Curcumin composition for osteoarthritis and joint health

Abstract

The present invention relates to a stable curcumin composition for the prevention, amelioration, and maintenance of arthritis, more particularly osteoarthritis. The stable curcumin composition is composed of curcuminoids alone or in combination with at least one pharmaceutically and / or nutraceutical acceptable excipient to form a stable curcumin composition with enhanced absorption and bioavailability. The composition is safe for human consumption and has enhanced stability and bioavailability due to the selected percentage of amorphous and crystalline polymorphic forms of curcumin, and selected excipients, such as pH adjusters and / or stabilizers, and optionally hydrophilic carriers, antioxidants, diluents, anti-caking agents, emulsifiers, fats, and surfactants, and when administered to a subject in need thereof, results in improvement of the symptoms of arthritis and related conditions.

Description

Technical Field

[0001] The present invention relates to a stable curcumin composition for the prevention, improvement, and maintenance of arthritis, more specifically osteoarthritis. More specifically, for enhanced bioavailability, it relates to a stable curcumin composition containing curcuminoids in different polymorphic forms such as amorphous and crystalline and / or in a selected ratio, with improved stability of curcumin in the alkaline pH environment of the intestine.

Background Art

[0002] Arthritis is a more specific term that means damage or inflammation in one or more joints. The condition is often manifested by pain, swelling, heat, redness, and restricted movement. Osteoarthritis of the knee is a type of disease caused by the breakdown of articular cartilage between bones. As a result of this cartilage reduction, more friction occurs between the bones, resulting in painful bone protrusions.

[0003] Curcumin is derived from the rhizome of turmeric and has traditionally been used in the treatment of inflammation, skin wounds, tumors, etc. Overall, curcumin is associated with several health claims, but its therapeutic use is limited due to its low bioavailability, poor water solubility, instability at neutral and basic pH, insufficient absorption, rapid metabolism, and short half-life. Curcumin is a class IV drug (low solubility and low permeability) based on the Biopharmaceutics Classification System (BCS). Many strategies have been developed to overcome these limitations, especially for oral delivery systems.

[0004] [[ID=ID=ID=20]]US2016089343A1 disclosed herein is a formulation for the topical delivery of a therapeutically effective dose of curcumin that provides serum levels of curcumin sufficient to treat diseases such as head and neck disorders and upper airway and gastrointestinal disorders.

[0005] EP3275430A1 relates to aqueous solutions for intravenous injection comprising curcumin and / or one or more curcumin derivatives, dimethyl sulfoxide, one or more solubilizers, sodium selenite, and a buffer in an aqueous injection medium, and to concentrates comprising curcumin and / or one or more curcumin derivatives, dimethyl sulfoxide, one or more solubilizers, and sodium selenite for the preparation thereof.

[0006] K.N. Pushpakumari, in the International Journal of Pharmaceutical Sciences and Research (impact factor 2019: 1.230, citation score 2017: 0.27), compared the bioabsorption of two turmeric preparations containing curcuminoids in diverse compositions with that of a standard turmeric extract. The document is completely silent on the effects of curcumin on osteoarthritis.

[0007] To find a satisfactory solution to the problem of persistence, there is a strong need to develop stable curcumin formulations for the treatment, prevention, and maintenance of arthritis, more specifically osteoarthritis. Stable curcumin compositions containing curcuminoids in different polymorphic forms such as amorphous and crystalline, and / or in selective ratios, for enhanced bioavailability. Stable curcumin compositions consist of curcuminoids alone or together with at least one pharmaceutically and / or nutritionally acceptable excipient to form a stable curcumin composition with enhanced absorption and bioavailability. [Overview of the Initiative]

[0008] Purpose of the invention The main objective of this invention is to develop a stable curcumin preparation for the prevention, improvement, and maintenance of arthritis.

[0009] Another object of the present invention is to provide a stable curcumin formulation with enhanced bioavailability that can be used in an orally administered form.

[0010] Another object of the present invention is to develop stable curcumin formulations containing curcuminoids in different polymorphic forms, such as amorphous and crystalline, and / or in selective ratios, for enhanced absorption and bioavailability.

[0011] Another object of the present invention is to develop a stable curcumin formulation containing curcuminoids, both alone and / or in combination with at least one pharmaceutically and / or nutritionally acceptable excipient to form a stable curcumin composition with enhanced absorption and bioavailability.

[0012] Another object of the present invention is to develop a stable curcumin formulation containing curcuminoids, which can be used alone and / or together with at least one pH adjuster and stabilizer to form a stable curcumin composition with enhanced absorption and bioavailability, and which may optionally be used together with a hydrophilic carrier, antioxidant, diluent, anti-caking agent, emulsifier, fat, and surfactant.

[0013] A further aspect of the present invention is to provide a process for preparing a stable curcumin composition suitable for formulation into tablets, capsules, blended powders, licaps, ointments, pastes, lotions, liniments, mouthwashes, gargles, dry syrups, liquid syrups, drinks, diet drinks, fruit juices, soft drinks, and the like.

[0014] Another object of the present invention is to provide a stable curcumin composition for use in preventing, improving, and maintaining arthritis, more specifically osteoarthritis, and related conditions such as articular cartilage, joint mobility, joint flexibility, reduction of joint stiffness, and reduction of inflammation.

[0015] A further object of the present invention is to provide a stable curcumin composition for use in improving muscle strength and performance, reducing muscle soreness and injury, and promoting faster muscle recovery.

