Compositions and methods using a combination of fisetin and quercetin for use in cartilage degeneration

Abstract

The present invention relates to the use of a composition comprising an effective amount of a combination of fisetin and / or its derivatives and quercetin and / or derivatives for maintaining joint health or preventing or treating joint disorders in an individual. In particular, the present invention relates to a composition comprising an effective amount of a combination of fisetin and / or its derivatives and quercetin and / or derivatives for use in preventing or treating cartilage degeneration in an individual.

Description

Technical Field

[0001] The present invention relates to joint health, and in particular to the use of a composition comprising a combination of fisetin and / or its derivatives and quercetin and / or its metabolites for the prevention or treatment of joint disorders or the maintenance of joint health.

Background Art

[0002] Osteoarthritis (OA) is a disease with a high morbidity rate that has a significant socioeconomic impact. Osteoarthritis is a degenerative disease of the articular cartilage of joints and is the most common form of arthritis, affecting 10% of the adult population. OA is a major cause of physical disability and medical costs in the elderly worldwide. Progressive degeneration and reduction of articular cartilage belong to the main features of the pathology, and are accompanied by other joint structure changes that all contribute to the clinical symptoms of OA, such as synovial proliferation, subchondral bone sclerosis and thickness, osteophyte formation around the joint, ligament relaxation, and muscle atrophy. These symptoms include severe pain, stiffness, reduced joint movement, and impairment. Since articular cartilage depends only on its resident cells, chondrocytes, for the maintenance of the extracellular matrix, impairment of chondrocyte function and survival leads to joint cartilage dysfunction.

[0003] Recent ex vivo studies have reported mitochondrial dysfunction in human OA chondrocytes, and analysis of mitochondrial electron transport chain activity in these cells has shown decreased activity of Complexes I, II, and III, as well as low ATP production, compared to normal chondrocytes. This mitochondrial dysfunction can affect several pathways involved in cartilage degeneration, such as oxidative stress, failure of chondrocyte biosynthesis and proliferation responses, increased cytokine-induced chondrocyte inflammation and matrix degradation, calcification of the cartilage matrix, and increased chondrocyte apoptosis (Blanco et al. “The role of mitochondria in osteoarthritis” Nat. Rev. Rheumatol. 7, 161 - 169 (2011)).

[0004] Mitochondria are the major source of aerobic energy production in mammalian cells and also maintain a large Ca2+ gradient across the inner membrane, providing the signaling potential of the molecule. Furthermore, mitochondrial Ca2+ may act in the regulation of ATP production in mitochondria and contribute to the orchestration of cellular metabolic homeostasis. (Glancy, B. and R. S. Balaban (2012). “Role of mitochondrial Ca2+ in the regulation of cellular energetics.” Biochemistry 51(14):2959 - 2973).

[0005] The number of individuals affected by OA is increasing, but there is still no cure, and current drug therapies still only exist as symptomatic treatments focused on alleviating symptoms. For example, pain and inflammation are treated using analgesics (such as acetaminophen) and non - steroidal anti - inflammatory drugs (NSAIDs). Furthermore, the use of these drugs often involves side effects such as gastrointestinal or cardiovascular risks. Since current treatments for OA do not prevent or cure OA, chondrocyte apoptosis is an effective target for regulating cartilage degeneration.

Summary of the Invention

[0006] The inventors have surprisingly demonstrated that the combination of fisetin and quercetin synergistically activates mitochondrial function at the cellular level via an increase in mitochondrial calcium.

[0007] Accordingly, an object of the present invention relates to providing a composition for use in improving joint health. In particular, an object of the present invention is to provide a composition that improves joint health by preventing or treating cartilage degeneration and solves the above-mentioned problems of the prior art regarding side effects such as gastrointestinal and / or cardiovascular risks.

[0008] Accordingly, one aspect of the present invention relates to a composition comprising an effective amount of a combination of fisetin and / or its metabolites and quercetin and / or its derivatives for use in preventing or treating cartilage degeneration in an individual.

[0009] Another aspect of the present invention relates to a method for manufacturing a composition for use according to the present invention.

[0010] In a last aspect, the present invention relates to a kit comprising an effective amount of a combination of fisetin and / or its derivatives and quercetin and / or its derivatives in one or more containers.

Effects of the Invention

[0011] Further features and advantages are described herein and will become apparent from the following drawings and the mode for carrying out the invention.

Brief Description of the Drawings

[0012]

Figure 1

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Figure 5

Mode for Carrying Out the Invention

[0013] Definitions Before discussing the present invention in more detail, the following terms and general technical common knowledge are first defined.

[0014] In the context of the present invention, unless otherwise specified, the percentages mentioned are weight / weight percentages.

[0015] The term "and / or" used in the context of "X and / or Y" should be interpreted as "X" or "Y", or "X and Y".

[0016] The numerical ranges used in this specification are intended to include all numbers and subsets of numbers included within the range, whether explicitly disclosed or not. Further, these numerical ranges should be construed to provide support for claims directed to any number or subset of numbers within the range. For example, the disclosure of 1 to 10 should be construed as corresponding to ranges such as 1 to 8, 3 to 7, 4 to 9, 3.6 to 4.6, 3.5 to 9.9, and the like.

