A collagen peptide composition for repairing joints and a method of preparing the same

Abstract

The present application relates to the field of biological medicine, and particularly relates to a collagen peptide composition for repairing joints and a preparation method thereof, mainly comprising the following raw materials: 20-25 parts of high-dispersion collagen peptide, 4-6 parts of ganoderma extract, 4-6 parts of lentinus edodes extract, 4-6 parts of composite embedded nano calcium, 2-4 parts of hyaluronic acid, 0.4-0.6 parts of vitamin K2, 0.2-0.4 parts of tea polyphenol, and 5-7 parts of pitaya fermentation original pulp; the high-dispersion collagen peptide is prepared through specific enzymolysis and ultrafiltration process, has a small molecular weight and good water solubility, and can promote joint cartilage repair and regeneration; the composite embedded nano calcium is designed through a double embedding structure, improves the bioavailability of calcium, promotes bone density increase and bone mineralization; the composition can efficiently repair joint cartilage, improve bone density and bone calcium content through synergistic action of multiple raw materials, has excellent antioxidant property, and is suitable for joint repair and bone health maintenance.

Description

Technical Field

[0001] This invention relates to the field of biomedicine, and in particular to a collagen peptide composition for repairing joints and its preparation method. Background Technology

[0002] With the aging population and changing lifestyles (such as prolonged sitting and excessive exercise), the incidence of degenerative joint diseases (such as osteoarthritis) and bone health problems (such as osteoporosis) is rising year by year, becoming a significant health hazard affecting the quality of life. Wear and tear of articular cartilage and decreased bone density are the core causes of these problems. The weak self-repair capacity of cartilage and accelerated calcium loss from bone tissue have made dietary supplementation with functional components to assist joint repair and bone health maintenance a hot research topic.

[0003] Chinese Patent Application No. CN202011445307.4 discloses a bovine bone collagen peptide composition, its preparation method, and its application. The composition comprises the following components and their weight parts: 14-28 parts bovine bone collagen peptide, 10-16 parts chondroitin sulfate, and 10-22 parts plant extracts, the plant extracts being composed of *Sinomenium acutum* extract and *Sagittaria sagittifolia* extract. Chinese Patent Application No. CN202210355647.0 discloses a composition for relieving bone and joint pain, its preparation method, and its application. This composition comprises the following components and their weight parts: 1-12 parts *Lycium barbarum*, 1-12 parts *Raspberry*, 1-15 parts *Polygonatum sibiricum*, 1-30 parts *Dioscorea opposita*, 1-9 parts *Panax ginseng*, 1-10 parts *Prunus persica* kernel, and 1-15 parts *Pueraria lobata* root. The preparation method of this composition includes the following steps: extracting, concentrating, drying, and pulverizing wolfberry, raspberry, polygonatum, yam, ginseng, peach kernel, and kudzu root with water; then mixing them evenly with collagen peptides, hydrolyzed egg yolk powder, bovine colostrum basic protein, and vitamin C, and adding excipients to prepare an oral preparation. However, both of the above schemes lack sufficient calcium sources and nutrients that promote calcium absorption and targeted deposition. Although the added collagen peptides are an important component of the bone organic matrix, helping to maintain bone dry weight and toughness, and vitamin C helps collagen synthesis and has a certain promoting effect on bone matrix formation, without the participation of calcium, it is impossible to effectively increase the mineral density and hardness of bones. Therefore, the effects of the above two compositions on improving bone health are limited.

[0004] Therefore, developing a joint repair collagen peptide composition with multidimensional synergistic effects, high stability and high bioavailability, and its preparation method, has become an urgent technical problem to be solved in this field. Summary of the Invention

[0005] To address the aforementioned problems, this invention provides a collagen peptide composition for joint repair and its preparation method. By mixing bovine bone collagen peptides with deionized water, followed by processes such as activated carbon purification, enzymatic hydrolysis with a complex protease, and ultrafiltration membrane separation, highly dispersed collagen peptides are obtained. By reacting casein phosphopeptides and sodium alginate with calcium carbonate, followed by encapsulation with hydroxypropyl-β-cyclodextrin, composite-encapsulated nano-calcium is obtained. Finally, by scientifically proportioning the highly dispersed collagen peptides, composite-encapsulated nano-calcium, chondroitin sulfate, hyaluronic acid, vitamin K2, Ganoderma lucidum / shiitake mushroom extract, and dragon fruit fermentation paste, a multi-target, synergistic joint repair composition is obtained, achieving integrated and efficient repair and protection of articular cartilage and bone.

[0006] The technical solution adopted by the present invention to achieve the above objectives is as follows:

[0007] A collagen peptide composition for repairing joints comprises the following raw materials in parts by weight: 20-25 parts of highly dispersed collagen peptides, 10-15 parts of chondroitin sulfate, 4-6 parts of Ganoderma lucidum extract, 4-6 parts of shiitake mushroom extract, 4-6 parts of composite-encapsulated nano-calcium, 2-4 parts of hyaluronic acid, 0.4-0.6 parts of vitamin K2, 0.2-0.4 parts of tea polyphenols, 5-7 parts of dragon fruit fermented pulp, 4-6 parts of maltodextrin, and 0.1-0.2 parts of ε-polylysine;

[0008] The preparation method of the highly dispersed collagen peptides is as follows:

[0009] Step S1: Add bovine bone collagen peptides to deionized water and stir at 40-50℃ for 20-25 minutes. Then add activated carbon and continue stirring for 30-40 minutes. After cooling to room temperature, filter to obtain the filtrate.

[0010] Step S2: Add the complex protease to the filtrate and hydrolyze it for 2-2.5 hours at 50-55℃ and pH 7.0-7.5. Then, cool it to room temperature and separate the permeate through an ultrafiltration membrane.

[0011] Step S3: Concentrate the permeate, add chitosan, and react at 55-65℃ and pH 7.8-8.2 for 1.5-2 hours. After post-processing, highly dispersed collagen peptides are obtained.

