Composition for promoting secretion of growth factors and proliferation of bone cells and preparation method thereof

Abstract

The invention discloses a composition for promoting secretion of growth factors and proliferation of bone cells and a preparation method of the composition, and belongs to the technical field of biological medicines and functional foods. Each unit dose of the combination group comprises 300 to 500 mg of calcium salt, 30 to 50 mg of magnesium salt, 2 to 5 mg of zinc salt, 250 to 350 mg of bovine bone protein peptide, 40 to 60 mg of hydrolyzed yolk powder, 8 to 15 [mu] g of vitamin D3, 5 to 20 [mu] g of vitamin K2 and 200 to 300 mg of an amino acid compound. Through cooperation of calcium, magnesium and zinc with bone-derived peptide and vitamin D / K2, calcium-sensitive receptor (CaSR) and BMP / Smad signaling pathways are activated, osteoblast proliferation rate and endogenous growth factor (BMP-2 and IGF-1) secretion amount are remarkably increased, and the bone-derived peptide composition is used for promoting height increase of children or teenagers, preventing or treating osteoporosis of various people, assisting fracture repair and bone regeneration, and has a good application prospect. The method has the advantages of high-efficiency collaboration, safety and controllability.

Description

TECHNICAL FIELD

[0001] The present application relates to the technical field of biological medicine and functional food, and particularly relates to a composition for promoting growth factor secretion and bone cell proliferation and a preparation method thereof. BACKGROUND

[0002] The height growth of children and adolescents depends on the proliferation and differentiation of growth plate chondrocytes, longitudinal bone growth and efficient regulation of the growth hormone (GH)-insulin-like growth factor 1 (IGF-1) axis. The existing intervention means for height promotion mainly include hormone therapy, nutritional supplementation and physical intervention, but there are still significant limitations: 1. Limitations and risks of growth hormone therapy Although exogenous recombinant human growth hormone (rhGH) can directly stimulate bone growth, its application faces the following problems: high cost, annual treatment cost of 100-300 thousand yuan, and long-term injection, heavy economic burden; side effect risk, which may cause joint pain, insulin resistance and scoliosis; applicability limitation, only approved for use in growth hormone deficiency (GHD) patients, and the effect on idiopathic short stature (ISS) is uncertain.

[0003] 2. Single targeting defect of traditional nutritional supplements Most of the commercially available "height increasing" products focus on a single mineral or vitamin (such as calcium, vitamin D), which can improve bone density, but cannot activate growth plate chondrocyte proliferation or regulate GH secretion: lack of multi-pathway synergy: the key window period before the closure of the growth plate requires simultaneous regulation of chondrocyte differentiation (such as through the BMP signaling pathway) and GH-IGF-1 axis activity, while traditional supplements only target mineralization; ignoring the synergistic effect of amino acids: specific ratios of amino acids such as lysine and arginine can stimulate pituitary GH secretion, but existing formulations have not optimized the ratio (such as the imbalance between gamma-aminobutyric acid (GABA) and arginine).

[0004] 3. Low efficiency and potential risks of plant extracts Some products add phytoestrogens (such as soy isoflavones) to mimic the effects of GH. Low bioavailability: after oral administration, the active ingredients are metabolized in the intestinal tract with an absorption rate of less than 5%. Risk of bone age acceleration: may induce premature closure of the growth plate, which in turn reduces adult height.

[0005] 4. Lack of synergy mechanism in existing technologies Longitudinal bone growth requires multi-component synergy: mineral-vitamin synergy deficiency: calcium, magnesium and zinc may be deposited in soft tissues rather than bone matrix if they lack the guidance of vitamin K2; lack of growth factor precursors: exogenous BMP-2 is costly and requires injection, and endogenous secretion regulation is not covered by existing nutritional programs.

[0006] In view of the above defects, it is urgent to develop a safe, economical and effective bone growth and repair technical solution. SUMMARY

[0007] The present application provides a composition for promoting growth factor secretion and bone cell proliferation and a preparation method thereof, to solve the defects of high cost, large side effects and insufficient bone growth effect in the prior art. Through the synergistic system of minerals, vitamins, peptides, amino acids and plant polyphenols, endogenous growth factor secretion and bone metabolism regulation pathways are activated, and the composition is suitable for height promotion of children and adolescents, prevention and treatment of osteoporosis, bone fracture repair and bone tissue engineering.