[0017] As used herein, the terms "prevent" and "prevention" mean administering to a subject that does not exhibit any symptoms of the condition a composition disclosed herein to suppress or prevent the occurrence of at least one symptom associated with the condition. Further, "prevention" includes a reduction in the risk, incidence, and / or severity of a condition or disorder.

[0018] As used herein, "effective amount" means an amount that treats or prevents a deficiency, treats or prevents a disease or medical condition, or more generally, reduces symptoms, manages disease progression, or provides a nutritional, physiological, or medical benefit to an individual.

[0019] "Animal" includes, but is not limited to, mammals such as rodents; aquatic mammals; domestic animals such as dogs, cats, and other pets; livestock such as sheep, pigs, cows, and horses; and humans. When the terms "animal," "mammal," or their plurals are used, these terms also apply to any animal to which the effects shown or intended, for example, by the benefit of the animal from the improvement of mitochondrial calcium transport, can be shown depending on the context. The term "individual" or "subject" is often used herein to refer to a human, but the disclosure is not so limited. Accordingly, the term "individual" or "subject" refers to any animal, mammal, or human that can benefit from the methods and compositions disclosed herein.

[0020] The term "pet" means any animal that can benefit from or tolerate the compositions provided by the present disclosure. For example, a pet may be an animal such as a bird, bovine, canine, equine, feline, goat, lupine, rodent, sheep, or pig, but a pet can be any suitable animal. The term "companion animal" means a dog or a cat.

[0021] A "subject" or "individual" is a mammal, preferably a human. The term "elderly" in the context of a human means an age of at least 60 years, preferably over 63 years, more preferably over 65 years, and most preferably over 70 years. The term "older adult" in the context of a human means a postnatal age of 45 years or older, preferably over 50 years, more preferably over 55 years, and includes the elderly. The term "older adult" in the context of a human means a postnatal age of 45 years or older, preferably over 50 years, more preferably over 55 years, and includes the elderly.

[0022] An "oral nutritional supplement" or "ONS" is a composition that contains at least one macronutrient and / or at least one micronutrient, and is, for example, in the form of a sterile liquid, semi-solid, or powder, and is intended to supplement other nutritional intakes such as nutritional intake from food. Non-limiting examples of commercially available ONS products include MERITENE®, BOOST®, NUTREN®, and SUSTAGEN®. In some embodiments, the ONS can be ingested without further addition of a liquid, for example, can be a beverage in liquid form where the liquid volume is for one serving of the composition.

[0023] "Kit" means that the components of the kit are physically associated in or with one or more containers and are considered as one unit for manufacturing, distribution, sale, or use. The containers include, but are not limited to, bags, boxes, cartons, bottles, packages of any type or design or material, overwraps, shrink wraps, attached components (e.g., stapled or glued ones), or combinations thereof.

[0024] Unless otherwise specified or clearly implied by the context in which it is mentioned as being incompatible with a single feature or a single limitation of the present invention, any reference to a single feature or a single limitation of the present invention shall be taken to include the corresponding plurality of features or plurality of limitations, and any reference to a plurality of features or plurality of limitations shall be taken to include the corresponding single feature or single limitation.

[0025] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art.

[0026] Composition for use Articular diseases may be accompanied by inflammation to varying degrees. In some diseases, for example, rheumatoid arthritis (RA), inflammation is the most important factor. In other diseases, such as OA, inflammation does not appear as prominently. However, all diseases have elements that show catabolic effects, by which the articular cartilage is destroyed.

[0027] The inventors have shown that the provision of a combination of fisetin and / or its derivatives and quercetin and / or derivatives synergistically improves the altered mitochondrial function, for example, in osteoarthritis.

[0028] That is, in a first aspect, the present invention relates to a composition comprising an effective amount of a combination of fisetin and / or its derivatives and quercetin and / or its derivatives for use, for example, in preventing or treating cartilage degeneration in an individual in order to improve joint health.

[0029] In another mode, this aspect of the present invention can be described as the use of an effective amount of a combination of fisetin and / or its derivatives and quercetin and / or its derivatives in the manufacture of a medicament for the prevention or treatment of cartilage degeneration in an individual.

[0030] The use for preventing or treating cartilage degeneration is synonymous with the use for inhibiting or reducing cartilage degeneration.

[0031] Accordingly, embodiments of the present invention include a composition comprising an effective amount of a combination of fisetin and / or its derivatives and quercetin and / or its derivatives for use in preventing or treating cartilage degeneration.

[0032] A further embodiment of the present invention includes a composition for use according to the present invention, the composition further comprising calcium.

[0033] Raw materials Main bioactive compounds Fisetin and quercetin are the main bioactive molecules according to the present invention.

[0034] Fisetin (7,3’,4’-flavon-3-ol) (see Figure 1) is a polyphenol found in many plants and is used as a yellow / ochre colorant. Fisetin is also found in many fruits and vegetables such as strawberries, apples, persimmons, grapes, onions, and cucumbers.

[0035] In one embodiment, at least a portion of the fisetin is obtained by known means such as, for example, extraction of fisetin from a plant source / vegetable source / fruit source. Additionally or alternatively, at least a portion of the fisetin and / or its derivatives can be obtained by chemical synthesis.

[0036] Non-limiting examples of suitable derivatives of fisetin include its glucuronidated form, its sulfated form, derivatives, and mixtures thereof. In a preferred embodiment, the derivative is gerardol.