[0012] Furthermore, in step S1, the bovine collagen peptide is added to deionized water to prepare a solution with a mass concentration of 7-9%, and the mass ratio of bovine collagen peptide to activated carbon is 1:0.0015-0.0025.

[0013] Furthermore, the complex protease in step S2 is composed of alkaline protease and papain in a mass ratio of 2.5-3.5:1, and the mass-volume ratio of the complex protease to the filtrate is 0.09-0.1 g / 100 mL.

[0014] Further, in step S3, the permeate is concentrated to a solid content of 9-11%, and the mass-to-volume ratio of chitosan to permeate is 0.035-0.045 g / 100 mL.

[0015] The preparation method of the composite-encapsulated calcium nanoparticles is as follows:

[0016] Step a: First, add casein phosphopeptide and sodium alginate to deionized water and stir for 30-40 minutes at 55-60℃ and pH 6.5-7.0 to obtain a composite encapsulation solution. Then add calcium carbonate and stir for 60-70 minutes at 40-45℃. After post-treatment, the initial encapsulated nano-calcium is obtained.

[0017] Step b: Add hydroxypropyl-β-cyclodextrin to deionized water and stir at 60-65℃ for 20-30 min to obtain a hydroxypropyl-β-cyclodextrin solution. Then cool the solution to 35-40℃, add the initially encapsulated nano-calcium, and stir at a constant temperature for 90-100 min. After post-treatment, obtain composite encapsulated nano-calcium.

[0018] Further, in step a, the mass ratio of casein phosphopeptide to sodium alginate is 1.5-2.5:1, the amount of sodium alginate added to deionized water is 1-1.5 g / 100 mL, and the mass ratio of calcium carbonate to composite embedding solution is 1:5-6.

[0019] Furthermore, the mass concentration of the hydroxypropyl-β-cyclodextrin solution in step b is 9.5-10.5%, and the mass ratio of the initially encapsulated nano-calcium to hydroxypropyl-β-cyclodextrin is 2.5-3.5:7.

[0020] The dragon fruit fermented pulp is made by blending red dragon fruit fermentation liquid and galactooligosaccharides in a mass ratio of 10:3-6.

[0021] A method for preparing a collagen peptide composition for repairing joints includes the following steps:

[0022] (1) The highly dispersed collagen peptides, chondroitin sulfate, Ganoderma lucidum extract, shiitake mushroom extract, composite encapsulated nano-calcium, hyaluronic acid, vitamin K2, tea polyphenols and ε-polylysine were sieved separately and set aside for later use;

[0023] (2) Add maltodextrin to deionized water, heat to 60-65℃ and stir to dissolve. After cooling to 30-35℃, add tea polyphenols, ε-polylysine, highly dispersed collagen peptides and chondroitin sulfate in sequence. Stir at 500-600 r / min for 20-30 min, add dragon fruit fermentation pulp and continue stirring for 10-15 min to form mixture A.

[0024] (3) Add Ganoderma lucidum extract, shiitake mushroom extract and hyaluronic acid to mixture A, and continue stirring for 20-30 minutes to form mixture B. Mix vitamin K2 and composite encapsulated nano-calcium evenly and add it to mixture B. After homogenization, the mixture is obtained.

[0025] The present invention has the following beneficial effects:

[0026] The collagen peptide composition for joint repair prepared in this invention achieves integrated repair of joints and bones through the synergistic effect of various raw materials. Its core lies in the "rigid yet flexible" bone repair foundation formed by highly dispersed collagen peptides and composite-encapsulated nano-calcium: the collagen peptides, as a high-quality raw material for the organic framework, ensure efficient absorption and targeted delivery due to their high dispersibility; while the double-encapsulated nano-calcium, as a calcium source, significantly improves absorption rate and stability. Under the precise regulation of vitamin K2, the two work synergistically, with vitamin K2 activating osteocalcin and guiding calcium ions to efficiently deposit in the collagen network, jointly and significantly increasing bone density, bone calcium content, and bone dry weight.

[0027] Simultaneously, chondroitin sulfate and hyaluronic acid directly replenish the cartilage matrix and lubricate the joints; Ganoderma lucidum and shiitake mushroom extracts, along with tea polyphenols, form a powerful antioxidant network that scavenge free radicals and reduce joint inflammation and damage. This network is further stabilized by the carrier protection effect of collagen peptides; fermented dragon fruit pulp possesses both antioxidant and prebiotic functions, working with ε-polylysine to regulate the intestinal microenvironment and indirectly promote nutrient absorption. These components, through synergistic effects, provide multiple supports required for joint repair, not only repairing cartilage and bone but also protecting joints from oxidative stress damage, significantly improving the repair effect. Therefore, the collagen peptide composition of this invention has significant advantages in joint repair and bone health. Attached Figure Description

[0028] Figure 1 Line graph showing DPPH free radical scavenging capacity;

[0029] Figure 2 Line graph showing the superoxide anion radical scavenging capacity;

[0030] Figure 3 This is a line graph showing the scavenging capacity of hydroxyl radicals. Detailed Implementation

[0031] The technical solutions in the embodiments of this application will be clearly and completely described below with reference to the embodiments of this application. Obviously, the described embodiments are only some embodiments of this application, and not all embodiments. Based on the embodiments of this application, all other embodiments obtained by those of ordinary skill in the art without creative effort are within the scope of protection of this application.