[0008] In a first aspect, the present application provides a composition, which includes, per unit dose: 300-500 mg of calcium salt, 30-50 mg of magnesium salt, 2-5 mg of zinc salt, 250-350 mg of bovine bone protein peptide, 40-60 mg of hydrolyzed egg yolk powder, 8-15 μg of vitamin D3, 5-20 μg of vitamin K2, and 200-300 mg of amino acid complex.

[0009] In the above composition formula, calcium can activate calcium-sensitive receptors (CaSR) and provide bone matrix raw materials; magnesium is a component of the skeletal crystal structure and can regulate the secretion of parathyroid hormone (PTH), cooperates with vitamin D to maintain blood calcium balance, and ensures the stability of the bone metabolism environment; zinc is a key coenzyme of alkaline phosphatase (ALP), which is responsible for creating a microenvironment suitable for mineral (hydroxyapatite) deposition in the bone matrix, and sufficient zinc can maximize the activity of ALP to ensure that calcium, phosphorus and other minerals can be successfully "solidified" on the skeleton.

[0010] Vitamin D up-regulates the expression of osteoblast VDR receptors, promotes the synthesis of calcium-binding proteins, and vitamin K2 guides the directional deposition of calcium in bone tissue, ensuring the directional deposition of calcium in long bones and reducing the risk of ectopic calcification. Both of them are classic calcium supplement adjuvants.

[0011] At the same time, hydrolyzed egg yolk powder is rich in Phosphovitin, which can effectively inhibit the activity of osteoclasts (cells responsible for "disassembling" old bone), thereby slowing down the decomposition and absorption of bone mass. The bone-forming function of calcium, magnesium and zinc in combination with hydrolyzed egg yolk powder to inhibit osteoclasts can maintain the net increase of bone mass.

[0012] On this basis, the bovine bone protein peptide contains a natural bone morphogenetic protein BMP-2 precursor fragment (molecular weight <10 kDa), which can promote chondrocyte proliferation through the Smad1 / 5 / 8 pathway, delay the closure of the growth plate, and directly promote the longitudinal growth of long bones.

[0013] Preferably, the amino acid complex includes not less than 45wt% of gamma-aminobutyric acid, and not less than 20wt% of lysine and L-arginine, and the weight ratio of the lysine and L-arginine is 4-6:2.5-3.5.

[0014] Gamma-aminobutyric acid (GABA) as an important neurotransmitter, can act on the hypothalamic GABA-B receptor, stimulate growth hormone releasing hormone (GHRH) neurons, thereby promoting the pituitary gland to secrete more growth hormone (GH) in a pulsatile manner at night.

[0015] Growth hormone (GH) stimulates the liver to synthesize and secrete insulin-like growth factor 1 (IGF-1). And a certain proportion (such as 5:3) of lysine and L-arginine can significantly enhance the liver's response to GH signal, thereby efficiently synthesizing IGF-1. IGF-1 is a key factor that directly acts on growth plate chondrocytes to promote their proliferation.

[0016] The above-mentioned amino acid complex can play a synergistic role in regulating the GH-IGF-1 axis, together with bovine bone protein peptides, to support bone growth and repair from the endocrine perspective.

[0017] Preferably, the amino acid complex includes 13-17 parts of gamma-aminobutyric acid, 4-6 parts of lysine, 2.5-3.5 parts of L-arginine, 1.5-2.5 parts of tyrosine, and 1.5-2.5 parts of glycine. More preferably, the weight ratio of lysine and arginine is 5:3.

[0018] Preferably, the calcium salt is a mixture of calcium citrate and calcium threonate, and the mass ratio of calcium citrate to calcium threonate is 1:0.5-2. The magnesium salt is a mixture of magnesium glycinate and magnesium gluconate, and the mass ratio of magnesium glycinate to magnesium gluconate is 1:0.5-3. The zinc salt is zinc gluconate, zinc citrate, or any combination thereof.