[0037] Quercetin is the aglycone form of many other flavonoid glycosides such as rutin and quercitrin found in citrus fruits, buckwheat, and onions. Quercetin is derived from quercitrin and rutin, which are glycosides accompanied by rhamnose and rutinose, respectively.

[0038] Similarly, guaijaverin is 3-O-arabinoside, hyperoside is 3-O-galactoside, isoquercetin is 3-O-glucoside, spireoside is 4’-O-glucoside. Michelianin is quercetin 3-O-β-D-glucuronopyranoside.

[0039] In a preferred embodiment, the derivative of quercetin can be selected from the group consisting of quercetin 3-O-galactoside, quercetin 3-O-glucoside (isoquercetin), quercetin 3-O-xyloside, quercetin 3-O-rhamnoside (quercitrin), quercetin 3-O-glucuronide, quercetin 7-O-glucoside, quercetin 3-O-diglucoside, quercetin 3,4’-diglucoside, quercetin 3-O-rhamnoside-7O-glucoside, quercetin 3-O-rutinose (rutin), quercetin 3-O-6’’-acetylglucoside, quercetin 3-methyl ether, quercetin 3,3’-dimethyl ether, isorhamnetin, and mixtures thereof.

[0040] Quercetin can be derived from any suitable raw material and may be isolated or chemically synthesized.

[0041] In preferred embodiments, fisetin, quercetin, and derivatives are obtained from plant sources. For example, fisetin can be obtained from strawberries, apples, persimmons, grapes, onions, cucumbers, etc. For example, quercetin can be obtained from onions, green tea, apples, berries, Ginkgo biloba, St. John’s wort, American ginseng, buckwheat tea, etc.

[0042] The effective amounts of each of fisetin and / or its derivatives and quercetin and / or its derivatives vary depending on the specific composition, the age and condition of the user, and the specific disorder or disease being treated. However, in general embodiments, 0.001 mg to 1.0 g per day, preferably 0.01 mg to 0.9 g per day, more preferably 0.1 mg to 750 mg per day, more preferably 0.5 mg to 500 mg per day, and most preferably 1.0 mg to 200 mg per day can be administered to an individual. Furthermore, the inventors have found that the active dose of fisetin or its derivative in the combination can be reduced with equivalent effectiveness.

[0043] In some embodiments, the combination of fisetin or its derivative and quercetin or its derivative is administered in a composition further comprising calcium. At least a portion of the calcium may be one or more calcium salts such as calcium acetate, calcium carbonate, calcium chloride, calcium citrate, calcium gluceptate, calcium gluconate, calcium lactate, or mixtures thereof. In general embodiments, 0.1 g to 1.0 g of calcium per day, preferably 125 mg to 950 g of calcium per day, more preferably 150 mg to 900 mg of calcium per day, more preferably 175 mg to 850 mg of calcium per day, and most preferably 200 mg to 800 mg of calcium per day are administered to an individual.

[0044] In alternative embodiments, the combination of fisetin and quercetin can be administered continuously with calcium in separate compositions. The term "continuously" means that at a first time, at least one of fisetin or its derivatives is administered without calcium, and at a second time (before or after the first time), calcium and at least one of fisetin or its derivatives are administered sequentially so that calcium is administered without the combination of fisetin and quercetin. The time between successive administrations can be, for example, one second or several seconds, minutes, or hours on the same day; one day or several days, or weeks in the same month; or one month or several months in the same year.

[0045] Fisetin or a derivative and quercetin or a derivative can be formulated in a specific ratio. In some embodiments, the formulation may contain these components in the following exemplary ratios: 1:1, 1:2, 1:3, 1:4, 1:5, 1:6, 1:7, 1:8, 1:9, 1:10, 1:20, and each of these ratios can be, in some embodiments, fisetin:quercetin, and in other embodiments, quercetin:fisetin. Preferably, the ratio is from 1:1 to 1:10.

[0046] In some embodiments, fisetin or its derivative and quercetin or its derivative are the only polyphenols in the composition and / or the only polyphenols administered to the individual.

[0047] The composition can contain an effective amount of at least one of fisetin or its derivatives. For example, one serving or administration of the composition can contain an effective amount, and the package can contain more than one serving or administration amount. Optionally, the composition may further contain calcium.

[0048] Raw materials Further bioactive compounds The composition for use according to the present invention may also contain at least one further bioactive compound selected from the group consisting of antioxidants, anti-inflammatory compounds, glycosaminoglycans, prebiotics, fibers, probiotics, fatty acids, enzymes, minerals, trace elements and / or vitamins.

[0049] In the context of this application, the term "bioactive" means that the compound contributes to the health of an individual or has an effect on the human body beyond meeting basic nutritional requirements.

[0050] At least one further bioactive compound may be of natural origin. That is, the compound may be obtained from extracts of plants, animals, fish, fungi, algae, or microbial fermentates. Minerals are also considered to be of natural origin within this definition.

[0051] In a preferred embodiment, the enzyme may be a protease such as trypsin or an enzyme extract such as bromelain.

[0052] Nutritional composition The composition for use according to the present invention may be a nutritional composition or a pharmaceutical composition, and may be for use in humans or livestock.

[0053] Therefore, in a preferred embodiment, the composition for use according to the present invention is a nutritional composition.

