Enzyme-containing composition for relieving arthralgia of dogs and cats

Abstract

The invention belongs to the technical field of pet nutrition and health care, and discloses an enzyme-containing composition for relieving arthralgia of dogs and cats, the composition is composed of a compound enzyme component, a joint repair nutrition component and a carrier, the compound enzyme component comprises bromelain, papain and hyaluronidase, the nutritional components comprise chondroitin sulfate, glucosamine, a curcumin extract and type II collagen. All the components have a synergistic effect, so that the composition has multiple effects of quickly relieving inflammation, relieving arthralgia and promoting cartilage repair. In order to improve the enzyme activity, a low-temperature vacuum drying technology is adopted for enzyme treatment, and meanwhile, liposome embedding is utilized for realizing targeted delivery, so that the bioavailability of the joint part is improved. The composition disclosed by the invention is good in stability, high in bioavailability and suitable for auxiliary nutrition intervention of arthritis and degenerative joint diseases of dogs and cats.

Description

Technical Field

[0001] This invention belongs to the field of pet nutrition and health care technology, and specifically relates to an enzyme-containing composition for relieving joint pain in dogs and cats. Background Technology

[0002] With the development of the pet economy, dogs and cats have gained increasing status as family companion animals, and pet health management has become increasingly important. Among older pets, the incidence of arthritis and osteoarthritis cartilage degeneration continues to rise, often manifesting as difficulty in movement, joint swelling, and persistent pain. This not only affects the animals' quality of life but also poses management challenges for owners. Currently available products for relieving joint pain in pets are mostly single-nutrient supplements such as glucosamine and chondroitin sulfate. While these have some effect on cartilage repair, their overall onset of action is slow, and their anti-inflammatory effects are weak, making it difficult to meet the comprehensive intervention needs for acute or persistent joint inflammation.

[0003] On the other hand, proteolytic enzymes such as bromelain and papain have been widely studied for their ability to decompose inflammatory exudates and alleviate soft tissue inflammation. However, their application in pet nutritional products is limited due to their unstable structure and easy degradation. Furthermore, some plant-based active ingredients have shown potential in anti-inflammatory and analgesic effects, but their low bioavailability and poor absorption limit their actual efficacy. Therefore, constructing a synergistic combination system with rapid anti-inflammatory effects, tissue repair, and good absorption in the management of joint pain in pets is a research hotspot and technical challenge in this field. Summary of the Invention

[0004] To address the shortcomings mentioned in the background art, the present invention aims to provide an enzyme-containing composition for relieving joint pain in dogs and cats. This composition utilizes bromelain, papain, and hyaluronidase for synergistic anti-inflammatory effects, and combines chondroitin sulfate, glucosamine, curcumin extract, and type II collagen to repair cartilage. Furthermore, liposome encapsulation enhances bioavailability. This composition has the advantages of rapid onset of action, synergistic function, and stable absorption, and is suitable for adjunctive intervention in canine and feline arthritis or osteoarthritis.

[0005] The objective of this invention can be achieved through the following technical solutions: An enzyme-containing composition for relieving joint pain in dogs and cats, comprising the following ingredients in parts by weight: (1) 3-8 parts of the complex enzyme component, including: a. 1-3 parts of bromelain; b. Papain 1.5–4 parts; c. Hyaluronidase 0.5–1.5 parts; (2) Joint repair nutrient components, 5-12 parts, including: a. Chondroitin sulfate 1.5–4 parts; b. 1.5 to 4 parts of glucosamine; c. Curcumin extract 0.5–2 parts; d. Type II collagen 1-2.5 parts; (3) Carrier raw materials, including: a. 20-35 parts of maltodextrin; b. 5-10 parts lactose; c. 0.5 to 1.5 parts magnesium stearate.

[0006] More preferably, the mass ratio of bromelain to papain in the complex enzyme component is 1:1.5.

[0007] More preferably, the type II collagen is a desalted and defatted type II collagen powder obtained by enzymatic hydrolysis of cartilage tissue.

[0008] More preferably, the curcumin extract is a total curcumin extract obtained by ethanol reflux extraction and spray drying, wherein the curcumin content is not less than 85%.