[0032] All raw materials used in the following examples are commercially available products. Chondroitin sulfate, particle size 80-100 mesh, active ingredient content 99%, purchased from Shandong Pingju Biotechnology Co., Ltd.; Ganoderma lucidum extract, specification 10:1, mesh size 80-100, purchased from Shanyang Lianfeng Biotechnology Co., Ltd.; Lentinus edodes extract, particle size 80 mesh, active ingredient content 99%, purchased from Sichuan Huanxu Biotechnology Co., Ltd.; Hyaluronic acid, active ingredient content 99%, purchased from Hebei Jiuxing Chemical Products Co., Ltd.; Vitamin K2, active ingredient content 99%, particle size 120 mesh, purchased from Hubei Haijia Biotechnology Co., Ltd.; Tea polyphenols, particle size 100 mesh, specification 10:1, active ingredient content 98%, purchased from Lanzhou Waterles Biotechnology Co., Ltd.; Maltodextrin, active ingredient content 99.9%, density 1.3-1.6. The following ingredients were purchased from various sources: ε-polylysine (97.2% effective ingredient content) from Shandong Yingsheng Chemical Co., Ltd.; bovine bone collagen peptide (99.2% effective ingredient content) from Sichuan Huanxu Biotechnology Co., Ltd.; activated carbon (325 mesh particle size) from Shijiazhuang Hongsen Activated Carbon Co., Ltd.; chitosan (99% effective ingredient content) from Xi'an Hongyao Pharmaceutical Excipients Co., Ltd.; alkaline protease from Tai'an Xindeli Bioengineering Co., Ltd.; papain from Nanning Dongheng Huadao Biotechnology Co., Ltd.; casein phosphopeptide (37.98% peptide content) from Hebei Hongtao Bioengineering Co., Ltd.; sodium alginate (99% effective ingredient content) from Xi'an Hongyao Pharmaceutical Excipients Co., Ltd.; and calcium carbonate (1250 mesh particle size, 99% effective ingredient content, density 1.7). Purchased from Henan Tuohong Chemical Co., Ltd.; Hydroxypropyl-β-cyclodextrin with an effective ingredient content of 99.5% was purchased from Shandong Tongwang Biotechnology Co., Ltd.; Galacto-oligosaccharide with an effective ingredient content of 99% was purchased from Sichuan Kangbairui Biotechnology Co., Ltd.

[0033] Example 1

[0034] A collagen peptide composition for repairing joints comprises the following raw materials in parts by weight: 20 parts highly dispersed collagen peptides, 10 parts chondroitin sulfate, 4 parts Ganoderma lucidum extract, 4 parts shiitake mushroom extract, 4 parts composite-encapsulated nano-calcium, 2 parts hyaluronic acid, 0.4 parts vitamin K2, 0.2 parts tea polyphenols, 5 parts dragon fruit fermented pulp, 4 parts maltodextrin, and 0.1 parts ε-polylysine;

[0035] The preparation method of the highly dispersed collagen peptides is as follows:

[0036] Step S1: Add bovine bone collagen peptides to deionized water and stir for 22 minutes at 45°C and 300 rpm. Then add activated carbon and continue stirring for 35 minutes. After naturally cooling to room temperature, filter the solution through a 0.22 μm pore size filter membrane to obtain the filtrate. The mass concentration of the solution prepared by adding bovine bone collagen peptides to deionized water is 8%, and the mass ratio of bovine bone collagen peptides to activated carbon is 1:0.002.

[0037] Step S2: Add the complex protease to the filtrate and hydrolyze it for 2.2 h at 52 °C and pH 7.0-7.5. After hydrolysis, inactivate the enzyme at 90 °C for 15 min, and then cool it naturally to room temperature. Separate the permeate through an ultrafiltration membrane with a molecular weight cutoff of 3000 Da. The complex protease is composed of alkaline protease and papain in a mass ratio of 3:1. The mass-volume ratio of the complex protease to the filtrate is 0.095 g / 100 mL.

[0038] Step S3: Concentrate the permeate, add chitosan, and react at 60℃ and pH 7.8-8.2 for 1.7h. Cool naturally to room temperature, then pre-freeze at -40℃ for 3h, and then sublimate and dry at a vacuum of 0.01-0.02MPa and a sublimation temperature of -10 to -5℃ for 10h to obtain highly dispersed collagen peptides. The permeate is concentrated to a solid content of 10%, and the mass-volume ratio of chitosan to permeate is 0.04g / 100mL.

[0039] The preparation method of the composite-encapsulated calcium nanoparticles is as follows:

[0040] Step a: First, add casein phosphopeptide and sodium alginate to deionized water and stir for 35 minutes at 57℃ and pH 6.5-7.0 to obtain a composite encapsulation solution. Then, add calcium carbonate and stir for 65 minutes at 42℃ and 400 rpm. Then, spray dry the solution with an inlet air temperature of 180-190℃, an outlet air temperature of 80-85℃, a feed rate of 15-20 mL / min, and an atomization pressure of 0.25-0.3 MPa to obtain primary encapsulated nano-calcium. The mass ratio of casein phosphopeptide to sodium alginate is 2:1, the amount of sodium alginate added to deionized water is 1.2 g / 100 mL, and the mass ratio of calcium carbonate to composite encapsulation solution is 1:5.5.

[0041] Step b: Add hydroxypropyl-β-cyclodextrin to deionized water and stir at 62℃ for 25 min to obtain a hydroxypropyl-β-cyclodextrin solution. Then cool to 37℃, add the initially encapsulated nano-calcium, and stir at a constant temperature of 300 rpm for 95 min. Allow it to cool naturally to room temperature, then pre-freeze at -40℃ for 3 h. Finally, sublimate and dry under vacuum of 0.01-0.02 MPa and sublimation temperature of -15 to -10℃ for 15 h to obtain composite encapsulated nano-calcium. The mass concentration of the hydroxypropyl-β-cyclodextrin solution is 10%, and the mass ratio of the initially encapsulated nano-calcium to hydroxypropyl-β-cyclodextrin is 3:7.

[0042] The dragon fruit fermented pulp is made by mixing red dragon fruit fermentation liquid and galactooligosaccharides in a mass ratio of 10:3.

[0043] The fermented liquid of red dragon fruit and its preparation method have been disclosed in the patent document with application number CN201810157407.3.

[0044] A method for preparing a collagen peptide composition for repairing joints includes the following steps:

[0045] (1) Highly dispersed collagen peptides, chondroitin sulfate, Ganoderma lucidum extract, shiitake mushroom extract, composite encapsulated nano-calcium, hyaluronic acid, vitamin K2, tea polyphenols, and ε-polylysine were sieved through a 60-mesh sieve and set aside for later use;

[0046] (2) Add maltodextrin to deionized water, heat to 60°C and stir to dissolve. After cooling to 30°C, add tea polyphenols, ε-polylysine, highly dispersed collagen peptides and chondroitin sulfate in sequence. Stir at 500 rpm for 20 min, add dragon fruit fermentation pulp and continue stirring for 10 min to form mixture A.