[0054] In the context of this application, the term "nutritional composition" means a composition that is a source of nutrition for an individual.

[0055] The nutritional preparation or nutritional composition of the present invention may be a complete source of nutrition or an incomplete source of nutrition.

[0056] As used herein, "complete nutrition" encompasses nutritional formulations and compositions that contain sufficient types and levels of macronutrients (protein, fat, and carbohydrates) and sufficient micronutrients to serve as the sole source of nutrition for the animal to which it is administered. A patient can obtain 100% of their nutritional requirements from such complete nutrition compositions.

[0057] As used herein, "incomplete nutrition" encompasses nutritional formulations or compositions that do not contain sufficient levels of macronutrients (protein, fat, and carbohydrates) or sufficient micronutrients to serve as the sole source of nutrition for the animal to which it is administered. Partially complete nutrition compositions or incomplete nutrition compositions can be used as nutritional supplements.

[0058] The combination of fisetin and quercetin can be administered in any composition suitable for human and / or animal consumption. In a preferred embodiment, such combination is administered orally or enterally (e.g., by tube feeding) to an individual. For example, such combination can be administered to an individual as a beverage, food product, capsule, tablet, powder, or suspension.

[0059] Non-limiting examples of suitable compositions include food compositions, dietary supplements, dietary supplements (e.g., liquid ONS), complete nutrition compositions, beverages, pharmaceuticals, oral nutritional supplements, medical foods, nutraceuticals, foods for special medical purposes (FSMP), powder nutritional formulations that are reconstituted with water or milk prior to ingestion, food additives, pharmaceuticals, drinks, pet foods, and combinations thereof.

[0060] Raw materials of nutritional compositions Protein source In one embodiment, the composition for use according to the present invention comprises a protein source. The protein source may be a food protein including, but not limited to, animal proteins (such as milk protein, meat protein, or egg protein), plant proteins (such as soy protein, wheat protein, rice protein, and pea protein), or combinations thereof. In one embodiment, the protein is selected from the group consisting of whey, chicken, corn, caseinate, wheat, flax, soy, lentils, peas, or combinations thereof.

[0061] Carbohydrate source In one embodiment, the composition comprises a carbohydrate source. Any suitable carbohydrate may be used in the composition of the present invention, examples of which include, but are not limited to, starch, sucrose, lactose, glucose, fructose, corn syrup solids, maltodextrin, modified starch, amylose starch, tapioca starch, corn starch, xylitol, sorbitol, or combinations thereof.

[0062] Fat source In one embodiment, the composition comprises a fat source. The fat source may include any suitable fat or fat mixture. For example, the fat source includes, but is not limited to, vegetable fats (such as olive oil, corn oil, sunflower oil, high oleic sunflower oil, rapeseed oil, canola oil, hazelnut oil, soybean oil, palm oil, coconut oil, blackcurrant seed oil, borage oil, and lecithin), animal fats (such as milk fat), or combinations thereof. The fat source may also be a crude or refined product of the above fats (such as olive oil containing polyphenols).

[0063] Flavorings, etc. In addition, the composition for use according to the present invention may also include natural or artificial flavors, such as fruit flavors like banana, orange, peach, pineapple or raspberry, or other plant flavors like vanilla, cocoa, coffee, etc.

[0064] Nutritional composition format In addition to the main bioactive ingredient, and any additional bioactive ingredients, and optionally one or more of a protein source, a carbohydrate source, and a fat source, the nutritional composition of the present invention may also contain, for example, one or more acidulants, additional thickeners, buffers or pH adjusters, chelating agents, colorants, emulsifiers, additives, flavorings, minerals, penetrants, pharmaceutically acceptable carriers, preservatives, stabilizers, saccharides, sweeteners, texturizers, and / or vitamins, etc., any number of optional additional food raw materials including conventional food additives (artificial or natural). The optional raw materials can be added in any suitable amount.

[0065] The nutritional composition of the present invention may be provided in any suitable format.

[0066] Examples of formats of nutritional compositions in which the composition for use according to the present invention can be supplied include solutions, ready-for-consumption compositions (e.g., ready-to-drink compositions or instant beverages), liquid foods, soft drinks, juices, sports drinks, milk beverages, milkshakes, yogurt drinks, soups, etc.

[0067] In other embodiments, the nutritional composition may be provided in the form of a concentrate, powder, or granules (e.g., effervescent granules), and can be diluted with water or other liquids, such as milk or fruit juice, etc., to make a ready-for-consumption composition.

[0068] Formats of nutritional compositions further include baked products, dairy products, desserts, confectionery products, cereal bars, and breakfast cereals. Examples of dairy products include milk and dairy products, yogurt, and other fermented dairy products, ice cream, and cheese. Examples of baked products include bread, biscuits, and cakes.

[0069] In one embodiment, the composition for use according to the invention may also be available in a variety of formats, particularly for animal food, especially for dogs or cats, regardless of whether it is in wet, semi-wet or dry form, particularly in the form of biscuits.

[0070] Route of administration The nutritional composition of the present disclosure may be administered in any suitable manner suitable for administration to humans, particularly to any part of the gastrointestinal tract. Enteral administration, oral administration, and administration through a tube or catheter are all included in the present disclosure. The nutritional composition of the present invention may also be administered by a method selected from oral, enteral, sublingual, sublabial, buccal, topical, etc.