[0009] More preferably, the complex enzyme components and joint repair nutrients are co-encapsulated in liposomes, wherein the liposomes use lecithin and cholesterol as the main film-forming materials, have a particle size of 100-300 nm, and an encapsulation rate of not less than 80%.

[0010] A method for preparing an enzyme-containing composition for relieving joint pain in dogs and cats includes the following steps: S1. Mix bromelain, papain and hyaluronidase, and perform enzyme activity protection treatment on the resulting complex enzyme components; S2. The treated complex enzyme components are mixed with chondroitin sulfate, glucosamine, curcumin extract and type II collagen to obtain a mixture of active ingredients; S3. The active ingredient mixture is encapsulated in liposomes with lecithin and cholesterol as the main film-forming materials; S4. The obtained liposomes are mixed evenly with maltodextrin, lactose and magnesium stearate to obtain the enzyme-containing composition.

[0011] More preferably, the enzyme activity protection treatment in step S1 adopts a low-temperature vacuum drying process, with the drying temperature not exceeding 40°C and the vacuum degree controlled between 0.06 and 0.09 MPa.

[0012] More preferably, the liposome encapsulation in step S3 is prepared by thin-film hydration-ultrasonic dispersion method, wherein the ultrasonic frequency is 20-40 kHz and the ultrasonic time is 3-10 minutes.

[0013] More preferably, the liposome encapsulation efficiency of the active ingredient mixture is determined by ultraviolet spectrophotometry at a wavelength of 420 nm.

[0014] More preferably, the composition is in the form of granules or compressible powder, and is suitable for oral nutritional supplements for dogs and cats.

[0015] The beneficial effects of this invention are: This invention utilizes a synergistic combination of bromelain, papain, and hyaluronidase in a specific ratio to rapidly act on the inflamed site, breaking down exudates, denatured proteins, and hyaluronic acid degradation products within the joint cavity. This significantly reduces local inflammation levels and improves synovial fluid flow, helping to alleviate swelling and pain. Simultaneously, this invention introduces chondroitin sulfate, glucosamine, curcumin extract, and highly active desalted and defatted type II collagen as nutritional repair modules into the combined system. This not only provides key raw materials for the synthesis of chondrocyte extracellular matrix but also promotes joint tissue regeneration and homeostasis through antioxidant, anti-apoptotic, and immunomodulatory mechanisms, making it suitable for long-term management of chronic degenerative arthritis. To improve in vivo absorption and component stability, this invention further employs a liposome encapsulation system constructed from lecithin and cholesterol to co-encapsulate the complex enzymes and nutritional factors, forming a delivery carrier with uniform particle size and high encapsulation efficiency. This provides excellent targeting and gastrointestinal protection, significantly enhancing the bioavailability of the active ingredients in vivo. By processing the complex enzyme components using low-temperature vacuum drying technology, the protease activity is significantly preserved, effectively avoiding the damage to the enzyme structure caused by traditional heat drying, and ensuring the stability of the product during storage and use. Attached Figure Description

[0016] The invention will now be further described with reference to the accompanying drawings.

[0017] Figure 1 The graphs show the joint inflammation relief curves after feeding in Examples 1-3 and Comparative Examples 1-2; Figure 2 Bar chart comparing the content of type II collagen (CII) and agglutinin in the synovial fluid of the knee joint after feeding in Examples 1-3 and Comparative Examples 1-2; Figure 3 Bar chart showing the comparison of peak concentrations of curcumin in the plasma of animals after feeding in Examples 1-3 and Comparative Examples 1-2. Detailed Implementation

[0018] The technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of the present invention, and not all embodiments. Based on the embodiments of the present invention, all other embodiments obtained by those skilled in the art without creative effort are within the scope of protection of the present invention.