[0019] The above-mentioned combination of calcium salts can provide high bioavailability calcium (absorption rate >40%) and is more conducive to synergistically stimulating the calcium-sensitive receptors of osteoblasts; the combination of magnesium salts can regulate PTH secretion and is more conducive to promoting collagen cross-linking.

[0020] Preferably, the composition further includes, per unit dose, 50-200 ug of vitamin A and 0.2-0.4 mg of vitamin B6.

[0021] Preferably, the composition further comprises 2000-3000 mg of a complex plant extract per unit dosage, the complex plant extract consisting of hawthorn concentrated juice, red date concentrated juice and apple concentrated juice, preferably in a mass ratio of 1 : 1.5-2.5: 1.

[0022] The complex plant extract provides polyphenols (such as proanthocyanidins, quercetin), inhibits the NF-κB pathway, reduces inflammatory factor IL-6 and osteoclast differentiation, and can also scavenge reactive oxygen species (ROS) in the growth plate area, reducing cartilage cell apoptosis caused by oxidative stress.

[0023] Preferably, the composition further comprises a functional adjuvant per unit dosage, the functional adjuvant being selected from at least one of the following: resistant dextrin 200-500 mg; fructooligosaccharide 1000-2000 mg; ferrous gluconate 1-5 mg.

[0024] The combination of fructooligosaccharide and resistant dextrin helps to regulate intestinal flora and improve calcium absorption rate.

[0025] Preferably, the composition is a medicine, food or health product; further preferably, the dosage form of the composition is an oral liquid, tablet, capsule or solid beverage.

[0026] Preferably, the composition comprises the following components per unit dosage: a) calcium salt 300-500 mg, the calcium salt being a mixture of calcium citrate and calcium threonate, wherein the mass ratio of calcium citrate to calcium threonate is 1 : 0.5-2; b) magnesium salt 30-50 mg, the magnesium salt being a mixture of magnesium glycinate and magnesium gluconate, the mass ratio of magnesium glycinate to magnesium gluconate being 1 : 0.5-3; c) zinc salt 2-5 mg, selected from zinc gluconate, zinc citrate or a combination thereof; d) bovine bone protein peptide 250-350 mg; e) hydrolyzed egg yolk powder 40-60 mg; f) vitamin D3 8-15 μg; g) vitamin K2 5-20 μg; h) vitamin A 50-200 ug; i) vitamin B6 0.2-0.4 mg; j) 200-300 mg of an amino acid complex consisting of gamma-aminobutyric acid, lysine, L-arginine, tyrosine and glycine in a mass ratio of 15±2 : 5±1 : 3±0.5 : 2±0.5 : 2±0.5; k) a pharmaceutically or dietetically acceptable carrier.

[0027] In a second aspect, the present application provides a preparation method of the above-mentioned composition, comprising the following steps: dissolving the calcium salt, the magnesium salt and the zinc salt in deionized water, adjusting the pH to 5.5-7.0 to obtain a mineral solution; adding the bovine bone protein peptide, the hydrolyzed egg yolk powder, the amino acid complex and the total vitamins into the mineral solution, and stirring and mixing at 40-60°C for 30-60 minutes to obtain a colloidal dispersion system; dry-mixing the colloidal dispersion system with the complex plant extract and the functional excipient.

[0028] In a third aspect, the present application provides any of the following applications of the above-mentioned composition: (1) promoting secretion of growth factors; (2) promoting proliferation and differentiation of osteoblasts; (3) improving bone density; (4) preparing a biomaterial for repairing bone defects.

[0029] The composition for promoting secretion of growth factors and proliferation of bone cells provided by the present application has at least the following advantages: Multi-channel synergistic mechanism: for the first time, the triple action of "GH-IGF-1 axis activation + growth plate targeting + bidirectional regulation of bone metabolism" is integrated, breaking through the limitation of single target intervention.

[0030] Endogenous regulation advantage: the host's own secretion of BMP-2 / GH is activated by the combination of bovine bone protein peptide and amino acids, avoiding the side effects of exogenous factors.

