New uses of whole molecular weight HA in the prevention and amelioration of osteoarthritis, compositions and applications and products thereof

Abstract

The application belongs to the field of food, and discloses a new use of hyaluronic acid (HA) with full molecular weight, which is used for preparing products for improving inflammation of arthritis, improving lubrication of joints, increasing moisture of connective tissue or restoring elasticity of connective tissue; the products are food, health food, special medical purpose formula food or medicine for preventing and / or improving osteoarthritis (OA). After further research on the HA with full molecular weight, it is found that the HA with full molecular weight has a certain improvement effect on OA and other related diseases. In addition, it is also found that when the HA with full molecular weight is combined with Q3C, the related effects of OA can be further improved, the inflammation can be greatly reduced, the joints can be lubricated, the moisture of connective tissue can be increased, and the elasticity of connective tissue can be restored, and thus the composition for treating and preventing OA has important practical significance. Meanwhile, the application also provides a related application of the composition.

Description

Technical Field

[0001] This invention relates to the pharmaceutical field, and more particularly to novel uses, compositions, applications, and products of full molecular weight HA in the prevention and improvement of osteoarthritis. Background Technology

[0002] With changes in modern lifestyles, such as prolonged sitting, desk work, and lack of exercise, the incidence of osteoarthritis (OA) and other degenerative joint diseases is increasing year by year, and showing a trend towards affecting younger people. These diseases often manifest as joint pain, swelling, stiffness, and limited mobility. In severe cases, they can affect normal walking, climbing stairs, and other daily activities, thus reducing quality of life. Their pathological basis typically involves wear and tear of articular cartilage, decreased joint lubrication, reduced water content and elasticity of connective tissue, and local inflammatory reactions.

[0003] Currently, interventions for osteoarthritis (OA) and other degenerative joint diseases mainly include drug intervention, physical therapy, and surgical treatment. Drug intervention often uses nonsteroidal anti-inflammatory drugs (NSAIDs) and analgesics, which can relieve pain symptoms to some extent, but long-term use may lead to gastrointestinal discomfort and increased burden on liver and kidney function. Physical therapy is mostly used as an adjunct therapy, with limited overall improvement. Surgical treatment has drawbacks such as significant trauma, long recovery period, and limited applicability. Therefore, developing functional products suitable for long-term consumption or continuous intervention, with high safety, and capable of acting on multiple aspects such as joint lubrication, connective tissue hydration, maintenance of tissue elasticity, and inflammation regulation has become an important research direction in this field.

[0004] Hyaluronic acid (HA) is a natural polysaccharide widely distributed in cartilage, synovial fluid, and other connective tissues. It possesses excellent water-retention, lubrication, and viscoelasticity, playing a crucial role in maintaining joint lubrication, preserving tissue hydration, and protecting cartilage surfaces. Based on this, there are existing reports on the use of HA or its derivatives for joint health maintenance, joint lubrication improvement, or inflammatory response modulation. For example, patent application JP2004531460A5 relates to the application of hyaluronic acid derivatives in inhibiting inflammatory arthritis; patent application JP2009521400A relates to oral joint function improvers and protectants containing hyaluronic acid phospholipid compounds.

[0005] However, current technologies regarding the application of HA in the joint field mostly focus on HA of a specific molecular weight or only cover the general uses of HA in general terms. Research on the application of full molecular weight HA in improving osteoarthritis remains insufficient. Unlike single molecular weight HA, full molecular weight HA has a wider molecular weight distribution, covering different molecular weight ranges simultaneously. Therefore, theoretically, it is expected to exhibit comprehensive advantages over single molecular weight HA in areas such as joint lubrication, tissue hydration, elasticity maintenance, and improvement of the joint microenvironment.

[0006] Furthermore, the pathogenesis of osteoarthritis (OA) is complex. Besides decreased joint lubrication, it is closely related to enhanced inflammation, cartilage matrix damage, and reduced tissue water content. Therefore, a single ingredient often cannot address all the needs for improvement. Current technologies lack sufficient research and clear explanation regarding whether combining full-molecular-weight HA with other active ingredients can produce better results in improving osteoarthritis, especially whether it can form a synergistic effect distinct from that of single-molecular-weight HA.

[0007] Therefore, this project further verified this effect during its development; and some new phenomena were discovered during the project's research and development process. Based on this, the present invention is proposed. Summary of the Invention

[0008] The purpose of this invention is to provide new uses for full molecular weight HA and compositions comprising full molecular weight HA and Q3C. Further research on full molecular weight HA revealed that it has certain therapeutic effects on OA and related conditions. Furthermore, this invention also found that when used in combination with Q3C, it can further improve the efficacy related to OA, effectively reducing inflammatory responses, lubricating joints, increasing connective tissue moisture, and restoring connective tissue elasticity. Therefore, compositions for the treatment and prevention of OA have significant practical implications.

[0009] In addition, the present invention also provides related applications of this composition.

[0010] To achieve the above objectives, this application discloses:

[0011] A novel use of full molecular weight HA, characterized in that the full molecular weight HA is used to prepare products that improve joint degenerative diseases such as osteoarthritis (OA).

