Bioconversion of oleuropein

Abstract

To provide a composition for the treatment and prevention of a condition selected from the group consisting of (i) loss of bone or cartilage metabolism balance, (ii) loss of bone or cartilage health, (iii) loss of mobility, (iv) synovitis, and (v) combinations thereof.SOLUTION: A composition comprises oleuropein and at least one probiotic or enzyme, wherein the at least one probiotic or enzyme has β-glycosidase activity, esterase activity, or both β-glycosidase activity and esterase activity.SELECTED DRAWING: Figure 2

Description

[Background technology]

[0001]

[0001] The present disclosure relates generally to compositions for maintaining or improving bone and / or cartilage health. The compositions may additionally or alternatively prevent, alleviate, and / or treat bone and / or cartilage disorders. More specifically, the present disclosure relates to the co-administration of oleuropein with probiotics or enzymes that biotransform oleuropein.

[0002]

[0002] Bone mass changes throughout life and is regulated by genetic, mechanical, and hormonal mechanisms. Bone mineral acquisition occurs during childhood and bone mass peaks around age 20. During this period, bone formation exceeds bone resorption. Later in life, especially around menopause or in older populations, bone turnover is higher and there is a decline in bone mass and quality due to excessive bone resorption leading to a gradual loss of bone mass, microarchitecture, structure, and strength.

[0003]

[0003] Restoration of the balance between bone formation and bone resorption is important for maintaining bone. This bone remodeling process is regulated at the cellular level of bone and involves a close interaction between bone-forming cells (osteoblasts) and bone-resorbing cells (osteoclasts). In healthy adults (humans or animals), the coordinated action of osteoblasts and osteoclasts maintains bone mass over time and at the same time ensures the remodeling of bone tissue by resorption and de novo synthesis of bone.

[0004]

[0004] In healthy adults, the formation rates of osteoclasts and osteoblasts achieve a balance between bone formation and bone resorption. However, in osteoporotic individuals, an imbalance in the bone remodeling process occurs, which causes bone loss to proceed more rapidly than the formation rate. This imbalance is present to some degree in most individuals as they age, but is more severe and occurs at a younger age in osteoporotic individuals. Thus, in humans and other mammals, various diseases are associated with abnormal metabolism of bone resorption and bone formation, resulting in an imbalance in metabolism or bone remodeling.

[0005]

[0005] Furthermore, there are many conditions in humans and animals that are characterized by the need to increase bone formation. For example, in fractures, bone growth needs to be stimulated to hasten the complete repair of the bone. Stimulation of bone growth is also necessary in periodontal disease, metastatic disease of the bone, osteolytic disease, and conditions where connective tissue repair is required, such as scarring or regeneration of cartilage defects or trauma. Stimulation of bone growth is also necessary in the case of primary and secondary hyperparathyroidism, as well as osteoporosis associated with diabetes and glucocorticoid-associated osteoporosis.

[0006]

[0006] Current treatments for stimulating bone formation and / or inhibiting bone resorption have had limited success.

[0007] Another joint disease is osteoarthritis, which is the leading cause of disability in the elderly. Osteoarthritis is a degenerative disease of the articular cartilage of the joints and is the most common form of arthritis, affecting 10% of the adult population. The main characteristic of osteoarthritis is the progressive destruction and loss of articular cartilage, accompanied by changes in other joint structures such as synovial proliferation, sclerosis and thickening of the subchondral bone, osteophyte formation at the joint margins, ligament laxity, and muscle atrophy, all of which contribute to the clinical symptoms of osteoarthritis. These symptoms include severe pain, stiffness, decreased joint motion, and disability.

[0008]

[0008] Currently, there is no cure for osteoarthritis, and therapies are only symptomatic, aimed at improving symptoms. For example, pain and inflammation are treated using analgesics (such as acetaminophen) and nonsteroidal anti-inflammatory drugs (NSAIDs). Furthermore, the use of these drugs is often accompanied by side effects, such as gastrointestinal or cardiovascular risks. Summary of the Invention [Problem to be solved by the invention]

[0009] Oleuropein is a polyphenol found in the fruit, root, stem, and more specifically in the leaves of plants belonging to the Oleaceae family, particularly olives (Olea europaea). The inventors have obtained significant positive results in in vitro data generated from the metabolism of chondrocytes, showing that the oleuropein metabolite hydroxytyrosol is more effective than oleuropein. Without being bound by theory, based on the literature, the inventors believe that the chemical structure of hydroxytyrosol suggests that it has a higher bioavailability than oleuropein. Also without being bound by theory, the inventors further believe that while a portion of the oleuropein dose may be absorbed, such as in the upper intestine, the remaining portion of the dose can reach the colon, where it will be mostly absorbed after bioconversion to hydroxytyrosol and / or intermediate metabolites, such as oleuropein aglycone or hydroxytyrosol acetate.

[0010]

[0010] Improved absorption of oleuropein's metabolites in the colon may increase the bioavailability of oleuropein, thereby enhancing its effectiveness. In this regard, co-administration of oleuropein with one or more probiotics having glycosidase and / or esterase activity can increase the presence of the probiotics in the colon, allowing for in situ degradation of oleuropein to optimize the absorption and resulting effects of oleuropein's metabolites. [Means for solving the problem]

[0011]

[0011] Thus, in a general embodiment, the present invention provides a composition for treating and preventing a condition in an individual, the condition being selected from the group consisting of: (i) loss of balance in bone or cartilage metabolism, (ii) loss of bone or cartilage health, (iii) reduced mobility, (iv) synovitis, and (v) combinations thereof, the composition comprising oleuropein and at least one probiotic or enzyme, the at least one probiotic or enzyme being selected from the group consisting of: (i) a probiotic having β-glycosidase activity (e.g., β-glucosidase activity), or a β-glycosidase (e.g., β-glucosidase), (ii) a probiotic having esterase activity, or an esterase, (iii) both β-glycosidase activity and esterase activity (e.g., β-glucosidase activity and esterase activity). In one embodiment, the composition comprises at least one probiotic selected from the group consisting of a probiotic having both β-glycosidase activity (e.g., β-glucosidase activity) and an enzyme having both β-glycosidase activity and esterase activity (e.g., both β-glucosidase activity and esterase activity), (iv) a first probiotic having β-glycosidase activity (e.g., β-glucosidase activity) and a second probiotic having esterase activity, (v) a probiotic having β-glycosidase activity (e.g., β-glucosidase activity) and an esterase, (vi) a probiotic having β-glycosidase (e.g., β-glucosidase) and esterase activity, and (vii) a β-glycosidase (e.g., β-glucosidase) and an esterase, wherein the composition comprises at least one of the probiotics or enzymes in an amount effective to achieve the effect. For example, the composition may comprise at least one probiotic selected from the group consisting of: (i) a probiotic having β-glycosidase activity (e.g., β-glucosidase activity), (ii) a probiotic having esterase activity, (iii) a probiotic having both β-glycosidase activity (e.g., β-glucosidase activity) and esterase activity, and (iv) a first probiotic having β-glycosidase activity (e.g., β-glucosidase activity) and a second probiotic having esterase activity.In a further example, the composition may be oleuropein, for example a composition that is co-administered with an effective amount of oleuropein.

[0012] For example, the present invention can provide a composition for treating and preventing a condition in an individual, the condition being selected from the group consisting of: (i) loss of balance in bone or cartilage metabolism, (ii) loss of bone or cartilage health, (iii) reduced mobility, (iv) synovitis, and (v) combinations thereof, the composition comprising a combination of oleuropein and at least one probiotic or enzyme, the at least one probiotic or enzyme being: (i) a probiotic having β-glycosidase activity, or a β-glycosidase, (ii) a probiotic having esterase activity, or an esterase. (iii) a probiotic having both β-glycosidase activity and esterase activity, or an enzyme having both β-glycosidase activity and esterase activity, (iv) a first probiotic having β-glycosidase activity and a second probiotic having esterase activity, (v) a probiotic having β-glycosidase activity and an esterase, (vi) a probiotic having β-glycosidase and esterase activity, and (vii) a β-glycosidase and an esterase, and the composition comprises the combination in an effective amount to obtain an effect. For example, the composition may comprise at least one probiotic selected from the group consisting of (i) a probiotic having β-glycosidase activity, (ii) a probiotic having esterase activity, (iii) a probiotic having both β-glycosidase activity and esterase activity, and (iv) a first probiotic having β-glycosidase activity and a second probiotic having esterase activity.

[0013] In one embodiment, the composition is for treating or preventing reduced mobility and the individual is an elderly individual having a condition selected from the group consisting of frailty, pre-frailty, sarcopenia, recovering from sarcopenia, osteoporosis, osteoarthritis, malnutrition, at risk of malnutrition, undergoing rehabilitation, scheduled to undergo rehabilitation within the next year, and combinations thereof. For example, the elderly individual may be an aged individual.

[0014]

[0014] In one embodiment, the composition is for treating or preventing loss of balance in bone metabolism in an individual having a condition selected from the group consisting of osteoporosis, Paget's disease, periprosthetic osteolysis, metastatic bone disease, hyperthyroidism, hypercalcemia due to cancer, multiple myeloma, periodontal disease, osteoarthritis, osteopenia, bone loss due to fracture, fracture healing, and combinations thereof.

[0015]

[0015] In one embodiment, the composition is for treating synovitis in an individual in need of treatment for synovitis, or degradation of articular cartilage following synovitis in an individual who has or is recovering from synovitis, wherein the synovitis is associated with a condition selected from the group consisting of lupus, gout, rheumatoid arthritis, osteoarthritis, osteochondritis disease, osteoarthritis, and combinations thereof.

