Fishbone extract composition containing plant extract and application of fishbone extract composition in bone joint health

Abstract

The invention provides a composition containing a plant extract and a fishbone extract and application of the composition in bone joint health. The composition can repair cartilage, promote secretion of bone joint lubricating liquid, remove inflammatory cells, inhibit inflammatory factors, delay cartilage degeneration, improve bone mineral density and bone strength, maintain bone mineral density, enhance bone firmness, improve osteoporosis and improve arthritis.

Description

Technical Field

[0001] The present invention belongs to the field of health care products, and relates to a composition comprising plant extracts and fish bone extracts and its use in bone and joint health. The composition can repair cartilage, promote the secretion of bone and joint lubricating fluid, remove inflammatory cells, inhibit inflammatory factors, delay cartilage degeneration, increase bone density and bone strength, maintain bone density, enhance bone firmness, improve osteoporosis, and improve arthritis. Background Art

[0002] Bones are important organs of the human body. Their structures include periosteum, bone and bone marrow. Periosteum contains abundant nerves and blood vessels, and bone includes compact bone and spongy bone. Bone mineral density (BMD) is the full name of bone mineral density, which is an important indicator of bone strength.

[0003] Osteoarthritis (OA) is a chronic joint disease characterized by degeneration, destruction and bone hyperplasia of articular cartilage. Clinically, joint swelling and pain, bone hyperplasia and limited mobility are the most common symptoms. Studies have shown that OA and osteoporosis are both age-related diseases, and the two often coexist. Some symptoms may be caused by osteoporosis. Calcium supplementation can promote the synthesis of articular cartilage matrix, delay the progression of OA, and is beneficial for OA prevention. Excessive calcium supplementation may promote bone hyperplasia, aggravate joint pain and joint movement disorders, so calcium supplementation should be moderate.

[0004] Articular cartilage is composed of bone matrix (collagen, proteoglycan, water) and chondrocytes. The progressive degeneration of articular cartilage is also an important sign of OA. With the increase of age, chondrocytes have metabolic abnormalities, their number gradually decreases, and the elasticity of articular cartilage decreases. Articular cartilage continues to degenerate and wear, and hard friction occurs between bones. Symptoms such as inflammation, pain, swelling, and bone friction sound appear in the joints, and osteoarthritis occurs. Therefore, it is necessary to repair cartilage, increase the secretion of osteoarticular lubricating fluid, remove inflammatory cells, and inhibit inflammatory factors, so as to delay cartilage degeneration.

[0005] In modern medical treatment of osteoarthritis, hormone-containing chemical drugs are often used to control the disease, or non-steroidal anti-inflammatory drugs are used, but the treatment effect is often poor and it is not suitable for long-term use. Therefore, it is of great significance to find a composition that can repair cartilage, promote the secretion of bone and joint lubricating fluid, remove inflammatory cells, inhibit inflammatory factors, delay cartilage degeneration, increase bone density and bone strength, maintain bone density, enhance bone firmness, improve osteoporosis, and improve arthritis. Summary of the invention

[0006] The purpose of the present invention is to provide a composition comprising plant extracts and fish bone extracts and its use in bone and joint health. The composition can repair cartilage, promote the secretion of bone and joint lubricating fluid, remove inflammatory cells, inhibit inflammatory factors, delay cartilage degeneration, increase bone density and bone strength, maintain bone density, enhance bone firmness, improve osteoporosis, and improve arthritis.

[0007] The present invention combines plant extracts with specific efficacy with fish bone extracts, which not only have good antioxidant and anti-inflammatory effects, but also can repair cartilage, promote the secretion of bone and joint lubricating fluid, remove inflammatory cells, inhibit inflammatory factors, and delay cartilage degeneration. The combined composition has a synergistic effect, significantly promotes the body's absorption, utilization and conversion of calcium, promotes collagen synthesis, increases bone strength, improves osteoporosis and arthritis caused by calcium deficiency, and has a good preventive and improving effect on various bone and joint diseases.

[0008] The present invention provides a fish bone extract composition comprising plant extracts, wherein the plant extracts at least comprise one or more of angelica extracts, frankincense extracts, and pepper extracts; and the content of the fish bone extract is ≥ 1 mg / portion. The composition provided by the present invention has the following advantages:

[0009] (1) It has good antioxidant and anti-inflammatory effects;

[0010] (2) It can repair cartilage, increase the secretion of lubricating fluid in bone joints, remove inflammatory cells, and inhibit inflammatory factors;

[0011] (3) Synergistically enhance the body's calcium absorption, utilization and conversion, and increase bone density and bone strength;

[0012] (4) It helps reduce bone loss and synthesize collagen. It can be taken for a long time and has a lasting effect.

[0013] The composition comprising plant extracts and fish bone extracts of the present invention, wherein the plant extracts at least comprise one or more of angelica extracts, frankincense extracts, and pepper extracts; and the content of the fish bone extracts is ≥1 mg / portion.

[0014] In one embodiment, in the composition of the present invention, the fish bone extract is derived from one or more of a freshwater fish bone extract and a marine fish bone extract. In a preferred embodiment, the fish bone extract is derived from a marine fish bone extract. In a more preferred embodiment, the fish bone extract is derived from a deep-sea fish bone extract.