[0047] (3) Add Ganoderma lucidum extract, shiitake mushroom extract and hyaluronic acid to mixture A, and continue stirring for 20 minutes to form mixture B. Mix vitamin K2 and composite embedded nano calcium evenly and add it to mixture B. Homogenize under a homogenization pressure of 30-35 MPa for 3 times to obtain the final product.

[0048] Example 2

[0049] A collagen peptide composition for repairing joints comprises the following raw materials in parts by weight: 25 parts highly dispersed collagen peptides, 15 parts chondroitin sulfate, 6 parts Ganoderma lucidum extract, 6 parts shiitake mushroom extract, 6 parts composite-encapsulated nano-calcium, 4 parts hyaluronic acid, 0.6 parts vitamin K2, 0.4 parts tea polyphenols, 7 parts dragon fruit fermented pulp, 6 parts maltodextrin, and 0.2 parts ε-polylysine;

[0050] The preparation methods for the highly dispersed collagen peptides and the composite encapsulated calcium nanoparticles are the same as in Example 1.

[0051] The dragon fruit fermented pulp is made by mixing red dragon fruit fermentation liquid and galactooligosaccharides in a mass ratio of 10:6.

[0052] The fermented liquid of red dragon fruit and its preparation method have been disclosed in the patent document with application number CN201810157407.3.

[0053] A method for preparing a collagen peptide composition for repairing joints includes the following steps:

[0054] (1) Highly dispersed collagen peptides, chondroitin sulfate, Ganoderma lucidum extract, shiitake mushroom extract, composite encapsulated nano-calcium, hyaluronic acid, vitamin K2, tea polyphenols, and ε-polylysine were sieved through a 60-mesh sieve and set aside for later use;

[0055] (2) Add maltodextrin to deionized water, heat to 65°C and stir to dissolve. After cooling to 35°C, add tea polyphenols, ε-polylysine, highly dispersed collagen peptides and chondroitin sulfate in sequence. Stir at 600 rpm for 30 min, add dragon fruit fermentation pulp and continue stirring for 15 min to form mixture A.

[0056] (3) Add Ganoderma lucidum extract, shiitake mushroom extract and hyaluronic acid to mixture A, and continue stirring for 30 minutes to form mixture B. Mix vitamin K2 and composite embedded nano calcium evenly and add it to mixture B. Homogenize under a homogenization pressure of 30-35 MPa for 3 times to obtain the final product.

[0057] Example 3

[0058] A collagen peptide composition for repairing joints comprises the following raw materials in parts by weight: 23 parts highly dispersed collagen peptides, 12 parts chondroitin sulfate, 5 parts Ganoderma lucidum extract, 5 parts shiitake mushroom extract, 5 parts composite-encapsulated nano-calcium, 3 parts hyaluronic acid, 0.5 parts vitamin K2, 0.3 parts tea polyphenols, 6 parts dragon fruit fermented pulp, 5 parts maltodextrin, and 0.15 parts ε-polylysine;

[0059] The preparation methods for the highly dispersed collagen peptides and the composite encapsulated calcium nanoparticles are the same as in Example 1.

[0060] The dragon fruit fermented pulp is made by mixing red dragon fruit fermentation liquid and galactooligosaccharides in a mass ratio of 10:5.

[0061] The fermented liquid of red dragon fruit and its preparation method have been disclosed in the patent document with application number CN201810157407.3.

[0062] A method for preparing a collagen peptide composition for repairing joints includes the following steps:

[0063] (1) Highly dispersed collagen peptides, chondroitin sulfate, Ganoderma lucidum extract, shiitake mushroom extract, composite encapsulated nano-calcium, hyaluronic acid, vitamin K2, tea polyphenols, and ε-polylysine were sieved through a 60-mesh sieve and set aside for later use;

[0064] (2) Add maltodextrin to deionized water, heat to 62°C and stir to dissolve. After cooling to 32°C, add tea polyphenols, ε-polylysine, highly dispersed collagen peptides and chondroitin sulfate in sequence. Stir at 550 rpm for 25 min, add dragon fruit fermentation pulp and continue stirring for 13 min to form mixture A.

[0065] (3) Add Ganoderma lucidum extract, shiitake mushroom extract and hyaluronic acid to mixture A, and continue stirring for 25 minutes to form mixture B. Mix vitamin K2 and composite embedded nano calcium evenly and add it to mixture B. Homogenize under a homogenization pressure of 30-35 MPa for 3 times to obtain the final product.

[0066] Example 4

[0067] A collagen peptide composition for repairing joints comprises the following raw materials in parts by weight: 23 parts highly dispersed collagen peptides, 12 parts chondroitin sulfate, 5 parts Ganoderma lucidum extract, 5 parts shiitake mushroom extract, 5 parts composite-encapsulated nano-calcium, 3 parts hyaluronic acid, 0.5 parts vitamin K2, 0.3 parts tea polyphenols, 6 parts dragon fruit fermented pulp, 5 parts maltodextrin, and 0.15 parts ε-polylysine;

[0068] The preparation method of the highly dispersed collagen peptides is as follows:

[0069] Step S1: Add bovine bone collagen peptides to deionized water and stir for 25 minutes at 50°C and 300 rpm. Then add activated carbon and continue stirring for 40 minutes. After naturally cooling to room temperature, filter the solution through a 0.22 μm pore size filter membrane to obtain the filtrate. The mass concentration of the solution prepared by adding bovine bone collagen peptides to deionized water is 9%, and the mass ratio of bovine bone collagen peptides to activated carbon is 1:0.0025.

[0070] Step S2: Add the complex protease to the filtrate and hydrolyze it for 2.5 h at 55 °C and pH 7.0-7.5. After hydrolysis, inactivate the enzyme at 90 °C for 15 min, and then cool it naturally to room temperature. Separate the permeate through an ultrafiltration membrane with a molecular weight cutoff of 3000 Da. The complex protease is composed of alkaline protease and papain in a mass ratio of 3.5:1. The mass-volume ratio of the complex protease to the filtrate is 0.1 g / 100 mL.