[0071] The nutritional composition of the present invention may be administered in a convenient dosage form in any known form, including, for example, tablets, capsules, liquids, chewables, soft gels, sachets, powders, syrups, suspensions, emulsions, and solutions. In soft capsules, it is preferred that the active raw material is dissolved or suspended in a suitable liquid such as fatty oil, paraffin oil, or liquid polyethylene glycol. If desired, stabilizers may be added.

[0072] When the nutritional composition is administered by tube feeding, the nutritional composition may be used for short-term or long-term tube feeding.

[0073] Inhibition or reduction of cartilage degeneration Cartilage degeneration can be the result of pathology (either chronic or acute), trauma, or a combination thereof.

[0074] Cartilage degeneration occurs in both disease states dominated by inflammation (e.g., rheumatoid arthritis) and disease states where inflammation is not so prominent (e.g., osteoarthritis).

[0075] Trauma can also initiate the cartilage degeneration process. For example, rupture of the ligaments in the knee leads to instability of the knee joint and the initiation of the degeneration process.

[0076] In the context of the present application, trauma refers to physiological damage caused by external factors such as impact due to, for example, a fall or a vehicle. Trauma may also be the accumulation of small damages over time, so-called "wear and tear".

[0077] Although it is often preferable to treat trauma surgically, in one embodiment, the present invention relates to a treatment method for treating trauma by surgery and by administering the composition of the present invention.

[0078] Accordingly, embodiments of the use according to the present invention include use for inhibiting or reducing cartilage degeneration, which is a result of pathology or trauma.

[0079] Examples of pathological conditions that include cartilage degeneration and thus for which the composition of the present invention may be useful are osteoarthritis, rheumatoid arthritis, gout and pseudogout, septic arthritis, ankylosing spondylitis, juvenile idiopathic arthritis, Still's disease, psoriasis (psoriatic arthritis), reactive arthritis, Ehlers-Danlos syndrome, hemochromatosis, hepatitis, Lyme disease, inflammatory bowel disease (including Crohn's disease and ulcerative colitis), Henoch-Schönlein purpura, hyperimmunoglobulinemia D with recurrent fever, sarcoidosis, TNF receptor-associated periodic syndrome, Wegener's granulomatosis (and many other vasculitis syndromes), familial Mediterranean fever, systemic lupus erythematosus.

[0080] In a preferred embodiment, the composition of the present invention is for use in inhibiting or reducing cartilage degeneration in RA and / or OA.

[0081] In a more preferred embodiment, the composition of the present invention is for use in inhibiting or reducing cartilage degeneration in OA.

[0082] Furthermore, although not wishing to be bound by theory, while inflammation often leads to cartilage degeneration in joints, it has been observed that cartilage degeneration occurs even in situations where the inflammatory component is much less, perhaps even negligible.

[0083] For example, trauma to a joint can sufficiently initiate cartilage degeneration without a significant inflammatory component such as that present in RA. Trauma can include, for example, a ligament tear or impact trauma to a joint (e.g., knee, finger).

[0084] In another example, OA is primarily a joint degenerative disease and is associated with a lesser inflammatory component.

[0085] Accordingly, in one embodiment, the present invention relates to a composition for use according to the present invention for inhibiting or reducing cartilage degeneration, wherein the cartilage degeneration occurs in relation to a condition with little or no inflammatory component such as trauma or OA.

[0086] Use for combating initial degenerative events Hypertrophy suggests the deregulated activity of chondrocytes that is not the normal phenotype.

[0087] Accordingly, in one embodiment, the present invention relates to a composition according to the present invention for use in inhibiting or reducing chondrocyte hypertrophy, which is one initial event indicating cartilage degeneration, and comprising a combination of fisetin and / or its derivatives and quercetin and / or derivatives.

[0088] Treatment or prevention of age-related mobility decline The composition for use according to the present invention has been shown to inhibit or reduce proteolytic activity.

[0089] Aging leads to cartilage degeneration.

[0090] Accordingly, the present invention relates to a composition of the present invention for use in inhibiting or reducing age-related cartilage degeneration.

[0091] In another embodiment, the present invention relates to the use of the composition of the present invention for inhibiting or reducing collagen denaturation in age-related cartilage degeneration, for example, for inhibiting or preventing collagen II denaturation in age-related cartilage.

[0092] Cartilage degeneration can contribute to joint stiffness and joint pain, resulting in a decrease in mobility in patients.

[0093] In other embodiments, the composition for use according to the present invention can be used for: i) maintaining or improving joint function including cartilage function during aging; ii) reducing joint pain including inflammatory pain and / or nociceptive pain.

[0094] In a further embodiment, the present invention relates to a composition for use according to the present invention for improving mobility in a subject, such as an adult or elderly mammal.

[0095] Thus, in a preferred embodiment, the composition according to the present invention can be used, for example, to improve the activity and / or mobility of an individual by preventing or treating osteoarthritis and / or by inhibiting or reducing cartilage degeneration.

[0096] Another preferred embodiment relates to a composition for use according to the present invention, the use of which is for preventing cartilage degeneration and thus maintaining healthy joints, or for preventing or reducing joint pain (inflammatory pain and / or nociceptive pain) in order to maintain or improve mobility. In a further embodiment, the composition of the present invention can be used to maintain the condition of cartilage.