[0016]

[0016] In one embodiment, at least one probiotic in the composition of the present invention forms an oleuropein metabolite selected from the group consisting of oleuropein aglycone, hydroxytyrosol acetate, hydroxytyrosol, elenoic acid, and mixtures thereof. For example, at least one probiotic in the composition of the present invention may form an oleuropein metabolite selected from the group consisting of oleuropein aglycone, elenoic acid, and mixtures thereof.

[0017] In another embodiment, the present invention provides a composition for enhancing the therapeutic and / or prophylactic effect of oleuropein in an individual, the composition comprising oleuropein and at least one probiotic or enzyme, the at least one probiotic or enzyme being selected from the group consisting of: (i) a probiotic having β-glycosidase activity, or a β-glycosidase; (ii) a probiotic having esterase activity, or an esterase; (iii) a probiotic having both β-glycosidase activity and esterase activity, or a β-glycosidase; - an enzyme having both glycosidase activity and esterase activity, (iv) a first probiotic having β-glycosidase activity and a second probiotic having esterase activity, (v) a probiotic having β-glycosidase activity and an esterase, (vi) a probiotic having β-glycosidase and esterase activity, and (vii) a β-glycosidase and an esterase, and the composition comprises the at least one probiotic or enzyme in an amount effective for an individual to obtain a benefit. For example, a composition for enhancing the therapeutic and / or preventive effect of oleuropein in an individual may comprise at least one probiotic selected from the group consisting of (i) a probiotic having β-glycosidase activity, (ii) a probiotic having esterase activity, (iii) a probiotic having both β-glycosidase activity and esterase activity, and (iv) a first probiotic having β-glycosidase activity and a second probiotic having esterase activity. At least one probiotic of the compositions of the present invention may form an oleuropein metabolite selected from the group consisting of oleuropein aglycone, hydroxytyrosol acetate, hydroxytyrosol, elenolic acid, and mixtures thereof. Oleuropein and oleuropein metabolites may provide therapeutic and / or prophylactic effects with a longer duration than would be obtained by administering the oleuropein metabolites themselves.

[0018]

[0018] The dietary benefits of olive polyphenols are enjoyed by healthy individuals. Olives have strong consumer acceptance as a health food. Producing olive polyphenol metabolites in foods and beverages is beneficial for healthy individuals, for example if the metabolites improve the bioavailability and efficacy of olive polyphenols. In a further embodiment, the present invention provides a non-therapeutic use of a composition comprising oleuropein and at least one probiotic or enzyme selected from the group consisting of: (i) a probiotic having β-glycosidase activity, or a β-glycosidase; (ii) a probiotic having esterase activity, or an esterase; (iii) a probiotic having both β-glycosidase and esterase activities, or an enzyme having both β-glycosidase and esterase activities; (iv) a first probiotic having β-glycosidase activity, and a second probiotic having esterase activity; (v) a probiotic having β-glycosidase activity, and an esterase; (vi) a probiotic having β-glycosidase and esterase activity, and (vii) a β-glycosidase and an esterase; the composition comprising the at least one probiotic or enzyme in an amount effective to obtain an effect. For example, the present invention may provide a non-therapeutic use of a composition comprising at least one probiotic selected from the group consisting of: (i) a probiotic having β-glycosidase activity, (ii) a probiotic having esterase activity, (iii) a probiotic having both β-glycosidase activity and esterase activity, and (iv) a first probiotic having β-glycosidase activity and a second probiotic having esterase activity.

[0019]

[0019] In one embodiment, the composition is co-administered with oleuropein, for example, an effective amount of oleuropein.

[0020]

[0020] In one embodiment, the composition includes oleuropein, for example in an effective amount.

[0021] In one embodiment, the probiotic having β-glucosidase activity has β-glucosidase activity.

[0022] In one embodiment, the β-glycosidase enzyme is a β-glucosidase.

[0023]

[0023] In one embodiment, the composition of the present invention is for oral administration. The composition may further comprise an excipient or a pharma- ceutically acceptable carrier.

[0024] In one embodiment, the at least one enzyme in the composition of the present invention comprises a β-glucosidase. For example, the at least one enzyme may be selected from the group consisting of enzymes without esterase activity, enzymes with esterase activity, and mixtures thereof.

[0025] In one embodiment, the at least one probiotic in the composition of the present invention comprises a probiotic having β-glucosidase activity. For example, the at least one probiotic having β-glucosidase activity may be selected from the group consisting of probiotics without esterase activity, probiotics with esterase activity, and mixtures thereof.

[0026] In one embodiment, the composition of the present invention is a food or beverage product.

[0027] In one embodiment, the composition is a dietary supplement.

[0028]

[0028] In one embodiment, the composition of the present invention further comprises a component selected from the group consisting of protein, carbohydrate, fat, and combinations thereof.

[0029]

[0029] In one embodiment, the composition of the present invention further comprises a food additive selected from the group consisting of acidulants, thickeners, pH adjusting buffers or agents, chelating agents, colorants, emulsifiers, flavorings, minerals, penetrants, preservatives, stabilizers, sugars, sweeteners, modifiers, vitamins, minerals, and combinations thereof.

[0030]

[0030] The present disclosure provides a method for treating or preventing movement disorders in elderly people, comprising orally administering to elderly people an effective amount of a composition comprising oleuropein and at least one probiotic or enzyme, the at least one probiotic or enzyme being selected from the group consisting of (i) a probiotic having β-glycosidase activity or β-glycosidase, (ii) a probiotic having esterase activity or esterase, (iii) a probiotic having both β-glycosidase activity and esterase activity or an enzyme having both β-glycosidase activity and esterase activity, (iv) a first probiotic having β-glycosidase activity and a second probiotic having esterase activity, (v) a probiotic having β-glycosidase activity and esterase, (vi) a probiotic having β-glycosidase and esterase activity, and (vii) a β-glycosidase and an esterase. For example, the method may include orally administering to an elderly person an effective amount of a composition comprising oleuropein and at least one probiotic or enzyme, the at least one probiotic or enzyme being selected from the group consisting of: (i) a probiotic having β-glycosidase activity, or a β-glycosidase; (ii) a probiotic having esterase activity, or an esterase; (iii) a probiotic having both β-glycosidase activity and esterase activity; (iv) a first probiotic having β-glycosidase activity and a second probiotic having esterase activity; (v) a probiotic having β-glycosidase activity and an esterase; (vi) a probiotic having β-glycosidase and esterase activity; and (vii) a β-glycosidase and an esterase.For example, the method may include orally administering to an elderly person an effective amount of a composition comprising at least one probiotic selected from the group consisting of: (i) a probiotic having β-glycosidase activity (preferably β-glucosidase activity), (ii) a probiotic having esterase activity, (iii) a probiotic having both β-glycosidase activity and esterase activity, and (iv) a first probiotic having β-glycosidase activity and a second probiotic having esterase activity. For example, the method may comprise orally administering to an elderly person an effective amount of a composition comprising oleuropein and at least one probiotic, the at least one probiotic being selected from the group consisting of: (i) a probiotic having β-glucosidase activity (preferably β-glucosidase activity), (ii) a probiotic having esterase activity, (iii) a probiotic having both β-glucosidase activity and esterase activity, and (iv) a first probiotic having β-glucosidase activity and a second probiotic having esterase activity.

[0031]

[0031] In one embodiment, the elderly is an aged individual.

[0032]

[0032] In one embodiment, the elderly person has a condition selected from the group consisting of frailty, pre-frailty, sarcopenia, recovering from sarcopenia, osteoporosis, osteoarthritis, malnutrition, at risk of malnutrition, undergoing rehabilitation, scheduled to undergo rehabilitation within the next year, and combinations thereof.

[0033] In one embodiment, the composition is administered daily for at least one month.

[0034]

[0034] In one embodiment, the at least one probiotic forms an oleuropein metabolite selected from the group consisting of oleuropein aglycone, hydroxytyrosol acetate, hydroxytyrosol, elenolic acid, and mixtures thereof.

[0035] In another embodiment, the present disclosure provides a method of stimulating bone formation and / or inhibiting bone resorption in an individual having a condition involving an imbalance between bone formation and bone resorption, comprising orally administering to the individual an effective amount of a composition comprising at least one probiotic or enzyme selected from the group consisting of: (i) a probiotic having β-glycosidase activity, or a β-glycosidase, (ii) a probiotic having esterase activity, or an esterase, (iii) a probiotic having both β-glycosidase activity and esterase activity, or an enzyme having both β-glycosidase activity and esterase activity, (iv) a first probiotic having β-glycosidase activity and a second probiotic having esterase activity, (v) a probiotic having β-glycosidase activity and an esterase, (vi) a probiotic having β-glycosidase and esterase activity, and (vii) a β-glycosidase and an esterase. For example, the method may include orally administering to an individual an effective amount of a composition comprising oleuropein and at least one probiotic or enzyme, the at least one probiotic or enzyme being selected from the group consisting of: (i) a probiotic having β-glycosidase activity, or a β-glycosidase; (ii) a probiotic having esterase activity, or an esterase; (iii) a probiotic having both β-glycosidase activity and esterase activity, or an enzyme having both β-glycosidase activity and esterase activity; (iv) a first probiotic having β-glycosidase activity and a second probiotic having esterase activity; (v) a probiotic having β-glycosidase activity and an esterase; (vi) a probiotic having β-glycosidase and esterase activity; and (vii) a β-glycosidase and an esterase.For example, the method may include orally administering to an individual an effective amount of a composition comprising at least one probiotic selected from the group consisting of: (i) a probiotic having β-glycosidase activity (preferably β-glucosidase activity), (ii) a probiotic having esterase activity, (iii) a probiotic having both β-glycosidase activity and esterase activity, and (iv) a first probiotic having β-glycosidase activity and a second probiotic having esterase activity. For example, the method may comprise orally administering to an individual an effective amount of a composition comprising oleuropein and at least one probiotic, the at least one probiotic being selected from the group consisting of: (i) a probiotic having β-glucosidase activity (preferably β-glucosidase activity), (ii) a probiotic having esterase activity, (iii) a probiotic having both β-glucosidase activity and esterase activity, and (iv) a first probiotic having β-glucosidase activity and a second probiotic having esterase activity.