[0012] The improvement of OA is specifically manifested in at least one of the following: improving joint inflammation response, increasing joint lubrication, increasing connective tissue moisture, restoring connective tissue elasticity, relieving joint discomfort, and improving joint mobility.

[0013] The product is food, health food, special medical purpose formula food, or medicine.

[0014] In the above-mentioned new applications, the weight-average molecular weight distribution range of the full molecular weight HA or its salt is 0.2 million to 1.5 million, and the molecular weight dispersion coefficient Mw / Mn≥5.

[0015] The full molecular weight HA salt is one or more of the following: full molecular weight HA sodium, full molecular weight HA zinc, full molecular weight HA potassium, and full molecular weight HA magnesium.

[0016] Preferably, such full molecular weight HA or its salts can be referred to the applicant's earlier application publication number CN113512134B, which is entitled a full molecular weight distribution HA sodium and its preparation method and application.

[0017] The present invention also discloses a composition comprising, by weight, the following components:

[0018] 0.5~1.25 parts of total molecular weight HA or its salt;

[0019] 0.2~0.5 parts Q3C (quercetin-3-caffeoyl robinoside).

[0020] The full molecular weight HA or its salts have a significant effect on improving joint lubrication, increasing tissue moisture, and maintaining tissue elasticity. Further research on Q3C has shown that it also has the effect of improving joint lubrication, increasing tissue moisture, and maintaining tissue elasticity. When the two are combined, they show a synergistic effect on improving OA-related indicators.

[0021] Preferably, the weight ratio of the total molecular weight HA to Q3C is 0.8-1.25 : 0.2-0.4.

[0022] Preferably, the above composition comprises:

[0023] 0.5~1.25 parts of total molecular weight HA or its salt;

[0024] 0.3~0.8 parts chitosan oligosaccharide;

[0025] 0.02~0.15 parts of non-denatured type II collagen;

[0026] 0.5 to 2 parts casein;

[0027] 0.5 to 2 parts soy protein;

[0028] 0.1~0.6 parts curcumin;

[0029] 0.05~0.5 parts of dihydroquercetin;

[0030] 0.5 to 2 parts calcium lactate;

[0031] 0.2~0.5 parts Q3C.

[0032] Although the non-denatured type II collagen peptides, chitosan oligosaccharides, curcumin and other components used in this invention have been proven to be beneficial to joint health, such as in publication number CN 114732131 A, which describes a compound HA composition for the prevention and relief of joint diseases, and publication number CN 118750587 A, which describes a type II collagen composition for increasing bone density and its preparation method.

[0033] However, the method of the present invention is not a simple superposition of existing technologies. The core innovation of the present invention is as follows:

[0034] 1. This invention has found that full molecular weight HA and its derivatives have positive effects on improving joint lubrication, increasing tissue moisture, and maintaining tissue elasticity, and show superior application potential in improving OA-related conditions compared to single molecular weight HA.

[0035] 2. The present invention further discovers that when Q3C is used in combination with full molecular weight HA and its derivatives, it can further improve joint lubrication, increase tissue moisture, and maintain tissue elasticity.

[0036] 3. This invention posits that the synergistic effect between full-molecular-weight HA and Q3C is not simply due to the superposition of their individual activities, but is closely related to the maintenance of the stability of HA's molecular weight distribution under OA conditions. Local inflammation and oxidative stress in OA joints can promote HA degradation and generate low-molecular-weight fragments, thereby weakening HA's original lubricating, water-retaining, and protective regulatory functions. Full-molecular-weight HA, with its broad molecular weight distribution, can simultaneously provide the viscoelastic lubrication and protective receptor regulation of high-molecular-weight components, as well as the superior interfacial hydration and matrix distribution advantages of intermediate-molecular-weight components, forming a multi-layered functional system not possessed by single-molecular-weight HA. Q3C, on the other hand, may slow down the rate of decrease in the molecular weight of full-molecular-weight HA by alleviating inflammation and oxidative stress-related damage and inhibiting the enzymatic and free radical degradation processes of HA, thus maintaining its broad molecular weight distribution structure and corresponding functional advantages. In contrast, single-molecular-weight HA, lacking distribution redundancy and functional complementarity, even with some inhibition of degradation, is unlikely to exhibit the same level of synergistic anti-OA effect when used in combination with Q3C as full-molecular-weight HA, even if its degradation process is somewhat inhibited.

[0037] 4. The present invention also found that, based on the core combination of full molecular weight HA and Q3C, the addition of calcium source, protein matrix and anti-inflammatory active ingredients can further enhance the improvement effect of the composition on OA-related indicators.

[0038] The above-mentioned efficacy has been fully demonstrated using a rat model.

[0039] In the above composition, the weight-average molecular weight distribution of the full molecular weight HA or its salt ranges from 0.2 million to 1.5 million; the molecular weight dispersion coefficient Mw / Mn is above 5.

[0040] In some embodiments of the present invention, the sodium HA of Examples 1 to 6 of the prior application were verified to exhibit good results.

[0041] In the above composition, the full molecular weight HA salt is one or a mixture of full molecular weight HA sodium, full molecular weight HA zinc, full molecular weight HA potassium, and full molecular weight HA magnesium.

