A composition for improving sarcopenia and a method of preparing the same

Abstract

The application discloses a composition for improving sarcopenia and a preparation method thereof. The composition comprises functional ingredients such as concentrated whey protein, curcumin, soybean peptide, branched-chain amino acid, calcium beta-hydroxy-beta-methylbutyrate (HMB-Ca) and fermented black tomato extract. The application adopts a composite fermentation system to improve the content of nicotinamide in the fermentation product, fully release active ingredients, and improve the stability and bioavailability of functional substances through HMB-Ca, concentrated whey protein and capsaicin. The composition has important application value for the prevention, improvement and rehabilitation of sarcopenia through multi-target synergistic effect, which comprehensively covers the key links of sarcopenia, including promoting protein synthesis, inhibiting protein decomposition, antioxidant and anti-inflammatory, inhibiting accumulation of harmful metabolites and improving neuromuscular function.

Description

Technical Field

[0001] This invention relates to the field of functional food technology, and more specifically to a method for preparing a composition that improves sarcopenia. Background Technology

[0002] Sarcopenia is a syndrome characterized by progressive reduction in skeletal muscle mass, decreased muscle strength, and impaired physical function, caused by a combination of multiple factors. Its core pathogenesis includes protein metabolism imbalance, neuronal degeneration, satellite cell dysfunction, inflammation and oxidative stress, and decreased hormone levels. It is also accompanied by the accumulation of harmful metabolites such as methylmalonic acid, total homocysteine, and kynurenine, as well as nicotinamide deficiency, severely impacting the quality of life of middle-aged and elderly individuals, with a particularly high prevalence among those over 60 years of age. Existing sarcopenia intervention products mostly target single pathogenic factors, resulting in limited effectiveness.

[0003] Sarcopenia is the result of multiple harmful factors working together, mainly including the following key links: protein metabolism imbalance, which is the core mechanism of sarcopenia; insufficient branched-chain amino acid intake, especially insufficient intake of leucine, which leads to the blockage of mTOR signaling and the inability to effectively initiate protein synthesis; weakened neuromuscular function and neuronal degeneration; satellite cell dysfunction, satellite cells are muscle stem cells, responsible for repairing damaged muscles and promoting muscle growth, and the activity of satellite cells decreases in the elderly, making them unable to effectively differentiate into mature muscle fibers; as well as inflammation and oxidative stress, accumulation of reactive oxygen species (ROS), and decreased hormone levels.

[0004] Curcumin, a natural polyphenolic compound extracted from turmeric, has shown clear potential value in the treatment of sarcopenia. Its mechanism of action is mainly achieved through several aspects: First, curcumin has a strong antioxidant capacity, which can improve the redox imbalance in aging skeletal muscle by increasing Nrf2 expression levels, activating the Keap1-Nrf2-ARE signaling pathway, and quenching reactive oxygen species through phenolic -OH groups; Second, it exerts an anti-inflammatory effect by activating Nrf2 to upregulate HO-1, inhibiting the NF-κB signaling pathway, reducing the accumulation of inflammatory factors such as IL-6 and TNF-α, and maintaining protein synthesis and metabolic homeostasis; Third, curcumin can regulate muscle protein metabolism by downregulating the expression levels of Atrogin-1 and MuRF-1 and upregulating the phosphorylation level of AKT protein in the mTOR signaling pathway, thereby promoting muscle protein synthesis and inhibiting protein breakdown; In addition, it can protect mitochondrial function, maintain normal mitochondria in muscle cells, and promote mitochondrial biogenesis by increasing cAMP expression levels; At the same time, curcumin can improve satellite cell function, increase the proportion of MyoD-positive satellite cells, and promote muscle regeneration. Animal studies have confirmed that curcumin can effectively prevent the development of sarcopenia, improve survival rates, and maintain muscle fiber size. However, more large-scale randomized controlled trials are needed in human clinical trials to determine the optimal route of administration, dosage, and duration of treatment. Despite challenges such as low bioavailability, curcumin, as a natural compound, remains a potential alternative therapy for sarcopenia.

[0005] Black tomatoes are rich in anthocyanins (delphinidin and its glycosides), lycopene, total phenols, chlorogenic acid, vitamin C, vitamin E, selenium, nitrates, and various amino acids. Based on this, fermentation with compound probiotics generates small-molecule active substances with core functions such as strong antioxidant, anti-inflammatory, mitochondrial protection, and regulation of muscle protein synthesis and degradation balance. Incorporating concentrated whey protein, curcumin, soy peptides, and other functional ingredients, a complex delivery system of capsaicin, β-hydroxy-β-methylbutyrate (HMB-Ca), and concentrated whey protein is developed. This allows for the creation of a nutritional supplement that simultaneously regulates muscle metabolism, inhibits the formation of harmful substances, and promotes nutrient absorption through multiple improvements, while simultaneously enhancing the stability and delivery efficiency of functional substances. Summary of the Invention

[0006] The technical problem to be solved: Current intervention products for sarcopenia mostly target a single pathogenesis stage, failing to cover the multiple pathogenesis stages of sarcopenia, thus limiting their effectiveness. This invention provides a composition that can improve sarcopenia with multiple targets and high efficiency. Through encapsulation and synergistic effects of multiple substances, it enhances the stability and delivery efficiency of functional substances, thereby improving bioavailability.

[0007] Technical solution: A composition for improving sarcopenia, comprising, by weight parts: 0.5-5 parts curcumin, 5-20 parts soybean peptides, 3-15 parts branched-chain amino acids, 10-30 parts concentrated whey protein, 10-30 parts HMB-Ca, 2-6 parts capsaicin, and 50-100 parts fermented black tomato extract.

[0008] Furthermore, the branched-chain amino acid is a mixture of leucine, isoleucine and valine in a mass ratio of 2:1:1.

[0009] Furthermore, the average molecular weight of the soybean peptide is less than 3000 Da.

[0010] Furthermore, the preparation method of the fermented black tomato extract is as follows: black tomato powder is mixed with sterile water, glucose, peptone, corn steep liquor and antimicrobial peptides are added, the pH value is adjusted, a compound probiotic suspension is inoculated, anaerobic fermentation is carried out, the pH value is adjusted again, fermentation is carried out under aerobic conditions, after inactivation, the supernatant is separated by centrifugation, and filtered through an ultrafiltration membrane to obtain the fermented black tomato extract.