[0036]

[0036] In one embodiment, the condition is selected from the group consisting of osteoporosis, Paget's disease, periprosthetic osteolysis, metastatic bone disease, hyperthyroidism, hypercalcemia due to cancer, multiple myeloma, periodontal disease, osteoarthritis, osteopenia, bone loss due to fracture, fracture healing, and combinations thereof.

[0037]

[0037] In another embodiment, the present disclosure provides a method for treating synovitis in an individual in need of treatment therefor, preventing synovitis in an individual at risk for synovitis, or treating or preventing articular cartilage degradation following synovitis in an individual who has or is recovering from synovitis. The method includes orally administering to an individual an effective amount of a composition comprising at least one probiotic or enzyme selected from the group consisting of: (i) a probiotic having β-glycosidase activity, or a β-glycosidase; (ii) a probiotic having esterase activity, or an esterase; (iii) a probiotic having both β-glycosidase activity and esterase activity, or an enzyme having both β-glycosidase activity and esterase activity; (iv) a first probiotic having β-glycosidase activity and a second probiotic having esterase activity; (v) a probiotic having β-glycosidase activity and an esterase; (vi) a probiotic having β-glycosidase and esterase activity; and (vii) a β-glycosidase and an esterase. For example, the method may include orally administering to an individual an effective amount of a composition comprising oleuropein and at least one probiotic or enzyme, the at least one probiotic or enzyme being selected from the group consisting of: (i) a probiotic having β-glycosidase activity, or a β-glycosidase; (ii) a probiotic having esterase activity, or an esterase; (iii) a probiotic having both β-glycosidase activity and esterase activity, or an enzyme having both β-glycosidase activity and esterase activity; (iv) a first probiotic having β-glycosidase activity and a second probiotic having esterase activity; (v) a probiotic having β-glycosidase activity and an esterase; (vi) a probiotic having β-glycosidase and esterase activity; and (vii) a β-glycosidase and an esterase.In a further example, the method may comprise orally administering to the individual an effective amount of a composition comprising at least one probiotic selected from the group consisting of: (i) a probiotic having β-glycosidase activity (preferably β-glucosidase activity), (ii) a probiotic having esterase activity, (iii) a probiotic having both β-glycosidase activity and esterase activity, and (iv) a first probiotic having β-glycosidase activity and a second probiotic having esterase activity. For example, the method may include orally administering to an individual an effective amount of a composition comprising oleuropein and at least one probiotic, the at least one probiotic selected from the group consisting of: (i) a probiotic having β-glycosidase activity, (ii) a probiotic having esterase activity, (iii) a probiotic having both β-glycosidase activity and esterase activity, and (iv) a first probiotic having β-glycosidase activity and a second probiotic having esterase activity.

[0038] In one embodiment, the synovitis is associated with a condition selected from the group consisting of lupus, gout, rheumatoid arthritis, osteoarthritis, osteochondritis disease, osteoarthritis, and combinations thereof.

[0039] In another embodiment, the present disclosure provides a method for preventing or treating cartilage destruction in an individual, comprising orally administering to the individual an effective amount of a composition comprising at least one probiotic or enzyme selected from the group consisting of: (i) a probiotic having β-glycosidase activity, or a β-glycosidase, (ii) a probiotic having esterase activity, or an esterase, (iii) a probiotic having both β-glycosidase activity and esterase activity, or an enzyme having both β-glycosidase activity and esterase activity, (iv) a first probiotic having β-glycosidase activity and a second probiotic having esterase activity, (v) a probiotic having β-glycosidase activity and an esterase, (vi) a probiotic having β-glycosidase and esterase activity, and (vii) a β-glycosidase and an esterase. For example, the method may include orally administering to an individual an effective amount of a composition comprising oleuropein and at least one probiotic or enzyme, the at least one probiotic or enzyme being selected from the group consisting of: (i) a probiotic having β-glycosidase activity, or a β-glycosidase; (ii) a probiotic having esterase activity, or an esterase; (iii) a probiotic having both β-glycosidase activity and esterase activity, or an enzyme having both β-glycosidase activity and esterase activity; (iv) a first probiotic having β-glycosidase activity and a second probiotic having esterase activity; (v) a probiotic having β-glycosidase activity and an esterase; (vi) a probiotic having β-glycosidase and esterase activity; and (vii) a β-glycosidase and an esterase.The method may comprise orally administering to the individual an effective amount of a composition comprising at least one probiotic selected from the group consisting of: (i) a probiotic having β-glycosidase activity (preferably β-glucosidase activity), (ii) a probiotic having esterase activity, (iii) a probiotic having both β-glycosidase activity and esterase activity, and (iv) a first probiotic having β-glycosidase activity and a second probiotic having esterase activity. For example, the method may comprise orally administering to an individual an effective amount of a composition comprising oleuropein and at least one probiotic, the at least one probiotic being selected from the group consisting of: (i) a probiotic having β-glucosidase activity (preferably β-glucosidase activity), (ii) a probiotic having esterase activity, (iii) a probiotic having both β-glucosidase activity and esterase activity, and (iv) a first probiotic having β-glucosidase activity and a second probiotic having esterase activity.

[0040] In another embodiment, the present disclosure provides a composition for co-administration with oleuropein, the composition comprising at least one probiotic selected from the group consisting of (i) a probiotic having β-glucosidase activity (preferably β-glucosidase activity), (ii) a probiotic having esterase activity, (iii) a probiotic having both β-glucosidase activity and esterase activity, and (iv) a first probiotic having β-glucosidase activity and a second probiotic having esterase activity. The composition comprises at least one probiotic in an amount effective to obtain an effect selected from the group consisting of (i) maintaining or restoring balance in bone or cartilage metabolism, (ii) maintaining or improving bone or cartilage health, (iii) maintaining or improving mobility in the elderly, (iv) treating or preventing synovitis, and (v) a combination thereof.

[0041] In another embodiment, the present disclosure provides a composition comprising a combination of oleuropein and at least one probiotic, the at least one probiotic being selected from the group consisting of (i) a probiotic having β-glucosidase activity (preferably β-glucosidase activity), (ii) a probiotic having esterase activity, (iii) a probiotic having both β-glucosidase activity and esterase activity, and (iv) a first probiotic having β-glucosidase activity and a second probiotic having esterase activity. The composition comprises the combination in an amount effective to obtain an effect selected from the group consisting of (i) maintaining or restoring balance in bone or cartilage metabolism, (ii) maintaining or improving bone or cartilage health, (iii) maintaining or improving mobility in the elderly, (iv) treating or preventing synovitis, and (v) combinations thereof.

[0042]

[0042] In another embodiment, the present disclosure provides a method for making a composition for obtaining an effect selected from the group consisting of (i) maintaining or restoring balance in bone or cartilage metabolism, (ii) maintaining or improving bone or cartilage health, (iii) maintaining or improving mobility in the elderly, (iv) treating or preventing synovitis, and (v) a combination thereof. The method includes adding an effective amount of a combination of oleuropein and at least one probiotic to at least one component selected from the group consisting of protein, carbohydrate, and fat, the at least one probiotic being selected from the group consisting of (i) a probiotic having β-glucosidase activity (preferably β-glucosidase activity), (ii) a probiotic having esterase activity, (iii) a probiotic having both β-glucosidase activity and esterase activity, and (iv) a first probiotic having β-glucosidase activity and a second probiotic having esterase activity.

[0043]

[0043] In another embodiment, the present disclosure provides a method for making a composition for obtaining an effect selected from the group consisting of (i) maintaining or restoring balance in bone or cartilage metabolism, (ii) maintaining or improving bone or cartilage health, (iii) maintaining or improving mobility in the elderly, (iv) treating or preventing synovitis, and (v) a combination thereof, which is co-administered with oleuropein. The method includes adding at least one probiotic selected from the group consisting of (i) a probiotic with β-glucosidase activity (preferably β-glucosidase activity), (ii) a probiotic with esterase activity, (iii) a probiotic with both β-glucosidase activity and esterase activity, and (iv) a first probiotic with β-glucosidase activity and a second probiotic with esterase activity, in an effective amount, to at least one component selected from the group consisting of protein, carbohydrate, and fat.

[0044]

[0044] In one embodiment, the method further includes the step of adding a food additive selected from the group consisting of acidulants, thickeners, pH adjusting buffers or agents, chelating agents, colorants, emulsifiers, excipients, flavorings, minerals, penetrants, pharma- ceutical acceptable carriers, preservatives, stabilizers, sugars, sweeteners, modifiers, vitamins, minerals, and combinations thereof to at least one of the ingredients.