[0042] In addition, the present invention also discloses the use of the composition described above to prepare products for treating and / or preventing osteoarthritis or degenerative joint diseases such as osteoarthritis (OA);

[0043] The product has at least one of the following functions: reducing inflammation, lubricating joints, increasing connective tissue moisture, and restoring connective tissue elasticity.

[0044] Finally, the present invention also discloses a product, which is a food, health food, special medical purpose formula food or medicine, and the product comprises the composition as described in any one of the above and pharmaceutically or food-grade excipients.

[0045] The product has at least one of the following functions: reducing inflammation, lubricating joints, increasing connective tissue moisture, and restoring connective tissue elasticity.

[0046] The products are tablets, capsules, powders, granules, pills, gummies, or beverages, etc.

[0047] This application has at least the following beneficial effects:

[0048] 1. The new research findings of this invention have revealed that full molecular weight HA and Q3C can be used to prepare drugs that reduce inflammation, lubricate joints, increase connective tissue moisture, and restore connective tissue elasticity.

[0049] 2. Further research of this invention has demonstrated that the combination of full molecular weight HA or its salts and Q3C has a more significant effect on reducing inflammatory response, lubricating joints, increasing connective tissue moisture, and restoring connective tissue elasticity;

[0050] 3. The preferred composition of the present invention contains non-denatured type II collagen, chitosan oligosaccharide, curcumin, dihydroquercetin, and other components, which can improve OA-related conditions from multiple aspects such as joint lubrication, cartilage matrix maintenance, inflammation regulation, and nutritional support. Specifically, full-molecular-weight HA lubricates joints and maintains connective tissue moisture and elasticity; chitosan oligosaccharide assists in cartilage repair; non-denatured type II collagen helps maintain cartilage matrix stability; curcumin and dihydroquercetin synergistically exert antioxidant and inflammation-regulating effects; calcium lactate provides a calcium source; casein and soy protein provide nutritional support; and Q3C can work in conjunction with the above components to further enhance the composition's effect on improving OA-related indicators.

[0051] In summary, the above-mentioned compositions, when used in combination in specific proportions, can better exert their activity and demonstrate significantly superior technical effects compared to single-component or simple compositions in the prevention and treatment of joint degenerative diseases such as osteoarthritis (OA). They also exhibit high safety, wide applicability in dosage forms, and broad application prospects. Detailed Implementation

[0052] The present invention will now be clearly and completely described in conjunction with embodiments thereof. It should be noted that, unless specific conditions are specified in the embodiments, conventional conditions or conditions recommended by the manufacturer shall apply. Reagents or instruments whose manufacturers are not specified are all commercially available conventional products.

[0053] Part 1: Applications of Q3C in the Treatment and Relief of Osteoarthritis (OA)

[0054] Experimental Example 1

[0055] An osteoarthritis (OA) rat model was established by intra-articular injection of 30 µl sodium iodoacetate (0.2 mg / rat). The specific method was as follows: Healthy male SD rats (200±20 g) were selected and acclimatized for 7 days. The ambient temperature was 22±2℃, relative humidity 50-60%, with a 12-hour light-dark cycle and free access to food and water. Under mild anesthesia, sodium iodoacetate solution was injected into the right knee joint cavity of the rats to establish the OA model. The model was considered successfully established when, 24 hours after model establishment, the rats exhibited significant lameness, reduced weight-bearing, or jumping behavior in the right hind limb. The rats were observed for 10 consecutive days after model establishment and then randomly divided into groups of 10 rats each according to Table 1. Each group received the corresponding treatment via gavage for 60 days. At the end of the experiment, the degree of joint swelling and pain threshold were measured.

[0056] The results are shown in Table 1. Compared with the normal group, the joint swelling of rats in the OA model group was significantly increased, while the mechanical tenderness threshold and thermal pain threshold were significantly reduced (P<0.05), indicating that the OA model was successfully established.

[0057] Compared with the OA model group, the Q3C treatment group significantly reduced joint swelling and significantly increased the pain threshold (P<0.05), indicating that Q3C has certain anti-inflammatory and analgesic effects and can relieve OA symptoms.

[0058] Table 1. Group settings and results of Experiment Example 1 (Mean±SD, n=10)

[0059] Group Intervention components (dosage) Joint swelling (%) Tenderness threshold (g) Thermal pain threshold (s) normal group PBS 0 466.35±25.61 18.30±1.05 OA Model Group PBS <![CDATA[55.80±6.50 # ]]> <![CDATA[221.43±35.82 # ]]> <![CDATA[5.44±3.12 # ]]> Positive drug group Celecoxib (100 mg / kg) <![CDATA[32.60±4.21 * ]]> <![CDATA[378.50±30.24 * ]]> <![CDATA[14.59±2.34 * ]]> Q3C Group Q3C (200 mg / kg) <![CDATA[43.43±5.41 * ]]> <![CDATA[308.37±32.28 * ]]> <![CDATA[10.82±1.91 * ]]>

[0060] Note: Compared with the normal group # P<0.05; compared with the OA model group, * P<0.05.

[0061] Part Two: Relevant Applications of Two-Component Compositions in the Treatment and Relief of Osteoarthritis (OA)

[0062] This section uses full molecular weight HA or its salts in combination with Q3C, and verifies the efficacy of both through rat experiments.

