Yak bone collagen peptide for improving osteoporosis as well as preparation and application of yak bone collagen peptide

Abstract

The invention discloses a yak bone collagen peptide for improving osteoporosis and preparation and application thereof, and relates to the field of biological medicine, the yak bone collagen peptide comprises collagen peptide with the molecular weight of 500-2000 Daltons, contains a glycine-proline-hydroxyproline tripeptide core structure, can promote osteoblast differentiation and inhibit osteoclast activity, and can be used for treating osteoporosis. The amino acid sequence of the collagen peptide is any one of the following sequences: glycine-proline-hydroxyproline-glycine-glutamic acid-arginine, and the amino acid sequence of the collagen peptide is any one of the following sequences: glycine-proline-hydroxyproline-glycine-glutamic acid-arginine; and the amino acid is glycine, proline, hydroxyproline, serine, leucine and alanine. The yak bone collagen peptide has a unique glycine-proline-hydroxyproline tripeptide structure, the molecular weight is controlled within the range of 500-2000 Daltons, the structure can promote osteoblast differentiation and inhibit osteoclast activity at the same time, and the problem that a traditional osteoporosis treatment medicine is single in function is solved.

Description

Technical Field

[0001] The invention discloses a yak bone collagen peptide for improving osteoporosis and a preparation method and application thereof, and relates to the field of biomedicine. Background Art

[0002] Osteoporosis is a metabolic bone disease characterized by decreased bone mass and destruction of bone microstructure. It is common in middle-aged and elderly women and postmenopausal women. It can easily lead to an increased risk of fractures and seriously affect the quality of life of patients. At present, the main drugs used in the clinical treatment of osteoporosis include bisphosphonates, estrogens, RANKL inhibitors, etc., but long-term use of these drugs may cause adverse reactions such as jaw necrosis, cardiovascular risks and gastrointestinal discomfort. In addition, traditional nutritional interventions (such as calcium and vitamin D supplements) have limited effects due to their low absorption rate and inability to directly regulate bone metabolic balance. Therefore, the development of safe, efficient and easily absorbed natural active ingredients has become a research hotspot.

[0003] Collagen peptides, due to their excellent biocompatibility and bone targeting, show potential in improving osteoporosis. Currently, collagen peptides are primarily derived from pigs, cattle, or fish. However, bovine sources may carry the risk of mad cow disease, while fish sources are prone to allergies, limiting their widespread application. Furthermore, existing collagen peptides are mostly mixed peptide segments with a wide molecular weight distribution and unclear activity. Most studies have focused solely on promoting bone formation while ignoring their ability to inhibit bone resorption, resulting in poor clinical efficacy.

[0004] Yak bones, as a by-product of plateau animal husbandry, are rich in high-quality collagen. However, due to the lack of high-value utilization technology, they are often discarded or used only as low-value-added raw materials. Yaks have long adapted to the high-altitude, hypoxic environment, and their bone collagen may have unique structural stability and biological activity, but there are few related studies, especially active peptides for osteoporosis, which have not been fully explored. Traditional collagen extraction processes (such as acid / alkali methods) easily destroy the natural structure, while peptides prepared by conventional enzymatic hydrolysis have low activity and strong bitterness, and lack precise separation technology, making it difficult to obtain high-purity active peptides.

[0005] Therefore, there is a need to develop collagen peptides with a safe source, clear activity, and efficient preparation to address the shortcomings of existing osteoporosis treatments and nutritional interventions. This invention uses yak bone as raw material, combining targeted enzymatic hydrolysis and activity-guided purification techniques to prepare collagen peptides with dual bone metabolism regulation functions, providing a new strategy for the prevention and treatment of osteoporosis. Summary of the Invention

[0006] The purpose of the present invention is to provide a yak bone collagen peptide for improving osteoporosis and its preparation and application, so as to solve the problem that the number of people with sub-healthy kidney deficiency mentioned in the above background technology continues to increase every year, and modern young people generally stay up late, which leads to endocrine disorders and affects kidney metabolism and repair function. Lack of sleep can also reduce immunity and aggravate kidney essence loss, and long-term sitting leads to poor blood circulation, insufficient blood supply to the kidneys, and accelerated renal function decline; lack of exercise affects the circulation of qi and blood throughout the body and aggravates the symptoms of kidney deficiency. Therefore, the demand for natural animal-derived health products among people with sub-healthy kidney deficiency, such as middle-aged and elderly people and those who are overly tired, is increasing.