[0011] Furthermore, in the preparation method of the fermented black tomato extract, the mass ratio of black tomato fruit powder to sterile water is 1:(5-10); the pH is adjusted to 6.0-7.0; Furthermore, the compound probiotic suspension contains Lactobacillus plantarum, Bifidobacterium, Saccharomyces cerevisiae, and Bacillus subtilis, and the inoculum amount of the compound probiotic suspension is 5-10 wt.%. Furthermore, the anaerobic fermentation temperature is 35-37℃, and the time is 24-36h; the pH value is adjusted again to 7.5-8.5; the aerobic fermentation temperature is 35-37℃, and the time is 24-36h. Furthermore, the amounts of glucose, peptone, corn steep liquor, and antimicrobial peptide added are 2-5 wt.%, 1-3 wt.%, 0.5-2 wt.%, and 0.05-0.1 wt.% of the black tomato fruit powder, respectively.

[0012] Furthermore, the concentration of *Lactobacillus plantarum* in the compound probiotic suspension is 1×10⁻⁶. 6 -1×10 8 CFU / mL, Bifidobacterium concentration 1×10 6 -1×10 8 CFU / mL, Saccharomyces cerevisiae concentration 1×10 5 -1×10 7 CFU / mL, Bacillus subtilis concentration was 1×10⁻⁶ 5 -1×10 7 CFU / mL.

[0013] The preparation method of the composition for improving sarcopenia described above includes the following steps: S1. Mix curcumin, soybean peptides, branched-chain amino acids and fermented black tomato extract to obtain a mixed functional liquid. Under constant temperature and stirring, slowly add concentrated whey protein and HMB-Ca, stir and mix, slowly add capsaicin solution, continue stirring, adjust the pH value, and homogenize to obtain a composite solution. S2. Dry the composite solution at low temperature, sieve it, add auxiliary raw materials and mix evenly to form the formulation.

[0014] Furthermore, in step S1, the solvent for the capsaicin solution is ethanol with a concentration of 10-15 wt.%; the temperature for constant temperature stirring is 35-40℃; the stirring speed is 500-800 rpm; the stirring and mixing time is 20-30 min; the stirring time is continued for 40-60 min; the pH value is adjusted to 7.0-7.5; and the homogenization pressure is 15-25 MPa.

[0015] Furthermore, the low-temperature drying temperature in step S2 is 45-60℃; the auxiliary raw materials are maltitol, microcrystalline cellulose, vitamin D, and sodium carboxymethyl cellulose.

[0016] The above-described composition for improving sarcopenia is used in the preparation of a medicament for the prevention or treatment of sarcopenia. Beneficial effects

[0017] This invention achieves comprehensive coverage of key aspects of sarcopenia pathogenesis through the scientific formulation of functional components. Branched-chain amino acids activate the mTOR signaling pathway, promoting muscle protein synthesis; HMB-Ca inhibits protein breakdown; soybean peptides provide a high-quality nitrogen source; curcumin improves the redox imbalance in aging skeletal muscle and exerts anti-inflammatory effects; in addition, substances such as fatty acids produced during fermentation may have an IDO inhibitory effect, reducing kynurenine accumulation and increasing muscle fiber size and muscle strength; capsaicin regulates neuromuscular function through TRPV1 receptors, and also improves the intestinal absorption of functional substances by inhibiting metabolic enzymes and transport proteins; the effect of improving sarcopenia is superior to that of a single component.

[0018] In this invention, curcumin demonstrates clear potential value in the treatment of sarcopenia. It can improve the redox imbalance in aging skeletal muscle by regulating signaling pathways and quenching reactive oxygen species through phenolic -OH groups. Furthermore, it reduces the accumulation of inflammatory factors such as IL-6 and TNF-α, maintains protein synthesis and metabolic homeostasis, regulates muscle protein metabolism, maintains normal mitochondria in muscle cells, and promotes muscle regeneration.

[0019] The raw materials of this invention are all natural food-grade ingredients, without any chemically synthesized additives. The fermentation process is mild and there are no irritating additives, making it suitable for long-term use. The composition improves muscle atrophy, enhances muscle strength and physical function, while supplementing high-quality protein, peptides, amino acids, vitamins and various plant active substances, addressing multiple health issues in the elderly such as nutritional deficiencies, muscle loss, and chronic low-grade inflammation.

[0020] This invention employs a complex fermentation system of Lactobacillus plantarum, Bifidobacterium, Saccharomyces cerevisiae, and Bacillus subtilis, which significantly increases nicotinamide production. The multiple microorganisms synergistically secrete proteases and cellulases, which efficiently degrade the cell walls of black tomatoes, fully releasing endogenous active ingredients such as anthocyanins, lycopene, and chlorogenic acid, producing functional secondary metabolites, reducing oxidative damage to muscle cells, and inhibiting the NF-κB pathway to alleviate chronic inflammatory responses. The nicotinamide produced during fermentation can reduce the risk of sarcopenia and improve muscle strength. The overall abundance of functional substances is far higher than that of traditional water extraction and alcohol extraction processes.

[0021] This invention constructs a novel composite delivery system of capsaicin-HMB-Ca-concentrated whey protein. It utilizes the film-forming properties of concentrated whey protein, the stability of HMB-Ca, and the enhanced bioavailability of capsaicin to effectively encapsulate active ingredients, as well as functional substances such as curcumin and soybean peptides, thereby reducing the degradation of functional substances during processing, storage, and in the gastrointestinal tract. Detailed Implementation

[0022] The present invention will be further described below with reference to embodiments. These embodiments are illustrative of the present invention, but the present invention is not limited to these embodiments: Example 1

[0023] A composition for improving sarcopenia, comprising, by weight: 3 parts curcumin, 10 parts soybean peptides, 10 parts branched-chain amino acids, 20 parts whey protein concentrate, 20 parts HMB-Ca, 3 parts capsaicin, and 80 parts fermented black tomato extract.