[0071] Step S3: Concentrate the permeate, add chitosan, and react at 65℃ and pH 7.8-8.2 for 2 hours. Cool naturally to room temperature, then pre-freeze at -40℃ for 3 hours. Then sublimate and dry under vacuum of 0.01-0.02MPa and sublimation temperature of -10 to -5℃ for 10 hours to obtain highly dispersed collagen peptides. The permeate is concentrated to a solid content of 11%, and the mass-volume ratio of chitosan to permeate is 0.045g / 100mL.

[0072] The preparation method of the composite-encapsulated calcium nanoparticles is as follows:

[0073] Step a: First, add casein phosphopeptide and sodium alginate to deionized water and stir for 40 min at 60℃ and pH 6.5-7.0 to obtain a composite encapsulation solution. Then, add calcium carbonate and stir for 70 min at 45℃ and 400 rpm. Then, spray dry the solution with an inlet air temperature of 180-190℃, an outlet air temperature of 80-85℃, a feed rate of 15-20 mL / min, and an atomization pressure of 0.25-0.3 MPa to obtain primary encapsulated nano-calcium. The mass ratio of casein phosphopeptide to sodium alginate is 2.5:1, the amount of sodium alginate added to deionized water is 1.5 g / 100 mL, and the mass ratio of calcium carbonate to composite encapsulation solution is 1:6.

[0074] Step b: Add hydroxypropyl-β-cyclodextrin to deionized water and stir at 65°C for 30 min to obtain a hydroxypropyl-β-cyclodextrin solution. Then cool to 40°C, add the initially encapsulated nano-calcium, and stir at a constant temperature of 300 rpm for 100 min. Allow it to cool naturally to room temperature, then pre-freeze at -40°C for 3 h. Finally, sublimate and dry under vacuum of 0.01-0.02 MPa and sublimation temperature of -15 to -10°C for 15 h to obtain composite encapsulated nano-calcium. The mass concentration of the hydroxypropyl-β-cyclodextrin solution is 10.5%, and the mass ratio of the initially encapsulated nano-calcium to hydroxypropyl-β-cyclodextrin is 3.5:7.

[0075] The dragon fruit fermented pulp is made by mixing red dragon fruit fermentation liquid and galactooligosaccharides in a mass ratio of 10:5.

[0076] The fermented liquid of red dragon fruit and its preparation method have been disclosed in the patent document with application number CN201810157407.3.

[0077] A method for preparing a collagen peptide composition for repairing joints includes the following steps:

[0078] (1) Highly dispersed collagen peptides, chondroitin sulfate, Ganoderma lucidum extract, shiitake mushroom extract, composite encapsulated nano-calcium, hyaluronic acid, vitamin K2, tea polyphenols, and ε-polylysine were sieved through a 60-mesh sieve and set aside for later use;

[0079] (2) Add maltodextrin to deionized water, heat to 62°C and stir to dissolve. After cooling to 32°C, add tea polyphenols, ε-polylysine, highly dispersed collagen peptides and chondroitin sulfate in sequence. Stir at 550 rpm for 25 min, add dragon fruit fermentation pulp and continue stirring for 13 min to form mixture A.

[0080] (3) Add Ganoderma lucidum extract, shiitake mushroom extract and hyaluronic acid to mixture A, and continue stirring for 25 minutes to form mixture B. Mix vitamin K2 and composite embedded nano calcium evenly and add it to mixture B. Homogenize under a homogenization pressure of 30-35 MPa for 3 times to obtain the final product.

[0081] Comparative Example 1

[0082] A collagen peptide composition for repairing joints comprises the following raw materials in parts by weight: 10 parts highly dispersed collagen peptides, 12 parts chondroitin sulfate, 5 parts Ganoderma lucidum extract, 5 parts shiitake mushroom extract, 1 part composite-encapsulated nano-calcium, 3 parts hyaluronic acid, 0.5 parts vitamin K2, 0.3 parts tea polyphenols, 1 part dragon fruit fermented pulp, 5 parts maltodextrin, and 0.15 parts ε-polylysine;

[0083] The preparation method of the highly dispersed collagen peptides is as follows:

[0084] Step S1: Add bovine bone collagen peptides to deionized water and stir for 10 minutes at 45°C and 300 rpm. Then add activated carbon and continue stirring for 35 minutes. After naturally cooling to room temperature, filter the solution through a 0.22 μm pore size filter membrane to obtain the filtrate. The mass concentration of the solution prepared by adding bovine bone collagen peptides to deionized water is 8%, and the mass ratio of bovine bone collagen peptides to activated carbon is 1:0.001.

[0085] Step S2: Add the complex protease to the filtrate and hydrolyze it for 2.2 h at 70 °C and pH 4.0-4.5. After hydrolysis, inactivate the enzyme at 90 °C for 15 min, and then cool it naturally to room temperature. Separate the permeate through an ultrafiltration membrane with a molecular weight cutoff of 3000 Da. The complex protease is composed of alkaline protease and papain in a mass ratio of 3:1. The mass-volume ratio of the complex protease to the filtrate is 0.095 g / 100 mL.

[0086] Step S3: Concentrate the permeate, add chitosan, and react at 60℃ and pH 7.8-8.2 for 1.7h. Cool naturally to room temperature, then pre-freeze at -40℃ for 3h, and then sublimate and dry at a vacuum of 0.01-0.02MPa and a sublimation temperature of -10 to -5℃ for 10h to obtain highly dispersed collagen peptides. The permeate is concentrated to a solid content of 10%, and the mass-volume ratio of chitosan to permeate is 0.01g / 100mL.