[0045]

[0045] In another embodiment, the present disclosure provides a method for enhancing the therapeutic and / or prophylactic effect of oleuropein in an individual. The method may include orally administering to the individual at least one probiotic selected from the group consisting of (i) a probiotic having β-glucosidase activity (preferably β-glucosidase activity), (ii) a probiotic having esterase activity, (iii) a probiotic having both β-glucosidase activity and esterase activity, and (iv) a first probiotic having β-glucosidase activity and a second probiotic having esterase activity. The at least one probiotic may be administered to the individual in the same composition as oleuropein, such that the at least one probiotic and oleuropein are administered simultaneously. The at least one probiotic may be administered to the individual in a separate composition from oleuropein.

[0046]

[0046] Preferably, at least one probiotic forms an oleuropein metabolite selected from the group consisting of oleuropein aglycone, hydroxytyrosol acetate, hydroxytyrosol, elenolic acid, and mixtures thereof, and the oleuropein and oleuropein metabolites provide a therapeutic and / or prophylactic effect with a longer duration than would be obtained by administering the oleuropein metabolite itself.

[0047]

[0047] An advantage of one or more embodiments provided by the present disclosure is mobility in aging, frail, or pre-frail individuals (e.g., such individuals are recovering from rehabilitation or are at risk of sarcopenia), particularly providing beneficial effects on bones and joints in such individuals.

[0048]

[0048] Another advantage of one or more embodiments provided by the present disclosure is treating or preventing osteopenia (mild loss of bone mass), promoting bone growth in young individuals, and / or treating or preventing diseases associated with an imbalance in the ratio between bone formation and bone resorption.

[0049]

[0049] Yet another advantage of one or more embodiments provided by the present disclosure is that it stimulates bone formation or inhibits bone resorption in subjects suffering from osteoporosis, periprosthetic osteolysis, periodontal disease, osteoarthritis, and / or osteopenia.

[0050]

[0050] Yet another advantage of one or more embodiments provided by the present disclosure is to enhance bone formation and / or cartilage assimilation; prevent or treat cartilage destruction; and / or limit synovitis and subsequent degradation of articular cartilage (osteoarthritis) during aging.

[0051] A further advantage of one or more embodiments provided by the present disclosure is to increase the potential efficacy of oleuropein after oral ingestion by increasing its bioavailability, thereby enhancing its absorption in the colon, thus providing efficacy from at least two absorption sites and at least two circulating compounds, i.e., the parent compound and one or more metabolites (e.g., dual efficacy), which results in a higher protective effect.

[0052]

[0052] Another advantage of one or more embodiments provided by the present disclosure is that after partial absorption of oleuropein in the small intestine, a portion of the oleuropein that reaches the colon undergoes bioconversion; higher absorption of 3,4-dihydroxyphenolethanol-elenolic acid (also known as oleuropein aglycone and 3,4-DHPEA-EA) and / or hydroxytyrosol and / or hydroxytyrosol acetate and / or elenolic acid in the colon is expected.

[0053] Yet another advantage of one or more embodiments provided by the present disclosure is that it may provide a longer duration of therapeutic effect from administration of oleuropein by extending the absorption over time that oleuropein or its metabolites are absorbed at two different levels (small intestine and colon). By co-administering oleuropein with probiotics or enzymes, the more rapid production of metabolites may result in a faster onset of their bioavailability, resulting in a longer overall absorption period.

[0054] Yet another advantage of one or more embodiments provided by the present disclosure is to enhance the microbiota in aging individuals.

[0055]

[0055] A further advantage of one or more embodiments provided by the present disclosure is that it reduces the dosage intake of oleuropein while achieving the same efficacy.

[0056]

[0056] Additional features and advantages are described herein, and will be apparent from the following drawings and detailed description of the invention.

[0057] [Brief description of the drawings]

[0057] [Figure 1] The chemical structure of oleuropein is shown. [Diagram 2] 1 shows biotransformation of oleuropein by microbiota. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0058]

[0059] definition

[0060] Some definitions are provided below. However, definitions may be found in the "Embodiments" section below, and the heading "Definitions" above does not imply that such disclosure in the "Embodiments" section is not a definition.

[0059]

[0061] Probiotics are microorganisms that, when administered in adequate amounts, confer a health benefit to the host.

[0060]

[0062] All percentages stated herein are by total weight of the composition unless otherwise indicated. As used herein, "about," "approximately," and "substantially" are understood to refer to numbers within a numerical range, e.g., within -10% to +10% of the referenced number, preferably within -5% to +5%, more preferably within -1% to +1% of the referenced number, and most preferably within -0.1% to +0.1% of the referenced number. All numerical ranges herein should be understood to include all integers or fractions within the range. Furthermore, these numerical ranges should be interpreted as being given to support claims directed to any number or subset of numbers within the range. For example, a disclosure of 1 to 10 should be interpreted to support ranges such as 1 to 8, 3 to 7, 1 to 9, 3.6 to 4.6, 3.5 to 9.9, etc.

[0061]

[0063] As used in this disclosure and the appended claims, the singular forms "a," "an," and "the" include plural references unless the context indicates otherwise. Thus, for example, reference to "a component" or "the component" includes two or more components.

[0062]

[0064] The terms "comprise", "comprises", and "comprising" are to be interpreted inclusively, not exclusively. Similarly, the terms "include", "including", and "or" are all to be interpreted as inclusive, unless such interpretation is clearly prevented by the context. However, the compositions disclosed herein may not include elements not specifically disclosed herein. Thus, disclosure of an embodiment using the term "comprising" includes disclosure of embodiments "consisting essentially of" and "consisting of" the specified component. A composition "consisting essentially of" means that the composition comprises at least 50% by weight of the referenced component, preferably at least 75% by weight of the referenced component, more preferably at least 85% by weight of the referenced component, and most preferably at least 95% by weight of the referenced component.

[0063]

[0065] The term "and / or" used in the context of "X and / or Y" should be interpreted as "X" or "Y" or "X and Y". As used herein, the terms "example" and "such as", particularly when followed by a listing of terms, are merely exemplary and illustrative and should not be considered exclusive or inclusive. As used herein, "associated with" and "linked with" mean occurring simultaneously, preferably caused by the same underlying condition, and most preferably, one of the specified conditions is caused by the other specified condition.

[0064]

[0066] In the context of the present invention, the term individual is not limited to humans. An individual may be, for example, a human, a livestock animal, or a companion animal.

[0065]

[0067] The terms "food," "food product," and "food composition" refer to a product or composition intended for ingestion by an individual, such as a human, and providing at least one nutrient to such an individual. The compositions of the present disclosure include many embodiments described herein and can comprise, consist of, or consist essentially of the elements disclosed herein, as well as any additional or optional ingredients, components, or elements described herein or otherwise useful in dietary therapy.

[0066]

[0068] "Prevention" includes reducing the risk and / or severity of a condition or disease. The terms "treatment", "treat" and "to alleviate" include both prophylactic or preventive treatments (which prevent and / or delay the onset of the targeted condition or disease) and curative, therapeutic or disease-modifying treatments, including therapeutic measures that cure, delay, attenuate the symptoms and / or halt the progression of a diagnosed condition or disease; as well as treatment of patients who are ill or diagnosed as suffering from a disease or medical condition, as well as patients who are at risk of contracting or suspected of having a disease. The terms do not necessarily imply that a subject is treated until cured. The terms "treatment" and "treating" also refer to maintaining and / or promoting the health of individuals who are not afflicted with a disease but are susceptible to developing an unhealthy condition. The terms "treatment", "treat" and "alleviating" are also intended to include the synergistic or otherwise potentiating action of one or more primary preventative or therapeutic measures. The terms "treatment", "treat" and "alleviating" are further intended to include the dietary management of a disease or condition, or for the prophylaxis or prevention of a disease or condition. Treatment may be patient-related or physician-related.

[0067]

[0069] The term "elderly" in the human context means an age of at least 60 years, preferably 63 years or older, more preferably 65 years or older, and most preferably 70 years or older. The term "older adult" in the human context means an age of at least 45 years, preferably 50 years or older, more preferably 55 years or older, and includes geriatric individuals.

[0068]

[0070] Sarcopenia is defined as the age-related decline in muscle mass and function, including muscle strength and walking speed.

[0069]

[0071] As used herein, "frailty" is defined as a clinically recognizable condition in which increasing vulnerability due to age-related declines in reserve and function across multiple physiological systems impairs the ability to cope with everyday or acute stressors. In the absence of established quantitative criteria, frailty has been operationally defined by Fried et al. as meeting three of five phenotypic criteria indicative of reduced energy: (1) weakness (grip strength in the bottom 20% of the population at baseline, adjusted for sex and body mass index), (2) reduced endurance and energy (self-reported fatigue associated with VO2 max), (3) slowness in walking speed (based on 15-foot walk time, bottom 20% of the population at baseline, adjusted for sex and height), (4) reduced physical activity (weighted score of kilocalories expended per week at baseline, bottom of sex-specific physical activity quintile; e.g., <383 kcal / week for men and <270 kcal / week for women), and / or unintentional weight loss (10 pounds in the past year). Fried LP, Tangen CM, Walston J, et al., "Frailty in older adults: evidence for a phenotype." J. Gerontol. A. Biol. Sci. Med. Sci. 56(3):M146-M156 (2001). The presence of one or two of these criteria identifies the pre-frailty stage as being at high risk for progression to frailty.

[0070]

[0072] As used herein, an "effective amount" is an amount that prevents a deficiency, treats a disease or medical condition in an individual, or more generally, alleviates symptoms, manages the progression of a disease, or provides a nutritional, physiological, or medical benefit to an individual. The relative terms "improved," "increased," "enhanced," and the like refer to the effect of a composition disclosed herein, i.e., a composition comprising oleuropein and a probiotic capable of biotransforming oleuropein, compared to an otherwise identical composition that does not contain the probiotic.