[0007] To achieve the above objectives, the present invention provides the following technical solutions: a yak bone collagen peptide for improving osteoporosis and its preparation and application, comprising a collagen peptide having a molecular weight of 500 to 2000 Daltons and containing a glycine-proline-hydroxyproline tripeptide core structure, which can promote osteoblast differentiation and inhibit osteoclast activity. The amino acid sequence of the collagen peptide is any one of the following: glycine-proline-hydroxyproline-glycine-glutamic acid-arginine; glycine-proline-hydroxyproline-serine-leucine-alanine. The preparation method comprises the following steps: The yak bones are defatted, demineralized, and crushed to obtain bone powder: pepsin is used to extract collagen under weakly acidic conditions; a composite protease is used to enzymatically hydrolyze under weakly alkaline conditions to obtain a collagen peptide mixture; and active peptides in the target molecular weight range are obtained through ultrafiltration and gel chromatography purification. Yak bone collagen peptides have a unique glycine-proline-hydroxyproline tripeptide structure with a molecular weight controlled in the range of 500-2000 Daltons. This structure can simultaneously promote osteoblast differentiation and inhibit osteoclast activity, solving the problem of the single function of traditional osteoporosis treatment drugs.

[0008] Preferably, the weakly acidic condition is pH 3 to 5, the extraction time is 24 to 48 hours, and the temperature is 4 to 25 degrees Celsius.

[0009] Preferably, the composite protease is a mixture of trypsin and alkaline protease, and the enzymolysis temperature is 50 to 60 degrees Celsius and the time is 2 to 4 hours.

[0010] Preferably, the content of the collagen peptide in the composition is 10% to 50%, and the excipients include microcrystalline cellulose, mannitol or trehalose.

[0011] Preferably, the yak collagen peptide and food-scientifically acceptable additives, and the functional food are powders, beverages or chewable tablets.

[0012] Preferably, the added amount of the collagen peptide is 1 to 10 grams per 100 grams of product, and the food further contains calcium, vitamin D or soy isoflavones.

[0013] Technical effects and advantages of the present invention:

[0014] 1. The yak bone collagen peptide of the present invention has a unique glycine-proline-hydroxyproline tripeptide structure with a molecular weight controlled within the range of 500-2000 Daltons. This structure can simultaneously promote osteoblast differentiation and inhibit osteoclast activity, solving the problem of single function of traditional osteoporosis treatment drugs. At the same time, its preparation method adopts a mild enzymatic hydrolysis process combined with ultrafiltration purification technology. Compared with traditional acid and alkaline extraction methods, the collagen activity retention rate has been significantly improved, and the peptide yield has also been correspondingly improved. In particular, by precisely controlling the enzymatic hydrolysis pH value of 3-5 and the temperature of 50-60°C, the destruction of active peptides is avoided, and the biological activity of the final product is greatly improved compared with conventional collagen peptides.

[0015] 2. The pharmaceutical composition of the present invention can be prepared into dosage forms such as tablets and capsules. When the peptide content is controlled at 10-50%, both the efficacy and the production cost are guaranteed. From the perspective of social benefits, the use of yak bones as raw materials realizes the high-value utilization of animal husbandry by-products. DETAILED DESCRIPTION

[0016] Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making any creative work shall fall within the scope of protection of the present invention.

[0017] The present invention provides a yak bone collagen peptide for improving osteoporosis and its preparation and application, including a collagen peptide with a molecular weight of 500 to 2000 Daltons and a glycine-proline-hydroxyproline tripeptide core structure, which can promote osteoblast differentiation and inhibit osteoclast activity. The amino acid sequence of the collagen peptide is any one of the following: glycine-proline-hydroxyproline-glycine-glutamic acid-arginine; glycine-proline-hydroxyproline-serine-leucine-alanine. The preparation method includes the following steps: defatting yak bones; , demineralization treatment, and then pulverization to obtain bone powder: pepsin is used to extract collagen under weak acidic conditions; composite protease is used to perform enzymatic hydrolysis under weak alkaline conditions to obtain a collagen peptide mixture; ultrafiltration and gel chromatography purification are used to obtain active peptides in the target molecular weight range. Yak bone collagen peptide has a unique glycine-proline-hydroxyproline tripeptide structure with a molecular weight controlled in the range of 500-2000 Daltons. This structure can simultaneously promote osteoblast differentiation and inhibit osteoclast activity, solving the problem of single function of traditional osteoporosis treatment drugs.