[0024] A method for preparing a composition for improving sarcopenia includes the following steps: S1. Select black tomatoes, disinfect them by rinsing, dry them at low temperature, pulverize them into ultrafine powder, and sieve them to obtain black tomato fruit powder. Mix 10g of black tomato fruit powder with 100g of sterile water, add 0.3g of glucose, 0.3g of peptone, 0.15g of corn steep liquor, and 0.08g of antimicrobial peptide, adjust the pH to 6.0, and inoculate with 8g of a compound probiotic suspension. The concentration of Lactobacillus plantarum in the compound probiotic suspension is 1×10⁻⁶. 6 -1×10 8 CFU / mL, Bifidobacterium concentration 1×10 6 -1×10 8 CFU / mL, Saccharomyces cerevisiae concentration 1×105 -1×10 7 CFU / mL, Bacillus subtilis concentration was 1×10⁻⁶ 5 -1×10 7 CFU / mL, anaerobic fermentation at 37℃ for 24h, pH adjusted to 8.0, fermentation at 37℃ under aerobic conditions for 36h, inactivation at 85℃ for 15min, then centrifugation at 8000rpm for 15min to separate the supernatant, and filtration through an ultrafiltration membrane to obtain fermented black tomato extract. S2. Fermented black tomato extract was mixed with curcumin, soybean peptides with a molecular weight of less than 3000 Da, and branched-chain amino acids to obtain a mixed functional liquid. The branched-chain amino acids were a mixture of leucine, isoleucine, and valine in a mass ratio of 2:1:1. The mixed functional liquid was stirred at 800 rpm under constant temperature of 35℃, and concentrated whey protein and HMB-Ca were slowly added and stirred for 20-30 min. Capsaicin was dissolved in ethanol to prepare a 10 wt.% capsaicin solution. The capsaicin solution was slowly added dropwise and stirred for 40 min. The pH was adjusted to 7.0 and homogenized at 20 MPa to obtain a composite solution. S3. Dry the composite solution at a low temperature of 45°C, sieve it, add maltitol, microcrystalline cellulose, vitamin D, and sodium carboxymethyl cellulose, mix evenly, and form the formulation. Example 2

[0025] A composition for improving sarcopenia, comprising, by weight: 3 parts curcumin, 10 parts soybean peptides, 10 parts branched-chain amino acids, 20 parts whey protein concentrate, 20 parts HMB-Ca, 3 parts capsaicin, and 80 parts fermented black tomato extract.

[0026] A method for preparing a composition for improving sarcopenia includes the following steps: S1. Select black tomatoes, disinfect them by rinsing, dry them at low temperature, pulverize them into ultrafine powder, and sieve them to obtain black tomato fruit powder. Mix 10g of black tomato fruit powder with 80g of sterile water, add 0.3g of glucose, 0.3g of peptone, 0.15g of corn steep liquor, and 0.08g of antimicrobial peptide, adjust the pH to 6.0, and inoculate with 8g of a compound probiotic suspension. The concentration of Lactobacillus plantarum in the compound probiotic suspension is 1×10⁻⁶. 6 -1×10 8 CFU / mL, Bifidobacterium concentration 1×10 6 -1×10 8 CFU / mL, Saccharomyces cerevisiae concentration 1×10 5 -1×10 7 CFU / mL, Bacillus subtilis concentration was 1×10⁻⁶ 5 -1×10 7CFU / mL, anaerobic fermentation at 37℃ for 24h, pH adjusted to 8.0, fermentation at 37℃ under aerobic conditions for 36h, inactivation at 85℃ for 15min, then centrifugation at 8000rpm for 15min to separate the supernatant, and filtration through an ultrafiltration membrane to obtain fermented black tomato extract. S2. Fermented black tomato extract was mixed with curcumin, soybean peptides with a molecular weight of less than 3000 Da, and branched-chain amino acids to obtain a mixed functional liquid. The branched-chain amino acids were a mixture of leucine, isoleucine, and valine in a mass ratio of 2:1:1. The mixed functional liquid was stirred at 800 rpm under constant temperature of 35℃, and concentrated whey protein and HMB-Ca were slowly added and stirred for 20-30 min. Capsaicin was dissolved in ethanol to prepare a 10 wt.% capsaicin solution. The capsaicin solution was slowly added dropwise and stirred for 40 min. The pH was adjusted to 7.0 and homogenized at 20 MPa to obtain a composite solution. S3. Dry the composite solution at a low temperature of 45°C, sieve it, add maltitol, microcrystalline cellulose, vitamin D, and sodium carboxymethyl cellulose, mix evenly, and form the formulation. Example 3

[0027] A composition for improving sarcopenia, comprising, by weight: 3 parts curcumin, 10 parts soybean peptides, 10 parts branched-chain amino acids, 20 parts whey protein concentrate, 20 parts HMB-Ca, 3 parts capsaicin, and 80 parts fermented black tomato extract.

[0028] A method for preparing a composition for improving sarcopenia includes the following steps: S1. Select black tomatoes, disinfect them by rinsing, dry them at low temperature, pulverize them into ultrafine powder, and sieve them to obtain black tomato fruit powder. Mix 10g of black tomato fruit powder with 100g of sterile water, add 0.3g of glucose, 0.3g of peptone, 0.15g of corn steep liquor, and 0.08g of antimicrobial peptides, adjust the pH to 6.0, and inoculate with 5g of a compound probiotic suspension. The concentration of Lactobacillus plantarum in the compound probiotic suspension is 1×10⁻⁶. 6 -1×10 8 CFU / mL, Bifidobacterium concentration 1×10 6 -1×10 8 CFU / mL, Saccharomyces cerevisiae concentration 1×10 5 -1×10 7 CFU / mL, Bacillus subtilis concentration was 1×10⁻⁶ 5 -1×10 7 CFU / mL, anaerobic fermentation at 37℃ for 24h, pH adjusted to 8.0, fermentation at 37℃ under aerobic conditions for 36h, inactivation at 85℃ for 15min, then centrifugation at 8000rpm for 15min to separate the supernatant, and filtration through an ultrafiltration membrane to obtain fermented black tomato extract. S2. Fermented black tomato extract was mixed with curcumin, soybean peptides with a molecular weight of less than 3000 Da, and branched-chain amino acids. The branched-chain amino acids were a mixture of leucine, isoleucine, and valine in a mass ratio of 2:1:1. The mixed functional solution was obtained. Under constant temperature of 35℃ and stirring at 800 rpm, concentrated whey protein and HMB-Ca were slowly added and stirred for 20-30 min. Capsaicin was dissolved in ethanol to prepare a 10 wt.% capsaicin solution. The capsaicin solution was slowly added dropwise and stirred for another 40 min. The pH was adjusted to 7.0 and homogenized at 20 MPa to obtain a composite solution. S3. Dry the composite solution at a low temperature of 45°C, sieve it, add maltitol, microcrystalline cellulose, vitamin D, and sodium carboxymethyl cellulose, mix evenly, and form the formulation. Example 4