[0087] The preparation method of the composite-encapsulated calcium nanoparticles is as follows:

[0088] Step a: First, add casein phosphopeptide and sodium alginate to deionized water and stir for 35 minutes at 57℃ and pH 6.5-7.0 to obtain a composite encapsulation solution. Then, add calcium carbonate and stir for 65 minutes at 42℃ and 400 rpm. Then, spray dry the solution with an inlet air temperature of 180-190℃, an outlet air temperature of 80-85℃, a feed rate of 15-20 mL / min, and an atomization pressure of 0.25-0.3 MPa to obtain primary encapsulated nano-calcium. The mass ratio of casein phosphopeptide to sodium alginate is 2:1, the amount of sodium alginate added to deionized water is 1.2 g / 100 mL, and the mass ratio of calcium carbonate to composite encapsulation solution is 1:1.

[0089] Step b: Add hydroxypropyl-β-cyclodextrin to deionized water and stir at 62℃ for 25 min to obtain a hydroxypropyl-β-cyclodextrin solution. Then cool to 37℃, add the initially encapsulated nano-calcium, and stir at a constant temperature of 300 rpm for 10 min. Allow to cool naturally to room temperature, then pre-freeze at -40℃ for 3 h. Finally, sublimate and dry under vacuum of 0.01-0.02 MPa and sublimation temperature of -15 to -10℃ for 15 h to obtain composite encapsulated nano-calcium. The mass concentration of the hydroxypropyl-β-cyclodextrin solution is 10%, and the mass ratio of the initially encapsulated nano-calcium to hydroxypropyl-β-cyclodextrin is 1:7.

[0090] The dragon fruit fermented pulp is made by mixing red dragon fruit fermentation liquid and galactooligosaccharides in a mass ratio of 10:1.

[0091] The fermented liquid of red dragon fruit and its preparation method have been disclosed in the patent document with application number CN201810157407.3.

[0092] A method for preparing a collagen peptide composition for repairing joints includes the following steps:

[0093] (1) Highly dispersed collagen peptides, chondroitin sulfate, Ganoderma lucidum extract, shiitake mushroom extract, composite encapsulated nano-calcium, hyaluronic acid, vitamin K2, tea polyphenols, and ε-polylysine were sieved through a 60-mesh sieve and set aside for later use;

[0094] (2) Add maltodextrin to deionized water and stir. Then add tea polyphenols, ε-polylysine, highly dispersed collagen peptides and chondroitin sulfate in sequence. Stir for 25 minutes at 550 rpm. Add dragon fruit fermentation pulp and continue stirring for 13 minutes to form mixture A.

[0095] (3) Add Ganoderma lucidum extract, shiitake mushroom extract and hyaluronic acid to mixture A, and continue stirring for 25 minutes to form mixture B. Mix vitamin K2 and composite embedded nano calcium evenly and add it to mixture B. Homogenize under a homogenization pressure of 30-35 MPa for 3 times to obtain the final product.

[0096] Comparative Example 2

[0097] In this comparative example, commercially available bovine bone collagen peptides were used instead of highly dispersed collagen peptides, and all other aspects were the same as in Example 1.

[0098] Comparative Example 3

[0099] In this comparative example, commercially available calcium carbonate was used instead of composite-encapsulated nano-calcium, and all other aspects were the same as in Example 1.

[0100] Comparative Example 4

[0101] In this comparative example, commercially available collagen peptides were used instead of highly dispersed collagen peptides, and commercially available calcium carbonate was used instead of composite encapsulated nano-calcium. All other aspects were the same as in Example 1.

[0102] (1) Antioxidant test

[0103] DPPH free radical scavenging ability: Samples from Examples 1-4 and Comparative Examples 1-4 were prepared into 0.5 mg / mL solutions, with concentrations of [missing information]. 2 mL of DPPH anhydrous ethanol solution was prepared and stored protected from light. 2 mL of the sample solution was mixed with 2 mL of DPPH anhydrous ethanol solution and shaken vigorously. The mixture was allowed to react at room temperature for 30 min, and the absorbance (Ai) was measured at 517 nm. The blank group was prepared using an equal volume of anhydrous ethanol solution instead of DPPH solution, and the control group was prepared using an equal volume of distilled water instead of the sample solution. The DPPH free radical scavenging rate was calculated using the following formula: In the formula, A0 represents the absorbance of the control group, Ai represents the absorbance of the sample group, and Aj represents the absorbance of the blank group.

[0104] Superoxide anion radical scavenging capacity: Under constant temperature of 25℃, take 3 mL of 50 mmol / L pH 8.2 Tris-HCl buffer solution (containing 1 mmol / L EDTA) and 10 μL of 50 mmol / L pyrogallol, mix rapidly, place in a 1 cm quartz cuvette, and measure the absorbance at 325 nm every 30 s. The reaction is stopped after 4.5 min. Plot absorbance against time, and the slope is the pyrogallol self-oxidation rate, A0. Add collagen peptide composition at a ratio of 0.5 mg / mL to 3 mL of pH 8.2, 50 mmol / L Tris-HCl buffer, and measure the pyrogallol oxidation rate As at 325 nm using the above method. The superoxide anion radical scavenging rate is calculated using the following formula: .

[0105] Hydroxyl radical scavenging ability: Take 0.1 mL of a solution with a concentration of 10 mmol / mL. The mixture was prepared by adding 0.3 mL of deoxyribose (10 mmol / mL), followed by 0.2 mL of the collagen peptide composition sample solution prepared in Examples 1-4 and Comparative Examples 1-4 with a concentration of 0.5 mg / mL. The volume was adjusted to 1.9 mL with 0.1 mol / L phosphate buffer (pH 7.4), and then 0.1 mL of H2O2 (10 mmol / mL) was added. After mixing, the mixture was placed in a 37°C water bath for 1 h. Then, 1 mL of 2.8% (w / w) trichloroacetic acid (TCA) solution and 1 mL of 1.0% (w / w) thiobarbituric acid (TBA) solution were added and mixed. The mixture was then reacted in a boiling water bath for 15 min. After cooling, the absorbance was measured at 532 nm. The absorbance without the addition of a scavenging agent is Ac. If the sample is added and has a scavenging effect on ·OH, it can inhibit the formation of oxidation products, resulting in a decrease in absorbance, which is measured as As. The actual blank absorbance is represented by A0. The formula for calculating the scavenging rate of hydroxyl radicals is: .