[0031] Cross-domain application adaptation: the same formula can be flexibly adapted to oral preparations, bone repair scaffolds and postoperative nutritional products, covering the whole life cycle of bone health needs. DETAILED DESCRIPTION

[0032] In order to make the objectives, technical solutions and advantages of the present application clearer, the following will clearly and completely describe the technical solutions in the present application. Obviously, the described embodiments are part of the embodiments of the present application, rather than all the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative labor fall within the scope of protection of the present application.

[0033] All raw materials used in the composition of this invention are known products and can be purchased commercially.

[0034] Example 1 This embodiment relates to a composition that promotes growth factor secretion and osteoblast proliferation, and its preparation method; the specific steps are as follows: Step 1 (Preparation of ion-chelated solution): Dissolve 200 mg calcium citrate, 200 mg calcium threonate, 20 mg magnesium glycine, 20 mg magnesium gluconate, and 3 mg zinc gluconate in deionized water, and adjust the pH to 6.0-6.5 to form an ion-chelated solution. Step 2 (Active Ingredient Integration): In the ion-chelated solution, add 300 mg bovine bone protein peptide, 50 mg hydrolyzed egg yolk powder, amino acid combination (150 mg γ-aminobutyric acid, 50 mg lysine, 30 mg L-arginine, 20 mg tyrosine, 20 mg glycine), and vitamin combination (10 μg vitamin K2, 100 μg vitamin A, 0.3 mg vitamin B6, 10 μg vitamin D). Stir at 45-50℃ for 30 minutes to form a colloidal dispersion system. Step 3 (Initial Product Formation): The colloidal dispersion system is mixed with 750mg hawthorn concentrate, 1375mg jujube concentrate, 1500mg fructooligosaccharide, and 750mg apple concentrate to obtain the initial product.

[0035] Step 4 (Preparation of final product): Add 250mL of purified water to the initial product to obtain the oral liquid final product.

[0036] The composition preparation method provided in this embodiment has significant advantages such as simple process, safe operation, high efficiency and energy saving, and suitability for large-scale production. The obtained composition can activate the calcium-sensitive receptor (CaSR) and BMP / Smad signaling pathway through calcium, magnesium, and zinc synergistic effects with bone-derived peptides and vitamin D / K2, significantly increasing osteoblast proliferation rate and endogenous growth factor (BMP-2, IGF-1) secretion. This demonstrates the feasibility of the composition and preparation method for promoting growth factor secretion and osteoblast proliferation.

[0037] Example 2 This embodiment relates to a composition that promotes growth factor secretion and osteoblast proliferation, and its preparation method; the specific steps are as follows: Step 1 (Preparation of ion-chelated solution): Dissolve 200 mg calcium citrate, 300 mg calcium threonate, 20 mg magnesium glycine, 20 mg magnesium gluconate, and 3 mg zinc gluconate in deionized water, and adjust the pH to 6.0-6.5 to form an ion-chelated solution. Step 2 (Active Ingredient Integration): In the ion-chelated solution, add 300 mg bovine bone protein peptide, 50 mg hydrolyzed egg yolk powder, amino acid combination (130 mg γ-aminobutyric acid, 50 mg lysine, 30 mg L-arginine, 20 mg tyrosine, 20 mg glycine), and vitamin combination (10 μg vitamin K2, 100 μg vitamin A, 0.3 mg vitamin B6, 10 μg vitamin D). Stir at 45-50℃ for 30 minutes to form a colloidal dispersion system. Step 3 (Initial Product Formation): The colloidal dispersion system is mixed with 750mg hawthorn concentrate, 1375mg jujube concentrate, 1500mg fructooligosaccharide, and 750mg apple concentrate to obtain the initial product.

[0038] Step 4 (Preparation of final product): Add 250mL of purified water to the initial product to obtain the oral liquid final product.

[0039] The composition preparation method provided in this embodiment has significant advantages such as simple process, safe operation, high efficiency and energy saving, and suitability for large-scale production. The obtained composition can activate the calcium-sensitive receptor (CaSR) and BMP / Smad signaling pathway through calcium, magnesium, and zinc synergistic effects with bone-derived peptides and vitamin D / K2, significantly increasing osteoblast proliferation rate and endogenous growth factor (BMP-2, IGF-1) secretion. This demonstrates the feasibility of the composition and preparation method for promoting growth factor secretion and osteoblast proliferation.