[0071]

[0073] Embodiment

[0074] Figure 1 shows the chemical structure of oleuropein. Oleuropein is a heterosidic ester of 3,4-dihydroxyphenylethanol (also known as hydroxytyrosol, labeled "A" in Figure 1) and elenolic acid (labeled "B" in Figure 1) containing a molecule of glucose (labeled "C" in Figure 1). The mechanism of oleuropein absorption is unclear. However, oleuropein resists degradation in the upper gastrointestinal tract and small intestine, and therefore at least a portion of the administered dose of oleuropein reaches the colon. Microflora in the colon can then carry out biotransformation on this oleuropein.

[0072]

[0075] As shown in Figure 2, deglycosylation converts oleuropein to oleuropein aglycone (also known as 3,4-dihydroxyphenylethanol-elenolic acid or "3,4-DHPEA-EA"), which can then be hydrolyzed (e.g., by an esterase) to form hydroxytyrosol acetate and / or elenolic acid, and further hydrolyzed to form hydroxytyrosol from hydroxytyrosol acetate.

[0073]

[0076] According to the literature, oleuropein aglycone is absorbed 2.2 times less than hydroxytyrosol, but 13.5 times more than oleuropein. Oleuropein aglycone is cleared slower from plasma than hydroxytyrosol and oleuropein. Furthermore, oleuropein aglycone has antioxidant activity equivalent to that of caffeic acid, oleuropein, and hydroxytyrosol.

[0074]

[0077] Hydroxytyrosol absorption is higher than that of oleuropein and oleuropein aglycone, and hydroxytyrosol acetate absorption is even higher than that of hydroxytyrosol.In this regard, hydroxytyrosol acetate is more soluble in the lipid phase than hydroxytyrosol due to the presence of ester groups in hydroxytyrosol acetate.The inventors believe that this improved lipid solubility suggests that hydroxytyrosol acetate is better absorbed across intestinal epithelial cell monolayers than free hydroxytyrosol.

[0075]

[0078] Oleuropein may be absorbed in the upper gastrointestinal tract, etc., or may reach the colon, where most of it will be absorbed after bioconversion to oleuropein aglycone, hydroxytyrosol acetate, hydroxytyrosol, and / or elenolic acid. However, the proportion of oleuropein that is not absorbed in the upper gastrointestinal tract and thus reaches the colon for further metabolism after bioconversion has not been fully elucidated. Therefore, without being bound by theory, the inventors believe that improved absorption of oleuropein or its derivatives in the colon can increase the potential efficacy of oleuropein after oral ingestion, thereby enhancing its bioavailability. In fact, improved absorption of oleuropein or its derivatives in the colon can provide at least two absorption sites and at least two circulating compounds, i.e., the parent compound and one or more metabolites (e.g., dual efficacy).

[0076]

[0079] Specifically, co-administration of oleuropein with one or more probiotics having β-glycosidase activity (preferably β-glucosidase activity) and / or esterase activity can increase the presence of probiotics in the colon, thus allowing the in situ degradation of oleuropein to optimize its absorption and resulting effect.For example, probiotics having β-glycosidase activity, preferably β-glucosidase activity (and preferably no esterase activity) can convert oleuropein to oleuropein aglycone in the colon. Probiotics having β-glycosidase activity (preferably β-glucosidase activity) and esterase activity can convert oleuropein to hydroxytyrosol acetate and / or hydroxytyrosol and / or elenolic acid in the colon. Establishing and / or increasing the presence of such microorganisms in the colon can enhance the effect of oleuropein administration, thereby prolonging the absorption of oleuropein over time (e.g., absorption at two different levels, the small intestine and the colon), e.g., absorption is prolonged compared to hydroxytyrosol itself, which results in a sharper absorption peak.

[0077]

[0080] Thus, one aspect of the present disclosure is a method of maintaining or restoring balance in bone metabolism by stimulating bone formation and / or preventing bone resorption. Another aspect of the present disclosure is a method of treating or preventing diseases related to cartilage turnover by stimulating cartilage anabolism by inhibiting or reducing cartilage destruction. Yet another aspect of the present disclosure is a method for preventing or treating bone diseases (e.g., diseases associated with an imbalance in the ratio of bone formation:bone resorption, such as osteoporosis) or maintaining bone health. Other aspects of the present disclosure include methods of stimulating bone formation and / or cartilage anabolism in adults (e.g., in pre- and post-menopausal women, healthy adults, healthy aging adults, and pre-osteoarthritic adults) to stimulate bone formation and / or cartilage anabolism during the growth period of young individuals and increase peak bone mass, methods of treating or preventing bone loss (osteopenia) that occurs with aging, methods of treating bone deficiency resulting from fractures, and methods of treating or preventing synovitis (e.g., synovitis associated with lupus, gout, or arthritis such as one or more of rheumatoid arthritis, osteoarthritis, osteochondritis disease, and osteoarthritis).

[0078]

[0081] In some embodiments, the compositions are administered to treat or prevent mobility impairment in elderly people, for example, by maintaining or improving joint function (e.g., bone function and / or cartilage function).Elderly people may have a condition selected from the group consisting of frailty, pre-frailty, sarcopenia, recovering from sarcopenia, osteoporosis, osteoarthritis, malnutrition, at risk of malnutrition, undergoing rehabilitation, scheduled to undergo rehabilitation (e.g., within the next year, preferably within 6 months, more preferably within the next month), and combinations thereof.

[0079]

[0082] The method comprises orally administering to an individual, preferably a human, an effective amount of a composition comprising oleuropein and at least one probiotic, the at least one probiotic being selected from the group consisting of: (i) a probiotic having β-glycosidase activity (preferably β-glucosidase activity) with or without esterase activity, (ii) a probiotic having esterase activity with or without β-glycosidase activity, (iii) a probiotic having both β-glycosidase activity and esterase activity, and (iv) a first probiotic having β-glycosidase activity with or without esterase activity, and a second probiotic having esterase activity with or without β-glycosidase activity. Optionally, the composition may contain an additional probiotic without β-glycosidase activity or esterase activity.

[0080]

[0083] The individual may be at risk for a disease or condition, in which case the effective amount of the composition is a prophylactically effective amount, or the individual may have a disease or condition, in which case the effective amount of the composition is a therapeutically effective amount, hi some embodiments, the method includes identifying the individual as having or at risk for the condition prior to administration.

[0081]

[0084] For example, the at least one probiotic may be a strain of the genus Lactobacillus, Bifidobacterium, Lactococcus or Streptococcus, such as Lactobacillus gasseri, Lactobacillus acidophilus, Lactobacillus salivarius, Lactobacillus rhamnosus, Lactobacillus paracasei (e.g., L. paracasei LMG 9192), Lactobacillus casei, Lactobacillus johnsonii, Lactobacillus plantarum (e.g., L. plantarum LMG 9192), Lactobacillus casei, Lactobacillus johnsonii, Lactobacillus plantarum (e.g., L. plantarum LMG 9192), Lactobacillus casei, Lactobacillus johnsonii, Lactobacillus plantarum (e.g., L. plantarum LMG 9192), Lactobacillus gasseri, Lactobacillus acidophilus, Lactobacillus salivarius, Lactobacillus rhamnosus, Lactobacillus paracasei (e.g., L. paracasei LMG 9192), Lactobacillus casei, Lactobacillus johnsonii, Lactobacillus plantarum (e.g., L. plantarum LMG 9192), Lactobacillus casei, ... 6907), Lactobacillus fermentum, Lactobacillus lactis, Lactobacillus delbrueckii, Lactobacillus helveticus, Lactobacillus brevis, Lactococcus lactis, Streptococcus salivarius, Streptococcus thermophilus, Bifidobacterium lactis, Bifidobacterium animalis, Bifidobacterium longum longum, Bifidobacterium breve, Bifidobacterium infantisThe bacteria may belong to the genus Bifidobacterium infantis, Bifidobacterium adolescentis, Bifidobacterium pseudocatenulatum, or a mixture thereof. In further examples, the probiotic may be selected from the group consisting of Lactobacillus gasseri, Lactobacillus acidophilus, Lactobacillus salivarius, Lactobacillus rhamnosus, Lactobacillus johnsonii, Lactobacillus fermentum, Lactobacillus lactis, Lactobacillus delbrueckii, Lactobacillus helveticus, Lactobacillus brevis, Lactococcus lactis, Streptococcus salivarius, Streptococcus thermophilus, Bifidobacterium lactis, Bifidobacterium animalis, Bifidobacterium longum, Bifidobacterium breve, Bifidobacterium infantis, Bifidobacterium alesentis, Bifidobacterium pseudocatenulatum, or mixtures thereof. Non-limiting examples of suitable strains include B. longum ATCC BAA-999, B. longum CNCM I-2618, B. animalis CNCM I-3446, S. thermophilus CNCM I-3915, Lactobacillus paracasei CNCM I-2116, Lactobacillus rhamnosus CGMCC 1.3724, Lactobacillus johnsonii CNCM I-1225, and Lactobacillus lactis CNCM I-4154.

[0082]

[0085] B. longum ATCC BAA-999 is publicly available from ATCC.

[0083]

[0086] B. longum CNCM I-2618, also designated NCC 2705, was deposited on January 29, 2001 at the Collection Nationale de Cultures de Microorganismes (CNCM), Institut Pasteur, 25 rue du Docteur Roux, F-75724 PARIS Cedex 15, France, and was given the deposit number I-2618.