[0029] A composition for improving sarcopenia, comprising, by weight: 3 parts curcumin, 10 parts soybean peptides, 10 parts branched-chain amino acids, 20 parts whey protein concentrate, 20 parts HMB-Ca, 3 parts capsaicin, and 80 parts fermented black tomato extract.

[0030] A method for preparing a composition for improving sarcopenia includes the following steps: S1. Select black tomatoes, disinfect them by rinsing, dry them at low temperature, pulverize them into ultrafine powder, and sieve them to obtain black tomato fruit powder. Mix 10g of black tomato fruit powder with 100g of sterile water, add 0.3g of glucose, 0.3g of peptone, 0.15g of corn steep liquor, and 0.08g of antimicrobial peptide, adjust the pH to 6.0, and inoculate with 8g of a compound probiotic suspension. The concentration of Lactobacillus plantarum in the compound probiotic suspension is 1×10⁻⁶. 6 -1×10 8 CFU / mL, Bifidobacterium concentration 1×10 6 -1×10 8 CFU / mL, Saccharomyces cerevisiae concentration 1×10 5 -1×10 7 CFU / mL, Bacillus subtilis concentration was 1×10⁻⁶ 5 -1×10 7 CFU / mL, anaerobic fermentation at 37℃ for 24h, pH adjusted to 8.0, fermentation at 37℃ under aerobic conditions for 24h, inactivation at 85℃ for 15min, then centrifugation at 8000rpm for 15min to separate the supernatant, and filtration through an ultrafiltration membrane to obtain fermented black tomato extract. S2. Fermented black tomato extract was mixed with curcumin, soybean peptides with a molecular weight of less than 3000 Da, and branched-chain amino acids. The branched-chain amino acids were a mixture of leucine, isoleucine, and valine in a mass ratio of 2:1:1. The mixed functional solution was obtained. Under constant temperature of 35℃ and stirring at 800 rpm, concentrated whey protein and HMB-Ca were slowly added and stirred for 20-30 min. Capsaicin was dissolved in ethanol to prepare a 10 wt.% capsaicin solution. The capsaicin solution was slowly added dropwise and stirred for another 40 min. The pH was adjusted to 7.0 and homogenized at 20 MPa to obtain a composite solution. S3. Dry the composite solution at a low temperature of 45°C, sieve it, add maltitol, microcrystalline cellulose, vitamin D, and sodium carboxymethyl cellulose, mix evenly, and form the formulation. Example 5

[0031] A composition for improving sarcopenia, comprising, by weight: 3 parts curcumin, 10 parts soybean peptides, 10 parts branched-chain amino acids, 20 parts whey protein concentrate, 20 parts HMB-Ca, 3 parts capsaicin, and 100 parts fermented black tomato extract.

[0032] A method for preparing a composition for improving sarcopenia includes the following steps: S1. Select black tomatoes, disinfect them by rinsing, dry them at low temperature, pulverize them into ultrafine powder, and sieve them to obtain black tomato fruit powder. Mix 10g of black tomato fruit powder with 100g of sterile water, add 0.3g of glucose, 0.3g of peptone, 0.15g of corn steep liquor, and 0.08g of antimicrobial peptide, adjust the pH to 6.0, and inoculate with 8g of a compound probiotic suspension. The concentration of Lactobacillus plantarum in the compound probiotic suspension is 1×10⁻⁶. 6 -1×10 8 CFU / mL, Bifidobacterium concentration 1×10 6 -1×10 8 CFU / mL, Saccharomyces cerevisiae concentration 1×10 5 -1×10 7 CFU / mL, Bacillus subtilis concentration was 1×10⁻⁶ 5 -1×10 7 CFU / mL, anaerobic fermentation at 37℃ for 24h, pH adjusted to 8.0, fermentation at 37℃ under aerobic conditions for 36h, inactivation at 85℃ for 15min, then centrifugation at 8000rpm for 15min to separate the supernatant, and filtration through an ultrafiltration membrane to obtain fermented black tomato extract. S2. Fermented black tomato extract was mixed with curcumin, soybean peptides with a molecular weight of less than 3000 Da, and branched-chain amino acids to obtain a mixed functional liquid. The branched-chain amino acids were a mixture of leucine, isoleucine, and valine in a mass ratio of 2:1:1. The mixed functional liquid was stirred at 800 rpm under constant temperature of 35℃, and concentrated whey protein and HMB-Ca were slowly added and stirred for 20-30 min. Capsaicin was dissolved in ethanol to prepare a 10 wt.% capsaicin solution. The capsaicin solution was slowly added dropwise and stirred for 40 min. The pH was adjusted to 7.0 and homogenized at 20 MPa to obtain a composite solution. S3. Dry the composite solution at a low temperature of 45°C, sieve it, add maltitol, microcrystalline cellulose, vitamin D, and sodium carboxymethyl cellulose, mix evenly, and form the formulation. Example 6

[0033] A composition for improving sarcopenia, comprising, by weight parts: 1 part curcumin, 15 parts soybean peptides, 10 parts branched-chain amino acids, 20 parts whey protein concentrate, 20 parts HMB-Ca, 3 parts capsaicin, and 80 parts fermented black tomato extract.