[0106] The results are shown in Table 1 and Figure 1-3 As shown.

[0107] Table 1. Antioxidant performance test of collagen peptide compositions used for joint repair.

[0108]

[0109] (2) Rat experiment

[0110] Female clean-grade Wistar rats weighing 180g-230g were selected and divided into 9 groups (one group each for Examples 1-4, Comparative Examples 1-4, and the solvent control group), with 10 rats in each group. The recommended human dose was 20g / day. Assuming an adult weight of 60kg, the recommended daily dose for rats was 0.33g / kg body weight (bw). Rats were administered 0.1mL / 10g body weight via gavage once daily. The solvent control group was administered distilled water via gavage. The experiment lasted 12 weeks. During this period, rats were housed individually, fed a formulated diet free of estrogen-active substances, and had free access to deionized water. At the end of the experiment, animals were euthanized by exsanguination via the femoral artery. Left femurs of rats were dried to constant weight in a 105℃ oven, weighed, and digested in Erlenmeyer flasks. Bone calcium content was determined by atomic absorption spectrometry, and bone mineral density was measured using a bone densitometer. The results are shown in Table 2.

[0111] Table 2. Performance tests of the collagen peptide composition for joint repair: femoral dry weight, bone mineral density, and bone calcium content.

[0112]

[0113] As shown in Table 1-2 and Figure 1-3 As shown, the collagen peptide compositions for joint repair prepared in Examples 1-4 are significantly superior to those in Comparative Example 1 in terms of antioxidant properties (DPPH free radical scavenging capacity, superoxide anion free radical scavenging capacity, hydroxyl free radical scavenging capacity), femoral dry weight, bone mineral density, and bone calcium content. This indicates that the collagen peptide compositions prepared in the examples can repair joints, improve bone mineral density, and maintain bone health. Therefore, it can be concluded that the ratio of the collagen peptide compositions proposed in this invention is optimal, and the femoral dry weight, bone mineral density, and bone calcium content are all superior to those of the solvent control group. This demonstrates that the compositions prepared in Examples 1-3 all have the effect of improving bone health.

[0114] As shown in Table 1-2 and Figure 1-3As shown, compared with the examples, the antioxidant properties and femoral-related data of the collagen peptide composition prepared in Comparative Example 2 decreased. This is because commercially available bovine bone collagen peptides were used instead of highly dispersed collagen peptides. Highly dispersed collagen peptides provide the most direct and essential building blocks for the repair of articular cartilage, bone, and connective tissue, namely collagen. Moreover, through a special preparation process, its molecular weight is smaller, its distribution is more concentrated, and its surface properties are improved, making it highly dispersed in aqueous solutions and body fluids and less prone to aggregation. This allows for faster and larger absorption by the intestines and more effective targeting and enrichment of the joints and bone tissues that need repair. The special preparation process also improves the biocompatibility of highly dispersed collagen peptides, enabling them to fully exert synergistic effects with other active ingredients (such as chondroitin sulfate, calcium, etc.). Composite encapsulation of nano-calcium can improve the stability and absorption rate of calcium, while highly dispersed collagen peptides provide precise anchoring points for absorbed calcium. The two work synergistically to ensure that calcium can be efficiently transported to the bone and deposited, thereby significantly increasing bone calcium content and bone density. Highly dispersed collagen peptides possess inherent antioxidant activity. The histidine, proline, and other amino acids and terminal amino acid residues in the collagen peptide chain have the ability to scavenge free radicals. Through the complex enzymatic hydrolysis in step S2, more active groups are exposed, enhancing their direct antioxidant capacity. In addition, the tea polyphenols, Ganoderma lucidum extract, and shiitake mushroom extract in the composition are potent but potentially unstable antioxidants. Highly dispersed collagen peptides can bind to these components through intermolecular interactions such as hydrogen bonds and hydrophobic interactions, achieving a protective effect and reducing their oxidative inactivation during processing and storage, ensuring that they can fully exert their effects after entering the body.

[0115] As shown in Table 1-2 and Figure 1-3As shown, compared with the examples, the antioxidant properties and femoral-related data of the collagen peptide composition prepared in Comparative Example 3 still decreased. This is because commercially available calcium carbonate was used instead of composite-encapsulated nano-calcium. Composite-encapsulated nano-calcium mainly achieves easier absorption and utilization in vivo by encapsulating the calcium source at the nanoscale. Through its combination with collagen peptides, composite-encapsulated nano-calcium can play a role in joint repair, enhancing bone mineralization and improving bone structure stability. Oxidative stress is an important cause of articular cartilage degeneration and bone matrix decomposition. Although composite-encapsulated nano-calcium is not a direct strong antioxidant, its encapsulation layer components (casein phosphopeptide, sodium alginate, hydroxypropyl-β-cyclodextrin) can form an antioxidant synergistic effect with highly dispersed collagen peptides and tea polyphenols through their own structure and activity, further reducing the damage of oxidative stress to articular cartilage and bone matrix, and providing a better microenvironment for joint repair. The calcium carbonate raw material used in the preparation of composite-encapsulated nano-calcium is nanoscale, with a huge specific surface area. Its contact area with digestive juices in the intestine is much larger than that of ordinary calcium supplements, making it easier to dissolve and ionize. This is the physical basis for its high absorption rate. The composite-encapsulated nano-calcium employs a double-encapsulation structure. The first layer consists of casein phosphopeptide (CPP) and sodium alginate. CPP is an excellent calcium absorption promoter; it can bind with calcium ions to form a soluble complex, effectively preventing calcium from binding with phosphate and phytate ions to form precipitates in the neutral or alkaline environment of the intestine. This prolongs the absorption time of calcium in the intestine and promotes its absorption through the small intestinal mucosal cells. The second layer consists of hydroxypropyl-β-cyclodextrin. Cyclodextrin has a special structure of being hydrophilic on the outside and hydrophobic on the inside, which can form inclusion complexes, further improving the stability and dispersibility of the initially encapsulated nano-calcium. It also helps it penetrate the intestinal mucus layer and make more thorough contact with the absorptive cells. Therefore, this double-encapsulation design makes the bioavailability of calcium much higher than that of ordinary calcium supplements, providing an ample source of calcium for increasing bone calcium content.