[0015] In one embodiment, in the composition of the present invention, the fish bone extract is derived from one or more of salmon bone extract, eel bone extract, cod bone extract, shark cartilage extract, sea bream bone extract, and tilapia bone extract. In a preferred embodiment, the fish bone extract is derived from one or more of salmon bone extract, eel bone extract, cod bone extract, and shark cartilage extract. In a more preferred embodiment, the fish bone extract is derived from one or more of salmon bone extract, eel bone extract, and cod bone extract. In a particularly preferred embodiment, the fish bone extract is derived from one or more of salmon bone extract and cod bone extract. In a particularly preferred embodiment, the fish bone extract is derived from salmon bone extract. In a particularly preferred embodiment, the fish bone extract is derived from cod bone extract. In a particularly preferred embodiment, the fish bone extract is derived from shark cartilage extract.

[0016] In one embodiment, the fish bone extract includes but is not limited to powder, liquid, and gel. The fish bone extract can be fish bone powder, and the fish bone is processed through a series of processes such as enzymolysis and drying to make fish bone extract powder. The fish bone extract can exist in liquid form, and the effective ingredients in the fish bone are dissolved in a suitable solvent (such as water or other organic solvents) through an extraction process. The fish bone extract can also exist in a gel form, and a suitable gelling agent is added during the extraction process so that the extract has the form of a gel. That is, the fish bone extract includes but is not limited to fish bone powder, fermented fish bone powder, fish bone powder extract, liquid fish bone extract, gel fish bone extract, and liquid collagen calcium.

[0017] In one embodiment, in the composition of the present invention, the plant extract comprises at least two or three of angelica extract, frankincense extract and pepper extract.

[0018] In a preferred embodiment, in the composition of the present invention, the plant extract is frankincense extract or pepper extract.

[0019] In one embodiment, the composition of the present invention, the effective ingredients of the plant extract include at least one or more of phytocannabinoids, purseratin, pursera alcohol, boswellic acid, sabinene, limonene, pinene, caryophyllene, and humulene.

[0020] In one embodiment, in the composition of the present invention, the boswellic acid includes one or more of ursolic acid type and oleanane type; further, the ursolic acid type includes β-boswellic acid, acetyl-β-boswellic acid, 3-acetyl-11-carbonyl-β-boswellic acid, 11-carbonyl-β-boswellic acid, acetyl-11-hydroxy-β-boswellic acid, acetyl-11α-methoxy-β-boswellic acid, 9,11-dehydro-β-boswellic acid, 3α-acetyl-9,11-dehydro-β-boswellic acid, one or more, 3-O-acetyl-11-keto-β-boswellic acid, and the oleanane type includes α-boswellic acid, acetyl-α-boswellic acid, 9,11-dehydro-α-boswellic acid, and 3-acetyl-9,11-dehydro-α-boswellic acid.

[0021] In one embodiment, the composition of the present invention, the content of the plant extract is ≥ 100 mg / portion, such as 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 230, 240, 250, 260, 270, 280, 290, 300, 310, 320, 330, 340, 350, 360, 370, 380, 390, 400, 410, 420, 430, 440, 450, 460, 470, 480, 490, 500 mg / portion.

[0022] In a preferred embodiment, in the composition of the present invention, the content of the plant extract is 100-500 mg / portion.

[0023] In a more preferred embodiment, the composition of the present invention, the content of the plant extract is 100-300 mg / serving.

[0024] In one embodiment, the preparation method of the plant extract is selected from enzymatic hydrolysis, water extraction, alcohol extraction, ultrasonic extraction, and supercritical fluid extraction.

[0025] In one embodiment, the method for preparing the plant extract specifically comprises: (1) selecting and processing plant raw materials, such as washing, cutting or crushing;

[0026] (2) Extract the active ingredients by enzymatic hydrolysis, water extraction, alcohol extraction, ultrasonic extraction, supercritical fluid extraction, etc.;

[0027] (3) Separate and purify the extract by filtration, centrifugation, chromatography, membrane separation and crystallization;

[0028] (4) Concentrate and dry the extract by techniques such as evaporation, freeze drying or spray drying.

[0029] In one embodiment, in the composition of the present invention, the content of 3-O-acetyl-11-keto-β-boswellic acid in the frankincense extract is 10-50% of the total mass of the frankincense extract. For example, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50%

[0030] In a preferred embodiment, in the composition of the present invention, the content of 3-O-acetyl-11-keto-β-boswellic acid in the frankincense extract accounts for 10-40% of the total mass of the frankincense extract.

[0031] In a more preferred embodiment, in the composition of the present invention, the content of 3-O-acetyl-11-keto-β-boswellic acid in the frankincense extract accounts for 20-40% of the total mass of the frankincense extract.

[0032] In a more preferred embodiment, in the composition of the present invention, the content of 3-O-acetyl-11-keto-β-boswellic acid in the frankincense extract accounts for 20-30% of the total mass of the frankincense extract.

[0033] In one embodiment, in the composition of the present invention, the pepper extract is black pepper extract.

[0034] In one embodiment, the content of β-caryophyllene in the black pepper extract is ≥ 20%.

[0035] In a preferred embodiment, the β-caryophyllene content in the black pepper extract is ≥ 25%.

[0036] In a more preferred embodiment, the β-caryophyllene content in the black pepper extract is ≥30%.

[0037] In one embodiment, the active ingredients in the black pepper extract further include α-pinene, β-pinene, humulene, limonene, and pinene.