[0034] A method for preparing a composition for improving sarcopenia includes the following steps: S1. Select black tomatoes, disinfect them by rinsing, dry them at low temperature, pulverize them into ultrafine powder, and sieve them to obtain black tomato fruit powder. Mix 10g of black tomato fruit powder with 100g of sterile water, add 0.3g of glucose, 0.3g of peptone, 0.15g of corn steep liquor, and 0.08g of antimicrobial peptide, adjust the pH to 6.0, and inoculate with 8g of a compound probiotic suspension. The concentration of Lactobacillus plantarum in the compound probiotic suspension is 1×10⁻⁶. 6 -1×10 8 CFU / mL, Bifidobacterium concentration 1×10 6 -1×10 8 CFU / mL, Saccharomyces cerevisiae concentration 1×10 5 -1×10 7 CFU / mL, Bacillus subtilis concentration was 1×10⁻⁶ 5 -1×10 7 CFU / mL, anaerobic fermentation at 37℃ for 24h, pH adjusted to 8.0, fermentation at 37℃ under aerobic conditions for 36h, inactivation at 85℃ for 15min, then centrifugation at 8000rpm for 15min to separate the supernatant, and filtration through an ultrafiltration membrane to obtain fermented black tomato extract. S2. Fermented black tomato extract was mixed with curcumin, soybean peptides with a molecular weight of less than 3000 Da, and branched-chain amino acids to obtain a mixed functional liquid. The branched-chain amino acids were a mixture of leucine, isoleucine, and valine in a mass ratio of 2:1:1. The mixed functional liquid was stirred at 800 rpm under constant temperature of 35℃, and concentrated whey protein and HMB-Ca were slowly added and stirred for 20-30 min. Capsaicin was dissolved in ethanol to prepare a 10 wt.% capsaicin solution. The capsaicin solution was slowly added dropwise and stirred for 40 min. The pH was adjusted to 7.0 and homogenized at 20 MPa to obtain a composite solution. S3. Dry the composite solution at a low temperature of 45°C, sieve it, add maltitol, microcrystalline cellulose, vitamin D, and sodium carboxymethyl cellulose, mix evenly, and form the formulation. Comparative Example 1

[0035] The difference between this comparative example and Example 1 is that fermentation is not performed, as detailed below: A composition for improving sarcopenia, comprising, by weight: 3 parts curcumin, 10 parts soybean peptides, 10 parts branched-chain amino acids, 20 parts whey protein concentrate, 20 parts HMB-Ca, 3 parts capsaicin, and 80 parts black tomato extract.

[0036] S1. Select black tomatoes, disinfect them by rinsing, dry them at low temperature, pulverize them into ultrafine powder, and sieve them to obtain black tomato fruit powder. Mix 10g of black tomato fruit powder with 100g of sterile water, add 0.3g of glucose, 0.3g of peptone, 0.15g of corn steep liquor and 0.08g of antimicrobial peptide, react in a water bath for 60min, then centrifuge at 8000rpm for 15min to separate the supernatant, filter it through an ultrafiltration membrane to obtain black tomato extract; S2. Black tomato extract was mixed with curcumin, soybean peptides with a molecular weight of less than 3000 Da, and branched-chain amino acids to obtain a mixed functional liquid. The branched-chain amino acids were a mixture of leucine, isoleucine, and valine in a mass ratio of 2:1:1. The mixed functional liquid was stirred at 800 rpm under constant temperature of 35℃, and concentrated whey protein and HMB-Ca were slowly added and stirred for 20-30 min. Capsaicin was dissolved in ethanol to prepare a 10 wt.% capsaicin solution. The capsaicin solution was slowly added dropwise and stirred for 40 min. The pH was adjusted to 7.0 and homogenized at 20 MPa to obtain a composite solution. S3. Dry the composite solution at a low temperature of 45°C, sieve it, add maltitol, microcrystalline cellulose, vitamin D, and sodium carboxymethyl cellulose, mix evenly, and form the formulation. Comparative Example 2

[0037] The difference between this comparative example and Example 1 is that Bacillus subtilis is not used, as detailed below: A composition for improving sarcopenia, comprising, by weight: 3 parts curcumin, 10 parts soybean peptides, 10 parts branched-chain amino acids, 20 parts whey protein concentrate, 20 parts HMB-Ca, 3 parts capsaicin, and 80 parts fermented black tomato extract.

[0038] S1. Select black tomatoes, disinfect them by rinsing, dry them at low temperature, pulverize them into ultrafine powder, and sieve them to obtain black tomato fruit powder. Mix 10g of black tomato fruit powder with 100g of sterile water, add 0.3g of glucose, 0.3g of peptone, 0.15g of corn steep liquor, and 0.08g of antimicrobial peptide, adjust the pH to 6.0, and inoculate with 8g of a compound probiotic suspension. The concentration of Lactobacillus plantarum in the compound probiotic suspension is 1×10⁻⁶. 6 -1×10 8 CFU / mL, Bifidobacterium concentration 1×10 6 -1×10 8 CFU / mL, Saccharomyces cerevisiae concentration 1×10 5 -1×10 7 CFU / mL, anaerobic fermentation at 37℃ for 24h, pH adjusted to 8.0, fermentation at 37℃ under aerobic conditions for 36h, inactivation at 85℃ for 15min, then centrifugation at 8000rpm for 15min to separate the supernatant, and filtration through an ultrafiltration membrane to obtain fermented black tomato extract. S2. Fermented black tomato extract was mixed with curcumin, soybean peptides with a molecular weight of less than 3000 Da, and branched-chain amino acids to obtain a mixed functional liquid. The branched-chain amino acids were a mixture of leucine, isoleucine, and valine in a mass ratio of 2:1:1. The mixed functional liquid was stirred at 800 rpm under constant temperature of 35℃, and concentrated whey protein and HMB-Ca were slowly added and stirred for 20-30 min. Capsaicin was dissolved in ethanol to prepare a 10 wt.% capsaicin solution. The capsaicin solution was slowly added dropwise and stirred for 40 min. The pH was adjusted to 7.0 and homogenized at 20 MPa to obtain a composite solution. S3. Dry the composite solution at a low temperature of 45°C, sieve it, add maltitol, microcrystalline cellulose, vitamin D, and sodium carboxymethyl cellulose, mix evenly, and form the formulation. Comparative Example 3

[0039] The difference between this comparative example and Example 1 is that the ingredients are mixed together, as detailed below: A composition for improving sarcopenia, comprising, by weight: 3 parts curcumin, 10 parts soybean peptides, 10 parts branched-chain amino acids, 20 parts whey protein concentrate, 20 parts HMB-Ca, 3 parts capsaicin, and 80 parts fermented black tomato extract.