[0019] Example 1 An enzyme-containing composition for relieving joint pain in dogs and cats comprises the following raw materials in parts by weight: 3 parts of a complex enzyme component, including 1 part of bromelain, 1.5 parts of papain, and 0.5 parts of hyaluronidase; 5 parts of a joint repair nutrient component, including 1.5 parts of chondroitin sulfate, 1.5 parts of glucosamine, 1 part of curcumin extract, and 1 part of type II collagen; and a carrier raw material, including 20 parts of maltodextrin, 5 parts of lactose, and 0.5 parts of magnesium stearate. The preparation steps of the enzyme-containing composition are as follows: S1. Take 30.3 g of bromelain, 45.5 g of papain and 15.2 g of hyaluronidase, add them to 100 mL of anhydrous ethanol and deionized water in a volume ratio of 1:9, stir at 4 ℃ for 20 minutes, pre-freeze the moistened enzyme mixture at -40 ℃ for 2 hours, and then freeze-dry it under vacuum of 0.08 MPa and drying temperature not exceeding 40 ℃ for 6 to 8 hours to obtain compound enzyme powder; S2. Add the compound enzyme powder to the mixing tank, and then add 45.5 g chondroitin sulfate, 45.5 g g glucosamine, 30.3 g curcumin extract and 30.3 g type II collagen in sequence. Stir at a constant temperature of 25 ℃ for 20 minutes, and control the stirring speed at 80 rpm to form a uniform active substance mixture system. S3. Dissolve 40 g of lecithin and 10 g of cholesterol in 200 mL of a mixed solvent of chloroform and methanol (volume ratio 2:1), and incubate at 60 °C in a water bath until completely dissolved. Then, evaporate the solvent at 45 °C and –0.08 MPa to form a homogeneous lipid membrane. Dissolve the active mixture obtained in step S2 in 200 mL of pH 7.0 PBS buffer, add it to a flask containing the lipid membrane, and hydrate it at 40 °C in a water bath for 30 minutes. Then, sonicate the solution to obtain a liposome solution. S4. 606 g of maltodextrin and 151.5 g of lactose were added to a high-speed mixer and dry-mixed for 10 minutes. The liposome solution prepared in step S3 was slowly added dropwise, controlling the material-to-liquid ratio to be approximately 1:1.5 (w / v). After uniform mixing, spray drying was performed under the following conditions: inlet air temperature 135 ℃, outlet air temperature 75 ℃, nozzle diameter 0.7 mm, and atomization pressure 0.35 MPa. After drying, 15.2 g of magnesium stearate was added to the resulting powder, and the mixture was mixed at low speed for 5 minutes. The mixture was then sieved through a 40-mesh sieve to obtain the enzyme-containing composition powder.

[0020] Example 2 An enzyme-containing composition for relieving joint pain in dogs and cats comprises the following raw materials in parts by weight: 8 parts of a complex enzyme component, including 3 parts of bromelain, 4.5 parts of papain, and 0.5 parts of hyaluronidase; 12 parts of a joint repair nutrient component, including 4 parts of chondroitin sulfate, 4 parts of glucosamine, 2 parts of curcumin extract, and 2 parts of type II collagen; and a carrier raw material, including 35 parts of maltodextrin, 10 parts of lactose, and 1.5 parts of magnesium stearate. The preparation steps of the enzyme-containing composition are as follows: S1. Weigh 90.9 g of bromelain, 136.4 g of papain, and 15.2 g of hyaluronidase, add them to 300 mL of 10% anhydrous ethanol, and stir at 4 ℃ for 20 minutes. After mixing, pre-freeze the mixture for 2 hours, and then freeze-dry it for 6-8 hours under a vacuum of 0.08 MPa and a drying temperature not exceeding 40 ℃ to obtain the compound enzyme powder. S2. Add 121.2 g chondroitin sulfate, 121.2 g g glucosamine, 60.6 g curcumin extract and 60.6 g type II collagen to the compound enzyme dry powder in sequence, and stir at low speed at 25 ℃ to form an active functional mixture. S3. Dissolve 120 g of lecithin and 30 g of cholesterol in a 600 mL mixture of chloroform and methanol (2:1 volume ratio) in a 60 °C water bath. Evaporate the solvent at 45 °C and –0.08 MPa to form a homogeneous lipid membrane. Dissolve the active mixture obtained in step S2 in 600 mL of pH 7.0 PBS buffer, add it to a flask containing the lipid membrane, and hydrate in a 40 °C water bath for 30 minutes. Then sonicate to obtain the liposome solution. S4. 1060.6 g of maltodextrin and 303.0 g of lactose were added to a high-speed mixer and dry-mixed for 10 minutes. Then, the liposome liquid obtained in S3 was added, and the mixture was uniformly humidified at a material-to-liquid ratio of 1:1.5 (w / v). The resulting wet mixture was granulated by spray drying. After drying, 45.5 g of magnesium stearate was added, and the mixture was mixed at low speed for 5 minutes. After sieving through a 40-mesh sieve, the enzyme-containing composition powder was obtained.