[0038] In one embodiment, in the composition of the present invention, the plant extract is frankincense extract or black pepper extract.

[0039] In a preferred embodiment, the composition of the present invention has an amount of frankincense extract of 100-200 mg / portion, such as 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200 mg / portion.

[0040] In a preferred embodiment, the composition of the present invention, the content of the black pepper extract is 10-100 mg / portion, such as 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 30, 40, 50, 60, 70, 80, 90, 100 mg / portion.

[0041] In one embodiment, in the composition of the present invention, the fish bone extract is prepared by enzymatic hydrolysis.

[0042] In one embodiment, the composition of the present invention, the enzymatic preparation steps of the fish bone extract are: (1) removing fish meat and fat from fish to obtain fish bones; (2) drying the fish bones, and then grinding them to obtain fish bone powder; (3) placing the obtained fish bone powder into an aqueous solution containing a composite protease for enzymatic hydrolysis to obtain fish bone material; (4) taking out the fish bone material after enzymatic hydrolysis, washing it, drying it again, and ultrafine grinding it.

[0043] In one embodiment, in the preparation steps of the fish bone extract, the drying temperature in step (2) is 50-90°C.

[0044] In a preferred embodiment, in the preparation steps of the fish bone extract, the drying temperature in step (2) is 75°C.

[0045] In one embodiment, in the preparation steps of the fish bone extract, the composite protease in step (3) is a composite of two or more selected from papain, trypsin, pepsin, neutral protease, alkaline protease and flavor protease, preferably a composite of alkaline protease and flavor protease.

[0046] In one embodiment, in the preparation steps of the fish bone extract, in step (3), the enzymatic hydrolysis temperature is 32-48° C., and the enzymatic hydrolysis time is 0.5-5 h.

[0047] In one embodiment, in the above-mentioned preparation steps of the fish bone extract, the temperature of the second drying in step (4) is 50-90°C.

[0048] In a preferred embodiment, in the above-mentioned preparation steps of the fish bone extract, the temperature of the second drying in step (4) is 65°C.

[0049] In one embodiment, in the composition of the present invention, the calcium content in the fish bone extract is 10-30% of the total weight of the fish bone extract; for example, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30%. The collagen content is 1-40% of the total weight of the fish bone extract; for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40%.

[0050] In a preferred embodiment, in the composition of the present invention, the calcium content in the fish bone extract accounts for 15-30% of the total weight of the fish bone extract; and the collagen content accounts for 5-40% of the total weight of the fish bone extract.

[0051] In a more preferred embodiment, in the composition of the present invention, the calcium content in the fish bone extract accounts for 15-30% of the total weight of the fish bone extract; and the collagen content accounts for 10-40% of the total weight of the fish bone extract.

[0052] In a more preferred embodiment, in the composition of the present invention, the calcium content in the fish bone extract accounts for 10-25% of the total weight of the fish bone extract; and the collagen content accounts for 10-35% of the total weight of the fish bone extract.

[0053] In a particularly preferred embodiment, in the composition of the present invention, the calcium content in the fish bone extract accounts for 15-25% of the total weight of the fish bone extract; and the collagen content accounts for 15-35% of the total weight of the fish bone extract.

[0054] In one embodiment, the composition of the present invention further comprises glucosamine.

[0055] In one embodiment, the composition of the present invention, the glucosamine content is ≥ 5 mg / portion, such as 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 40, 50, 60, 70, 80, 90, 100 mg / portion.

[0056] In a preferred embodiment, the glucosamine content is 5-100 mg / portion. Further, the glucosamine content is 5-50 mg / portion. Further, the glucosamine content is 10-50 mg / portion.

[0057] In one embodiment, the composition of the present invention further comprises chondroitin sulfate.

[0058] In one embodiment, the composition of the present invention has a chondroitin sulfate content of ≥1 mg / portion, such as 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20 mg / portion.

[0059] In a preferred embodiment, the content of the chondroitin sulfate is 1-10 mg / portion.

[0060] In one embodiment, the composition of the present invention further comprises vitamins.

[0061] In one embodiment, the vitamins in the composition of the present invention include but are not limited to vitamin D, vitamin C, and vitamin E.

[0062] Bones are made up of bone matrix and bone minerals. The main components of bone matrix are collagen and collagen, which act as adhesives and scaffolds. Collagen can help calcium salts deposit. Vitamin C is an antioxidant that can reduce oxidative stress and the generation of free radicals, which can attack or destroy joint tissue. The synthesis of collagen requires the participation of vitamin C. Vitamin C helps reduce bone loss, enhance bone and joint strength, and reduce joint load.

[0063] In one embodiment, the vitamin of the present invention is vitamin C.

[0064] In one embodiment, the composition of the present invention has a vitamin C content of ≥5 mg / portion, such as 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 mg / portion.

[0065] In a preferred embodiment, the vitamin C content is 5-20 mg / serving.

[0066] In one embodiment, the composition of the present invention further comprises hyaluronic acid.

[0067] In one embodiment, the composition of the present invention has a hyaluronic acid content of ≥5 mg / portion, such as 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25 mg / portion.

[0068] In a more preferred embodiment, the composition of the present invention has a hyaluronic acid content of 5-20 mg / portion.