[0040] S1. Select black tomatoes, disinfect them by rinsing, dry them at low temperature, pulverize them into ultrafine powder, and sieve them to obtain black tomato fruit powder. Mix 10g of black tomato fruit powder with 100g of sterile water, add 0.3g of glucose, 0.3g of peptone, 0.15g of corn steep liquor, and 0.08g of antimicrobial peptide, adjust the pH to 6.0, and inoculate with 8g of a compound probiotic suspension. The concentration of Lactobacillus plantarum in the compound probiotic suspension is 1×10⁻⁶. 6 -1×10 8 CFU / mL, Bifidobacterium concentration 1×10 6 -1×10 8 CFU / mL, Saccharomyces cerevisiae concentration 1×10 5 -1×10 7 CFU / mL, Bacillus subtilis concentration was 1×10⁻⁶ 5 -1×10 7 CFU / mL, anaerobic fermentation at 37℃ for 24h, pH adjusted to 8.0, fermentation at 37℃ under aerobic conditions for 36h, inactivation at 85℃ for 15min, then centrifugation at 8000rpm for 15min to separate the supernatant, and filtration through an ultrafiltration membrane to obtain fermented black tomato extract. S2. Fermented black tomato extract, curcumin, soybean peptides with a molecular weight of less than 3000 Da, branched-chain amino acids, concentrated whey protein, HMB-Ca, and capsaicin solution are stirred and mixed for 20-30 minutes. Capsaicin is dissolved in ethanol to prepare a 10 wt.% capsaicin solution. The pH value is adjusted to 7.0, and the solution is homogenized at 20 MPa to obtain a composite solution. S3. Dry the composite solution at a low temperature of 45°C, sieve it, add maltitol, microcrystalline cellulose, vitamin D, and sodium carboxymethyl cellulose, mix evenly, and form the formulation. Comparative Example 4

[0041] The difference between this comparative example and Example 1 is that calcium β-hydroxy-β-methylbutyrate (HMB-Ca) is not added, as detailed below: A composition for improving sarcopenia, comprising, by weight parts: 3 parts curcumin, 10 parts soybean peptides, 10 parts branched-chain amino acids, 20 parts whey protein concentrate, 3 parts capsaicin, and 80 parts black tomato extract.

[0042] S1. Select black tomatoes, disinfect them by rinsing, dry them at low temperature, pulverize them into ultrafine powder, and sieve them to obtain black tomato fruit powder. Mix 10g of black tomato fruit powder with 100g of sterile water, add 0.3g of glucose, 0.3g of peptone, 0.15g of corn steep liquor, and 0.08g of antimicrobial peptide, adjust the pH to 6.0, and inoculate with 8g of a compound probiotic suspension. The concentration of Lactobacillus plantarum in the compound probiotic suspension is 1×10⁻⁶. 6 -1×10 8 CFU / mL, Bifidobacterium concentration 1×10 6-1×10 8 CFU / mL, Saccharomyces cerevisiae concentration 1×10 5 -1×10 7 CFU / mL, Bacillus subtilis concentration was 1×10⁻⁶ 5 -1×10 7 CFU / mL, anaerobic fermentation at 37℃ for 24h, pH adjusted to 8.0, fermentation at 37℃ under aerobic conditions for 36h, inactivation at 85℃ for 15min, then centrifugation at 8000rpm for 15min to separate the supernatant, and filtration through an ultrafiltration membrane to obtain fermented black tomato extract. S2. Fermented black tomato extract was mixed with curcumin, soybean peptides with a molecular weight of less than 3000 Da, and branched-chain amino acids to obtain a mixed functional liquid. The branched-chain amino acids were a mixture of leucine, isoleucine, and valine in a mass ratio of 2:1:1. The mixed functional liquid was stirred at 800 rpm at a constant temperature of 35°C, and concentrated whey protein was slowly added and stirred for 20-30 minutes. Capsaicin was dissolved in ethanol to prepare a 10 wt.% capsaicin solution. The capsaicin solution was slowly added dropwise and stirred for another 40 minutes. The pH was adjusted to 7.0 and homogenized at 20 MPa to obtain a composite solution. S3. Dry the composite solution at a low temperature of 45°C, sieve it, add maltitol, microcrystalline cellulose, vitamin D, and sodium carboxymethyl cellulose, mix evenly, and form the formulation. Comparative Example 5

[0043] The difference between this comparative example and Example 1 is that capsaicin is not added, as detailed below: A composition for improving sarcopenia, comprising, by weight: 3 parts curcumin, 10 parts soy peptides, 10 parts branched-chain amino acids, 20 parts whey protein concentrate, 20 parts HMB-Ca, and 80 parts black tomato extract.

[0044] S1. Select black tomatoes, disinfect them by rinsing, dry them at low temperature, pulverize them into ultrafine powder, and sieve them to obtain black tomato fruit powder. Mix 10g of black tomato fruit powder with 100g of sterile water, add 0.3g of glucose, 0.3g of peptone, 0.15g of corn steep liquor, and 0.08g of antimicrobial peptide, adjust the pH to 6.0, and inoculate with 8g of a compound probiotic suspension. The concentration of Lactobacillus plantarum in the compound probiotic suspension is 1×10⁻⁶. 6 -1×10 8 CFU / mL, Bifidobacterium concentration 1×10 6 -1×10 8 CFU / mL, Saccharomyces cerevisiae concentration 1×10 5 -1×10 7 CFU / mL, Bacillus subtilis concentration was 1×10⁻⁶ 5 -1×10 7CFU / mL, anaerobic fermentation at 37℃ for 24h, pH adjusted to 8.0, fermentation at 37℃ under aerobic conditions for 36h, inactivation at 85℃ for 15min, then centrifugation at 8000rpm for 15min to separate the supernatant, and filtration through an ultrafiltration membrane to obtain fermented black tomato extract. S2. Fermented black tomato extract was mixed with curcumin, soybean peptides with a molecular weight of less than 3000 Da, and branched-chain amino acids to obtain a mixed functional liquid. The branched-chain amino acids were a mixture of leucine, isoleucine, and valine in a mass ratio of 2:1:1. The mixed functional liquid was stirred at 800 rpm under constant temperature of 35℃, and concentrated whey protein and HMB-Ca were slowly added. The mixture was stirred and mixed for 20-30 min, the pH was adjusted to 7.0, and the mixture was homogenized at 20 MPa to obtain a composite solution. S3. Dry the composite solution at a low temperature of 45°C, sieve it, add maltitol, microcrystalline cellulose, vitamin D, and sodium carboxymethyl cellulose, mix evenly, and form the formulation.