[0016] Summary of the present invention According to one aspect of the present invention, a stable curcumin composition having enhanced bioavailability is provided; i) Curcumin containing curcuminoids in crystalline polymorphisms ranging from 0 to 15%, and curcuminoids in amorphous polymorphisms ranging from 85 to 100%, and ii) At least one pharmaceutically or nutritionally acceptable material component selected from hydrophilic carriers, antioxidants, stabilizers, pH adjusters, solubilizers, fats, and anti-caking agents. [Brief explanation of the drawing]

[0017] [Figure 1] This is an X-ray diffraction (XRD) graph of the composition prepared according to Example 01. [Figure 2] This is an X-ray diffraction (XRD) graph of the composition prepared according to Example 02. [Figure 3] This is an X-ray diffraction (XRD) graph of the composition prepared according to Example 03. [Figure 4] This is an X-ray diffraction (XRD) graph of the composition prepared according to Example 04. [Figure 5] This study shows the effect of curcumin (Cur) on the expression of knee joint proteins IL-β (A), IL-6 (B), TNF-α (C), and NF-κβ (D) in monosodium iodoacetate (MIA)-induced osteoarthritis (OA) rats. Bars indicate standard deviation. The blot was repeated at least three times (n=3), and a representative blot is shown. Asterisks above the lines indicate statistical differences between groups (ANOVA and Tukey post-hoc test; **P<0.01; ***P<0.001; ****P<0.0001). OA, osteoarthritis; Cur, curcumin; MIA, monosodium iodoacetate; IL-1β, interleukin-1β; IL-6, interleukin-6; TNF-α, tumor necrosis factor α; NF-κβ, nuclear factor κβ. [Figure 6]This image shows the effect of curcumin (Cur) on the expression of knee joint proteins at the levels of type II collagen (A), MMP-3 (B), COX-2 (C), and LOX-5 (D) in rats with monosodium iodoacetate (MIA)-induced osteoarthritis (OA). Bars indicate standard deviation. The blot was repeated at least three times (n=3), and a representative blot is shown. Asterisks above the lines indicate statistical differences between groups (ANOVA and Tukey post-hoc tests; **P<0.01; ***P<0.001; ****P<0.0001). OA, osteoarthritis; Cur, curcumin; MIA, monosodium iodoacetate; MMP-3, matrix metalloproteinase-3; COX-2, cyclooxygenase-2; LOX-5, 5-lipoxygenase. [Figure 7] This study demonstrates the effect of curcumin (Cur) on the knee joint in rats with monosodium iodoacetate (MIA)-induced osteoarthritis (OA). Representative X-ray images obtained at the end of the experiment are shown (A). The mean Kellgren-Lawrence score is shown using ± standard deviation (B). Asterisks indicate statistical differences in Kellgren-Lawrence scores between groups, represented by bars (Kruskal-Wallis, followed by Mann-Whitney U; *P<0.05**P<0.01; compared to the OA group). [Figure 8] This study demonstrates the effect of curcumin (Cur) on the histopathological diagnosis of the knee joint in rats with monosodium iodoacetate (MIA)-induced osteoarthritis (OA). Representative histopathological images obtained at the end of the experiment, stained with hematoxylin-eosin (A) and toluidine blue (B), are shown. The mean Mankin score is shown using ± standard deviation. Asterisks above the lines indicate statistical differences in Mankin scores between groups, represented by bars (C) (Kruskal-Wallis, followed by Mann-Whitney U; *P<0.05; compared to the OA group). [Figure 9]Shows the effects of curcumin (Cur) on knee swelling (A), left knee joint diameter (B), right knee joint diameter (C), and the ratio of right-to-left diameter values (D) in rats with monosodium iodoacetate (MIA)-induced osteoarthritis (OA). Asterisks above the lines indicate statistical differences between groups (ANOVA and Tukey post hoc test; **P < 0.01; ***P < 0.001; ****P < 0.0001). [Figure 10] Shows the effects of curcumin (Cur) on foot area (B) and step length (D) in rats with monosodium iodoacetate (MIA)-induced osteoarthritis (OA). Representative measurements of foot area (A) and step length (C) are shown. Asterisks above the lines indicate statistical differences between groups (ANOVA and Tukey post hoc test; *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001). [Figure 11] Shows the plasma total curcuminoid concentration of the curcumin composition vs. reference.

Mode for Carrying Out the Invention

[0018] According to an embodiment of the present invention, a stable curcumin formulation contains curcuminoids in different polymorphic forms such as amorphous and crystalline at a selective ratio for enhancing bioavailability. According to a further embodiment of the present invention, a stable curcumin formulation contains curcuminoids alone and / or together with at least one excipient for forming a stable curcumin composition with enhanced absorption and bioavailability for the prevention, improvement, and maintenance of arthritis, more specifically, osteoarthritis.

[0019] In the context of this invention, the technical term "curcumin composition" is generally used herein to refer to a composition containing curcuminoids in different polymorphic forms such as amorphous and crystalline at a selective ratio, either alone and / or together with at least one excipient for forming a stable curcumin composition with enhanced absorption and bioavailability.

[0020] Curcumin (1,7-bis(4-hydroxy-3-methoxyphenyl)-1,6-heptadiene-3,5-dione) is the main curcuminoid of turmeric, a well-known Indian spice and perennial herb commonly known as turmeric, or ginger (Zingiberaceae). It should be recognized that the term "curcumin" may be interpreted as falling within the range of curcuminoids, and may generally include curcumin, demethoxycurcumin, and bisdemethoxycurcumin. Products that may be called "curcumin" may contain these components along with other components belonging to the curcuminoid class. Curcumin extracts contain total curcuminoids in the range of 40-99%.

[0021] According to a further embodiment of the present invention, the curcumin composition contains curcuminoids present in the range of 10-90% w / w of the composition.