[0021] Example 3 An enzyme-containing composition for relieving joint pain in dogs and cats comprises the following raw materials in parts by weight: 6.5 parts of a complex enzyme component, including 2 parts of bromelain, 3 parts of papain, and 1.5 parts of hyaluronidase; 8 parts of a joint repair nutrient component, including 2.75 parts of chondroitin sulfate, 2.75 parts of glucosamine, 1.25 parts of curcumin extract, and 1.75 parts of type II collagen; and a carrier raw material, including 27.5 parts of maltodextrin, 7.5 parts of lactose, and 1 part of magnesium stearate. The preparation steps of the enzyme-containing composition are as follows: S1. Weigh 60.6 g of bromelain, 90.9 g of papain, and 45.45 g of hyaluronidase, add them to 250 mL of 10% anhydrous ethanol, and stir at 4 °C for 20 minutes. Pre-freeze the mixture for 2 hours, then freeze-dry it under a vacuum of 0.08 MPa and a drying temperature not exceeding 40 °C for 6–8 hours to obtain the compound enzyme powder. S2. Add the compound enzyme powder into a mixing tank, add 83.3 g chondroitin sulfate, 83.3 g g glucosamine, 37.9 g curcumin extract and 53.0 g type II collagen, and stir at low speed at room temperature to obtain an active functional complex. S3. Dissolve 180 g of lecithin and 20 g of cholesterol in 400 mL of chloroform-methanol mixed solvent in a 60 °C water bath. Evaporate the solvent at 45 °C and –0.08 MPa to form a homogeneous lipid membrane. Dissolve the active mixture obtained in step S2 in 400 mL of pH 7.0 PBS buffer to hydrate the lipid membrane at 40 °C for 30 minutes, followed by sonication to obtain a liposome solution. S4. 963.6 g of maltodextrin and 227.3 g of lactose were dry-mixed in a high-speed mixer for 10 minutes. Then, the liposome liquid obtained in S3 was added, and the mixture was mixed at a material-to-liquid ratio of 1:1.5 (w / v). The resulting wet mixture was then granulated by spray drying. After drying, 30.3 g of magnesium stearate was added, and the mixture was mixed for 5 minutes. After sieving through a 40-mesh sieve, the enzyme-containing composition powder was obtained.

[0022] Comparative Example 1 An enzyme-containing composition for relieving joint pain in dogs and cats comprises the following raw materials in parts by weight: 3.5 parts of a complex enzyme component, including 2 parts of bromelain and 1.5 parts of hyaluronidase; 8 parts of a joint repair nutrient component, including 2.75 parts of chondroitin sulfate, 2.75 parts of glucosamine, 1.25 parts of curcumin extract, and 1.75 parts of type II collagen; and a carrier raw material, including 27.5 parts of maltodextrin, 7.5 parts of lactose, and 1 part of magnesium stearate. The preparation process of the enzyme-containing composition is the same as that in Example 3, except that papain is not added in step S1.