[0040] Example 3 Step 1 (Preparation of ion-chelated solution): Dissolve 250 mg calcium citrate, 250 mg calcium threonate, 20 mg magnesium glycine, 20 mg magnesium gluconate, and 3 mg zinc gluconate in deionized water, and adjust the pH to 6.0-6.5 to form an ion-chelated solution. Step 2 (Active Ingredient Integration): In the ion-chelated solution, add 300 mg bovine bone protein peptide, 50 mg hydrolyzed egg yolk powder, amino acid combination (130 mg γ-aminobutyric acid, 40 mg lysine, 30 mg L-arginine, 25 mg tyrosine, 25 mg glycine), and vitamin combination (10 μg vitamin K2, 100 μg vitamin A, 0.3 mg vitamin B6, 10 μg vitamin D). Stir at 45-50℃ for 30 minutes to form a colloidal dispersion system. Step 3 (Initial Product Formation): The colloidal dispersion system is mixed with 750mg hawthorn concentrate, 1375mg jujube concentrate, 1500mg fructooligosaccharide, and 750mg apple concentrate to obtain the initial product.

[0041] Step 4 (Preparation of final product): Add 250mL of purified water to the initial product to obtain the oral liquid final product.

[0042] The composition preparation method provided in this embodiment has significant advantages such as simple process, safe operation, high efficiency and energy saving, and suitability for large-scale production. The obtained composition can activate the calcium-sensitive receptor (CaSR) and BMP / Smad signaling pathway through calcium, magnesium, and zinc synergistic effects with bone-derived peptides and vitamin D / K2, significantly increasing osteoblast proliferation rate and endogenous growth factor (BMP-2, IGF-1) secretion. This demonstrates the feasibility of the composition and preparation method for promoting growth factor secretion and osteoblast proliferation.

[0043] Example 4 Step 1 (Preparation of Ion-chelated Solution) This embodiment relates to a composition that promotes growth factor secretion and osteoblast proliferation, and its preparation method; the specific steps are as follows: Dissolve 200 mg calcium citrate, 200 mg calcium threonate, 20 mg magnesium glycine, 20 mg magnesium gluconate, and 3 mg zinc gluconate in deionized water, and adjust the pH to 6.0-6.5 to form an ion-chelated solution. Step 2 (Active Ingredient Integration): In the ion-chelated solution, add 300 mg bovine bone protein peptide, 50 mg hydrolyzed egg yolk powder, amino acid combination (150 mg γ-aminobutyric acid, 50 mg lysine, 30 mg L-arginine, 20 mg tyrosine, 20 mg glycine), and vitamin combination (10 μg vitamin K2, 100 μg vitamin A, 0.3 mg vitamin B6, 10 μg vitamin D). Stir at 45-50℃ for 30 minutes to form a colloidal dispersion system. Step 3 (Initial Product Formation): The colloidal dispersion system is mixed with 750mg hawthorn concentrate, 1375mg jujube concentrate, 1500mg fructooligosaccharide, and 750mg apple concentrate to obtain the initial product.

[0044] Step 4 (Preparation of final product): Spray drying (inlet air temperature 160℃, outlet air temperature 75℃), control the moisture content ≤5%, obtain powder matrix, and obtain the solid beverage final product.

[0045] The preparation method provided in this embodiment has significant advantages such as simple process, safe operation, high efficiency and energy saving, and suitability for large-scale production. The obtained composition can activate the calcium-sensitive receptor (CaSR) and BMP / Smad signaling pathway through calcium, magnesium, and zinc synergistic effects with bone-derived peptides and vitamin D / K2, significantly increasing osteoblast proliferation rate and endogenous growth factor (BMP-2, IGF-1) secretion. This demonstrates the feasibility of the composition and preparation method for promoting growth factor secretion and osteoblast proliferation.