[0084]

[0087] B. animalis CNCM I-3446, also designated NCC 2818, was deposited on June 7, 2005 at the Collection Nationale de Cultures de Microorganismes (CNCM), Institut Pasteur, 25 rue du Docteur Roux, F-75724 PARIS Cedex 15, France, and received the deposit number I-3446.

[0085]

[0088] S. thermophilus CNCM I-3915, also designated NCC 2496, was deposited on February 5, 2008 at the Collection Nationale de Cultures de Microorganismes (CNCM), Institut Pasteur, 25 rue du Docteur Roux, F-75724 PARIS Cedex 15, France, and received the deposit number I-3915.

[0086]

[0089] Lactobacillus paracasei CNCM I-2116, also designated NCC 2461, was deposited on January 12, 1999 at the Collection Nationale de Cultures de Microorganismes (CNCM), Institut Pasteur, 25 rue du Docteur Roux, F-75724 PARIS Cedex 15, France, and received the deposit number I-2116.

[0087]

[0090] Lactobacillus rhamnosus CGMCC 1.3724, also designated NCC 4007, was deposited with the China General Microbiological Culture Collection Centre, Beijing in October 2004 and was given the deposit number 1.3724.

[0088]

[0091] Lactobacillus johnsonii CNCM I-1225, also designated La1, was deposited on June 30, 1992 at the Collection Nationale de Cultures de Microorganismes (CNCM), Institut Pasteur, 25 rue du Docteur Roux, F-75724 PARIS Cedex 15, France, and was given the deposit number I-1225.

[0089]

[0092] Lactobacillus lactis CNCM I-4154, also designated NCC 2287, was deposited on April 24, 2009 at the Collection Nationale de Cultures de Microorganismes (CNCM), Institut Pasteur, 25 rue du Docteur Roux, F-75724 PARIS Cedex 15, France, and received the deposit number I-4154.

[0090]

[0093] The present disclosure is not limited to a particular embodiment of at least one probiotic, and the probiotic can be any non-harmful microorganism with β-glycosidase activity and / or esterase activity. Such microorganisms can be identified by one or more of: identifying probiotic strains using genome in silico analysis of multiple strains, evaluating in vitro oleuropein bioconversion (microbial deglycosylation and / or hydrolysis) by probiotic strains, determining whether the probiotic strains improve the bioavailability of oleuropein (and optionally determining the effect of a carrier matrix), evaluating in vitro efficacy using primary chondrocyte and osteoblast cultures, and / or using clinical trials investigating the effect of oleuropein and probiotic strains on bone and joint health outcomes.

[0091]

[0094] The at least one probiotic forms an oleuropein metabolite selected from the group consisting of oleuropein aglycone, hydroxytyrosol acetate, hydroxytyrosol, elenolic acid, and mixtures thereof. In some embodiments, the at least one probiotic comprises a first and a second probiotic that form different oleuropein metabolites and / or different amounts of one or more oleuropein metabolites.

[0092]

[0095] In one embodiment, the composition is administered to the individual for a period of at least one month, preferably at least two months, more preferably at least three, four, five or six months, and most preferably at least one year. During this period, the composition can be administered to the individual at least one day per week, preferably at least two days per week, more preferably at least three, four, five or six days per week, and most preferably seven days per week. The composition can be administered once a day or in multiple individual doses per day.

[0093]

[0096] In alternative embodiments, oleuropein and probiotics can be administered consecutively in separate compositions.The term "consecutively" means that oleuropein and probiotics are administered sequentially, such that oleuropein is administered at a first time without probiotics, and probiotics is administered at a second time following the first time without oleuropein.The time between consecutive administrations can be, for example, one or a few seconds, minutes, or hours on the same day; one or a few days, or weeks on the same month; or one or a few months on the same year.The order of consecutive administrations can also be reversed.

[0094]

[0097] Further in this regard, "co-administration" of oleuropein and at least one probiotic or enzyme means that at least one probiotic or enzyme is administered to an individual who has consumed oleuropein and / or will consume oleuropein, not necessarily in the same composition at the same time. Although co-administration is actually preferred, the present disclosure is not limited to this embodiment. When oleuropein and probiotic or enzyme are included in the same composition, oleuropein can be separated from probiotic or enzyme to prevent fermentation of oleuropein during storage. For example, probiotic or enzyme can be encapsulated separately from oleuropein. Preferably, probiotic or enzyme only reacts with oleuropein after administration, for example in the digestive tract of an individual.

[0095]

[0098] In some embodiments, the composition is used in one of the methods disclosed in U.S. Patent Application Publication Nos. 2016 / 0045519 and 2016 / 0120891, and WO 2015 / 055468, which are incorporated by reference in their entireties.

[0096]

[0099] The effective amount of the composition will vary depending on the particular composition, the age and condition of the patient, and the particular disease or disorder being treated. However, in a typical embodiment, the composition may be administered to an individual in an amount that provides from 0.01 mg to 2 g of oleuropein per day, preferably from 0.1 mg to 1 g of oleuropein per day, more preferably from 1 mg to 200 g of oleuropein per day, and the composition may be administered in an amount that provides from 10 mg to 200 g of oleuropein per day. 5 ~10 12 Colony forming units (cfu) of probiotics, preferably 10 per day 7 ~10 11 The probiotic may be administered to an individual in an amount that provides cfu of probiotics.

[0097]

[0100] Some individuals may have a microbiome that contains probiotics that already have β-glycosidase and / or esterase activity, and thus, in some cases, administration of the composition enhances the therapeutic and / or prophylactic effects of these probiotics by providing more bacteria.

[0098]

[0101] In one embodiment, at least a portion of the oleuropein is obtained by extraction, for example from one or more of the stems, leaves, fruits, or kernels of a plant belonging to the family Oleaceae, preferably from a plant belonging to the family Oleaceae, such as olive (olive tree), plants of the genus Ligustrum, plants of the genus Syringa, plants of the genus Fraximus, plants of the genus Jasminum, plants of the genus Osmanthus, etc. Additionally or alternatively, at least a portion of the oleuropein can be obtained by chemical synthesis.

[0099]

[0102] Another aspect of the present disclosure is a method of making a composition to obtain an effect selected from the group consisting of: (i) maintaining or restoring balance to bone or cartilage metabolism, (ii) maintaining or improving bone or cartilage health, (iii) maintaining or improving mobility in the elderly, (iv) treating or preventing synovitis, and (v) a combination thereof. The composition and method may additionally or alternatively prevent, alleviate, and / or treat bone and / or cartilage disorders. The composition is preferably a food product.

[0100]

[0103] The method includes adding oleuropein and probiotics capable of biotransforming oleuropein to ingredients selected from the group consisting of proteins, carbohydrates, lipids, and combinations thereof. The composition (e.g., food product) can be made prior to administration (e.g., the composition is made, packaged, and then purchased by a consumer who administers the composition to themselves or another individual) or can be made substantially simultaneously with administration (the composition is made by an individual who administers the composition to themselves or another individual less than 30 minutes prior to administration, preferably less than 15 minutes prior to administration, more preferably less than 10 minutes prior to administration, and most preferably less than 5 minutes prior to administration).

[0101]

[0104] The composition can include an effective amount of a combination of oleuropein and probiotics, for example, a serving or dose of the composition can include an effective amount, and a package can include one or more servings or doses.

[0102]

[0105] The composition may include a food additive selected from the group consisting of an acidulant, a thickener, a buffer or agent for adjusting pH, a chelating agent, a colorant, an emulsifier, an excipient, a flavoring, a mineral, an osmotic agent, a pharma- ceutical acceptable carrier, a preservative, a stabilizer, a sugar, a sweetener, a modifier, a vitamin, a mineral, and combinations thereof.

[0103]

[0106] The composition may include additional components for bone quality, such as protein, vitamin C, vitamin D, vitamin K2, calcium, phosphorus, magnesium, zinc, hesperidin (flavanone), or combinations thereof. The composition may include additional components for joint quality, including at least one component for short-term joint quality, such as glucosamine (e.g., glucosamine sulfate), chondroitin (e.g., chondroitin sulfate), hyaluronic acid (e.g., hyaluronic acid-rich rooster comb extract), or combinations thereof (preferably at least hyaluronic acid), and / or at least one component for long-term joint quality, such as vitamin C, another polyphenol (e.g., curcumin, quercetin, and / or rutin), omega-3 fatty acid, or combinations thereof. Non-limiting examples of other suitable additional components for joint quality include collagen, hydrolyzed collagen, Boswellia serrata, rose hips, and combinations thereof.

[0104]

[0107] The protein may be whey, such as native whey, intact non-hydrolyzed whey, whey protein concentrate, whey protein isolate, acid whey, sweet whey, modified sweet whey (sweet whey with caseinoglycomacropeptide removed), whey protein fraction, or whey protein hydrolysate; casein; vegetable protein, such as soy protein; and combinations thereof. Casein may be provided in free form or in the form of a salt, such as a sodium, calcium, or potassium salt. The protein may include vegetable protein, although in some embodiments the composition is gluten-free.

[0105]

[0108] The protein may be a highly hydrolyzed protein hydrolysate prepared from animal and vegetable proteins treated with acid or enzymes, such as casein hydrolysate, whey hydrolysate, casein / whey hydrolysate, soy hydrolysate, and mixtures thereof. By "highly hydrolyzed" protein hydrolysate, it is meant that the native protein has been hydrolyzed into peptide fragments, with the majority of the peptide fragments having a molecular weight of less than 1,000 Daltons, preferably at least about 75% of the peptide fragments, and most preferably at least about 95% of the peptide fragments having a molecular weight of less than about 1,000 Daltons. Free amino acids and synthetic short chain peptides may be substituted for or added to the protein hydrolysate.