[0022] According to further embodiments of the present invention, a stable curcumin formulation comprises a curcuminoid alone and / or together with at least one excipient to form a stable curcumin composition with enhanced absorption and bioavailability. More preferably, the stable curcumin composition is formulated using excipients selected from the group of hydrophilic carriers, antioxidants, solvents, emulsifiers, surfactants, solubilizers, stabilizers, pH adjusters, binders, anticaking agents, fats and / or combinations thereof.

[0023] In another embodiment, the solid hydrophilic carrier used in the preparation of the curcumin composition is selected from the group, for example, but is not limited to, cellulose derivatives, polyacrylates, polyethylene glycol, povidone, starch and starch derivatives, gums, sugars, pectin, plant polysaccharides, proteins, such as casein, whey protein, soy protein, pea protein, sugar alcohols, glucose, polyvinylpyrrolidone, and the like.

[0024] According to a preferred embodiment, the hydrophilic carrier is hydroxypropyl methylcellulose.

[0025] According to further embodiments, the hydrophilic carrier is present in the range of 1-90% w / w of the composition.

[0026] In another embodiment, the antioxidant in the preparation of the curcumin composition is selected from, for example, but is not limited to, natural tocopherols, mixed tocopherols, ascorbyl palmitate, rosemary extract, epigallocatechin gallate, catechin, and ascorbic acid.

[0027] According to further embodiments, antioxidants are present in the composition in the range of 0.1–10% w / w.

[0028] In another embodiment, the solvent in the preparation of the stable curcumin composition is selected from the group of, for example, isopropyl alcohol, acetone, methanol, alcohol, ethyl acetate, ethanol, methylene chloride, water, and mixtures thereof, but is not limited to these. The temperature maintained to obtain a homogeneous mass may be in the range of ambient ~80°C; preferably 25°C to 60°C.

[0029] According to further embodiments, the solvent is present in the range of 40-98% w / w of the composition.

[0030] In another embodiment, the emulsifier in the preparation of a stable curcumin composition is selected from, for example, polysorbates, sugar alcohols, glycerol and its derivatives, but is not limited to these.

[0031] According to further embodiments, the emulsifier is present in the composition in the range of 0.1–10% w / w.

[0032] In another embodiment, the surfactant in the preparation of a stable curcumin composition is selected from, for example, but is not limited to, sugar alcohols, sugar esters (e.g., sucrose stearate), and esters of sugar alcohols (e.g., sorbitan). According to a further embodiment, the surfactant is present in the composition at a concentration of 0.1-10% w / w.

[0033] In another embodiment, the solubilizer in the preparation of the curcumin composition is selected from the group, for example, propylene glycol alginate, sugar alcohols, sugar esters, phospholipids, lecithin, d-limonene, vitamin E TPGS (d-α-tocopherol polyethylene glycol 1000 succinic acid), sodium lauryl sulfate, and β-cyclodextrin, although this group is not limited to these.

[0034] According to further embodiments, the solubilizer is present in the composition in the range of 0.1–40% w / w.

[0035] In another embodiment, the stabilizer in the preparation of the curcumin composition is selected from, for example, but is not limited to, glyceryl monostearate, sugar alcohols, triglycerides, antioxidants, monoglycerides, and phospholipids.

[0036] According to further embodiments, the stabilizer is present in the composition in the range of 0.1–10% w / w.

[0037] In another embodiment, the pH adjuster in the preparation of the curcumin composition is selected from the group consisting of, for example, but not limited to, citric acid, trisodium citrate, lactic acid, L-arginine, calcium carbonate, and magnesium carbonate.

[0038] According to further embodiments, the pH adjuster is present in the composition in the range of 0.1–10% w / w.

[0039] In another embodiment, the binder in the preparation of the curcumin composition is selected from, for example, but is not limited to, hydroxypropyl cellulose, pregelatinized cellulose, and starch.

[0040] According to further embodiments, the binder is present in the composition in the range of 0.1–10% w / w.

[0041] In another embodiment, the anti-caking agent in the preparation of the curcumin composition is selected from, for example, but is not limited to, colloidal silicon dioxide, magnesium stearate, stearic acid, and mannitol.

[0042] According to further embodiments, the anti-solidification agent is present in the composition in the range of 0.1–10% w / w.

[0043] In another embodiment, the fat used in the preparation of the curcumin composition is selected from, for example, but is not limited to, medium-chain triglycerides, long-chain triglycerides, vegetable oils, fatty acid esters, hydrocarbons, such as terpenes, and monoglycerides.

[0044] According to further embodiments, fat is present in the composition in the range of 0.1–40% w / w.

[0045] According to further embodiments of the present invention, the stable curcumin compositions described herein exhibit enhanced bioavailability, and the compositions can be obtained in orally administrative solid, semi-solid, or liquid forms, selected from, but not limited to, formulations such as powders, granules, pellets, beadlets, caplets, tablets, capsules, softgel capsules, solutions, emulsions, suspensions, and dispersions.

[0046] According to another embodiment of the present invention, the curcumin composition is in the form of granules.

[0047] According to another embodiment of the present invention, the curcumin composition is in the form of a powder.

[0048] In some embodiments, a process for preparing a stable curcumin composition with enhanced stability and bioavailability for use in osteoarthritis includes the following: (i) Dissolve one or more excipients in a solvent or a mixture thereof in a solvent with continuous stirring for 10-15 minutes at 50-80°C; (ii) Dissolve curcumin in the solvent or a mixture thereof at 50-80°C for 5-10 minutes with continuous stirring, and then cool to a temperature of 30-40°C; and (iii) Adding other excipients such as hydrophilic carriers, antioxidants, diluents, stabilizers, pH adjusters, anti-caking agents, fats, emulsifiers, and surfactants to the solvent to form a uniform mass; (iv) Remove the solvent by evaporation to form a dry mass; and (v) To pulverize the dry mass to form a fine powder.