[0063] (I) Preparation method of two-component composition

[0064] The preparation methods for Examples 1-5 and Comparative Examples 1-8 are as follows:

[0065] Grind: Grind the HA and Q3C of each group listed in Table 2 below thoroughly and filter through an 80-100 mesh sieve;

[0066] Mixing: Weigh the composition of each group according to the components and weight parts of the formula, place them in a three-dimensional mixer, mix and stir for 30 minutes until the color is uniform, and make each group of powder.

[0067] Table 2 Formulation Tables for Examples 1-5 and Comparative Examples 1-8

[0068] Composition of the composition (parts by weight) HA Q3C Example 1 0.5 (HA1) 0.5 Example 2 1 (HA1) 0.3 Example 3 1.25 (HA1) 0.2 Example 4 1 (HA2) 0.3 Example 5 1 (HA3) 0.3 Comparative Example 1 1 (HA1) 0 Comparative Example 2 0 0.3 Comparative Example 3 1 (HA4) 0.3 Comparative Example 4 1 (HA5) 0.3 Comparative Example 5 1 (HA6) 0.3 Comparative Example 6 1 (HA4) 0 Comparative Example 7 1 (HA5) 0 Comparative Example 8 1 (HA6) 0

[0069] The information about HA used in this invention is as follows:

[0070] HA1: CN113512134B, Example 6.

[0071] HA2: CN113512134B, Example 1;

[0072] HA3: CN113512134B, Example 3;

[0073] All of the above HA1 to HA3 were provided by Shandong Guantianxia Biotechnology Co., Ltd.

[0074] HA4 to HA6 are all single molecular weight HA; their molecular weights are as follows:

[0075] HA4: 300,000;

[0076] HA5: 600,000;

[0077] HA6: 1.4 million.

[0078] (II) Effect of Two-Component Composition on OA

[0079] Experiment Example 2

[0080] Male SD rats (200±20 g) were selected to establish an osteoarthritis (OA) model, using the same method as in Experiment 1. After modeling, rats were randomly divided into groups according to Table 2, with 10 rats in each group receiving the drug via gavage once daily for 60 consecutive days. After the last administration, the degree of joint swelling and pain threshold-related indicators were measured in the rats, and the test results are shown in Table 3.

[0081] Table 3. Group settings and results of Experiment Example 2 (Mean±SD, n=10)

[0082] Group Intervention components (dosage) Joint swelling (%) Tenderness threshold (g) Thermal pain threshold (s) normal group PBS 0 470.25±28.14 19.65±1.22 OA Model Group PBS <![CDATA[52.32±6.81 # ]]> <![CDATA[220.18±34.56 # ]]> <![CDATA[5.62±2.95 # ]]> Positive drug group Celecoxib (100 mg / kg) <![CDATA[28.86±4.32 * ]]> <![CDATA[431.47±31.26 * ]]> <![CDATA[17.83±2.41 * ]]> Example 1 Example 1 Formulation (30 mg / kg) <![CDATA[39.22±4.94 * ]]> <![CDATA[349.63±30.18 * ]]> <![CDATA[13.08±2.03 * ]]> Example 2 Example 2 Formulation (30 mg / kg) <![CDATA[33.40±3.48 * ]]> <![CDATA[394.72±28.63 * ]]> <![CDATA[15.58±2.19 * ]]> Example 3 Example 3 Formulation (30 mg / kg) <![CDATA[38.13±4.27 * ]]> <![CDATA[361.84±29.36 * ]]> <![CDATA[13.94±2.11 * <!-- 5 -->]]> Example 4 Example 4 Formulation (30 mg / kg) <![CDATA[35.96±4.11 * ]]> <![CDATA[371.26±27.71 * ]]> <![CDATA[14.29±2.06 * ]]> Example 5 Example 5 Formulation (30 mg / kg) <![CDATA[36.84±4.39 * ]]> <![CDATA[367.93±28.18 * ]]> <![CDATA[14.07±2.14 * ]]> Comparative Example 1 Comparative Example 1 formulation (30 mg / kg) 43.78±5.63 301.47±26.34 10.23±1.86 Comparative Example 2 Comparative Example 2 formulation (30 mg / kg) 45.64±5.30 319.28±27.76 11.04±1.94 Comparative Example 3 Comparative Example 3 formulation (30 mg / kg) 45.91±5.16 328.64±28.07 11.76±2.04 Comparative Example 4 Comparative Example 4 Formulation (30 mg / kg) 43.85±4.96 339.82±28.48 12.31±2.01 Comparative Example 5 Comparative Example 5 formulation (30 mg / kg) 42.90±4.80 344.67±29.06 12.68±2.09 Comparative Example 6 Comparative Example 6 Formulation (30 mg / kg) 46.03±5.92 286.14±25.73 9.79±1.74 Comparative Example 7 Comparative Example 7 Formulation (30 mg / kg) 43.57±5.07 294.86±26.44 10.08±1.81 Comparative Example 8 Comparative Example 8 formulation (30 mg / kg) 43.96±5.54 299.41±26.88 10.34±1.87

[0083] Note: Compared with the normal group # P<0.05; compared with the OA model group, * P<0.05.

