.02 kg of sodium hyaluronate are put into the powder hopper and mixed in the powder hopper. The mixed powder is added to suspension B1 to obtain suspension B2. Stir at 1000 rpm for 15 min until HMB and vitamin minerals are evenly dispersed in the wall material. Then, it is temporarily stored in a vacuum mixing tank through a vacuum conveying system.

[0271] 5) Preparation of total mixture: Mix milk A1, oil solution A2 and suspension B2. After all the mixing is completed, the total mixture C is obtained. Based on the total weight of the total mixture C, the dry matter content of milk A1 is 75 wt%, the content of oil solution A2 is 0.2 wt%, and the dry matter content of suspension B2 is 24.8 wt%.

[0272] 6) The total mixture C is subjected to the following steps: homogenization at a pressure of 140-160 bar for 3 hours, with a homogenization flow rate of 30-40 m³ / h. 3 / h; Sterilize in DSI sterilizer at 85-95℃ for 30-60s; Flash evaporation in flash evaporator, followed by filtration through dual filter and into falling film evaporator; After filtration, spray-dry in drying tower by high pressure pump at 160℃ and 80℃ to obtain material D1;

[0273] 7) Mixing heat-sensitive materials and packaging: Mix approximately 92 kg of material D1 with 0.092 kg of colostrum basic protein evenly, pass metallization and other tests, and then package and seal it in a suitable container to obtain the modified milk powder described in this invention.

[0274] Comparative Example 1:

[0275] Commercially available whole milk powder (composed of raw cow's milk) was dry-mixed with calcium β-hydroxy-β-methylbutyrate (CaHMB), colostrum basic protein (CBP), and sodium hyaluronate (HA) so that the mass ratio and content of CaHMB, CBP, and HA were the same as those in Example 1 of the preparation of modified milk powder.

[0276] The whey protein-rich modified milk powders containing calcium β-hydroxy-β-methylbutyrate (CaHMB), colostrum basic protein (CBP), and sodium hyaluronate (HA) prepared in Preparation Examples 1-3 were compared with Comparative Example 1 (dry mix of whole milk powder) for reconstitution and sensory evaluation tests. Twenty experienced evaluators were randomly selected to test the performance of the above modified milk powders.

[0277] Sensory evaluation was conducted according to Section 3.2 of GB19644 National Food Safety Standard for Milk Powder and Modified Milk Powder, including tests for color, taste, odor, and state. The sensory evaluation results are shown in Table 2 below, where "+" indicates that the sensory requirements are met, and "-" indicates that the sensory results are unsatisfactory.

[0278] Method for evaluating reconstitution property: Add 25 g of formulated milk powder to 180 mL of water, and stir with a magnetic stirrer at 900 rpm while adding. Stir for 5 min, control the powder adding time within 15 - 20 s. After adding the powder, continue to stir for 30 s. Let it stand at room temperature of 25 °C for 5 min, observe the milk liquid layering situation. Take 5 mL of milk liquid and slowly pour it onto the center of a black rubber plate, let the milk liquid spread naturally into a thin liquid film, and count the number of small white dots, white patches and lumps in the undissolved particles. The evaluation criteria are shown in Table 1 below. Among them, "small white dots" usually refer to protein denaturation points, with diameter R: R ≤ 0.5 mm; insoluble fine salt substances (observable under strong light) and air bubbles with particle size ≤ 0.5 mm during the operation are not included. "White patches" usually refer to undissolved powder or flocs of protein denaturation, with diameter R: 0.5 mm < R ≤ 3 mm. "Lumps" usually refer to aggregated blocks of undissolved powder or protein denaturation, with diameter R: R > 3 mm. The schematic diagram of the evaluation criteria is as Figure 11 shown.

[0279] Table 1:

[0280]

[0281] Table 2: Results of reconstitution property and sensory evaluation

[0282]

[0283] It can be seen that the preparation method provided by the present invention achieves the technical effects of improving the stability of milk powder, enhancing the taste and improving the reconstitution property without any food thickeners and edible flavors.

Claims

1. A nutritional composition, characterized in that, The nutritional composition includes calcium β-hydroxy-β-methylbutyrate, milk-derived basic protein, and sodium hyaluronate; wherein the milk-derived basic protein includes colostrum basic protein, the mass ratio of calcium β-hydroxy-β-methylbutyrate to colostrum basic protein is 80:1-5:1, and the mass ratio of calcium β-hydroxy-β-methylbutyrate to sodium hyaluronate is 430:1-22.5:

1.