[0049] The solvent in step (ii) can be removed by vacuum distillation, evaporation, or spray drying. The resulting dry mass is then finely ground using, for example, a mortar and pestle, a mixer-grinder, a multi-mill, a ball mill, or a jet mill.

[0050] A further embodiment of the present invention provides a stable curcumin composition for use in preventing, improving, and maintaining arthritis, more specifically osteoarthritis.

[0051] A further embodiment of the present invention provides a stable curcumin composition for use in preventing, improving, and maintaining osteoarthritis.

[0052] A further embodiment of the present invention provides a stable curcumin composition for use in maintaining joint health.

[0053] According to further embodiments of the present invention, a stable curcumin is provided that exhibits increased joint mobility, sufficient movement to facilitate daily activities, improved walking ability, increased joint flexibility, reduced joint stiffness, reduced joint discomfort / pain, reduced inflammation, and reduced cartilage destruction.

[0054] A further embodiment of the present invention provides a stable curcumin composition that can be used to improve muscle strength and muscle performance, reduce muscle soreness and muscle damage, and promote faster muscle recovery.

[0055] Example 01: [Table 1] The process for preparing the composition is defined step by step as follows: i) Dissolve anhydrous citric acid, glyceryl monostearate, and lecithin in a mixture of isopropyl alcohol and methylene chloride under heating at 55-60°C. Add the curcumin extract, containing curcumin, demethoxycurcumin, and bisdemethoxycurcumin, under heating at 55-60°C and stir for 25-40 minutes until completely dissolved. Cool the above solution to 30-40°C. ii) Hydroxypropyl methylcellulose and sodium lauryl sulfate are further added under stirring conditions. iii) Add medium-chain triglyceride oil, d-limonene, and tocopherol to the solution from step (ii) while stirring. Stir the solution for 35-45 minutes to obtain a homogeneous solution. iv) A homogeneous solution was used as the feed liquid for a spray dryer. The powder obtained from the spray dryer was further dried in a vacuum dryer at a temperature of 50-90°C, then sorted, and blended after the addition of colloidal silicon dioxide. The resulting product was a yellowish-orange, freely flowing homogeneous powder. v) Figure 01 shows the X-ray powder diffraction (XRD) of a curcumin composition having 1.2% crystalline polymorph and 98.8% amorphous polymorph curcuminoids.

[0056] Example 02: [Table 2] The process for preparing the composition is defined step by step as follows: i) Dissolve anhydrous citric acid, glyceryl monostearate, and lecithin in a mixture of isopropyl alcohol and methylene chloride under heating at 55-60°C. Add the curcumin extract, containing curcumin, demethoxycurcumin, and bisdemethoxycurcumin, under heating at 55-60°C and stir for 25-40 minutes until completely dissolved. Cool the above solution to 30-40°C. ii) Add hydroxypropyl methylcellulose under stirring conditions. iii) Add medium-chain triglyceride oil and tocopherol to the solution from step (ii) while stirring. Stir the solution for 35-45 minutes to obtain a homogeneous solution. iv) A homogeneous solution was used as the feed liquid for a spray dryer. The powder obtained from the spray dryer was further dried in a vacuum dryer at a temperature of 50-90°C, then sorted, and blended after the addition of colloidal silicon dioxide. The resulting product was a yellowish-orange, freely flowing homogeneous powder. v) Figure 02 shows the X-ray powder diffraction (XRD) of a curcumin composition having 1.2% crystalline polymorph and 98.8% amorphous polymorph curcuminoids.

[0057] Example 03: [Table 3] i) Dissolve anhydrous citric acid, glyceryl monostearate, and lecithin in a mixture of ethanol and ethyl acetate under heating at 55-60°C. Add curcumin extract under heating at 55-60°C and stir for 25-40 minutes until completely dissolved. Cool the above solution to 30-40°C. ii) Add hydroxypropyl methylcellulose under stirring conditions. iii) Add medium-chain triglyceride oil, d-limonene, and tocopherol to the solution from step (ii) while stirring. Stir the solution for 35-45 minutes to obtain a homogeneous solution. iv) A homogeneous solution was used as the feed liquid for a spray dryer. The powder obtained from the spray dryer was further dried in a vacuum dryer at a temperature of 50-90°C, then sorted, and blended after the addition of colloidal silicon dioxide. The resulting product was a yellowish-orange, freely flowing homogeneous powder. v) Figure 03 shows the X-ray powder diffraction (XRD) of a curcumin composition having 2% crystalline polymorph and 98% amorphous polymorph curcuminoids.

[0058] Example 04: [Table 4] i) Dissolve anhydrous citric acid, glyceryl monostearate, and lecithin in a mixture of ethanol and ethyl acetate under heating at 55-60°C. Add curcumin extract under heating at 55-60°C and stir for 25-40 minutes until completely dissolved. Cool the above solution to 30-40°C. ii) Add hydroxypropyl methylcellulose under stirring conditions. iii) Add medium-chain triglyceride oil and tocopherol to the solution from step (ii) while stirring. Stir the solution for 35-45 minutes to obtain a homogeneous solution. iv) A homogeneous solution was used as the feed liquid for a spray dryer. The powder obtained from the spray dryer was further dried in a vacuum dryer at a temperature of 50-90°C, then sorted, and blended after the addition of colloidal silicon dioxide. The resulting product was a yellowish-orange, freely flowing homogeneous powder. v) Figure 04 shows the X-ray powder diffraction (XRD) of a curcumin composition having 4% crystalline polymorph and 96% amorphous polymorph curcuminoids.