[0084] Results analysis:

[0085] The test results are shown in Table 3. Compared with the normal group, the joint swelling of rats in the OA model group was significantly increased, while the tenderness threshold and thermal pain threshold were significantly decreased. The differences were statistically significant (P<0.05), indicating that the OA model was successfully constructed.

[0086] Compared to the OA model group, all other intervention groups showed varying degrees of reduction in joint swelling and improvement in tenderness and thermal pain thresholds. Comparative Example 1 (HA1 only) and Comparative Example 2 (Q3C only) showed improvement in related symptoms compared to the OA model group, indicating that both have a certain effect in improving OA. Furthermore, the combination of HA2 and HA3 with Q3C in this invention showed a better improvement effect, suggesting that full-molecular-weight HAs with different distribution characteristics have the potential to improve OA when used in combination with Q3C.

[0087] Of the formulations in Examples 1 to 3, the formulation in Example 2 showed the best improvement in osteoarthritis (OA) (P<0.05). Further analysis revealed that Example 2 showed better improvement in joint swelling, tenderness threshold, and thermal pain threshold than Comparative Example 1 (HA1 only) and Comparative Example 2 (Q3C only), indicating that the combination of Q3C and full molecular weight HA1 had a better improvement effect than any single component, suggesting that the combination of the two has a good synergistic effect.

[0088] On the other hand, compared with the corresponding single molecular weight HA group (comparative examples 6-8), the single molecular weight HA and Q3C formulation group (comparative examples 3-5) showed similar changes in joint swelling, tenderness threshold and thermal pain threshold, and did not show the same advantages as the full molecular weight HA and Q3C combination. This indicates that the combination of single molecular weight HA and Q3C does not have a good synergistic effect on OA improvement.

[0089] In summary, the results show that the combined use of full-molecular-weight HA and Q3C achieves better improvement in OA-related indicators than using either component alone; compared to the single-molecular-weight HA system, the full-molecular-weight HA system exhibits a more significant gain effect when combined with Q3C. Therefore, full-molecular-weight HA and Q3C constitute the core combination for achieving the optimal technical effect in this invention.

[0090] Part Three: Relevant Applications of the Complete Formulation Composition in the Treatment and Relief of OOA

[0091] This section introduces auxiliary ingredients to construct a complete formulation composition based on the two-component composition in Part Two, further enhancing the anti-inflammatory and joint-protective effects.

[0092] (I) Preparation method of the complete formulation composition

[0093] Grinding: Grind and crush each composition of the group described in Table 4, and filter through an 80-100 mesh sieve;

[0094] Mixing: Weigh the composition of each group according to the components and weight parts of the formula, place them in a three-dimensional mixer, mix and stir for 30 minutes until the color is uniform, and make each group of powder.

[0095] Table 4 Formulations of Examples 6-10 and Comparative Examples 9-19

[0096] Composition of the composition (parts by weight) HA Chitosan oligosaccharide Type II collagen Casein Soy protein Curcumin Dihydroquercetin Calcium lactate Q3C Example 6 1.0 (HA1) 0.65 0.08 1 1 0.3 0.1 1 0.3 Example 7 1.0 (HA2) 0.65 0.08 1 1 0.3 0.1 1 0.3 Example 8 1.0 (HA3) 0.65 0.08 1 1 0.3 0.1 1 0.3 Example 9 0.8 (HA1) 0.65 0.08 1 1 0.3 0.1 1 0.3 Example 10 1.25 (HA1) 0.65 0.08 1 1 0.3 0.1 1 0.3 Comparative Example 9 0 0.65 0.08 1 1 0.3 0.1 1 0.3 Comparative Example 10 1.0 (HA1) 0.65 0.08 1 1 0.3 0.1 1 0 Comparative Example 11 0 0.65 0.08 1 1 0.3 0.1 1 0 Comparative Example 12 1.0 (HA4) 0.65 0.08 1 1 0.3 0.1 1 0.3 Comparative Example 13 1.0 (HA5) 0.65 0.08 1 1 0.3 0.1 1 0.3 Comparative Example 14 1.0 (HA6) 0.65 0.08 1 1 0.3 0.1 1 0.3 Comparative Example 15 1.0 (HA1) 0 0.08 1 1 0.3 0.1 1 0.3 Comparative Example 16 1.0 (HA1) 0.65 0 1 1 0.3 0.1 1 0.3 Comparative Example 17 1.0 (HA1) 0.65 0.08 0 0 0.3 0.1 1 0.3 Comparative Example 18 1.0 (HA1) 0.65 0.08 1 1 0 0.1 1 0.3 Comparative Example 19 1.0 (HA1) 0.65 0.08 1 1 0.3 0.1 0 0.3

[0097] (II) The effect of the complete formulation composition on OA

[0098] Experimental Example 3

[0099] Male SD rats (200±20 g) were selected to establish an osteoarthritis (OA) model, using the same method as in Experiment 1. After modeling, rats were randomly divided into groups according to Table 5 and administered the drug via gavage, with 10 rats in each group, once daily for 60 consecutive days. After the last administration, the degree of joint swelling and pain threshold-related indicators were measured in the rats.