[0023] Comparative Example 2 An enzyme-containing composition for relieving joint pain in dogs and cats comprises the following raw materials in parts by weight: 6.5 parts of a complex enzyme component, including 2 parts of bromelain, 3 parts of papain, and 1.5 parts of hyaluronidase; 8 parts of a joint repair nutrient component, including 2.75 parts of chondroitin sulfate, 2.75 parts of glucosamine, 1.25 parts of curcumin extract, and 1.75 parts of type II collagen; and a carrier raw material, including 27.5 parts of maltodextrin, 7.5 parts of lactose, and 1 part of magnesium stearate. The preparation steps of the enzyme-containing composition are as follows: S1. Weigh 60.6 g of bromelain, 90.9 g of papain, and 45.45 g of hyaluronidase, add them to 250 mL of 10% anhydrous ethanol, and stir at 4 °C for 20 minutes. Pre-freeze the mixture for 2 hours, then freeze-dry it under a vacuum of 0.08 MPa and a drying temperature not exceeding 40 °C for 6–8 hours to obtain the compound enzyme powder. S2. Add the compound enzyme powder into a mixing tank, add 83.3 g chondroitin sulfate, 83.3 g g glucosamine, 37.9 g curcumin extract and 53.0 g type II collagen, and stir at low speed at room temperature to obtain an active functional complex. S3. 963.6 g of maltodextrin and 227.3 g of lactose were added to a high-speed mixer and dry-mixed for 10 minutes. Then, the active functional complex obtained in S2 was added, and the mixture was further mixed for 5 minutes under low-speed stirring. Then, 30.3 g of magnesium stearate was added, and the mixture was mixed for 5 minutes. After sieving through a 40-mesh sieve, the enzyme-containing composition powder was obtained.

[0024] Performance testing 1. Test on the relief effect of joint inflammation A soft tissue arthritis model was established in healthy adult dogs using collagen-induced controlled inflammation in the knee joint. After successful modeling, the dogs were randomly divided into five groups, receiving the oral compositions of Examples 1-3 and Comparative Examples 1-2, respectively, once daily for 14 consecutive days. The diameter of joint swelling was measured before administration (day 0) and on days 3, 7, and 14. Inflammation scores (0-5 points) were assigned by professional evaluators based on the degree of redness and swelling, tenderness, and limitation of movement. All animals were kept under identical housing conditions. The results are shown in Table 1 below.

[0025] Table 1 Results of joint inflammation relief As shown in Table 1, the joint inflammation scores of Examples 1-3 all showed a significant decreasing trend at each observation time point, especially Example 2, which saw a decrease of up to 78% on day 14, demonstrating superior anti-inflammatory effects. In contrast, Comparative Example 1, which removed papain, showed a significantly weaker decrease in inflammation score, only 40%, indicating that the synergistic effect of papain in the complex enzyme played a crucial role in anti-inflammatory activity. Comparative Example 2, lacking liposome encapsulation, resulted in decreased absorption efficiency of the active substance, with a decrease of only 45% on day 14. Overall, the synergistic effect of the complex enzyme ratio and the liposome delivery system in this invention significantly promote the speed of anti-inflammatory onset and overall efficacy.

[0026] 2. Cartilage Repair and Tissue Reconstruction Capacity Test Dogs with soft tissue arthritis were selected and fed the enzyme-containing compositions of Examples 1-3 and Comparative Examples 1-2 for 14 consecutive days. Synovial fluid and articular cartilage tissue samples were collected from the knee joint to detect the levels of cartilage repair-related markers. The content of type II collagen (CII) and agglutinin in the synovial fluid was measured by ELISA. Histological sections were stained with hematoxylin and eosin (HE) and analyzed by immunohistochemistry to observe the cartilage layer thickness and structural integrity. The degree of cartilage tissue repair was assessed using a scoring system. The results are shown in Table 2 below.

[0027] Table 2 Results of cartilage repair capacity As shown in Table 2, Examples 1-3 were superior to Comparative Examples 1-2 in terms of CII content, aggregated proteoglycan expression, cartilage layer thickness, and tissue repair score, indicating that the compositions of the present invention have significant advantages in promoting cartilage regeneration and joint tissue reconstruction. Example 2 achieved a CII content of 145 ng / mL, a cartilage layer thickness of 160 μm, and a tissue repair score of 4.5, indicating that it had the best effect in enhancing cartilage matrix synthesis and structural repair. Comparative Example 1 lacked the key protease component, resulting in insufficient promotion of cartilage repair, with a tissue score of only 2.6; Comparative Example 2 did not undergo liposome encapsulation, leading to decreased absorption efficiency and localization ability of the active substances, and its repair effect was also unsatisfactory.