[0069] The present invention provides a composition comprising a plant extract and a fish bone extract, wherein the composition is added with pharmaceutically acceptable excipients to prepare a preparation form that is easily accepted by the human body, including but not limited to tablets, capsules, powders, oral liquids, sprays, and soft capsules. Furthermore, the preparation form of the composition is a soft capsule.

[0070] The present invention also provides use of any of the compositions comprising plant extracts and fish bone extracts in bone and joint health.

[0071] Furthermore, the present invention also provides the use of any of the compositions comprising plant extracts and fish bone extracts in improving osteoporosis, improving arthritis, increasing bone density, increasing bone strength, maintaining bone density, and enhancing bone firmness.

[0072] Furthermore, the present invention also provides use of any of the compositions comprising plant extracts and fish bone extracts in arthritis.

[0073] The present invention provides a composition of plant extracts and fish bone extracts, which can repair cartilage, promote the secretion of bone joint lubricating fluid, remove inflammatory cells, inhibit inflammatory factors, delay cartilage degeneration, improve bone density and bone strength, maintain bone density, enhance bone firmness, improve osteoporosis and improve arthritis.

[0074] Explanation of terms

[0075] The "fish bone extract" refers to a calcium-containing carrier extracted from fish bones, including but not limited to salmon bone extract, eel bone extract, cod bone extract, shark cartilage extract, sea bream bone extract, and tilapia bone extract.

[0076] The "plant extract" is a plant extract with anti-inflammatory effect. Angelica extract contains many types of compounds, the main chemical components of which are volatile oils, sugars, trace elements, and various organic acids and amino acids, etc., which can relieve inflammatory reactions, resist oxidation, inhibit extracellular matrix degradation, regulate chondrocyte metabolism, and reduce cell metabolic degeneration, thereby effectively protecting cartilage. In the present invention, the angelica extract also contains decursinol as the main active ingredient, which is a phenolic compound and also includes decursinol, which has antioxidant, anti-inflammatory, anti-tumor, antibacterial and other effects. The main components of frankincense extract are boswellic acid and terpenoid antioxidants. Boswellic acid inhibits the production of inflammatory reaction substances and inflammatory substances such as TNF-α and IL-6 in the body by inhibiting the activity of 5-lipoxygenase (5-LO), and promotes the production of proteins that constitute proteins. Frankincense extract is an effective anti-arthritic and immunomodulator (Kimmatkar et al.; Phytomedicine. 2003 Jan; 10(1): 3-7; Pungle et al.; Indian J Exp Biol. 2003 Dec; 41(12): 1460-1462). It has been continuously used for the treatment of pain and inflammation and its analgesic effect. Black pepper extract contains piperine, phytocannabinoids, β-caryophyllene, etc., which can improve the bioavailability of nutrients and reduce inflammation and relieve pain.

[0077] The "glucosamine" is a compound composed of glucose and amino groups. It is a derivative of glucose. It has the functions of maintaining the health and elasticity of articular cartilage, promoting the growth and repair of chondrocytes, and slowing down the degeneration and damage of articular cartilage. It also has a certain anti-inflammatory effect and helps to reduce the inflammatory response caused by arthritis.

[0078] The "chondroitin sulfate" is a polysaccharide compound, which mainly exists in cartilage tissue, helps maintain the structure and elasticity of cartilage, helps maintain the health of articular cartilage and slow down cartilage degeneration, promotes the growth and repair of chondrocytes, and has a certain anti-inflammatory effect, can increase the water content in articular cartilage, and improve the elasticity of cartilage. Note: The above explanation is only for further explanation and is not a limitation of the present invention. BRIEF DESCRIPTION OF THE DRAWINGS

[0079] Figure 1 The gait score observation results of rats in each group

[0080] Figure 2 The joint pain score observation results of rats in each group

[0081] Figure 3 The results of the observation of autonomous pain scores of rats in each group

[0082] Figure 4 The macroscopic scoring results of the knee joints of rats in each group

[0083] Figure 5 The results of IL1β detection in the serum of rats in each group

[0084] Figure 6 The results of IL6 detection in the serum of rats in each group are shown in Figure 2

[0085] Figure 7 The results of IL1β gene detection in the knee joints of rats in each group

[0086] Figure 8 The results of IL-6 gene detection in the knee joints of rats in each group

[0087] Fig. 9 This is the result of human body test DETAILED DESCRIPTION

[0088] In order to facilitate the understanding of those skilled in the art, the present invention is further described below in conjunction with embodiments, and the contents mentioned in the implementation modes are not intended to limit the present invention.

[0089] Example

[0090] Example 1

[0091] Remove the fish meat and fat from the fish to obtain the skull and spine, dry the fish bones, and then grind them to obtain fish bone powder. Put the obtained fish bone powder into a composite protease aqueous solution with alkaline protease and flavor protease, and perform enzymolysis at 35°C for 30 minutes to obtain fish bone material. Filter, take out the enzymolyzed fish bone material, wash it, dry it again, and grind it into ultrafine powder to obtain fish bone extract.

[0092] Example 2

[0093] The plant raw materials are selected and processed, such as washing, cutting or crushing. The active ingredients are then extracted by enzymatic hydrolysis, water extraction, alcohol extraction, ultrasonic extraction, supercritical fluid extraction, etc. The extracts are separated and purified by filtration, centrifugation, chromatography, membrane separation and crystallization. The extracts are concentrated and dried by evaporation, freeze drying or spray drying to obtain the corresponding plant extracts.