[0045] Performance testing 1. Antioxidant activity assay (1) Determination of DPPH free radical scavenging ability Take 2 mL of test sample solutions of different concentrations, add 2 mL of 0.2 mmol / L DPPH ethanol solution, mix well, and react in the dark for 30 min. Measure the absorbance A1 at 517 nm. Use an equal volume of anhydrous ethanol instead of DPPH solution as a blank control and measure the absorbance A0. Use an equal volume of distilled water instead of the sample solution as a negative control and measure the absorbance A2. The formula for calculating the DPPH free radical scavenging rate is: Clearance rate (%) = [1 - (A1 - A0) / A2] × 100% (2) Determination of ABTS cationic free radical scavenging ability Mix equal volumes of 7 mmol / L LABTS solution and 2.45 mmol / L potassium persulfate solution, and react at room temperature in the dark for 12-16 h to generate the ABTS⁺ working solution. Before use, dilute with phosphate buffer (pH 7.4) to an absorbance of 0.70 ± 0.02 (734 nm). Take 0.1 mL of the sample solution to be tested, add 3.9 mL of the ABTS⁺ working solution, mix well, and react at room temperature for 6 min. Measure the absorbance at 734 nm. Use distilled water as a control instead of the sample. The ABTS⁺ scavenging rate is calculated using the following formula: Clearance rate (%) = (Control A - Sample A) / Control A × 100% As shown in Table 1, the DPPH radical scavenging rate and ABTS cationic radical scavenging rate of each embodiment of the present invention are all above 87%, significantly better than those of the comparative examples, indicating that the compositions of the present invention have excellent in vitro antioxidant activity. Example 5 showed a DPPH radical scavenging rate of 93.8% and an ABTS cationic radical scavenging rate of 95.4%, comparable to the positive control VC. Comparative Example 1 showed the lowest radical scavenging rates, only 68.4% and 70.5%, respectively, indicating that the fermentation process significantly improved the antioxidant activity of black tomatoes.

[0046] Table 1. Results of antioxidant activity assays for each example and comparative example.

[0047] Stability test The composition of the present invention was stored at 37±2℃ and 75±5% RH under accelerated conditions. Samples were taken after 0 and 3 months, respectively, and the retention rates of nicotinamide, lycopene and curcumin were determined by high performance liquid chromatography (HPLC).

[0048] As shown in Table 2, the compositions of the present invention exhibit good stability under accelerated storage conditions, with the average retention rate of each active ingredient exceeding 83% after 3 months. Comparative Example 3 showed the worst stability, with an average retention rate of only 63.2%, while Example 1 had an average retention rate of 85.8%, a significant difference. This fully demonstrates that the capsaicin-HMB-Ca-whey protein concentrate complex delivery system constructed in this invention has excellent protective effects on functional substances, effectively preventing oxidation, photolysis, and thermal degradation of active ingredients during storage. The stability of Comparative Example 1 (average retention rate 70.3%) was significantly lower than that of Example 1, indicating that certain components produced during fermentation may contribute to improving the stability of the composition. Comparative Example 4 lacked calcium β-hydroxy-β-methylbutyrate (HMB-Ca), resulting in reduced stability and a lower retention rate. The stability of Comparative Example 5 was significantly lower than that of Example 1, indicating that capsaicin plays a key stabilizing role in the complex delivery system and may form a tighter complex structure with whey protein concentrate and HMB-Ca.

[0049] Table 2. Retention rate of active ingredients (%) of each embodiment and comparative example after accelerated storage

[0050] Sarcopenia animal model experiment Twelve-month-old male C57BL / 6J mice (natural aging model) with a body weight of 35±5 g were provided by Beijing Vital River Laboratory Animal Technology Co., Ltd., license number: SCXK (Beijing) 2021-0006. After 1 week of adaptive feeding, the mice were randomly divided into groups of 10. Except for the blank control group, the mice in the other groups were injected subcutaneously with D-galactose and fed a low-protein diet to establish a sarcopenia model. The blank control group was gavaged with an equal volume of distilled water, the model control group was gavaged with an equal volume of distilled water, the experimental group was gavaged with the prepared composition at a dose of 500 mg / kg·bw; the positive control group was gavaged with a commercially available sarcopenia nutritional supplement (the main components were whey protein + HMB) at a dose of 500 mg / kg·bw.

[0051] A mouse grip strength tester was used to measure the forelimb grip strength of the mice. Each measurement was taken 3 times, and the maximum value was recorded, with the unit of g. For the measurement of exercise endurance, a rotarod fatigue tester was used, and the rotation speed was set at 20 rpm. The time from the start of the rotarod to the mouse falling off was recorded, with the unit of s. Each measurement was taken 3 times, and the average value was recorded.

[0052] As can be seen from Table 3, the grip strength and exercise endurance of the mice in the model control group were significantly decreased, decreasing by 35.3% and 50.3% respectively compared with the blank control group, indicating that the sarcopenia model mice had obvious muscle dysfunction. Each administration group could improve the grip strength and exercise endurance to varying degrees. Among them, the group of Example 1 had the best effect, with both the grip strength and the rotarod time increased, significantly better than other administration groups. The grip strength and rotarod time of Comparative Example 1 were significantly lower than those of Example 1, further proving the important role of compound probiotic fermentation in improving muscle function.