[0100] The test results are shown in Table 5.

[0101] Table 5. Group settings and results of Experiment Example 3 (Mean±SD, n=10)

[0102] Group Intervention components (dosage) Joint swelling (%) Tenderness threshold (g) Thermal pain threshold (s) normal group PBS 0 469.84±27.91 18.57±1.24 OA Model Group PBS <![CDATA[55.28±6.77 # ]]> <![CDATA[221.36±34.28 # ]]> <![CDATA[5.71±2.88 # ]]> Positive drug group Celecoxib (100 mg / kg) <![CDATA[28.74±4.26 * ]]> <![CDATA[402.15±30.94 * ]]> <![CDATA[16.88±2.37 * ]]> Example 2 Example 2 Formulation (30 mg / kg) <![CDATA[35.78±3.56 * ]]> <![CDATA[362.84±28.47 * ]]> <![CDATA[13.12±2.16 * ]]> Example 6 Example 6 Formulation (30 mg / kg) <![CDATA[31.18±3.94 * ]]> <![CDATA[387.86±29.72 * ]]> <![CDATA[15.54±2.21 * ]]> Example 7 Example 7 Formulation (30 mg / kg) <![CDATA[34.26±4.03 * ]]> <![CDATA[371.95±29.18 * ]]> <![CDATA[14.03±2.13 * ]]> Example 8 Example 8 Formulation (30 mg / kg) <![CDATA[34.58±4.08 * ]]> <![CDATA[369.84±29.36 * ]]> <![CDATA[13.95±2.18 * ]]> Example 9 Example 9 Formulation (30 mg / kg) <![CDATA[35.92±4.22 * ]]> <![CDATA[367.72±29.88 * ]]> <![CDATA[13.41±2.24 * ]]> Example 10 Example 10 Formulation (30 mg / kg) <![CDATA[36.47±4.31 * ]]> <![CDATA[363.41±29.64 * ]]> <![CDATA[13.62±2.20 * ]]> Comparative Example 9 Comparative Example 9 Formulation (30 mg / kg) 41.82±4.90 337.94±28.84 11.28±2.01 Comparative Example 10 Comparative Example 10 Formulation (30 mg / kg) 41.08±4.82 341.26±28.96 11.46±2.03 Comparative Example 11 Comparative Example 11 Formulation (30 mg / kg) 43.63±5.01 323.68±28.76 10.04±1.98 Comparative Example 12 Comparative Example 12 Formulation (30 mg / kg) <![CDATA[38.12±4.37 * ]]> <![CDATA[352.76±28.95 * ]]> <![CDATA[12.88±2.06 * ]]> Comparative Example 13 Comparative Example 13 Formulation (30 mg / kg) <![CDATA[38.05±4.28 * ]]> <![CDATA[357.83±29.06 * ]]> <![CDATA[13.16±2.10 * ]]> Comparative Example 14 Comparative Example 14 Formulation (30 mg / kg) <![CDATA[37.18±4.21 * ]]> <![CDATA[361.95±29.18 * ]]> <![CDATA[13.29±2.12 * ]]> Comparative Example 15 Comparative Example 15 Formulation (30 mg / kg) <![CDATA[36.98±4.34 * ]]> <![CDATA[355.82±29.06 * ]]> <![CDATA[13.10±2.08 * ]]> Comparative Example 16 Comparative Example 16 Formulation (30 mg / kg) <![CDATA[37.18±4.36 * ]]> <![CDATA[354.26±29.10 * ]]> <![CDATA[13.03±2.10 * ]]> Comparative Example 17 Comparative Example 17 Formulation (30 mg / kg) <![CDATA[36.74±4.29 * ]]> <![CDATA[357.64±29.14 * ]]> <![CDATA[13.21±2.11 * ]]> Comparative Example 18 Comparative Example 18 Formulation (30 mg / kg) <![CDATA[37.42±4.41 * ]]> <![CDATA[352.94±29.22 * ]]> <![CDATA[12.94±2.12 * ]]> Comparative Example 19 Comparative Example 19 Formulation (30 mg / kg) <![CDATA[37.36±4.39 * ]]> <![CDATA[353.41±29.16 * ]]> <![CDATA[12.98±2.09 * ]]>

[0103] Note: Compared with the normal group # P<0.05; compared with the OA model group, * P<0.05.

[0104] Compared with the normal group, the OA model group rats showed significantly increased joint swelling, while the tenderness threshold and thermal pain threshold were significantly reduced, with statistically significant differences (P<0.05), indicating that the OA model was successfully established.

[0105] Compared with the OA model group, the positive drug group, Example 2 group, Examples 6-10 and Comparative Examples 12-19 all reduced joint swelling and increased the tenderness threshold and thermal pain threshold to varying degrees, indicating that the above compositions all have certain anti-inflammatory and analgesic effects.

[0106] Further comparison revealed that Example 6 showed a further improvement in three indicators—joint swelling, tenderness threshold, and thermal pain threshold—compared to Example 2, and its overall effect was close to that of the positive drug group. This indicates that by further introducing auxiliary components such as chitosan oligosaccharide, type II collagen, protein matrix, curcumin, and calcium lactate on the basis of the core ratio of full molecular weight HA1 and Q3C, the anti-inflammatory, analgesic, and joint-protective effects of the composition can be further enhanced. In other words, the effect of the complete formula is better than that of the two-component composition of HA1 and Q3C.