[0018] The weak acidic conditions are pH 3 to 5, the extraction time is 24 to 48 hours, the temperature is 4 to 25 degrees Celsius, the composite protease is a mixture of trypsin and alkaline protease, the enzymatic hydrolysis temperature is 50 to 60 degrees Celsius, the time is 2 to 4 hours, the gentle extraction, pepsin and weak acid conditions avoid the destruction of collagen structure by strong acid / alkali, the content of collagen peptide in the composition is 10% to 50%, the excipients include microcrystalline cellulose, mannitol or trehalose, composite enzymatic hydrolysis, trypsin and alkaline protease improve the hydrolysis efficiency, reduce the generation of bitter peptides, yak bone collagen peptide, and food science The functional food is in the form of powder, beverage or chewable tablet. Pharmaceutical compositions such as tablets and capsules are suitable for clinical treatment. Functional food, beverage and chewable tablet are suitable for daily prevention to meet different needs. The amount of collagen peptide added is 1 to 10 grams per 100 grams of product. The food also contains calcium, vitamin D or soy isoflavones. The pharmaceutical composition can be made into tablets, capsules and other dosage forms. When the peptide content is controlled at 10-50%, it can both ensure efficacy and reduce production costs. From the perspective of social benefits, the use of yak bones as raw materials realizes the high-value utilization of livestock by-products.

[0019] Specific Example 1: Raw material pretreatment: Fresh yak bones were taken and crushed into 3-5 cm bone blocks after cleaning. A degreasing process (n-hexane, 50°C) was used for 6 hours, and the degreasing rate reached over 95%. Subsequently, 0.5 mol / L hydrochloric acid solution was used for demineralization for 48 hours, and the demineralization rate reached 85%. Collagen extraction was then carried out. The defatted and demineralized bone blocks were crushed to 80 mesh, and 0.5 mol / L acetic acid solution (containing 1% pepsin) was added at a solid-liquid ratio of 1:15. The collagen extract was stirred and extracted at 15°C for 36 hours. The extract was centrifuged (4000 rpm, 15 min) and the supernatant was collected to obtain the collagen extract. The collagen extract was then subjected to directed enzymatic hydrolysis. The extract pH was adjusted to 8.0, and a composite protease (trypsin and alkaline protease mixed in a 1:1 ratio) was added at an enzyme dosage of 2% of the substrate weight. The collagen extract was hydrolyzed at 55°C for 3 hours. After enzymatic hydrolysis, the product was inactivated at 90°C for 10 minutes and then separated and purified. The hydrolyzate was first filtered through a 10kDa ultrafiltration membrane to remove large proteins, and then filtered through a 3kDa ultrafiltration membrane to collect the target peptides. Further purification was performed using dextran gel chromatography to collect active peptide fractions with molecular weights of 500-2000 Da. The final product was freeze-dried, yielding 6.8% of the original bone.

[0020] Specific embodiment 2: The formula composition comprises 50 g of yak bone collagen peptide powder (molecular weight 800-1500 Da), 20 g of calcium carbonate, 0.01 g of vitamin D3, 25 g of microcrystalline cellulose, 4.9 g of xylitol, and 0.09 g of strawberry essence. The preparation process is then carried out: the yak bone collagen peptide, calcium carbonate, and vitamin D3 are pre-mixed uniformly, microcrystalline cellulose and xylitol are added, and the mixture is continued for 30 minutes. The essence is added and mixed for 10 minutes, and the mixture is pressed into a 1 g / tablet chewable tablet with a hardness controlled at 5-7 kg / cm 2 Available in aluminum-plastic packaging, 30 tablets per box. Each tablet contains 500mg of active peptides, 100mg of calcium, and 2.5μg of vitamin D3. It has a sweet and sour taste with no fishy smell and a shelf life of 24 months at room temperature. It should be taken twice daily, one tablet at a time, chewed. Clinical trials have shown that after three months of continuous use, subjects' bone density T-scores increased by an average of 0.12, and serum bone formation marker levels increased by 15%.