[0097] Subject group The target group of the composition for use according to the present invention can be any mammal that exhibits cartilage degeneration due to suffering from one or more of the pathological conditions involving cartilage degeneration mentioned herein. Cartilage degeneration can be detected by visual means such as radiography. Alternatively, products due to cartilage degeneration can be detected in body fluids. For example, one or more collagen II epitopes such as (Coll2-1, Coll2-1 NO2, CTX-II) can be detected in a sample such as a plasma sample or a urine sample.

[0098] Another target group can be any mammal that does not yet exhibit cartilage degeneration but is at risk of cartilage degeneration, for example, having a risk of OA, RA, or any of the pathological conditions involving cartilage degeneration mentioned herein. In a preferred embodiment, the present invention relates to a composition according to the present invention comprising a combination of fisetin or its derivative and quercetin or a derivative for use in inhibiting or reducing early degeneration of cartilage, the composition being administered to this target group.

[0099] Certain embodiments of the present invention relate to a composition for use in improving the activity and / or mobility of an individual, for example, by preventing or treating osteoarthritis and / or by inhibiting or reducing cartilage degeneration in the elderly or aged individual.

[0100] In a further embodiment, the composition for use according to the present invention may be for use in mammals such as humans or pets. Examples of pets include cats, dogs, and horses.

[0101] The present invention may be useful in many different age groups, but in a preferred embodiment, the composition for use in improving mobility according to the present invention is targeted at the elderly population, particularly healthy elderly and / or aged mammals.

[0102] Method for manufacturing the nutritional composition of the present invention In a further aspect, the present invention is a method for manufacturing a nutritional composition for use according to the present invention, A step of preparing raw materials for a nutritional composition containing a combination of fisetin and / or its derivatives and quercetin and / or its derivatives, and a step of mixing so that the nutritional composition contains a combination of fisetin and / or its derivatives and quercetin and / or its derivatives.

[0103] Pharmaceutical composition for use In a further embodiment, the present invention relates to a composition for use in inhibiting or preventing cartilage degeneration according to the present invention, which is a pharmaceutical composition.

[0104] By making it a pharmaceutical, unlike a nutritional composition, it means a composition in which a substance is used as a drug on or in the body to prevent, diagnose, relieve, treat or cure diseases in humans or animals. According to the present invention, the present pharmaceutical can be used to suppress or reduce cartilage degeneration.

[0105] The pharmaceutical may be for human use. Alternatively, it may be a veterinary composition suitable for, for example, dogs, cats or horses, particularly thoroughbred horses.

[0106] In a preferred embodiment, the pharmaceutical composition of the present invention contains a combination of fisetin or its derivative and quercetin or its derivative.

[0107] In another preferred embodiment, the pharmaceutical composition of the present invention contains fisetin or its derivative, quercetin or its derivative, and curcumin.

[0108] The present invention further relates to the use of the pharmaceutical according to the present invention as described herein as the use of the composition of the present invention.

[0109] A pharmaceutical composition for use according to the present invention, comprising fisetin or its derivative and quercetin or its derivative and / or curcumin, in combination with at least one additive selected from the group consisting of pharmaceutically acceptable additives. Procedures for the preparation of the pharmaceutical composition according to the present invention can be readily found by those skilled in the art, for example, in the handbook Remington’s Pharmaceutical Sciences, Mid. Publishing Co, Easton, Pa., USA. Also, physiologically acceptable additives, vehicles and adjuvants are described in the handbook entitled "Handbook of Pharmaceutical Excipients, Second edition, American Pharmaceutical Association, 1994". In order to formulate the pharmaceutical composition according to the present invention, it would be advantageous for those skilled in the art to refer to the latest edition of the European Pharmacopoeia or the United States Pharmacopoeia (USP). In particular, it would be advantageous for those skilled in the art to refer to the 4th edition "2002" of the European Pharmacopoeia or the USP 25-NF 20th edition of the United States Pharmacopoeia (U.S.Pharmacopoeia).

[0110] Advantageously, the pharmaceutical composition as defined above is suitable for oral administration, parenteral administration or intravenous administration. When the pharmaceutical composition for use according to the present invention contains at least one pharmaceutically acceptable or physiologically acceptable additive, in particular, the additive is suitable for administration of the composition by the oral route, or the additive is suitable for administration of the composition by the parenteral route.

[0111] The pharmaceutical composition for use according to the present invention is available in solid or liquid form without distinction. For oral administration, solid pharmaceutical compositions in the form of tablets, capsules, or gelatin capsules would be preferred.

[0112] In liquid form, the pharmaceutical composition would preferably be in the form of an aqueous or non-aqueous suspension, or in the form of an oil-in-water or water-in-oil emulsion.

[0113] Solid pharmaceutical forms may include, as vehicle, adjuvant or additive, at least one diluent, one flavoring agent, one solubilizing agent, one lubricant, one suspending agent, one binder, one disintegrating agent and one encapsulating agent. Such compounds are, for example, magnesium carbonate, magnesium stearate, talc, lactose, pectin, dextrin, starch, gelatin, cellulose-based materials, cocoa butter and the like. Compositions in liquid form may include water, optionally as a mixture with propylene glycol or polyethylene glycol, and may optionally also include coloring agents, flavoring agents, stabilizers and thickening agents.