[0046] Example 5 Step 1 (Preparation of Ion-chelated Solution) This embodiment relates to a composition that promotes growth factor secretion and osteoblast proliferation, and its preparation method; the specific steps are as follows: Dissolve 200 mg calcium citrate, 200 mg calcium threonate, 20 mg magnesium glycine, 20 mg magnesium gluconate, and 3 mg zinc gluconate in deionized water, and adjust the pH to 6.0-6.5 to form an ion-chelated solution. Step 2 (Active Ingredient Integration): In the ion-chelated solution, add 300 mg bovine bone protein peptide, 50 mg hydrolyzed egg yolk powder, amino acid combination (150 mg γ-aminobutyric acid, 50 mg lysine, 30 mg L-arginine, 20 mg tyrosine, 20 mg glycine), and vitamin combination (10 μg vitamin K2, 100 μg vitamin A, 0.3 mg vitamin B6, 10 μg vitamin D). Stir at 45-50℃ for 30 minutes to form a colloidal dispersion system. Step 3 (Initial Product Formation): The colloidal dispersion system is mixed with 750mg hawthorn concentrate, 1375mg jujube concentrate, 1500mg fructooligosaccharide, and 750mg apple concentrate to obtain the initial product.

[0047] Step 4 (Preparation of final product): Spray drying (inlet air temperature 160℃, outlet air temperature 75℃), control the moisture content ≤5%, obtain powder matrix, and encapsulate to obtain capsule final product.

[0048] The composition preparation method provided in this embodiment has significant advantages such as simple process, safe operation, high efficiency and energy saving, and suitability for large-scale production. The obtained composition can activate the calcium-sensitive receptor (CaSR) and BMP / Smad signaling pathway through calcium, magnesium, and zinc synergistic effects with bone-derived peptides and vitamin D / K2, significantly increasing osteoblast proliferation rate and endogenous growth factor (BMP-2, IGF-1) secretion. This demonstrates the feasibility of the composition and preparation method for promoting growth factor secretion and osteoblast proliferation.

[0049] Example 6 This embodiment relates to a composition that promotes growth factor secretion and osteoblast proliferation, and its preparation method; the specific steps are as follows: Step 1 (Preparation of ion-chelated solution): Dissolve 200 mg calcium citrate, 200 mg calcium threonate, 20 mg magnesium glycine, 20 mg magnesium gluconate, and 3 mg zinc gluconate in deionized water, and adjust the pH to 6.0-6.5 to form an ion-chelated solution. Step 2 (Active Ingredient Integration): In the ion-chelated solution, add 300 mg bovine bone protein peptide, 50 mg hydrolyzed egg yolk powder, amino acid combination (150 mg γ-aminobutyric acid, 50 mg lysine, 30 mg L-arginine, 20 mg tyrosine, 20 mg glycine), and vitamin combination (10 μg vitamin K2, 100 μg vitamin A, 0.3 mg vitamin B6, 10 μg vitamin D). Stir at 45-50℃ for 30 minutes to form a colloidal dispersion system. Step 3 (Initial Product Formation): The colloidal dispersion system is mixed with 750mg hawthorn concentrate, 1375mg jujube concentrate, 1500mg fructooligosaccharide, and 750mg apple concentrate to obtain the initial product.

[0050] Step 4 (Preparation of final product): Spray drying (inlet air temperature 160℃, outlet air temperature 75℃), control the moisture content ≤5%, obtain powder matrix, compress into tablets, and obtain the final tablet product.

[0051] The composition preparation method provided in this embodiment has significant advantages such as simple process, safe operation, high efficiency and energy saving, and suitability for large-scale production. The obtained composition can activate the calcium-sensitive receptor (CaSR) and BMP / Smad signaling pathway through calcium, magnesium, and zinc synergistic effects with bone-derived peptides and vitamin D / K2, significantly increasing osteoblast proliferation rate and endogenous growth factor (BMP-2, IGF-1) secretion. This demonstrates the feasibility of the composition and preparation method for promoting growth factor secretion and osteoblast proliferation.