[0028] 3. Bioavailability and delivery efficiency testing After establishing an arthritis model in healthy dogs, they were fed the enzyme-containing compositions of Examples 1-3 and Comparative Examples 1-2, respectively, and curcumin was used as a representative marker component to evaluate in vivo delivery efficiency. Peripheral blood was collected at 0.5, 1, 2, 4, 8, 12, and 24 hours after administration using fluorescently labeled curcumin or ultraviolet spectrophotometry to detect plasma curcumin concentration. The main pharmacokinetic parameters—peak concentration, time to peak concentration, and area under the plasma concentration-time curve (AUC)—were calculated using WinNonlin. The results are shown in Table 3 below.

[0029] Table 3 Bioavailability and Pharmacokinetic Parameters As shown in Table 3, Examples 1-3 outperformed Comparative Examples 1-2 in pharmacokinetic parameters such as AUC, peak concentration, and time to peak concentration, indicating that the enzyme-containing composition of the present invention has higher bioavailability and better delivery efficiency in vivo. Example 2 showed the best performance, with an AUC of 335 μg·h / mL, a peak concentration of 19.2 μg / mL, and a time to peak concentration of 2.5 h, demonstrating that curcumin was fully absorbed and released stably in vivo, exhibiting a longer duration of action. Comparative Example 1, lacking a key enzyme component, had a lower overall absorption efficiency; while the active ingredient in Comparative Example 2 was metabolized more rapidly in vivo, reaching peak concentration earlier at 1.4 h, and had an AUC of only 205 μg·h / mL.

[0030] 4. Palatability and compliance testing To evaluate the practical suitability of the compositions of the present invention in dogs and cats, a free-feeding behavior observation experiment was conducted on 10 adult dogs. The enzyme-containing compositions prepared in Examples 1-3 and Comparative Examples 1-2 were administered, respectively. Each composition was mixed into regular feed at the standard single-dose recommended dose. The feeding time was limited to 15 minutes, and the actual amount of food consumed, the time of administration, and whether there were any instances of picky eating, refusal to eat, or food residue were recorded. During the experiment, the animals' feeding conditions, hunger state, and environment were kept consistent. The experiment lasted for 3 days, and the average values ​​were used for statistical analysis to calculate the feeding completion rate, average feeding time, and refusal to eat rate. The results are shown in Table 4 below.

[0031] Table 4. Results of palatability and compliance As shown in Table 4, Examples 1-3 prepared in this invention exhibited excellent palatability and compliance during canine feeding, with a feeding completion rate exceeding 93% and an average feeding time of approximately 6 minutes, without any instances of food refusal. This indicates that the combined action of the complex enzyme components and joint nutrients, through liposome encapsulation and a palatable excipient system, significantly improved the flavor and smoothness of the composition, enabling animals to quickly and autonomously complete their feeding. In contrast, Comparative Examples 1 and 2 showed feeding completion rates of 81% and 84%, respectively, with significantly increased food refusal rates and prolonged average feeding times. This suggests that the lack of papain or the absence of liposome encapsulation leads to poor palatability or insufficient powder dispersibility, thereby affecting the animals' voluntary intake.

[0032] 5. Enzyme activity retention rate test To verify the protective effect of low-temperature vacuum freeze-drying on protease activity, a composite enzyme raw material (prepared from a mixture of bromelain, papain, and hyaluronidase) from the same batch and in the same proportion was selected. No other functional components were introduced, and the raw materials were divided into three groups, differing only in the drying method. The experimental group underwent low-temperature vacuum freeze-drying, while the control group underwent hot air drying and spray drying, respectively. After drying, equal volumes of enzyme powder were dissolved in phosphate buffer at pH 7.0. Using casein as a substrate, the enzymatic hydrolysis rate was determined colorimetrically. The relative enzyme activity retention rate of each group was calculated using the initial enzyme activity of the undried composite enzyme as 100% as the baseline. The results are shown in Table 5 below.

[0033] Table 5 Enzyme activity retention rate As shown in Table 5, the composite enzyme system treated with the low-temperature vacuum freeze-drying process proposed in this invention exhibits an enzyme activity retention rate as high as 91.8%, significantly higher than samples obtained using hot air drying (72.3%) and spray drying (66.5%). This indicates that low-temperature freeze-drying effectively avoids thermal denaturation and structural damage during enzyme treatment, maximizing the preservation of protease bioactivity. In contrast, while hot air drying, with its relatively high temperature, still results in enzyme activity loss, and spray drying, due to the more severe impact of instantaneous high temperature, leads to a more pronounced decrease in enzyme activity.