[0107] Compared with Examples 7 and 8, Example 6 shows better results, indicating that HA of different molecular weights can form effective combinations with Q3C and auxiliary components, with HA1 showing better compatibility. Example 6 also shows better performance than Examples 9 and 10, indicating that in the complete formulation system of this invention, the optimal two-component ratio of HA and Q3C in Part Two yields the best results, further demonstrating that the complete formulation has good inheritance of the core synergistic system in Part Two.

[0108] Example 6 showed better results compared to Comparative Examples 12-14, indicating that using full molecular weight HA in the complete formulation system is better than using single molecular weight HA. Although single molecular weight HA can improve relevant indicators to a certain extent, its overall effect is still weaker than that of the full molecular weight HA system.

[0109] Comparative Examples 9, 10, and 11 represent formulations with HA removed, Q3C removed, and both HA and Q3C removed, respectively. The effects of all these groups were significantly lower than those of Example 6, with Comparative Example 11 showing the weakest improvement. This indicates that full-molecular-weight HA and Q3C remain the core active combination in the complete formulation of this invention. When both are absent, relying solely on other auxiliary components can still produce some improvement, but the overall effect decreases, making it difficult to achieve the optimal level of the complete formulation.

[0110] Comparative Examples 15-19 removed chitosan oligosaccharide, type II collagen, protein matrix, curcumin, and calcium lactate, respectively. The results were all lower than those of Example 6, but were still better than those of Comparative Examples 9-11, which removed the core components. This shows that the introduction of auxiliary components can further enhance the overall effect of the composition of the present invention, while full molecular weight HA and Q3C are the necessary core components to achieve the best technical effect.

[0111] In summary, the effects of complete formulation compositions are superior to those of two-component compositions; complete formulations containing full molecular weight HA are superior to those containing single molecular weight HA; and full molecular weight HA and Q3C are the core combination for achieving the best technical effect, and neither can be omitted. On this basis, the addition of auxiliary ingredients such as chitosan oligosaccharide, type II collagen, protein matrix, curcumin, dihydroquercetin and calcium lactate can further enhance the composition's effect on improving OA.

[0112] Part Four: Application of Complete Formulation Compositions in Products for the Treatment and Relief of OA-Related Issues

[0113] (a) Application Example 1

[0114] Tablet preparation

[0115] This application example provides a tablet with the formulation shown in Table 6: Table 6 Tablet Formulation

[0116] Raw material name Dosage (g) Example 6 Formula Powder 85 microcrystalline cellulose 12 Cross-linked carboxymethyl cellulose sodium 2 magnesium stearate 0.5 Micronized silicone 0.5 total 100

[0117] The tablets are prepared using a direct powder compression process, and the specific steps are as follows:

[0118] (1) Raw material pretreatment: Microcrystalline cellulose, cross-linked sodium carboxymethyl cellulose, magnesium stearate and micronized silica gel are passed through a 40-80 mesh sieve for later use;

[0119] (2) Mixing: Weigh the powder of the formulation in Example 6 with microcrystalline cellulose and croscarmellose sodium according to the formulation ratio, and mix them in a three-dimensional mixer for 20-30 min; add micronized silica gel and continue mixing for 5 min; finally add magnesium stearate and mix for 3-5 min until the mixture is uniform.

[0120] (3) Tableting: The uniformly mixed material is fed into a tablet press and tableted. The weight of a single tablet is controlled to be 0.99~1.01 g, the tablet hardness is 50~70 N, and the thickness is 3~5 mm, thus obtaining the tablet.

[0121] (II) Application Example 2

[0122] Preparation of capsules

[0123] This application example provides a capsule formulation, the formula of which is shown in Table 7: Table 7 Capsule Formulation

[0124] Raw material name Dosage (g) Example 6 Formula Powder 85 microcrystalline cellulose 12 magnesium stearate 1 Micronized silicone 2 total 100

[0125] The specific preparation steps for capsules are as follows:

[0126] (1) Raw material pretreatment: The powder and each excipient of the formulation in Example 6 are passed through a 40-80 mesh sieve for later use;

[0127] (2) Mixing: Mix the powder of the formulation in Example 6 with microcrystalline cellulose evenly; add micronized silica gel and mix for 5 min; then add magnesium stearate and mix for 3-5 min until evenly distributed;

[0128] (3) Filling: Use a capsule filling machine to fill the uniformly mixed powder into the empty capsule shell, and control the amount of each capsule to be 400~600 mg;

[0129] (4) Inspection and packaging: The capsules are inspected for appearance and weight difference. After passing the inspection, they are packaged in a light-proof and sealed manner.