[0052] Comparative Example 1 Step 1 (Preparation of ion-chelated solution): Dissolve 200 mg calcium citrate, 200 mg calcium threonate, 20 mg magnesium glycine, 20 mg magnesium gluconate, and 3 mg zinc gluconate in deionized water, and adjust the pH to 6.0-6.5 to form an ion-chelated solution. Step 2 (Active Ingredient Integration): In the ion-chelated solution, add 600 mg bovine bone protein peptide, 50 mg hydrolyzed egg yolk powder, amino acid combination (200 mg γ-aminobutyric acid, 20 mg lysine, 30 mg L-arginine, 20 mg tyrosine, 10 mg glycine), and vitamin combination (10 μg vitamin K2, 100 μg vitamin A, 0.3 mg vitamin B6, 10 μg vitamin D). Stir at 45-50℃ for 30 minutes to form a colloidal dispersion system. Step 3 (Initial Product Formation): The colloidal dispersion system is mixed with 750mg hawthorn concentrate, 1375mg jujube concentrate, 1500mg fructooligosaccharide, and 750mg apple concentrate to obtain the initial product.

[0053] Step 4 (Preparation of final product): Add 250mL of purified water to the initial product to obtain the oral liquid final product.

[0054] The only difference between this comparative example and Example 1 is that the amount of lysine added is 20 mg per serving.

[0055] The composition preparation method provided in this comparative example has significant advantages such as simple process, safe operation, high efficiency and energy saving, and suitability for large-scale production. The obtained composition cannot activate the calcium-sensitive receptor (CaSR) and BMP / Smad signaling pathways through calcium, magnesium, and zinc synergistic effects with bone-derived peptides and vitamin D / K2, and cannot increase osteoblast proliferation rate or endogenous growth factor (BMP-2, IGF-1) secretion. This indicates that the composition and its preparation method that promote growth factor secretion and osteoblast proliferation under these conditions are not feasible.

[0056] Comparative Example 2 Step 1 (Preparation of ion-chelated solution): Dissolve 200 mg calcium citrate, 600 mg calcium threonate, 20 mg magnesium glycine, 50 mg magnesium gluconate, and 3 mg zinc gluconate in deionized water, and adjust the pH to 6.0-6.5 to form an ion-chelated solution. Step 2 (Active Ingredient Integration): In the ion-chelated solution, add 300 mg bovine bone protein peptide, 50 mg hydrolyzed egg yolk powder, amino acid combination (150 mg γ-aminobutyric acid, 50 mg lysine, 30 mg L-arginine, 20 mg tyrosine, 20 mg glycine), and vitamin combination (10 μg vitamin K2, 100 μg vitamin A, 0.3 mg vitamin B6, 10 μg vitamin D). Stir at 45-50℃ for 30 minutes to form a colloidal dispersion system. Step 3 (Initial Product Formation): The colloidal dispersion system is mixed with 750mg hawthorn concentrate, 1375mg jujube concentrate, 1500mg fructooligosaccharide, and 750mg apple concentrate to obtain the initial product.

[0057] Step 4 (Preparation of final product): Add 250mL of purified water to the initial product to obtain the oral liquid final product.

[0058] The only difference between this comparative example and Example 1 is that the amount of calcium threonate added is 600 mg per serving, and the amount of magnesium gluconate added is 50 mg per serving.

[0059] The composition preparation method provided in this comparative example has significant advantages such as simple process, safe operation, high efficiency and energy saving, and suitability for large-scale production. The obtained composition cannot activate the calcium-sensitive receptor (CaSR) and BMP / Smad signaling pathways through calcium, magnesium, and zinc synergistic effects with bone-derived peptides and vitamin D / K2, and cannot increase osteoblast proliferation rate or endogenous growth factor (BMP-2, IGF-1) secretion. This indicates that the composition and its preparation method that promote growth factor secretion and osteoblast proliferation under these conditions are not feasible.

[0060] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, and not to limit them; although the present invention has been described in detail with reference to the foregoing embodiments, those skilled in the art should understand that modifications can still be made to the technical solutions described in the foregoing embodiments, or equivalent substitutions can be made to some of the technical features; and these modifications or substitutions do not cause the essence of the corresponding technical solutions to deviate from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A composition, characterized in that, Each unit dose of the combination group includes: 300-500 mg of calcium salt, 30-50 mg of magnesium salt, 2-5 mg of zinc salt, 250-350 mg of bovine bone protein peptide, 40-60 mg of hydrolyzed egg yolk powder, 8-15 μg of vitamin D3, 5-20 μg of vitamin K2, and 200-300 mg of amino acid complex.