[0034] In the description of this specification, references to terms such as "an embodiment," "example," "specific example," etc., indicate that a specific feature, structure, material, or characteristic described in connection with that embodiment or example is included in at least one embodiment or example of the invention. In this specification, illustrative expressions of the above terms do not necessarily refer to the same embodiment or example. Furthermore, the specific features, structures, materials, or characteristics described may be combined in any suitable manner in one or more embodiments or examples.

[0035] The foregoing has shown and described the basic principles, main features, and advantages of the present invention. Those skilled in the art should understand that the present invention is not limited to the above embodiments. The embodiments and descriptions in the specification are merely illustrative of the principles of the invention. Various changes and modifications can be made to the invention without departing from its spirit and scope, and all such changes and modifications fall within the scope of the claimed invention.

Claims

1. An enzyme-containing composition for relieving joint pain in dogs and cats, characterized in that, Includes the following raw materials by weight: (1) 3-8 parts of the complex enzyme component, including: a. 1-3 parts of bromelain; b. Papain 1.5–4 parts; c. Hyaluronidase 0.5–1.5 parts; (2) Joint repair nutrient components, 5-12 parts, including: a. Chondroitin sulfate 1.5–4 parts; b. 1.5 to 4 parts of glucosamine; c. Curcumin extract 0.5–2 parts; d. Type II collagen 1-2.5 parts; (3) Carrier raw materials, including: a. 20-35 parts of maltodextrin; b. 5-10 parts lactose; c. 0.5 to 1.5 parts magnesium stearate.

2. The enzyme-containing composition according to claim 1, characterized in that, The mass ratio of bromelain to papain in the complex enzyme component is 1:1.

5.

3. The enzyme-containing composition according to claim 1, characterized in that, The type II collagen is a desalted and defatted type II collagen powder obtained by enzymatic hydrolysis of cartilage tissue.

4. The enzyme-containing composition according to claim 1, characterized in that, The curcumin extract is a total curcumin extract obtained by ethanol reflux extraction and spray drying, wherein the curcumin content is not less than 85%.

5. The enzyme-containing composition according to any one of claims 1 to 4, characterized in that, The complex enzyme components and joint repair nutrients are co-encapsulated in liposomes. The liposomes use lecithin and cholesterol as the main film-forming materials, with a particle size of 100-300 nm and an encapsulation rate of not less than 80%.

6. A method for preparing an enzyme-containing composition for relieving joint pain in dogs and cats, wherein the enzyme-containing composition is as described in any one of claims 1 to 5, characterized in that, Includes the following steps: S1. Mix bromelain, papain and hyaluronidase, and perform enzyme activity protection treatment on the resulting complex enzyme components; S2. The treated complex enzyme components are mixed with chondroitin sulfate, glucosamine, curcumin extract and type II collagen to obtain a mixture of active ingredients; S3. The active ingredient mixture is encapsulated in liposomes with lecithin and cholesterol as the main film-forming materials; S4. The obtained liposomes are mixed evenly with maltodextrin, lactose and magnesium stearate to obtain the enzyme-containing composition.

7. The method for preparing the enzyme-containing composition according to claim 6, characterized in that, In step S1, the enzyme activity protection treatment adopts a low-temperature vacuum drying process, with the drying temperature not exceeding 40°C and the vacuum degree controlled between 0.06 and 0.09 MPa.

8. The method for preparing the enzyme-containing composition according to claim 6, characterized in that, In step S3, the liposomes are embedded using a thin-film hydration-ultrasonic dispersion method, wherein the ultrasonic frequency is 20–40 kHz and the ultrasonic time is 3–10 minutes.

9. The method for preparing the enzyme-containing composition according to claim 6, characterized in that, The liposome encapsulation efficiency of the active ingredient mixture was determined by ultraviolet spectrophotometry at a wavelength of 420 nm.

10. The enzyme-containing composition according to claim 1, characterized in that, The composition is available in granule or compressible powder form and is suitable for oral nutritional supplements for dogs and cats.

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