[0059] Example 05 [Table 5] i) Dissolve anhydrous citric acid, glyceryl monostearate, and lecithin in a mixture of ethanol and ethyl acetate under heating at 55-60°C. Add curcumin extract under heating at 55-60°C and stir for 25-40 minutes until completely dissolved. Cool the above solution to 30-40°C. ii) Add hydroxypropyl methylcellulose under stirring conditions. iii) Add medium-chain triglyceride oil, d-limonene, and tocopherol to the solution from step (ii) while stirring. Stir the solution for 35-45 minutes to obtain a homogeneous solution. iv) A homogeneous solution was used as the feed liquid for a spray dryer. The powder obtained from the spray dryer was further dried in a vacuum dryer at a temperature of 50-90°C, then sorted, and blended after the addition of colloidal silicon dioxide. The resulting product was a yellowish-orange, freely flowing homogeneous powder.

[0060] It should be understood at the outset that the following description merely illustrates specific forms of the invention. However, such specific forms are merely illustrative embodiments and are not intended to be constrained in any way that implies limitations to the scope of the invention.

[0061] Clinical Experimental Study 1: Objective: This study evaluated the efficacy of a curcumin composition (test sample) in monosodium iodoacetate (MIA)-induced osteoarthritis.

[0062] Test sample: Curcumin composition containing curcuminoids, manufactured by OmniActive Health Technologies Limited, India.

[0063] Dosage 1: A formulation containing 100 mg / kg (20 mg / kg of total curcuminoids) (Example 1: 1.2% crystalline polymorph and 98.8% amorphous polymorph curcuminoids)

[0064] Dosage 2: A formulation containing 200 mg / kg (40 mg / kg of total curcuminoids) (Example 1: 1.2% crystalline polymorph and 98.8% amorphous polymorph curcuminoids)

[0065] Experimental design: Female Wistar rats (8 weeks) were randomly assigned to the following groups (n=7 each): I. Normal control group II. Osteoarthritis Group III. Osteoarthritis, curcumin composition dose 1 (100 mg / kg preparation) group and IV. Osteoarthritis, curcumin composition dose 2 (200 mg / kg preparation) group.

[0066] To induce osteoarthritis in a rat model, rats were anesthetized with xylazine (10 mg / kg) and ketamine hydrochloride (50 mg / kg), then their right knees were shaved and disinfected with 70% alcohol. 3 mg of MIA was dissolved in 50 μL of saline and injected into the right knee joint via the infrapatellar ligament using a 0.3 ml insulin syringe fitted with a 29-G needle. The control group received 50 μL of saline. Two weeks after MIA injection, formulations 1 and 2 were administered orally for 4 weeks, dissolved in 1 mL of saline. All rats were observed every other day to assess knee joint swelling.

[0067] Four weeks later, the rats were sacrificed, and blood and knee joint specimens were collected for follow-up experiments. Blood samples were centrifuged at 3,000 rpm for 10 minutes, and the recovered serum was maintained at -20°C until the day of analysis.

[0068] Sample preparation: The curcumin composition according to the present invention is as follows: Test samples for 100 mg / kg and 200 mg / kg were prepared using the provided process parameters in the example. 0 It was prepared according to step 1.

[0069] biochemical analysis Serum MDA (malondialdehyde) was analyzed by HPLC. Antioxidant enzymes (SOD, CAT, GSHPx), IL-1β, IL-6, IL-10, and TNF-α were measured according to enzyme-linked immunosorbent assay (ELISA) using relevant commercially available kits.

[0070] Western blot analysis Articular cartilage samples were analyzed using Western blotting techniques for the expression of TNF-α, IL-6, IL-1β, nuclear factor κB, cyclooxygenase-2 (COX-2), type II collagen, CRP, 5-LOX, MMP3, and cartilage oligomeric matrix protein (COMP).

[0071] Histological analysis The effect of the product on cartilage degeneration in the knee joints of MIA-induced osteoarthritis rats was confirmed by evaluating histological changes. After rat sacrifice, each knee joint was fixed in 10% formalin at 4°C for 24 hours and decalcified with 5% hydrochloric acid at 4°C for 4 days. After decalcification, the specimens were dehydrated in stepwise acetone and embedded in paraffin. Sections (thickness 2-3 μm) were stained with 0.2% hematoxylin and 1% eosin for 5 minutes and 3 minutes, respectively. Histological preparations were analyzed and photographed under a microscope using a digital image acquisition camera.

[0072] Example 1 The objective of the study was to demonstrate the effect of the curcumin composition of this invention on serum inflammatory markers.

[0073] Methods: Animals were sacrificed, serum samples were collected, and serum biomarkers were evaluated using relevant commercially available kits according to enzyme-linked immunosorbent assay (ELISA), as shown in Table 1 and provided illustratively in Figure 5.

[0074] [Table 6] Abbreviation: OA: Osteoarthritis Cur1: Curcumin composition dose 1 (100 mg / kg preparation) Cur2: Curcumin composition dose 2 (200 mg / kg preparation) TNF-α: Tumor necrosis factor α; IL-1β: Interleukin-1β; IL-6: Interleukin-6; IL-10: Interleukin-10; COMP: Cartilage oligomeric matrix protein; CRP: C-reactive protein.

[0075] Statistical comparisons are indicated using different superscripts (ad) within the same column (P<0.05; * (ANOVA and Tukey post-hoc test). The mean of each item is shown using ± standard deviation.

[0076] Conclusion: Table 1 and Figure 5 show that serum inflammatory markers such as TNFα, IL1β, IL-6, COMP, and CRP, which contribute to the induction of inflammation and related pathological conditions, increased during OA, and that treatment with curcumin composition 1 and curcumin composition 2 reduced these levels in a statistically significant manner. The cytokine IL-10 level, which was reduced during OA, was restored by treatment.

[0077] Example 2 The objective of the study was to demonstrate the effect of a curcumin composition on serum antioxidant markers.