[0106]

[0109] In one embodiment, the protein comprises whey protein micelles as described in US 2009 / 0035437 and its corresponding EP 1839492(A1) and further characterized in C. Schmitt et al., Soft Matter 6:4876-4884 (2010), herein referred to as whey protein microgels (WPM). In particular, "whey protein micelles" are micelles contained in a whey protein micelle concentrate obtained by the process disclosed in US 2009 / 0035437 and its corresponding EP 1839492(A1). In this disclosure, the process for producing the whey protein micelle concentrate includes: a) adjusting the pH of the aqueous whey protein solution to a value between 3.0 and 8.0; and b) subjecting the aqueous solution to a temperature between 80 and 98°C. and c) concentrating the dispersion obtained in step b). The micelles prepared by this process have a very narrow size distribution, with more than 80% of the micelles having a diameter of less than 1 μm, preferably between 100 nm and 900 nm. The whey protein micelles can also be in liquid concentrate or powder form. Importantly, the basic micellar structure of whey protein is maintained whether in liquid concentrate form, powder form, or when reconstituted from the powder, for example with water. The whey protein micelles are physically stable in the dispersion as a powder, and are also physically stable during spray drying or freeze drying.

[0107]

[0110] Non-limiting examples of suitable carbohydrates include starch, sucrose, lactose, glucose, fructose, corn syrup solids, maltodextrin, modified starch, amylose starch, tapioca starch, corn starch, xylitol, sorbitol, or combinations thereof. Non-limiting examples of suitable lipids include vegetable fats (e.g., 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, lecithin, etc.), animal fats (e.g., dairy fat), or combinations thereof. The fat source may also be less refined versions of these fats (e.g., olive oil for polyphenol content).

[0108]

[0111] The composition can be in any oral nutritional form, for example, as a health drink, as a ready-made drink, optionally as a soft drink including juice, milkshake, yogurt drink, smoothie, or soy-based drink; it can be a food bar; or it can be dispersed in any type of food such as baked products, cereal bars, dairy bars, snack foods, soups, breakfast cereals, muesli, candy, tubs, cookies, biscuits, crackers (such as rice crackers), and dairy products.

[0109]

[0112] The compositions may be in the form of, for example, a tablet, capsule, lozenge, or liquid. The compositions may further contain protective hydrocolloids (gums, proteins, modified starches, etc.), binders, film-forming agents, encapsulating agents / materials, wall / shell materials, matrix compounds, coatings, emulsifiers, surfactants, solubilizers (oils, fats, waxes, lecithin, etc.), adsorbents, carriers, fillers, co-compounds, dispersants, wetting agents, processing aids (solvents), flow agents, flavoring agents, bulking agents, jellifying agents, and gel forming agents.

[0110]

[0113] Aspects of the subject matter described herein are set forth in the following numbered sections.

[0111] 1. A method of treating or preventing movement disorders in an elderly person, the method comprising orally administering to the elderly person an effective amount of a composition comprising oleuropein and at least one probiotic or enzyme, the at least one probiotic or enzyme being selected from the group consisting of: (i) a probiotic having β-glycosidase activity, or a β-glycosidase; (ii) a probiotic having esterase activity, or an esterase; (iii) a probiotic having both β-glycosidase activity and esterase activity; (iv) a first probiotic having β-glycosidase activity and a second probiotic having esterase activity; (v) a probiotic having β-glycosidase activity and an esterase; (vi) a probiotic having β-glycosidase and esterase activity; and (vii) a β-glycosidase and an esterase.

[0112] 2. The method of claim 1, wherein the elderly person is an aged individual.

[0113] 3. The method of clause 1, wherein the older adult has a condition selected from the group consisting of frailty, pre-frailty, sarcopenia, recovering from sarcopenia, osteoporosis, osteoarthritis, malnutrition, at risk of malnutrition, undergoing rehabilitation, scheduled to undergo rehabilitation in the next year, and combinations thereof.

[0114] 4. The method of clause 1, wherein the composition is administered once daily for at least one month.

[0115] 5. The method of clause 1, wherein the at least one probiotic comprises a probiotic having β-glucosidase activity.

[0116] 6. The method of clause 1, wherein the at least one probiotic forms an oleuropein metabolite selected from the group consisting of oleuropein aglycone, hydroxytyrosol acetate, hydroxytyrosol, elenolic acid, and mixtures thereof.

[0117] 7. A method of stimulating bone formation and / or inhibiting bone resorption in an individual having a condition involving an imbalance between bone formation and bone resorption, comprising orally administering to the individual an effective amount of a composition comprising oleuropein and at least one probiotic or enzyme, wherein the at least one probiotic or enzyme is selected from the group consisting of: (i) a probiotic having β-glycosidase activity, or a β-glycosidase; (ii) a probiotic having esterase activity, or an esterase; (iii) a probiotic having both β-glycosidase activity and esterase activity; (iv) a first probiotic having β-glycosidase activity and a second probiotic having esterase activity; (v) a probiotic having β-glycosidase activity and an esterase; (vi) a probiotic having β-glycosidase and esterase activity; and (vii) a β-glycosidase and an esterase.

[0118] 8. The method of clause 7, wherein the condition is selected from the group consisting of osteoporosis, Paget's disease, periprosthetic osteolysis, metastatic bone disease, hyperthyroidism, hypercalcemia due to cancer, multiple myeloma, periodontal disease, osteoarthritis, osteopenia, bone loss due to fracture, fracture healing, and combinations thereof.

[0119] 9. The method of clause 7, wherein the composition is administered once a day for at least one month.

[0120] 10. The method of clause 7, wherein the at least one probiotic having β-glycosidase activity is selected from the group consisting of probiotics without esterase activity, probiotics with esterase activity, and mixtures thereof.

[0121] 11. A method of treating synovitis in an individual in need thereof, preventing synovitis in an individual at risk for synovitis, or treating or preventing degradation of articular cartilage following synovitis in an individual having or recovering from synovitis, comprising orally administering to the individual an effective amount of a composition comprising oleuropein and at least one probiotic or enzyme, wherein the at least one probiotic or enzyme is selected from the group consisting of: (i) a probiotic having β-glycosidase activity, or a β-glycosidase; (ii) (iii) a probiotic having both β-glycosidase activity and esterase activity, (iv) a first probiotic having β-glycosidase activity and a second probiotic having esterase activity, (v) a probiotic having β-glycosidase activity and an esterase, (vi) a probiotic having β-glycosidase and esterase activity, and (vii) a β-glycosidase and an esterase.

[0122] 12. The method of clause 11, wherein the synovitis is associated with a condition selected from the group consisting of lupus, gout, rheumatoid arthritis, osteoarthritis, osteochondritis disease, osteoarthritis, and combinations thereof.

[0123] 13. The method of clause 11, wherein the composition is administered once a day for at least one month.

[0124] 14. The method of clause 11, wherein the at least one probiotic comprises a probiotic having β-glucosidase activity.

[0125] 15. A method of preventing or treating cartilage destruction in an individual, the method comprising orally administering to the individual an effective amount of a composition comprising oleuropein and at least one probiotic or enzyme, the at least one probiotic or enzyme being selected from the group consisting of: (i) a probiotic having β-glycosidase activity, or a β-glycosidase; (ii) a probiotic having esterase activity, or an esterase; (iii) a probiotic having both β-glycosidase activity and esterase activity; (iv) a first probiotic having β-glycosidase activity and a second probiotic having esterase activity; (v) a probiotic having β-glycosidase activity and an esterase; (vi) a probiotic having β-glycosidase and esterase activity; and (vii) a β-glycosidase and an esterase.

[0126] 16. The method of clause 15, wherein the individual is an aged individual.

[0127] 17. The method of clause 15, wherein the composition is administered once a day for at least one month.

[0128] 18. The method of clause 15, wherein the at least one probiotic comprises a probiotic having β-glucosidase activity.

[0129] 19. A composition comprising a combination of oleuropein and at least one probiotic or enzyme, the at least one probiotic or enzyme being: (i) a probiotic having β-glycosidase activity or a β-glycosidase; (ii) a probiotic having esterase activity or an esterase; (iii) a probiotic having both β-glycosidase activity and esterase activity; (iv) a first probiotic having β-glycosidase activity and a second probiotic having esterase activity; (v) a β-glycosidase or a probiotic having β-glycosidase activity; (vi) a probiotic with β-glycosidase and esterase activity, and (vii) a probiotic with β-glycosidase and esterase activity, the composition comprising the combination in an amount effective to obtain an effect selected from the group consisting of (i) maintaining or restoring balance in bone or cartilage metabolism, (ii) maintaining or improving bone or cartilage health, (iii) maintaining or improving mobility in the elderly, (iv) treating or preventing synovitis, and (v) combinations thereof.

[0130] 20. The composition of clause 19, wherein the composition is a food product comprising components selected from the group consisting of proteins, carbohydrates, fats, and combinations thereof.