[0053] Table 3 Changes in grip strength and exercise endurance of mice in each group

[0054] For muscle weight measurement, after the experiment, the mice were sacrificed by cervical dislocation. The bilateral gastrocnemius, tibialis anterior, and soleus muscles were quickly dissected, blotted dry on the surface with filter paper, and weighed. The relative muscle weight (muscle weight / body weight × 100%) was calculated.

[0055] As can be seen from Table 4, the body weight of the mice in the model control group continued to decrease during the experiment, with the body weight decreasing by 18.9% compared with the initial body weight, indicating that the sarcopenia model was successfully established. The body weight of the mice in the blank control group increased normally by 11.1%. The decrease in body weight of the mice in the group of Example 1 was significantly lower than that of the model control group, with the body weight only decreasing by 5.1% and the body weight maintenance rate being 94.9%, significantly better than each comparative example group. This indicates that the composition of the present invention can effectively slow down the body weight decrease caused by sarcopenia.

[0056] All treatment groups showed varying degrees of improvement in muscle atrophy, with the most significant effect observed in Example 1 group. The relative weight of the gastrocnemius muscle increased by 36.2% compared to the model control group (P<0.01), significantly superior to all comparative groups. The relative weight of the gastrocnemius muscle in Example 1 group reached 425.3 mg / 100g, an increase of 36.2% compared to the model control group, recovering to 92.7% of the blank control group. The relative weight of the gastrocnemius muscle in Comparative Example 1 group was 368.4 mg / 100g, 13.4% lower than Example 1 group, demonstrating that the compound probiotic fermentation is a key process for enhancing the composition's effect on sarcopenia. In Comparative Example 2 group (lacking Bacillus subtilis), the relative weight of the gastrocnemius muscle was 392.7 mg / 100g, 7.6% lower than Example 1 group, indicating a synergistic effect of the compound fermentation system of the four probiotics. In Comparative Example 3 group, the relative weight of the gastrocnemius muscle was only 352.7 mg / 100g, fully demonstrating the significant impact of the compound delivery system constructed in this invention on improving sarcopenia.

[0057] Table 4 Comparison of relative muscle mass in mice of different groups (mg / 100g body weight)

[0058] The above description is merely a preferred embodiment of the present invention and is not intended to limit the present invention in any way. Any person skilled in the art can make many possible variations and modifications to the technical solutions of the present invention, or modify them into equivalent embodiments, without departing from the spirit and technical essence of the present invention. Therefore, any simple modifications, equivalent substitutions, equivalent changes, and modifications made to the above embodiments based on the technical essence of the present invention, without departing from the content of the technical solutions of the present invention, shall still fall within the scope of protection of the present invention.

Claims

1. A composition for improving sarcopenia, characterized in that, The ingredients, by weight, include: 0.5-5 parts curcumin, 5-20 parts soybean peptides, 3-15 parts branched-chain amino acids, 10-30 parts concentrated whey protein, 10-30 parts HMB-Ca, 2-6 parts capsaicin, and 50-100 parts fermented black tomato extract.

2. The composition for improving sarcopenia according to claim 1, characterized in that: The branched-chain amino acids are a mixture of leucine, isoleucine and valine in a mass ratio of 2:1:

1.

3. The composition for improving sarcopenia according to claim 1, characterized in that: The average molecular weight of the soybean peptide is less than 3000 Da.

4. The composition for improving sarcopenia according to claim 1, characterized in that: The preparation method of the fermented black tomato extract is as follows: black tomato powder is mixed with sterile water, glucose, peptone, corn steep liquor and antimicrobial peptides are added, the pH value is adjusted, a compound probiotic suspension is inoculated, anaerobic fermentation is carried out, the pH value is adjusted again, fermentation is carried out under aerobic conditions, after inactivation, the supernatant is separated by centrifugation, and filtered through an ultrafiltration membrane to obtain the fermented black tomato extract.

5. The composition for improving sarcopenia according to claim 4, characterized in that: The preparation method of the fermented black tomato extract includes a compound probiotic suspension containing Lactobacillus plantarum, Bifidobacterium, Saccharomyces cerevisiae, and Bacillus subtilis. The inoculation amount of the compound probiotic suspension is 5-10 wt.%. The anaerobic fermentation time is 24-36 h, and the aerobic fermentation time is 24-36 h.

6. The composition for improving sarcopenia according to claim 4, characterized in that: The concentration of *Lactobacillus plantarum* in the compound probiotic suspension was 1×10⁻⁶. 6 -1×10 8 CFU / mL, Bifidobacterium concentration 1×10 6 -1×10 8 CFU / mL, Saccharomyces cerevisiae concentration 1×10 5 -1×10 7 CFU / mL, Bacillus subtilis concentration was 1×10⁻⁶ 5 -1×10 7 CFU / mL.

7. A method for preparing a composition for improving sarcopenia according to any one of claims 1-6, characterized in that, Includes the following steps: S1. Mix curcumin, soybean peptides, branched-chain amino acids and fermented black tomato extract to obtain a mixed functional liquid. Under constant temperature and stirring, slowly add concentrated whey protein and HMB-Ca, stir and mix, slowly add capsaicin solution, continue stirring, adjust the pH value, and homogenize to obtain a composite solution. S2. Dry the composite solution at low temperature, sieve it, add auxiliary raw materials and mix evenly to form the formulation.

8. A method for preparing a composition for improving sarcopenia according to claim 7, characterized in that: In step S1, the constant temperature stirring temperature is 35-40℃; the stirring time is 20-30 min; the stirring time is 40-60 min; the pH value is adjusted to 7.0-7.5; the solvent of capsaicin solution is ethanol with a concentration of 10-15 wt.%; and the homogenization pressure is 15-25 MPa.

9. A method for preparing a composition for improving sarcopenia according to claim 7, characterized in that: The low-temperature drying temperature in step S2 is 45-60℃; the auxiliary raw materials are maltitol, microcrystalline cellulose, vitamin D, and sodium carboxymethyl cellulose.

10. Use of the composition for improving sarcopenia according to any one of claims 1-6 in the preparation of a medicament for the prevention or treatment of sarcopenia.

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