[0078] Methods: Animals were sacrificed, serum samples were collected, and serum antioxidant markers were evaluated. Serum from MDA was analyzed by HPLC. Antioxidant enzymes (SOD, CAT, GSHPx) were measured according to enzyme-linked immunosorbent assay (ELISA) using relevant commercially available kits, and the data are shown in the table below. [Table 7] MDA: Malondialdehyde; SOD: Superoxide dismutase; GSH-Px: Glutathione peroxidase; CAT: Catalase. Statistical comparisons are indicated using different superscripts (ad) in the same column (P<0.05;* (ANOVA and Tukey post-hoc test). The mean value of the markers is shown using ± standard deviation.

[0079] Conclusion: MDA, a marker of oxidative stress that plays a crucial role in inducing the pathogenesis of OA, was significantly increased during OA and significantly decreased during treatment. Similarly, treatment significantly improved levels of antioxidant enzymes, SOD, CAT, and GSH-Px, as shown in Table 2.

[0080] Example 3 The objective of the study was to demonstrate the effect of a curcumin composition on inflammatory markers in synovial tissue.

[0081] Methods: Animals were sacrificed, synovial tissue samples were collected, and evaluated for inflammatory markers by Western blotting as shown in Figure 05.

[0082] Conclusion: The protein levels of the inflammatory markers shown below—IL-β (A), IL-6 (B), TNF-α (C), and NF-κβ (D)—were found to be elevated in OA rats, as measured by Western blotting followed by relative intensity concentration analysis with a control group after β-actin normalization to ensure equal protein loading. The blot was repeated at least three times (n=3), and a representative blot (E) is shown.

[0083] Treatment with curcumin compositions at two doses (100 and 200 mg / kg) was observed to reduce levels of inflammatory markers in joint tissue in a statistically significant manner.

[0084] Example 4 The objective of the study was to demonstrate the effects of a curcumin composition on cartilage degradation and inflammatory markers in synovial tissue.

[0085] Methods: Animals were sacrificed, synovial tissue samples were collected, and evaluated for markers of cartilage degradation and inflammation by Western blotting as shown in Figure 06.

[0086] Conclusion: Protein levels of collagen degradation markers such as type II collagen (A), MMP-3 (B), and inflammatory markers COX-2 (C) and LOX-5 (D), as shown below, were found to be increased in OA rats, as measured by Western blotting followed by relative intensity concentration analysis with a control group after β-actin normalization to ensure equal protein loading. The blot was repeated at least three times (n=3), and a representative blot (E) is shown.

[0087] Treatment with two doses of curcumin composition was found to reduce levels of markers of cartilage degradation and inflammation in joint tissue in a statistically significant manner.

[0088] Example 5 The objective of this study was to demonstrate the effect of a curcumin composition on the articular structure of synovial joints, as evaluated by X-ray image analysis.

[0089] Methods: The structural integrity of the synovial joints in the animals was evaluated by radiographic analysis, as shown in Figure 07 and Table 04. [Table 8]

[0090] Conclusion: The induction of osteoarthritis (OA) was found to be associated with a loss of joint structural integrity observed in the X-ray image (A) obtained at the end of the experiment. Significant improvements in joint structure were observed after treatment with curcumin composition 1 and curcumin composition 2. These results were further validated by measuring the mean ± standard deviation of the Kellgren-Lawrence score (B).

[0091] Example 6 The objective of this study was to demonstrate the effect of curcumin compositions on the morphological analysis of synovial tissue.

[0092] Methods: Animals were sacrificed, synovial tissue samples were collected, fixed in 10% formalin at 4°C for 24 hours, decalcified with 5% hydrochloric acid at 4°C for 4 days, dehydrated stepwise in acetone, and embedded in paraffin. Sections 2–3 μm thick were stained with 0.2% hematoxylin and 1% eosin for 5 minutes and 3 minutes, respectively. Histological preparations were analyzed and photographed under a microscope using a digital image acquisition camera, as shown in Figure 8.

[0093] Conclusion: As observed after the histopathological findings obtained at the end of the experiment using hematoxylin-eosin (A) and toluidine blue (B) staining, significant loss of joint structure and inflammatory infiltration of the joints were observed in the OA state. This was further confirmed by the mean Mankin score (C).

[0094] Example 7 The objective of this study was to demonstrate the effect of curcumin compositions on the morphological analysis of synovial tissue.

[0095] Methods: The knee was visually evaluated for morphological changes in joint pathology, such as knee swelling, knee joint diameter, and the ratio of right-to-left diameter values, as shown in Figure 09.

[0096] Conclusion: Swelling of both the right and left knee joints was observed when osteoarthritis (OA) was induced in rats, and this was significantly reduced by treatment with curcumin composition 1 and curcumin composition 2 (A). This was further confirmed by measuring the left knee joint diameter (B), the right knee joint diameter (C), and the ratio of the right to left diameter values, and a significant therapeutic effect was observed with curcumin composition 1 and curcumin composition 2.

[0097] Example 8 The objective of this study was to demonstrate the effects of curcumin composition on foot area and stride length.

[0098] Method: Ink was brushed onto the hind legs of rats, and the animals were made to run on a 60 cm long, 7 cm wide track covered with white paper. A dark room was placed at the end of the track to lure the rats into. After the test was completed, the paper was scanned at 300 dpi. The dimensions of the foot circumference were measured as the foot area (cm²). 2 The distance between the first and fifth toes was defined as the foot width (cm), the distance between the same hind limbs over two steps was defined as the stride length (cm), the horizontal distance between the left and right feet was defined as the base (cm), the distance between the third toe and the heel was defined as the foot length (cm), and the foot angle was defined as the angle (°) through which the hind limb passes. The footprint measurements were quantified using ImageJ software as shown in Figure 10.