2. The nutritional composition according to claim 1, characterized in that, The nutritional composition consists of calcium β-hydroxy-β-methylbutyrate, milk-derived basic protein, and sodium hyaluronate. The milk source includes sheep milk and / or cow milk.

3. A formulation comprising the nutritional composition as described in claim 1 or 2.

4. A product comprising the nutritional composition as described in claim 1 or 2, or the formulation as described in claim 3.

5. The product as described in claim 4, characterized in that, The product is food or a food additive.

6. The product as described in claim 4, characterized in that, The product in question is a health food product.

7. A pharmaceutical composition, characterized in that, The pharmaceutical composition comprises a nutritional composition comprising β-hydroxy-β-methylbutyric acid or a salt thereof, milk-derived basic protein, and sodium hyaluronate; wherein the milk-derived basic protein includes colostrum basic protein, the mass ratio of β-hydroxy-β-methylbutyric acid or a salt thereof to colostrum basic protein is 80:1-5:1, and the mass ratio of calcium β-hydroxy-β-methylbutyrate to sodium hyaluronate is 430:1-22.5:

1.

8. The pharmaceutical composition according to claim 7, characterized in that, The nutritional composition comprises β-hydroxy-β-methylbutyric acid or its salt, milk-derived basic protein, and sodium hyaluronate; and / or The milk source includes sheep milk and / or cow milk.

9. The pharmaceutical composition according to claim 7, characterized in that, β-hydroxy-β-methylbutyrate is a salt formed by β-hydroxy-β-methylbutyric acid and a metal cation, wherein the metal cation includes one or more of sodium ions, potassium ions, lithium ions, calcium ions, magnesium ions, and zinc ions.

10. The pharmaceutical composition according to claim 7, characterized in that, The pharmaceutical composition also contains pharmaceutically acceptable excipients.

11. Use of the nutritional composition as claimed in claim 1 or 2, or the formulation as claimed in claim 3: (1) Uses in food preparation (2) Use in the preparation of pharmaceutical compositions for the prevention and / or treatment of diseases, said diseases being senile degenerative sarcopenia or senile osteoporosis, and (3) Use in the preparation of formulations that alleviate muscle loss, muscle function decline, reduce bone loss and alter bone microstructure.

12. The use as described in claim 11, characterized in that, (1) Used in the preparation of health food products.

13. The use as described in claim 11, characterized in that, In (2), the use includes one or more of the following uses: (i) reducing the content of bone resorption markers, (ii) increasing the content of bone formation markers, (iii) regulating myo-myogenic factors and bone factors, including increasing Irisin content and / or decreasing MSTN content, (iv) regulating bone factors, increasing OCN content and / or decreasing SOST content, (v) activating IGF-1, and (vi) improving myo-myogenic axis interaction.

14. Use of the nutritional composition of claim 1 or 2, the formulation of claim 3, or the product of any one of claims 4-6 for non-therapeutic purposes, including one or more of the following uses: (1) alleviating muscle loss, alleviating muscle function decline, reducing bone loss and altering bone microstructure, and (2) regulating bone metabolism.

15. A method for preparing the product according to any one of claims 4-6, characterized in that, The method includes the steps of: mixing calcium β-hydroxy-β-methylbutyrate and organic or inorganic acid to obtain a suspension and an aqueous phase; wherein the particle size of the calcium salt in the suspension is less than or equal to 60 μm.

16. The method as described in claim 15, characterized in that, The method satisfies one or more of the following characteristics: Adding wall material to the suspension yields an aqueous phase; The calcium salt is an organic acid calcium salt or an inorganic acid calcium salt; The suspension is sheared and homogenized at a shear rate of 2000-5000 r / min and / or for a shearing time of 1-5 hours. The wall material includes starch products and / or dietary fiber; and The product in question is a food product.

17. The method as described in claim 15, characterized in that, The method satisfies one or more of the following characteristics: Adding wall material to the suspension yields an aqueous phase; The calcium salt is an organic acid calcium salt or an inorganic acid calcium salt; The suspension is subjected to shear homogenization at a shear rate of 2000-5000 r / min and / or a shearing time of 4-6 hours; The wall material includes starch products and / or dietary fiber; and The product in question is a food product.