[0094] Example 3

[0095] 1 part of fish bone extract, 30 parts of frankincense extract, 10 parts of black pepper extract, 4 parts of glucosamine, 1 part of chondroitin sulfate, 2 parts of vitamin C and 2 parts of hyaluronic acid were added to obtain a solid composition.

[0096] Example 4

[0097] 1 part of fish bone extract, 25 parts of frankincense extract, 10 parts of black pepper extract, 4 parts of glucosamine, 1 part of chondroitin sulfate, 3 parts of vitamin C and 2 parts of hyaluronic acid were added to obtain a solid composition.

[0098] Example 5

[0099] 1 part of fish bone extract, 25 parts of frankincense extract, 10 parts of black pepper extract, 4 parts of glucosamine, 1 part of chondroitin sulfate, 2 parts of vitamin C and 2 parts of hyaluronic acid were added to obtain a solid composition.

[0100] Example 6

[0101] 1 part of fish bone extract, 30 parts of frankincense extract, 10 parts of black pepper extract, 4 parts of glucosamine, 1 part of chondroitin sulfate and 2 parts of vitamin C were added to obtain a solid composition.

[0102] Example 7

[0103] 1 part of fish bone extract, 30 parts of frankincense extract, 10 parts of black pepper extract, 4 parts of glucosamine, 2 parts of vitamin C and 2 parts of hyaluronic acid were added to obtain a solid composition.

[0104] Example 8

[0105] 1 part of fish bone extract, 30 parts of frankincense extract, 10 parts of black pepper extract, 1 part of chondroitin sulfate, 2 parts of vitamin C and 2 parts of hyaluronic acid were added to obtain a solid composition.

[0106] Example 9

[0107] 1 part of fish bone extract, 20 parts of frankincense extract, 10 parts of black pepper extract, 4 parts of glucosamine, 1 part of chondroitin sulfate, 2 parts of vitamin C and 2 parts of hyaluronic acid were added to obtain a solid composition.

[0108] Example 10

[0109] 1 part of fish bone extract, 40 parts of frankincense extract, 10 parts of black pepper extract, 4 parts of glucosamine, 1 part of chondroitin sulfate, 2 parts of vitamin C and 2 parts of hyaluronic acid were added to obtain a solid composition.

[0110] Embodiment 11

[0111] 1 part of fish bone extract, 35 parts of frankincense extract, 10 parts of black pepper extract, 4 parts of glucosamine, 1 part of chondroitin sulfate, 2 parts of vitamin C and 2 parts of hyaluronic acid were added to obtain a solid composition.

[0112] Example 12

[0113] 1 part of fish bone extract, 30 parts of frankincense extract, 15 parts of black pepper extract, 4 parts of glucosamine, 1 part of chondroitin sulfate, 2 parts of vitamin C and 2 parts of hyaluronic acid were added to obtain a solid composition.

[0114] Embodiment 13

[0115] 1 part of fish bone extract, 30 parts of frankincense extract, 15 parts of black pepper extract, 4 parts of glucosamine, 1 part of chondroitin sulfate, 2 parts of vitamin C and 2 parts of hyaluronic acid are added to obtain a solid composition, and the composition and food-acceptable auxiliary materials are respectively crushed, sieved and further mixed to obtain a uniformly mixed pre-capsule filling material, which is filled into capsule shells to obtain capsules.

[0116] Embodiment 14

[0117] Add 1 part of fish bone extract, 30 parts of frankincense extract, 15 parts of black pepper extract, 4 parts of glucosamine, 1 part of chondroitin sulfate, 2 parts of vitamin C and 2 parts of hyaluronic acid to obtain a solid composition, and grind the composition and auxiliary materials separately, sieve them, further mix them, press them into tablets and coat them.

[0118] Embodiment 15

[0119] 1 part of fish bone extract, 28 parts of frankincense extract, 15 parts of black pepper extract, 4 parts of glucosamine, 1 part of chondroitin sulfate, 2 parts of vitamin C and 2 parts of hyaluronic acid are added to obtain a solid composition, the composition is mixed with auxiliary materials, a capsule shell is prepared by using the auxiliary materials (such as gelatin, etc.), a mixture obtained by mixing the composition and the auxiliary materials is filled into the capsule shell by a dripping method or a pressing method, and the capsule is pressed into a pill, dried, and the pills are inspected to obtain a soft capsule.

[0120] Example 16

[0121] 5 mg of fish bone extract, 130 mg of frankincense extract, 50 mg of black pepper extract, 20 mg of glucosamine, 5 mg of chondroitin sulfate, 12 mg of vitamin C and 10 mg of hyaluronic acid were added to obtain a solid composition.

[0122] Embodiment 17

[0123] 5 mg of fish bone extract, 150 mg of frankincense extract, 50 mg of black pepper extract, 20 mg of glucosamine, 5 mg of chondroitin sulfate, 12 mg of vitamin C and 10 mg of hyaluronic acid were added to obtain a solid composition.

[0124] Embodiment 18

[0125] 5 mg of fish bone extract, 100 mg of frankincense extract, 70 mg of black pepper extract, 20 mg of glucosamine, 5 mg of chondroitin sulfate, 12 mg of vitamin C and 12 mg of hyaluronic acid were added to obtain a solid composition.