[0099] Conclusion: Foot area and stride length were significantly reduced due to osteoarthritis. Significant improvements were observed in both foot area and stride length with curcumin composition 1 and curcumin composition 2.

[0100] Clinical Experimental Study 2: Objective: The objective of this study was to evaluate the plasma pharmacokinetic profile of a curcumin composition in adult male Sprague Dolly's genus.

[0101] Dosage selection A dose of 1000 mg / kg body weight (200 mg / kg total curcuminoids) was selected for oral administration.

[0102] Test sample: Curcumin composition according to Example 01 Reference: Curcumin extract (curcumin extract without any formulation or polymorphic form) Curcumin extract (regular curcumin) is an extract that is not in any formulation or polymorph, and is generally prepared by drying turmeric rhizomes extracted with a solvent. It is evaporated to form oleoresin turmeric, which is then crystallized using a suitable solvent. The resulting crystals are dried and powdered, and contain 95% curcuminoids. Time points (hours): 0.00, 0.25, 0.50, 1.00, 2.00, 3.00, 4.00, 6.00, 8.00, and 24.00 [Table 9]

[0103] Conclusion: The curcumin composition showed a 64.71 times higher AUC compared to the reference curcumin extract.

[0104] Based on the results and conclusions provided regarding the osteoarthritis model, we conclude here that the same curcumin composition also has beneficial effects on muscle health. Cell line study models further elucidated by the research protocol are described below for studying the efficacy on muscle health.

[0105] Further research to demonstrate the effects of curcumin compositions on muscle health is as follows:

[0106] In vitro research: Objective: To demonstrate the ergogenic potential of curcumin compositions (test samples) to support muscle development (preventing muscle protein loss as seen in age-related sarcopenia), enhance athletic performance, alleviate muscle injury and inflammation (DOMS), and improve physiological or metabolic responses. Cell model: C2C12 mouse myoblasts Experimental method: The study was conducted based on a basic assay to generate more information and identify the selective material components for development. C2C12 myoblasts were cultured at 37°C in humidified 5% CO2, seeded in 24-well plates, incubated for 24 hours, and then treated. 1. Cytotoxicity assay 2. Mitochondrial density assay: The effect of material components on mitochondrial biogenesis is analyzed by measuring cellular respiration accompanied by a calculated increase in mitochondrial mass. 3. Antioxidant capacity: After 24 hours of treatment, the total antioxidant capacity was determined in the C2C12 homogenate. 4. Lactate dehydrogenase and creatine kinase activity: LDH and CK activity are used as indicators of muscle cell damage and are evaluated by determining the release of the cytosolic enzyme lactate dehydrogenase (LDH) and creatine kinase (CK) activity. 5. IGF-1 Elisa Assay: IGF-I exerts rapid anabolic effects on protein and carbohydrate metabolism by increasing the cellular uptake of amino acids in cells, and plays a major role in regulating skeletal muscle growth.

[0107] In vivo research: Objective: To evaluate the effects of a curcumin composition (test sample) on endurance, grip strength, and muscle metabolism in rats.

[0108] Animal and experimental design: Twenty-one male Wistar albino or Sprague Dolly rats (20% extra animals), aged 8 weeks and weighing 180 ± 20 g, were housed in a controlled environment with a 12:12 light-dark cycle at 22°C and given free access to solid feed and water.

[0109] The rats were randomly divided into three treatment groups, each containing seven animals. 1. Control group (normal, with movement) 2. Curcumin composition 1 (100 mg / kg preparation) 3. Curcumin composition 2 (200 mg / kg preparation)

[0110] During the screening period, grip strength was tested on the third day. A treadmill test was conducted on the second day.

[0111] Grip strength test: The grip strength of rats was evaluated using a force measurement system. Combined forelimb and hindlimb grip strength, as well as forelimb grip strength, were measured at the end of the oral administration period. The system has an electronic digital force meter to determine peak force. Each rat was held by its tail until the pull bar was released. Five consecutive tests were performed for each rat to obtain peak values.

[0112] Treadmill test: Running endurance in rats was measured using an animal treadmill. Rats were fasted overnight, sacrificed by cervical dislocation, and blood and calf muscle tissue were collected. Serum samples were obtained by centrifuging blood samples (5000 rpm at 4°C for 10 minutes) and then transferring them to gel biochemistry tubes. Liver and calf muscle samples (obtained from approximately the same location at each time) were immediately removed, placed on ice, and maintained at -80°C until analysis. For biochemical assays, tissue samples were homogenized within 10 minutes in 10 volumes of cold Tris 10 mM (pH 7.4). Muscle homogenates were centrifuged at 4000 × g at 4°C for 10 minutes to obtain a slow-speed supernatant fraction, which was used for lipid peroxidation analysis.

[0113] Biochemical analysis: Serum glucose, lipid profiles, aspartate aminotransferase (AST), alanine aminotransferase (ALT), urea, and creatinine levels are analyzed using a portable automated chemical analyzer (Samsung LABGEO PT10V, Samsung Electronics Co., Suwon, South Korea). Serum lactate concentration is measured by enzyme-linked immunosorbent assay (ELISA, Elx-800, Bio-Tek Instruments Inc, Vermont, USA) using a rat lactate assay kit (Cayman Chemical Co., Ann Arbor, MI, USA). ELISA (MyBioSource, San Diego, CA, USA) is also used to measure serum myoglobin concentration.

[0114] Malondialdehyde (MDA) levels in muscle tissue were measured by high-performance liquid chromatography (Shimadzu, Tokyo, Japan) using a Shimadzu UV-vis SPD-10 AVP detector and a C18 ODS-3, 5 μm, 4.6 mm × 250 mm column. The activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GSH-Px) were determined using commercially available kits (Cayman Chemical, Ann Arbor, MI, USA) according to the manufacturer's procedures.