[0116] As shown in Table 1-2 and Figure 1-3 As shown, compared with the examples, the collagen peptide composition prepared in Comparative Example 4 had the worst overall performance. This is because commercially available bovine bone collagen peptides replaced highly dispersed collagen peptides, and commercially available calcium carbonate replaced composite-encapsulated nano-calcium. Highly dispersed collagen peptides provide structural support for joints and bones and promote repair, while composite-encapsulated nano-calcium promotes bone mineralization and enhances bone structural strength by providing an effective calcium source. The combination of the two can effectively promote bone matrix repair, improve bone density, enhance antioxidant capacity, and enhance bone calcium deposition. Therefore, highly dispersed collagen peptides and composite-encapsulated nano-calcium are indispensable synergistic core components in the preparation of this collagen composition.

[0117] Although embodiments of this application have been shown and described, it will be understood by those skilled in the art that various changes, modifications, substitutions and variations can be made to these embodiments without departing from the principles and spirit of this application, the scope of which is defined by the appended claims and their equivalents.

Claims

1. A collagen peptide composition for repairing joints, characterized in that, It is composed of the following raw materials in parts by weight: 20-25 parts of highly dispersed collagen peptides, 10-15 parts of chondroitin sulfate, 4-6 parts of Ganoderma lucidum extract, 4-6 parts of shiitake mushroom extract, 4-6 parts of composite encapsulated nano-calcium, 2-4 parts of hyaluronic acid, 0.4-0.6 parts of vitamin K2, 0.2-0.4 parts of tea polyphenols, 5-7 parts of dragon fruit fermented pulp, 4-6 parts of maltodextrin, and 0.1-0.2 parts of ε-polylysine; The preparation method of the highly dispersed collagen peptides is as follows: Step S1: Add bovine bone collagen peptides to deionized water and stir at 40-50℃ for 20-25 minutes. Then add activated carbon and continue stirring for 30-40 minutes. After cooling to room temperature, filter to obtain the filtrate. Step S2: Add the complex protease to the filtrate and hydrolyze it for 2-2.5 hours at 50-55℃ and pH 7.0-7.

5. Then, cool it to room temperature and separate the permeate through an ultrafiltration membrane. The complex protease is composed of alkaline protease and papain in a mass ratio of 2.5-3.5:

1. Step S3: Concentrate the permeate, add chitosan, and react at 55-65℃ and pH 7.8-8.2 for 1.5-2 hours. After post-treatment, highly dispersed collagen peptides are obtained. The preparation method of the composite-encapsulated calcium nanoparticles is as follows: Step a: First, add casein phosphopeptide and sodium alginate to deionized water and stir for 30-40 minutes at 55-60℃ and pH 6.5-7.0 to obtain a composite encapsulation solution. Then add calcium carbonate and stir for 60-70 minutes at 40-45℃. After post-treatment, the initial encapsulated nano-calcium is obtained. Step b: Add hydroxypropyl-β-cyclodextrin to deionized water and stir at 60-65℃ for 20-30 min to obtain hydroxypropyl-β-cyclodextrin solution. Then cool to 35-40℃, add the initially encapsulated nano-calcium, stir at constant temperature for 90-100 min, and obtain composite encapsulated nano-calcium after post-treatment. The dragon fruit fermented pulp is made by blending red dragon fruit fermentation liquid and galactooligosaccharides in a mass ratio of 10:3-6.

2. The collagen peptide composition for repairing joints according to claim 1, characterized in that, The bovine collagen peptides added to deionized water in step S1 have a solution concentration of 7-9%, and the mass ratio of bovine collagen peptides to activated carbon is 1:0.0015-0.0025.

3. The collagen peptide composition for repairing joints according to claim 1, characterized in that, The mass-to-volume ratio of the complex protease and the filtrate in step S2 is 0.09-0.1 g / 100 mL.

4. The collagen peptide composition for repairing joints according to claim 1, characterized in that, In step S3, the permeate is concentrated to a solid content of 9-11%, and the mass-to-volume ratio of chitosan to permeate is 0.035-0.045 g / 100 mL.

5. The collagen peptide composition for repairing joints according to claim 1, characterized in that, In step a, the mass ratio of casein phosphopeptide to sodium alginate is 1.5-2.5:1, the amount of sodium alginate added to deionized water is 1-1.5 g / 100 mL, and the mass ratio of calcium carbonate to composite embedding solution is 1:5-6.

6. The collagen peptide composition for repairing joints according to claim 1, characterized in that, The mass concentration of the hydroxypropyl-β-cyclodextrin solution in step b is 9.5-10.5%, and the mass ratio of the initially encapsulated nano-calcium to hydroxypropyl-β-cyclodextrin is 2.5-3.5:

7.

7. The method for preparing the collagen peptide composition for joint repair according to any one of claims 1-6, characterized in that, Includes the following steps: (1) The highly dispersed collagen peptides, chondroitin sulfate, Ganoderma lucidum extract, shiitake mushroom extract, composite encapsulated nano-calcium, hyaluronic acid, vitamin K2, tea polyphenols and ε-polylysine were sieved separately and set aside for later use; (2) Add maltodextrin to deionized water, heat to 60-65℃ and stir to dissolve. After cooling to 30-35℃, add tea polyphenols, ε-polylysine, highly dispersed collagen peptides and chondroitin sulfate in sequence. Stir at 500-600 r / min for 20-30 min, add dragon fruit fermentation pulp and continue stirring for 10-15 min to form mixture A. (3) Add Ganoderma lucidum extract, shiitake mushroom extract and hyaluronic acid to mixture A, and continue stirring for 20-30 minutes to form mixture B. Mix vitamin K2 and composite encapsulated nano-calcium evenly and add it to mixture B. After homogenization, the mixture is obtained.

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