[0126] Biological test cases

[0127] Test Example 1

[0128] Experimental animals: 130 SPF grade, 6-week-old, male, SD rats. Purchased from Shanghai Slake Experimental Animal Co., Ltd., experimental animal license number: SCXK (Shanghai) 2022-0004. All experimental animals were adaptively fed for one week before the formal experiment began. All rats were kept in a clean animal laboratory, and the environmental conditions were maintained at a 12-h day and night cycle, constant temperature (21°C ~ 23°C), and humidity of 55% ~ 58%. The formal experiment began after one week of adaptive feeding.

[0129] Instruments, consumables and reagents: electronic balance (Mettler-Toledo, ME204TE); multifunctional vortex mixer (JOANLAB, VM-500pro); desktop high-speed refrigerated centrifuge (Eppendorf, 5425R).

[0130] Sodium iodoacetate (MIA) (S104897, Aladdin), rat interleukin 1β (IL-1β) ELISA kit (ml037361, ELISA), rat interleukin 6 (IL6) ELISA kit (ml002859, ELISA), rat type II collagen (ColⅡ) ELISA kit, rat bone collagen cross-linking ELISA kit (ml028473, ELISA), bone tissue protein extraction kit (100T) (EX1270, Solarbio).

[0131] 1. Test Method

[0132] (1) Normal group G1 (n=10): After the adaptive feeding, the mice were gavaged with the same volume of 0.5% CMC-Na once a day for 2 weeks, and then 50 μL of sterile saline was injected into the left knee joint. The gavage was continued for 4 weeks, during which the mice were allowed to eat and drink freely.

[0133] (2) Model group G2 (n=10): After the adaptive feeding, the mice were gavaged with the same volume of 0.5% CMC-Na once a day for 2 consecutive weeks, and then 50 μL MIA was injected into the left knee joint for another 4 weeks. The mice were allowed to eat and drink freely during this period.

[0134] (3) Positive drug group G3 (n=10): After the adaptive feeding, the mice were gavaged with a mixture of glucosamine (156.3 mg / kg) and chondroitin sulfate (20.8 mg / kg) once a day for 2 consecutive weeks. Then, 50 μL of MIA was injected into the left knee joint for another 4 weeks. The mice were allowed to eat and drink freely during this period.

[0135] (4) Calcium carbonate group G4 (n=10): After adaptive feeding, calcium carbonate (153.6 mg / kg) was gavaged once a day for 2 weeks, and then 50 μL MIA was injected into the left knee joint for another 4 weeks. The mice were allowed to eat and drink freely.

[0136] (5) Calcium citrate group G5 (n=10): After the adaptive feeding, calcium citrate (292.7 mg / kg) was gavaged once a day for 2 weeks, and then 50 μL MIA was injected into the left knee joint for another 4 weeks. The mice were allowed to eat and drink freely.

[0137] (6) Sample 1 Group G6 (n=10): After the adaptive feeding, calcium citrate (232.5 mg / kg) and salmon bone extract (66.7 mg / kg) were gavaged once a day for 2 weeks. Then, 50 μL MIA was injected into the left knee joint and gavage was continued for 4 weeks. The rats were allowed to eat and drink freely during this period.

[0138] (7) Sample 2 Group G7 (n=10): After the adaptive feeding, calcium citrate (465.0 mg / kg) and salmon bone extract (133.3 mg / kg) were gavaged once a day for 2 weeks. Then, 50 μL MIA was injected into the left knee joint and gavage was continued for 4 weeks. The rats were allowed to eat and drink freely during this period.

[0139] (8) Sample Group 3 (n=10) G8: After the adaptive feeding, calcium citrate (465.0 mg / kg) and salmon bone extract (266.7 mg / kg) were gavaged once a day for 2 weeks. Then, 50 μL MIA was injected into the left knee joint and gavage was continued for 4 weeks. The mice were allowed to eat and drink freely during this period.

[0140] (9) Sample 4 Group G9 (n=10): After the adaptive feeding, the mice were gavaged with salmon bone extract (156.3 mg / kg), frankincense extract (10.4 mg / kg) and black pepper extract (5.2 mg / kg) once a day for 2 consecutive weeks. Then, 50 μL MIA was injected into the left knee joint for another 4 weeks. The mice were allowed to eat and drink freely during this period.

[0141] (10) Sample 5 Group G10 (n=10): After the adaptive feeding, the mice were gavaged with salmon bone extract (546.7 mg / kg), frankincense extract (156.3 mg / kg) and black pepper extract (78.1 mg / kg) once a day for 2 consecutive weeks. Then, 50 μL MIA was injected into the left knee joint and gavage was continued for 4 more weeks. The mice were allowed to eat and drink freely during this period.

[0142] (11) Sample 6 Group G11 (n=11): After the adaptive feeding, the mice were gavaged with a combination of salmon bone extract (6 mg / kg), frankincense extract (156.3 mg / kg), black pepper extract (78.1 mg / kg), glucosamine (24 mg / kg), chondroitin sulfate (6 mg / kg), vitamin C (15 mg / kg), and hyaluronic acid (12 mg / kg) once a day for 2 consecutive weeks. Then, 50 μL of MIA was injected into the left knee joint and gavage was continued for 4 weeks. The mice were allowed to eat and drink water freely during this period.