[0115] It will be apparent to those skilled in the art that the present invention is not limited to the details of the exemplary embodiments, and that the present invention may be embodied in other specific forms without departing from its essential attributes. Therefore, it is desirable to consider these embodiments and examples as illustrative and non-limiting in all respects, and that the appended claims, rather than the above description, should be referred to; therefore, all modifications within the meaning and scope of the equivalent claims are intended to be encompassed within the claims.

Claims

1. A stable curcumin composition having enhanced bioavailability for use in methods for preventing, improving, and / or maintaining osteoarthritis in a subject, i) Curcumin comprising (a) curcumin in crystalline polymorphic form ranging from 0% w / w to 15% w / w of total curcuminoids and (b) curcumin in amorphous form ranging from 85% w / w to 100% w / w of total curcuminoids, ii) At least one pharmaceutically or nutritionally acceptable material component selected from the group consisting of hydrophilic carriers, antioxidants, stabilizers, pH adjusters, solubilizers, fats, and anti-caking agents. Includes, The use described above includes administering the composition to the subject, A stable curcumin composition.

2. The stable curcumin composition according to claim 1, wherein the curcuminoid is selected from the group consisting of curcumin, demethoxycurcumin, bisdemethoxycurcumin, and mixtures thereof.

3. The stable curcumin composition according to claim 1, wherein the curcuminoid is present in an amount ranging from 10% w / w to 90% w / w of the composition.

4. The stable curcumin composition according to claim 1, wherein the composition has a bioavailability that is 60 to 70 times greater than that of the reference.

5. The stable curcumin composition according to claim 1, wherein the hydrophilic carrier is selected from the group consisting of cellulose, starch, and derivatives thereof, and is present in the composition in an amount of 1% w / w to 90% w / w.

6. The stable curcumin composition according to claim 1, wherein the antioxidant is selected from the group consisting of tocopherol, mixed tocopherol, ascorbyl palmitate, catechin, and ascorbic acid, and is present in the composition at a concentration of 0.1% w / w to 10% w / w.

7. The stable curcumin composition according to claim 1, wherein the stabilizer is selected from the group consisting of glyceryl monostearate, sugar alcohols, and triglycerides, and is present in the composition at a concentration of 0.1% w / w to 10% w / w.

8. The stable curcumin composition according to claim 1, wherein the pH adjusting agent is selected from the group consisting of citric acid, trisodium citrate, lactic acid, and magnesium carbonate, and is present in the composition at a concentration of 0.1% w / w to 10% w / w.

9. The stable curcumin composition according to claim 1, wherein the solubilizing agent is selected from the group consisting of lecithin, d-limonene, propylene glycol alginate, and sodium lauryl sulfate, and is present in the composition in an amount of 0.1% w / w to 40% w / w.

10. The stable curcumin composition according to claim 1, wherein the fat is selected from the group consisting of medium-chain triglycerides, terpenes, and vegetable oils, and is present in the composition at a concentration of 0.1% w / w to 40% w / w.

11. The stable curcumin composition according to claim 1, wherein the anti-solidification agent is selected from the group consisting of colloidal silicon dioxide, magnesium stearate, stearic acid, and mannitol, and is present in the composition in an amount of 0.1% w / w to 10% w / w.

12. The stable curcumin composition according to claim 1, wherein the composition is in a form selected from the group consisting of tablets, capsules, blended powders, ricups, ointments, pastes, lotions, liniments, mouthwashes, gargles, dry syrups, liquid syrups, drinks, diet drinks, fruit juices, soft drinks, and the like.

13. The stable curcumin composition according to claim 1, wherein the composition is in a form selected from the group consisting of granules, powders, and mixtures thereof.

14. The stable curcumin composition according to claim 1, wherein the administration is by oral administration.

15. The stable curcumin composition according to claim 1, wherein the use results in the following in the subject: improved joint mobility, improved joint flexibility, reduced joint stiffness, improved walking ability, reduced cartilage destruction, and / or reduced inflammation.

16. The aforementioned use in the subject (a) reducing one or more inflammatory markers selected from the group consisting of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-6 (IL-6), NF-κβ, type II collagen, matrix metalloproteinase-3 (MMP3), cyclooxygenase-2 (COX-2), 5-lipoxygenase (5-LOX), cartilage oligomeric matrix protein (COMP); C-reactive protein (CRP), and malondialdehyde (MDA), and / or (b) Increase one or more antioxidant markers selected from the group consisting of interleukin-10 (IL-10), superoxide dismutase (SOD), glutathione peroxidase (GSH-Px), and catalase (CAT). A stable curcumin composition according to claim 1.

17. The stable curcumin composition according to claim 1, wherein the use results in improved muscle strength, improved muscle performance, reduced muscle soreness, reduced muscle damage, and / or faster muscle recovery in the subject.

18. The stable curcumin composition according to claim 1, wherein the subject is a human and / or an animal.

19. The stable curcumin composition according to claim 1, in an administration form, the dose being 10 to 1000 mg / kg.

20. A stable curcumin composition having enhanced bioavailability for use in methods for preventing, improving, and / or maintaining osteoarthritis in a subject, (a) Curcuminoids in crystalline polymorphic forms ranging from 0% w / w to 15% w / w of total curcuminoids, and (b) Amorphous curcuminoids in the range of 85% w / w to 100% w / w of total curcuminoids Includes, A stable curcumin composition whose use comprises orally administering the composition to the subject.

21. The stable curcumin composition according to claim 1 or 20, wherein crystalline polymorphic curcuminoids are present in an amount of 1.2% w / w of the total curcuminoids, and amorphous curcuminoids are present in an amount of 98.8% w / w of the total curcuminoids.

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