[0130] (III) Application Example 3

[0131] Preparation of granules

[0132] This application example provides a granule formulation, the formulation of which is shown in Table 8: Table 8 Granule Formulation

[0133] Raw material name Dosage (g) Example 6 Formula Powder 50 maltodextrin 25 Fructooligosaccharides 20 Sodium carboxymethyl cellulose 3 Citric acid 0.5~1.5 essence 0.5~1.5 total 100

[0134] The specific preparation steps for granules are as follows:

[0135] (1) Raw material pretreatment: The powder, maltodextrin, fructooligosaccharide and sodium carboxymethyl cellulose of the formulation in Example 6 were passed through a 40-80 mesh sieve for later use;

[0136] (2) Mixing: Weigh the powder of the formulation in Example 6, maltodextrin, and fructooligosaccharides in proportion, place them in a mixer and mix for 10-20 minutes until they are evenly mixed;

[0137] (3) Preparation of soft material: Slowly add purified water to the above mixture while stirring. The amount of purified water added is 8% to 15% of the total weight of the material to obtain soft material.

[0138] (4) Granulation: Granulate the soft material through a 14-20 mesh sieve;

[0139] (4) Drying and granulation: Drying at 50~60℃ with a moisture content of no more than 5%, and then granulating through a 16~24 mesh sieve to obtain uniform particles;

[0140] (5) Flavoring and packaging: Add citric acid and flavoring, mix evenly at low speed, and then package into granules.

[0141] (iv) Application Example 4

[0142] Preparation of gummies

[0143] This application example provides a gummy candy with the following formula as shown in Table 9: Table 9 Gummy Recipes

[0144] Raw material name Dosage (g) Example 6 Formula Powder 200 gelatin 130 Sorbitol liquid 150 glycerin 20 Citric acid 0.5~2 juice 20~80 Purified water 50~150 Vitamin C 2

[0145] The specific preparation steps are as follows:

[0146] (1) Preparation of gelatin solution: Soak gelatin in purified water for 20-30 min, then heat it in a water bath at 60-70℃ until it is completely dissolved to make a uniform and transparent gelatin solution, and keep it warm for later use;

[0147] (2) Mixing the main ingredients: Mix the sorbitol solution and glycerin evenly; disperse or suspend the powder of the formulation in Example 6 and vitamin C in the juice and stir evenly; then mix the two parts to form a uniform slurry;

[0148] (3) Mixing and molding: Slowly add the above slurry to the gelatin solution and mix evenly at 50~60℃; after degassing, add citric acid and continue to stir evenly; pour the mixture into the soft candy mold and cool for 20~30 min until solidified.

[0149] (4) Demolding and packaging: After demolding, sand can be applied, and after drying, it can be packaged in aluminum foil bags and sealed for storage.

[0150] The above description is merely an embodiment of this application and is not intended to limit the scope of this application. Various modifications and variations can be made to this application by those skilled in the art. Any modifications, equivalent substitutions, improvements, etc., made within the spirit and principles of this application should be included within the scope of the claims of this application.

Claims

1. A novel application of full molecular weight HA, characterized in that, This product is used to prepare a product that improves joint inflammation response, enhances joint lubrication, increases connective tissue moisture, or restores connective tissue elasticity; the product is a food, health food, special medical purpose formula food, or medicine used to prevent and / or improve OA.

2. The novel application according to claim 1, characterized in that, The weight-average molecular weight distribution range of the full molecular weight HA or its salt is 0.2 million to 1.5 million, and the molecular weight dispersion coefficient Mw / Mn≥5. The full molecular weight HA salt is one or more of the following: full molecular weight HA sodium, full molecular weight HA zinc, full molecular weight HA potassium, and full molecular weight HA magnesium.

3. A composition, characterized in that, By weight, it includes the following components: 0.5~1.25 parts of total molecular weight HA or its salt; 0.2~0.5 parts Q3C; The combination of full molecular weight HA and Q3C exhibits a synergistic effect in improving OA-related indicators.

4. The composition according to claim 3, characterized in that, The weight ratio of the total molecular weight HA to Q3C is 0.8–1.25 : 0.2–0.

4.

5. The composition according to claim 3 or 4, characterized in that, The weight-average molecular weight distribution of the full molecular weight HA or its salts ranges from 0.2 million to 1.5 million, and the molecular weight dispersion coefficient Mw / Mn ≥ 5.

6. The composition according to any one of claims 3 to 5, characterized in that, It also includes the following components by weight: 0.3~0.8 parts chitosan oligosaccharide; 0.02~0.15 parts of non-denatured type II collagen; 0.5 to 2 parts casein; 0.5 to 2 parts soy protein; 0.1~0.6 parts curcumin; 0.05~0.5 parts of dihydroquercetin; 0.5 to 2 parts calcium lactate.

7. The composition according to claim 6, characterized in that, The full molecular weight HA salt is one or more of the following: full molecular weight HA sodium, full molecular weight HA zinc, full molecular weight HA potassium, and full molecular weight HA magnesium.

8. Use of the composition according to any one of claims 3 to 7 in the preparation of products for the prevention and / or improvement of OA.

9. A product characterized in that, The product is a food, health food, special medical purpose formula food or medicine, and the product contains the composition of any one of claims 3 to 7, as well as pharmaceutically or food-grade excipients. The product has at least one of the following functions: reducing inflammation, lubricating joints, increasing connective tissue moisture, and restoring connective tissue elasticity.

10. The product according to claim 9, characterized in that, The product is in the form of tablets, capsules, powders, granules, pills, gummies, or beverages.