18. The method as described in claim 15 or 16, characterized in that, The preparation method of the product includes the following steps: (1) Preparation of milk liquid: Sterilize and cool raw cow's milk or raw sheep's milk to obtain milk liquid; (2) Preparation of suspension: β-hydroxy-β-methylbutyrate calcium and organic acid or inorganic acid are mixed, sheared and homogenized to obtain a suspension, wherein the particle size of the organic acid calcium or inorganic acid calcium in the suspension is less than or equal to 60 μm; (3) Preparation of aqueous phase: Mix the raw material protein and wall material to obtain a mixture, and add the mixture to the suspension in step (2) to obtain an aqueous phase; (4) Mix the milk and aqueous phase, homogenize, sterilize, concentrate and dry to obtain powder; and (5) The powder is mixed with milk-derived basic protein to obtain the product.

19. The method as described in claim 18, characterized in that, The method satisfies one or more of the following characteristics: In step (1), the sterilization is pasteurization; The method further includes step (1.5) between steps (1) and (2), which includes: mixing phospholipids and oils to obtain an oil phase; and step (4) is: mixing the milk, aqueous phase and oil phase, homogenizing, sterilizing, concentrating and drying to obtain powder. In step (2), the acidity of the organic or inorganic acid is greater than that of β-hydroxy-β-methylbutyric acid; In step (2), the molar ratio of calcium β-hydroxy-β-methylbutyrate to organic or inorganic acid depends on the number of basic acids the organic or inorganic acid is. If the organic or inorganic acid is a monobasic acid, calcium β-hydroxy-β-methylbutyrate and organic or inorganic acid are added in a molar ratio of <1:

2. If the organic or inorganic acid is a tribasic acid, calcium β-hydroxy-β-methylbutyrate and organic or inorganic acid are added in a molar ratio of <3:

2. In step (2), the rate of shearing homogenization is 2000-5000 r / min, and / or the time of shearing homogenization is 1-5 hours; In step (3), stirring is used to assist mixing. The stirring time is 10-15 min, and / or the stirring speed is 800-1000 rpm. In step (3), the raw material protein includes milk protein or immunoglobulin; In step (4), the dry matter content in the aqueous phase is 5-30 wt% based on the total weight of the mixture. In step (4), the homogenization pressure is 140-160 bar, and / or the homogenization time is 2-3 hours, and / or the homogenization flow rate is 30-40 m³ / h. 3 / h; In step (4), the sterilization temperature is 85-95℃, and / or the sterilization time is 30-60s; In step (4), the concentration is achieved by flash evaporation; and In step (4), the drying includes spray drying.

20. The method as described in claim 18, characterized in that, The method satisfies one or more of the following characteristics: In step (1), the sterilization is pasteurization; The method further includes step (1.5) between steps (1) and (2), which includes: mixing phospholipids and oils to obtain an oil phase; and step (4) is: mixing the milk, aqueous phase and oil phase, homogenizing, sterilizing, concentrating and drying to obtain powder. In step (2), the acidity of the organic or inorganic acid is greater than that of β-hydroxy-β-methylbutyric acid; In step (2), the molar ratio of calcium β-hydroxy-β-methylbutyrate to organic or inorganic acid depends on the number of basic acids the organic or inorganic acid is. If the organic or inorganic acid is a monobasic acid, calcium β-hydroxy-β-methylbutyrate and organic or inorganic acid are added in a molar ratio of <1:

2. If the organic or inorganic acid is a tribasic acid, calcium β-hydroxy-β-methylbutyrate and organic or inorganic acid are added in a molar ratio of <3:

2. In step (2), the rate of shearing homogenization is 2000-5000 r / min, and / or the time of shearing homogenization is 4-6 hours; In step (3), stirring is used to assist mixing. The stirring time is 10-15 min, and / or the stirring speed is 800-1000 rpm. In step (3), the raw material protein includes milk protein or immunoglobulin; In step (4), the dry matter content in the aqueous phase is 5-30 wt% based on the total weight of the mixture. In step (4), the homogenization pressure is 140-160 bar, and / or the homogenization time is 2-3 hours, and / or the homogenization flow rate is 30-40 m³ / h. 3 / h; In step (4), the sterilization temperature is 85-95℃, and / or the sterilization time is 30-60s; In step (4), the concentration is achieved by flash evaporation; and In step (4), the drying includes spray drying.

21. The method as described in claim 19, characterized in that, The raw material proteins include plant proteins.

22. The method as described in claim 19, characterized in that, The raw material protein includes lactoferrin.

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