[0114] Combination with known treatments In the absence of such disease-modifying OA drugs (DMOADs), alternative treatments and OA prevention through nutrition are also possible.

[0115] From histological data, it can be understood that oleuropein has a greater impact on the OA score than compounds that mainly affect degeneration. Thus, the effectiveness of oleuropein may be due to its combined effect on inflammation and degeneration. Accordingly, in a preferred embodiment, the compositions of the present invention, which have been shown to inhibit or reduce degeneration, can be combined with a treatment for inhibiting or reducing inflammation.

[0116] Methods of treatment The present invention also relates to a method for preventing or treating cartilage degeneration, such as a condition in which cartilage degeneration occurs or an injury associated with cartilage degeneration, the method comprising administering to an individual in need thereof an effective amount of a composition according to the present invention. For example, the method comprises administering an effective amount of a composition comprising a combination of fisetin or a derivative thereof and quercetin or a derivative thereof.

[0117] As used herein, "effective amount" means an amount that prevents a deficiency, treats an individual's disease or medical condition, or more generally, alleviates symptoms, manages the progression of a disease, or provides nutritional, physiological or medical benefits to an individual.

[0118] The effective amount of the composition of the present invention required to achieve a therapeutic effect will, of course, vary depending on the specific composition, route of administration, age and condition of the user, and the specific disorder or disease to be treated.

[0119] The present invention further provides a method for preventing or treating a condition associated with cartilage degeneration, such as OA or RA, inhibiting or reducing cartilage degeneration, inhibiting or reducing collagen degeneration in cartilage, or inhibiting or reducing type II collagen degeneration in cartilage, the method comprising administering to an individual an effective amount of a composition for use according to the present invention.

[0120] In one embodiment, the treatment method according to the present invention relates to the prevention or treatment of osteoarthritis.

[0121] The treatment method according to the present invention may be in mammals, such as humans or pets, such as dogs, cats and / or horses.

[0122] In certain embodiments, the composition of the present invention administered in the treatment method may be one or more nutritional compositions of the present invention and / or pharmaceutical compositions of the present invention.

[0123] Kit The present disclosure also provides a kit comprising a combination of fisetin and / or its derivatives and a combination of quercetin and / or derivatives in one or more containers. In an embodiment of the kit, the one or more containers include at least one first container that stores fisetin and / or derivatives separately from quercetin and / or derivatives stored in at least one second container, and the kit further includes instructions for mixing fisetin with quercetin to form a unit dosage form.

[0124] In one embodiment of the kit, the combination may be provided together as one or more pre-packaged unit dosage forms. For example, each container may be provided as a separate container containing a dry powder, such that each container contains one pre-packaged unit dosage form.

[0125] In another embodiment, the kit can include a plurality of compositions for mixing together to form one or more of the compositions disclosed herein. For example, the kit can contain two or more dry powders in separate containers that are related to each other, and each of these separate powders contains a part of the final unit dosage form. As a non-limiting example of such an embodiment, the kit can include one or more first containers containing fisetin, and can also include one or more second containers containing quercetin. The contents of one of the first containers can be admixed with one of the second containers to form at least a part of the unit dosage form of the composition.

[0126] The above dosage examples do not require continuous daily administration without interruption. Rather, there may be several short-term interruptions in administration, such as an interruption for 2 to 4 days during the administration period. The ideal duration of administration of the compositions of the present invention can be determined by those skilled in the art.

[0127] The disclosed combination It should be noted that the embodiments and features described in the context of one aspect of the present invention also apply to other aspects of the present invention.

[0128] The compositions for use according to the present invention are described herein in terms of different parameters, such as raw materials, form of the nutritional composition, use, target population, etc. It should be noted that the embodiments and features described in one situation of the parameters of the compositions for use according to the present invention may also be combined with other embodiments and features described in another situation of the parameters, unless explicitly stated otherwise.

[0129] All patent documents and non-patent documents cited in this application are hereby incorporated by reference in their entirety into this specification.

[0130] The present invention will be described in more detail by the following non-limiting examples.

Example

[0131] The following non-limiting examples show experimental data that support the compositions and methods disclosed herein.

[0132] Example 1 To test the effects of fisetin, quercetin, and combinations thereof in living cells, the inventors measured mitochondrial calcium elevation in HeLa cells. HeLa cells were purchased from ATCC. HeLa cells were seeded in 96-well plates at a density of 50,000 cells / well in minimum essential medium (DMEM, Gibco), high glucose, +10% fetal bovine serum. Mitochondrial calcium measurements were performed using HeLa cells (manufactured by Sirion biotech) infected with an adenovirus expressing mitochondrial-targeted calcium sensor mitochondrial-targeted aequorin (Montero et al., 2004). For aequorin reconstitution, 1 μM of wild-type coelenterazine was added to the standard medium (145 mM NaCl, 5 mM KCl, 1 mM MgCl 2 , 1 mM CaCl 2, in 10 mM glucose and 10 mM Hepes, pH 7.4), the cells were incubated at room temperature (22 ± °C) for 2 hours. During the treatment, the compound was added directly to the cell culture or myotube culture 2 hours before the measurement. Luminescence was measured with FLIPR Tetra Aequorin (Molecular Devices). The increase in mitochondrial calcium was obtained by stimulating the cells with 100 μM histamine. The conversion of luminescence data to calcium concentration (Calibration) was performed using the algorithm as described above (Alvarez & Montero, 2002). For quantification, custom module analysis based on Excel (Microsoft) and GhaphPad Prism 7.02 (GrhaphPad) software was used.