2. The composition according to claim 1, characterized in that, The amino acid complex includes not less than 45 wt% γ-aminobutyric acid, and not less than 20 wt% lysine and L-arginine, wherein the weight ratio of lysine to L-arginine is 4-6:2.5-3.

5.

3. The composition according to claim 2, characterized in that, The amino acid complex comprises: 13-17 parts of γ-aminobutyric acid, 4-6 parts of lysine, 2.5-3.5 parts of L-arginine, 1.5-2.5 parts of tyrosine, and 1.5-2.5 parts of glycine; Preferably, the weight ratio of lysine to arginine is 5:

3.

4. The composition according to claim 1, characterized in that, The calcium salt is a mixture of calcium citrate and calcium threonate, wherein the mass ratio of calcium citrate to calcium threonate is 1:0.5~2; The magnesium salt is a mixture of magnesium glycinate and magnesium gluconate, with a mass ratio of magnesium glycinate to magnesium gluconate of 1:0.5~3. The zinc salt is zinc gluconate, zinc citrate, or any combination thereof.

5. The composition according to any one of claims 1-4, characterized in that, Each unit dose of the composition further includes 50-200 μg of vitamin A and 0.2-0.4 mg of vitamin B6; and / or, each unit dose of the composition further includes 2000-3000 mg of a compound plant extract, wherein the compound plant extract includes hawthorn concentrate, jujube concentrate and apple concentrate, preferably in a mass ratio of 1:1.5-2.5:

1.

6. The composition according to any one of claims 1-5, characterized in that, Each unit dose of the composition further includes a functional excipient selected from at least one of the following: Resistant dextrin 200-500 mg; Fructooligosaccharides 1000-2000 mg; Ferrous gluconate 1-5 mg.

7. The composition according to any one of claims 1-6, characterized in that, The composition is a drug, food, or health product; preferably, the dosage form of the composition is an oral liquid, tablet, capsule, or solid beverage.

8. The composition according to claim 1, characterized in that, Each unit dose contains the following components: a) 300-500 mg of calcium salt, wherein the calcium salt is a mixture of calcium citrate and calcium threonate, wherein the mass ratio of calcium citrate to calcium threonate is 1:0.5-2; b) 30-50 mg of magnesium salt, wherein the magnesium salt is a mixture of magnesium glycinate and magnesium gluconate, and the mass ratio of magnesium glycinate to magnesium gluconate is 1:0.5-3; c) 2-5 mg of zinc salt, selected from zinc gluconate, zinc citrate, or a combination thereof; d) Bovine bone protein peptides 250-350 mg; e) 40-60 mg of hydrolyzed egg yolk powder; f) Vitamin D3 8-15 μg; g) Vitamin K2 5-20 μg; h) Vitamin A 50-200 ug; i) Vitamin B6 0.2-0.4mg; j) 200-300 mg of an amino acid complex, wherein the amino acid complex is composed of γ-aminobutyric acid, lysine, L-arginine, tyrosine and glycine in a mass ratio of 15±2 :5±1 :3±0.5 :2±0.5 :2±0.5; k) A pharmaceutically or food-grade acceptable carrier.

9. A method for preparing the composition according to any one of claims 1-8, characterized in that, Includes the following steps: The calcium, magnesium, and zinc salts are dissolved in deionized water, and the pH is adjusted to 5.5-7.0 to obtain a mineral solution. Bovine bone protein peptides, hydrolyzed egg yolk powder, amino acid complex and all vitamins are added to the mineral solution and stirred at 40-60℃ for 30-60 minutes to obtain a colloidal dispersion system. The colloidal dispersion system was dry-mixed with the compound plant extract and functional excipients.

10. Any of the following applications of the composition according to any one of claims 1-8: (1) Promotes the secretion of growth factors; (2) Promotes osteoblast proliferation and differentiation; (3) Increase bone density; (4) Preparation of biomaterials for bone defect repair.