[0021] Its preparation process can stably obtain highly active peptides, solving the problems of low activity and poor yield of traditional collagen peptides; functional foods have confirmed that the peptide can effectively improve osteoporosis in real applications, and it has both scientific and commercial potential. The combination of the two fully demonstrates the value of the entire chain from laboratory research and development to industrial implementation.

[0022] The working principle of the present invention: Osteoporosis is a disease caused by bone metabolism imbalance, resulting in bone loss and destruction of bone tissue structure. The yak bone collagen peptide provided by the present invention effectively improves this pathological process through a multi-target and multi-pathway mechanism of action. Its core working principle is mainly reflected in promoting bone formation. The specific tripeptide structure contained in the collagen peptide can directly act on the bone formation process: activate the bone cell growth-related signal pathway, promote the proliferation and differentiation of osteoblasts; increase the activity of key enzymes in osteoblasts, accelerate the mineralization process of bone matrix; provide the necessary raw materials for bone collagen synthesis, directly participate in the construction of bone matrix, and improve bone microstructure. At the same time, it inhibits bone loss. Small molecule collagen peptides regulate osteoclast activity through multiple mechanisms: blocking the conduction of key signal pathways for osteoclast differentiation; reducing the activity of bone resorption-related enzymes in osteoclasts; and regulating the balance between bone formation-promoting factors and bone resorption-promoting factors in the bone microenvironment. At the same time, it has a comprehensive regulatory effect, improving the intestinal absorption and utilization of minerals such as calcium and phosphorus, promoting local blood circulation in the bones, providing adequate nutrition for bone tissue, enhancing the antioxidant capacity of bone tissue, and reducing the damage of oxidative stress to bones; synergistic enhancement mechanism, synergistically working with nutrients such as vitamin D and calcium to form multiple protections, by regulating the endocrine system, improving the overall bone metabolism environment, and enhancing the self-repair ability of bone tissue.

[0023] This multi-target, multi-level comprehensive regulatory mechanism enables yak collagen peptide to improve osteoporosis symptoms more comprehensively and effectively, while avoiding the side effects of traditional drug treatments, providing a new solution for the prevention and treatment of osteoporosis.

[0024] Finally, it should be noted that the above is only a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art can still modify the technical solutions described in the aforementioned embodiments or make equivalent substitutions for some of the technical features therein. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the scope of protection of the present invention.

Claims

1. A yak collagen peptide for improving osteoporosis and its preparation and application, characterized in that: The collagen peptide has a molecular weight of 500 to 2000 Daltons and contains a glycine-proline-hydroxyproline tripeptide core structure, which can promote osteoblast differentiation and inhibit osteoclast activity. The amino acid sequence of the collagen peptide is any one of the following: glycine-proline-hydroxyproline-glycine-glutamic acid-arginine; glycine-proline-hydroxyproline-serine-leucine-alanine.

2. The yak collagen peptide for improving osteoporosis and its preparation and use according to claim 1, characterized in that: The following steps are involved: The yak bones were defatted, demineralized, and pulverized to obtain bone powder. Collagen was extracted using pepsin under weakly acidic conditions. The collagen peptide mixture is obtained by enzymatic hydrolysis using a composite protease under weak alkaline conditions; active peptides within the target molecular weight range are obtained through ultrafiltration and gel chromatography purification.

3. The yak collagen peptide for improving osteoporosis and its preparation and use according to claim 2, characterized in that: The weakly acidic conditions are pH 3 to 5, the extraction time is 24 to 48 hours, and the temperature is 4 to 25 degrees Celsius.

4. The yak collagen peptide for improving osteoporosis and its preparation and use according to claim 2, characterized in that: The compound protease is a mixture of trypsin and alkaline protease, and the enzymolysis temperature is 50 to 60 degrees Celsius and the time is 2 to 4 hours.

5. The yak collagen peptide for improving osteoporosis and its preparation and use according to claim 1, characterized in that: The content of the collagen peptide in the composition is 10% to 50%, and the auxiliary materials include microcrystalline cellulose, mannitol or trehalose.

6. The yak collagen peptide for improving osteoporosis and its preparation and use according to claim 1, characterized in that: The yak bone collagen peptide and food science acceptable additives, and the functional food are powder, beverage or chewable tablets.

7. The yak collagen peptide for improving osteoporosis and its preparation and use according to claim 6, characterized in that: The added amount of the collagen peptide is 1 to 10 grams per 100 grams of the product, and the food further contains calcium, vitamin D or soy isoflavones.