[0133] As shown in Figure 2, for the activation of mitochondria via the increase in mitochondrial Ca2+ in HeLa cells, the effect of the combination of phycetin and quercetin is greater than the effect of phycetin or quercetin alone. As shown in Figure 3, phycetin synergistically acts with quercetin to activate mitochondria via the increase in mitochondrial Ca2+ in HeLa cells.

[0134] Example 2 To verify the synergistic effect of the combination of quercetin + phycetin on a cell model of osteoarthritis chondrocytes (chondrocytes), the inventors measured mitochondrial calcium in SW1353 cells treated with the pro-inflammatory cytokine interleukin-1β. SW1353 cells were purchased from ATCC. SW1353 cells were seeded in 96-well plates at a density of 10,000 cells / well in a 100 mm dish. The cells were cultured in high-glucose minimum essential medium (DMEM, Gibco) containing 10% fetal bovine serum and 1% penicillin-streptomycin. SW1353 cells were treated with 10 ng / ml of the pro-inflammatory cytokine interleukin-1β for 48 hours as a mimic of osteoarthritis.

[0135] Measurement of mitochondrial calcium in SW1353 cells was performed using the same procedure as that used for HeLa cells. The synergistic effect of quercetin and fisetin in the osteoarthritis chondrocyte model was quantified as described for Hela cells.

[0136] Results As shown in Figure 4, in the described cell model of osteoarthritis chondrocytes (SW1353 cells treated with IL-1β), quercetin acts synergistically with fisetin to activate mitochondria via an increase in mitochondrial Ca 2+ As shown in Figure 5, in the described cell model of osteoarthritis chondrocytes, several combinations of quercetin + fisetin act synergistically to activate mitochondria via an increase in mitochondrial Ca2+.

Claims

1. A composition comprising an effective amount of a combination of fisetin and / or its derivatives and quercetin and / or its derivatives for use in the prevention or treatment of cartilage degeneration in an individual.

2. The composition for use according to claim 1, wherein the derivative of fisetin is selected from the group consisting of its glucuronidated form, its sulfated form, derivatives, and mixtures thereof, and is preferably geraldol.

3. The composition for use according to claim 1, wherein the quercetin derivative is selected from the group consisting of quercetin 3-O-galactoside, quercetin 3-O-glucoside (isoquercetin), quercetin 3-O-xyloside, quercetin 3-O-rhamnoside (quercitrin), quercetin 3-O-glucuronide, quercetin 7-O-glucoside, quercetin 3-O-diglucoside, quercetin 3,4'-diglucoside, quercetin 3-O-rhamnoside-7O-glucoside, quercetin 3-O-rutinoside (rutin), quercetin 3-O-6''-acetylglucoside, quercetin 3-methyl ether, quercetin 3,3'-dimethyl ether, isorhamnetin, and mixtures thereof.

4. The composition for use according to claim 1, further comprising calcium.

5. The composition for use according to claim 1, further comprising at least one compound selected from the group consisting of antioxidants, anti-inflammatory compounds, glycosaminoglycans, prebiotics, fibers, probiotics, fatty acids, enzymes, minerals, trace elements, and / or vitamins.

6. A composition for use according to claim 1, selected from the group consisting of food compositions, dietary supplements, nutritional compositions, oral nutritional supplements, medical foods, nutraceuticals, beverages, powdered nutritional preparations that are reconstituted with water or milk before ingestion, food additives, foods for special medical purposes (FSMPs), pharmaceuticals, drinks, pet food, and combinations thereof.

7. A composition for use according to claim 1, which is in the form of a solid powder, powder stick, capsule, or solution.

8. The composition for use according to claim 1, wherein the use is i) to maintain or improve joint function, including cartilage function, in the event of aging, and ii) to reduce joint pain, including inflammatory pain and / or nociceptive pain.

9. The composition for use according to claim 1, wherein the use is for improving the activity and / or mobility of the individual.

10. The composition for use according to claim 1, wherein the use is for inhibiting or reducing cartilage degeneration in osteoarthritis.

11. The composition for use according to claim 1, wherein the individual is a middle-aged or elderly person.

12. A method for producing a nutritional composition for use according to any one of claims 1 to 11, comprising the steps of: preparing one or more raw materials for the nutritional composition; fisetin or a derivative thereof; quercetin and / or a derivative thereof; and optionally further calcium; and mixing them.

13. A kit comprising one or more containers containing a combination of fisetin and / or its derivatives and quercetin and / or its derivatives.

14. The kit according to claim 13, wherein the one or more containers include at least one first container for storing the fisetin and / or derivative thereof separately from the quercetin and / or derivative thereof stored in at least one second container, and the kit further includes instructions for mixing the fisetin with the quercetin to form a unit dosage form.

15. The kit according to claim 13, wherein each of the one or more containers contains a unit dosage form of a combination of fisetin and / or a derivative thereof and quercetin and / or a derivative thereof.

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