[0143] (12) Calcium fructose borate group (n=10): After the adaptive feeding, calcium fructose borate (156.3 mg / kg) was gavaged once a day for 2 weeks, and then 50 μL MIA was injected into the left knee joint for another 4 weeks. During this period, the mice were allowed to eat and drink freely.

[0144] (13) Hyaluronic acid group (n=10): After adaptive feeding, mice were gavaged with hyaluronic acid (12 mg / kg) once a day for 2 weeks. Then, 50 μL MIA was injected into the left knee joint for another 4 weeks. During this period, the mice were allowed to eat and drink freely.

[0145] 2 Model construction and scoring criteria

[0146] 2.1 Construction of rat osteoarthritis model

[0147] After the adaptive feeding, 130 rats were randomly divided into 13 groups. Except for the normal control group, which was given an equal volume of 0.5% CMC-Na by gavage, the other groups were given samples of corresponding concentrations by gavage, once a day, for 2 consecutive weeks. After 2 weeks of preventive administration, except for the normal control group, the knee arthritis rat model of rats in the other groups was established by intra-articular injection of sodium iodoacetate (MIA) to induce osteoarthritis with MIA. Specific operation process: SD rats were deeply anesthetized by intraperitoneal injection of chloral hydrate, the hair around the knees was removed, and routine disinfection was performed. 50 μL of sodium iodoacetate was injected into the left knee with an injection needle to induce an osteoarthritis model.

[0148] 2.2 Biological sample collection

[0149] Blood was collected from the inguinal artery of rats without anticoagulation. After the whole blood was allowed to stand for 30 minutes, it was centrifuged at 3500 rpm / min for 10 minutes, and the supernatant blood was drawn, labeled, and stored in a -80°C refrigerator for testing.

[0150] 2.3 Behavioral indicators of rats

[0151] ①Gait scoring

[0152] The test environment was an open field of 3m×3m. The rats were guided to move freely for 5 minutes, and their walking gait was observed.

[0153] ② Joint pain score

[0154] Fix the head of the rat, let its hind legs and tail extend out of the fixing tube, and measure after it stabilizes for 5 minutes. Slowly and gently bend the joints of the rat, once every 5 seconds, for a total of 5 times, and add up the scores of the 5 times as the final score.

[0155] ③Autonomous pain score

[0156] The rats' behavior was observed daily for 5 min / time.

[0157] The scoring criteria are as follows: 0 points for normal behavior; 1 point for toe flexion; 2 points for foot eversion; 3 points for partial weight-bearing: the limb on the model side cannot completely contact the ground; 4 points for no weight-bearing and protection: the limb on the model side cannot completely contact the ground; 5 points for avoiding contact with the model joint: when the limb on the model side is lightly touched, there is obvious avoidance behavior.

[0158] 3 Experimental results analysis

[0159] 3.1 Rats’ gait scoring

[0160] The results are as follows Figure 1 As shown in the figure, compared with the normal control group, the gait score of the rats in the model group increased significantly and was statistically significant, indicating that the osteoarthritis model was successfully established. Sample group 6 can significantly reduce the gait score of the osteoarthritis rats, that is, sample group 6 can effectively control and improve the osteoarthritis of rats.

[0161] 3.2 Joint pain scores in rats

[0162] The results are as follows Figure 2 As shown, sample group 6 can significantly reduce the joint pain scores of rats with osteoarthritis, that is, sample group 6 can effectively control and improve the osteoarthritis condition of rats.

[0163] 3.3 Autonomous pain scores in rats

[0164] The results are as follows Figure 3As shown in Figure 4, sample group 6 can significantly reduce the autonomous pain scores of rats with osteoarthritis, that is, sample group 6 can effectively control and improve the osteoarthritis condition of rats.

[0165] 3.4 Macroscopic scoring results of rat knee joints

[0166] At the end point, the knee joints of rats in each group were dissected, muscles and ligament tissues were removed, and macroscopic scores were performed. Figure 4 As shown. Compared with the normal control group, the macroscopic score of the knee joint of the rats in the model group increased significantly and was statistically significant, indicating that the osteoarthritis model was successfully established. Sample group 6 can significantly reduce the macroscopic score of the knee joint of the osteoarthritis rats, that is, sample group 6 can effectively control and improve the osteoarthritis of the rats.

[0167] 3.5 Detection results of IL-1β and IL-6 levels in rat serum

[0168] In osteoarthritis, the increased levels of interleukin 1β (IL-1β) and interleukin 6 (IL6) can stimulate synovial cells, chondrocytes and other immune cells to produce more inflammatory mediators, such as prostaglandin E2 and nitric oxide, thereby aggravating joint inflammation. At the same time, the increase in inflammatory factors can directly act on chondrocytes and induce their apoptosis, which will lead to a decrease in the number of chondrocytes, a decrease in repair ability, and accelerated cartilage degeneration. In addition, inflammatory factors may also participate in the pain of osteoarthritis patients by stimulating nerve endings or affecting the release of pain-related mediators.

[0169] The results of IL-1β and IL6 in rat serum are shown in Figure 5 , Figure 6 As shown in the figure, compared with the normal control group, the levels of IL-1β and IL6 in the serum of the model group rats increased significantly and were statistically significant, indicating that the osteoarthritis model was successfully established. Sample group 6 was able to reduce the levels of IL-1β and IL6 in the knee joints of osteoarthritis rats, that is, the experimental results showed that sample group 6 had a good anti-inflammatory effect, could improve the repair ability, reduce cartilage degeneration, and relieve the pain of osteoarthritis patients.

[0170] 3.6 Detection results of IL-1β and IL-6 gene expression in rat knee joints

[0171] The gene expression levels of IL-1β and IL-6 in the knee joint were detected. Figure 7 , Figure 8As shown in the figure, compared with the normal control group, the gene expression of IL-1β and IL-6 in the knee joints of rats in the model group increased significantly, and it was statistically significant, indicating that the osteoarthritis model was successfully established. Sample group 6 can significantly reduce the gene expression of IL-1β and IL-6 in the knee joints of rats with osteoarthritis. That is, the experimental results show that sample group 6 has a good anti-inflammatory effect, can improve the repair ability, reduce cartilage degeneration, and relieve the pain of patients with osteoarthritis.

[0172] Test Example 2 Human body test

[0173] Thirteen people with joint discomfort were recruited and had the volunteers take the composition of the present invention (such as Example 15) daily, and their joint discomfort relief was statistically analyzed.

[0174] The results show that after taking the composition of the present invention, joint discomfort is significantly alleviated.

[0175] It will be understood by those skilled in the art that, unless otherwise defined, all terms (including technical and scientific terms) used herein have the same meaning as generally understood by those skilled in the art in the art to which the present invention belongs. It should also be understood that terms such as those defined in general dictionaries should be understood to have meanings consistent with the meanings in the context of the prior art, and will not be interpreted with idealized or overly formal meanings unless specifically defined as herein.

[0176] It should be understood that the detailed description of the technical solutions of the present invention by means of the preferred embodiments is illustrative rather than restrictive. A person skilled in the art may modify the technical solutions described in the embodiments, or replace some of the technical features by equivalents, based on reading the specification of the present invention; and these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. A composition comprising a plant extract and a fish bone extract, wherein the plant extract comprises at least one or more of an angelica extract, an frankincense extract, and a pepper extract; and the fish bone extract has a content of ≥1 mg / portion.

2. The composition according to claim 1, characterized in that The plant extracts are frankincense extract and pepper extract.

3. The composition according to claim 1, characterized in that The effective components of the plant extract at least include one or more of phytocannabinoids, purseratin, pursera alcohol, boswellic acid, sabinene, limonene, pinene, caryophyllene, and humulene.

4. The composition according to claim 3, wherein the boswellic acid comprises one or more of ursolic acid type and oleanane type; further, the ursolic acid type comprises β-boswellic acid, acetyl-β-boswellic acid, 3-acetyl-11-carbonyl-β-boswellic acid, 11-carbonyl-β-boswellic acid, acetyl-11-hydroxy-β-boswellic acid, acetyl-11α-methoxy-β-boswellic acid, 9,11-dehydro-β-boswellic acid, 3α-acetyl-9,11-dehydro-β-boswellic acid, or one or more of 3-O-acetyl-11-keto-β-boswellic acid, and the oleanane type comprises α-boswellic acid, acetyl-α-boswellic acid, 9,11-dehydro-α-boswellic acid, and 3-acetyl-9,11-dehydro-α-boswellic acid.

5. The composition according to claim 2, characterized in that The content of the plant extract in the composition is ≥100 mg / portion.

6. The composition according to claim 2, characterized in that The content of 3-O-acetyl-11-keto-β-boswellic acid in the frankincense extract accounts for 10-50% of the total mass of the frankincense extract.

7. The composition according to claim 2, characterized in that The pepper extract is a black pepper extract; further, the β-caryophyllene content in the black pepper extract is ≥20%.

8. The composition according to claim 1, characterized in that The composition also includes glucosamine.

9. The composition according to claim 8, characterized in that The glucosamine content is ≥5 mg / portion, and further, the glucosamine content is 5-100 mg / portion.

10. The composition according to claim 1, characterized in that The composition also includes chondroitin sulfate.

11. The composition according to claim 10, characterized in that The content of the chondroitin sulfate is ≥1 mg / portion; further, the content of the chondroitin sulfate is 1-10 mg / portion.

12. The composition according to claim 1, characterized in that The composition also includes vitamins; further, the vitamins include one or more of vitamin D, vitamin C, and vitamin E, and further, the vitamin is vitamin C.

13. The composition according to claim 12, wherein the vitamin C content is ≥5 mg / portion, and further, the vitamin C content is 5-20 mg / portion.

14. The composition according to claim 1, characterized in that The composition also includes hyaluronic acid.

15. The composition according to claim 14, characterized in that The hyaluronic acid content is ≥5 mg / portion.

16. The composition according to any one of claims 1 to 15, characterized in that The composition is added with pharmaceutically acceptable excipients to prepare into one or more of tablets, soft capsules, powders, oral liquids, sprays, and capsules.

17. The composition according to claim 16, characterized in that The dosage form is a soft capsule.

18. The composition according to any one of claims 1 to 15, for use in bone and joint health.

19. The composition according to any one of claims 1 to 15, for use in improving osteoporosis, improving arthritis, increasing bone density, increasing bone strength, maintaining bone density, and enhancing bone firmness.

20. The composition according to any one of claims 1 to 15, for use in improving arthritis.

Images