[0131] 21. A method for making a composition for obtaining an effect selected from the group consisting of (i) maintaining or restoring balance in bone or cartilage metabolism, (ii) maintaining or improving bone or cartilage health, (iii) maintaining or improving mobility in the elderly, (iv) treating or preventing synovitis, and (v) a combination thereof, comprising the step of adding an effective amount of a combination of oleuropein and at least one probiotic or enzyme to at least one component selected from the group consisting of protein, carbohydrate, and fat, wherein the at least one probiotic or enzyme (i) has β-glycosidase activity. (ii) a probiotic with esterase activity or an esterase, (iii) a probiotic with both β-glycosidase activity and esterase activity, (iv) a first probiotic with β-glycosidase activity and a second probiotic with esterase activity, (v) a probiotic with β-glycosidase activity and an esterase, (vi) a probiotic with β-glycosidase and esterase activity, and (vii) a β-glycosidase and an esterase.

[0132] 22. The method of clause 21, further comprising adding a food additive to the at least one ingredient selected from the group consisting of acidulants, thickening agents, pH adjusting buffers or agents, chelating agents, colorants, emulsifiers, excipients, flavorings, minerals, penetrants, pharma- ceutical acceptable carriers, preservatives, stabilizers, sugars, sweeteners, modifiers, vitamins, minerals, and combinations thereof.

[0133] 23. A method for enhancing the therapeutic and / or prophylactic effect of oleuropein in an individual, the method comprising orally administering to the individual at least one probiotic or enzyme selected from the group consisting of: (i) a probiotic having β-glycosidase activity, or a β-glycosidase; (ii) a probiotic having esterase activity, or an esterase; (iii) a probiotic having both β-glycosidase activity and esterase activity; (iv) a first probiotic having β-glycosidase activity and a second probiotic having esterase activity; (v) a probiotic having β-glycosidase activity and an esterase; (vi) a probiotic having β-glycosidase and esterase activity; and (vii) a β-glycosidase and an esterase.

[0134] 24. The method of clause 23, wherein the at least one probiotic is administered to the individual in the same composition as oleuropein, such that the at least one probiotic and oleuropein are administered simultaneously.

[0135] 25. The method of claim 23, wherein at least one probiotic is administered to the individual in a composition separate from the oleuropein.

[0136] 26. The method of claim 23, wherein at least one probiotic forms an oleuropein metabolite selected from the group consisting of oleuropein aglycone, hydroxytyrosol acetate, hydroxytyrosol, elenolic acid, and mixtures thereof, and the oleuropein and the oleuropein metabolite provide a therapeutic and / or prophylactic effect with a longer duration than would be obtained by administering the oleuropein metabolite itself.

[0137]

[0114] It should be understood that various changes and modifications to the preferred embodiments of the invention described herein will be apparent to those skilled in the art. Such changes and modifications can be made without departing from the spirit and scope of the subject matter of the invention and without diminishing its intended advantages. Accordingly, such changes and modifications are intended to be encompassed within the scope of the appended claims.

[0138] Further embodiments are as follows. [Embodiment 1] A composition for treating and preventing a condition in an individual, the condition being selected from the group consisting of (i) loss of balance in bone or cartilage metabolism, (ii) loss of bone or cartilage health, (iii) reduced mobility, (iv) synovitis, and (v) combinations thereof, the composition comprising oleuropein and at least one probiotic or enzyme, the at least one probiotic or enzyme being (i) a probiotic having β-glycosidase activity or a β-glycosidase, (ii) a probiotic having esterase activity or an esterase, (iii) a probiotic having β-glycosidase activity and an esterase, (iv) a first probiotic with β-glycosidase activity and a second probiotic with esterase activity, (v) a probiotic with β-glycosidase activity and an esterase, (vi) a probiotic with β-glycosidase and esterase activity, and (vii) a β-glycosidase and an esterase, wherein the composition comprises at least one of the probiotics or enzymes in an amount effective to achieve the effect. [Embodiment 2] The composition of embodiment 1 for treating or preventing reduced mobility, wherein the individual is an elderly person with a condition selected from the group consisting of frailty, pre-frailty, sarcopenia, recovering from sarcopenia, osteoporosis, osteoarthritis, malnutrition, at risk of malnutrition, undergoing rehabilitation, scheduled to undergo rehabilitation in the next year, and combinations thereof. [Embodiment 3] The composition of embodiment 1 for treating or preventing loss of balance in bone metabolism, wherein the individual has a condition selected from the group consisting of osteoporosis, Paget's disease, periprosthetic osteolysis, metastatic bone disease, hyperthyroidism, hypercalcemia due to cancer, multiple myeloma, periodontal disease, osteoarthritis, osteopenia, bone loss due to fracture, fracture healing, and combinations thereof. [Embodiment 4] 2. The composition of embodiment 1 for treating synovitis in an individual in need thereof, or degradation of articular cartilage following synovitis in an individual having or recovering from synovitis, wherein the synovitis is associated with a condition selected from the group consisting of lupus, gout, rheumatoid arthritis, osteoarthritis, osteochondritis disease, osteoarthritis, and combinations thereof. [Embodiment 5] 1. A composition for enhancing the therapeutic and / or prophylactic effect of oleuropein in an individual, the composition comprising at least one probiotic or enzyme selected from the group consisting of: (i) a probiotic having β-glycosidase activity, or a β-glycosidase; (ii) a probiotic having esterase activity, or an esterase; (iii) a probiotic having both β-glycosidase activity and esterase activity, or an enzyme having both β-glycosidase activity and esterase activity; (iv) a first probiotic having β-glycosidase activity, and a second probiotic having esterase activity; (v) a probiotic having β-glycosidase activity, and an esterase; (vi) a probiotic having β-glycosidase and esterase activity; and (vii) a β-glycosidase and an esterase, the composition comprising the at least one probiotic or enzyme in an amount effective for the individual to obtain the benefit. [Embodiment 6] The composition of any one of embodiments 1 to 5, which is co-administered with oleuropein. [Embodiment 7] 6. The composition of any one of embodiments 1-5, further comprising an effective amount of oleuropein. [Embodiment 8] The composition of any one of the preceding embodiments, wherein the at least one probiotic forms an oleuropein metabolite selected from the group consisting of oleuropein aglycone, hydroxytyrosol acetate, hydroxytyrosol, elenolic acid, and mixtures thereof. [Embodiment 9] The composition according to any one of embodiments 1 to 8, wherein the composition is for oral administration. [Embodiment 10] The composition of any one of embodiments 1-9, wherein the composition is administered once a day for at least one month. [Embodiment 11] 11. The composition of any one of the preceding claims, wherein the at least one probiotic comprises a probiotic with β-glucosidase activity selected from the group consisting of probiotics without esterase activity, probiotics with esterase activity, and mixtures thereof. [Embodiment 12] 12. The composition of any one of the preceding embodiments, wherein the composition is a food product or a beverage product. [Embodiment 13] 13. The composition of embodiment 12, wherein the composition further comprises a component selected from the group consisting of protein, carbohydrate, fat, and combinations thereof. [Embodiment 14] 14. The composition of embodiment 12 or 13, further comprising a food additive selected from the group consisting of acidulants, thickeners, pH adjusting buffers or agents, chelating agents, colorants, emulsifiers, flavorings, minerals, osmotic agents, preservatives, stabilizers, sugars, sweeteners, modifiers, vitamins, minerals, and combinations thereof. [Embodiment 15] 1. A method for treating or preventing movement disorders in an elderly person, the method comprising orally administering to the elderly person an effective amount of a composition comprising oleuropein and at least one probiotic or enzyme, wherein the at least one probiotic or enzyme is selected from the group consisting of: (i) a probiotic having β-glycosidase activity, or a β-glycosidase; (ii) a probiotic having esterase activity, or an esterase; (iii) a probiotic having both β-glycosidase activity and esterase activity; (iv) a first probiotic having β-glycosidase activity and a second probiotic having esterase activity; (v) a probiotic having β-glycosidase activity and an esterase; (vi) a probiotic having β-glycosidase and esterase activity; and (vii) a β-glycosidase and an esterase. [Embodiment 16] The method of embodiment 15, wherein the elderly person has a condition selected from the group consisting of frailty, pre-frailty, sarcopenia, recovering from sarcopenia, osteoporosis, osteoarthritis, malnutrition, at risk of malnutrition, undergoing rehabilitation, scheduled to undergo rehabilitation within the next year, and combinations thereof. [Embodiment 17] 1. A method of stimulating bone formation and / or inhibiting bone resorption in an individual having a condition involving an imbalance between bone formation and bone resorption, the method comprising orally administering to the individual an effective amount of a composition comprising oleuropein and at least one probiotic or enzyme, the at least one probiotic or enzyme being selected from the group consisting of: (i) a probiotic having β-glycosidase activity, or a β-glycosidase; (ii) a probiotic having esterase activity, or an esterase; (iii) a probiotic having both β-glycosidase activity and esterase activity; (iv) a first probiotic having β-glycosidase activity and a second probiotic having esterase activity; (v) a probiotic having β-glycosidase activity and an esterase; (vi) a probiotic having β-glycosidase and esterase activity; and (vii) a β-glycosidase and an esterase. [Embodiment 18] 18. The method of embodiment 17, wherein the condition is selected from the group consisting of osteoporosis, Paget's disease, periprosthetic osteolysis, metastatic bone disease, hyperthyroidism, hypercalcemia due to cancer, multiple myeloma, periodontal disease, osteoarthritis, osteopenia, bone loss due to fracture, fracture healing, and combinations thereof. [Embodiment 19] 1. A method of treating synovitis in an individual in need thereof, preventing synovitis in an individual at risk for synovitis, or treating or preventing articular cartilage degradation following synovitis in an individual having or recovering from synovitis, the method comprising orally administering to the individual an effective amount of a composition comprising oleuropein and at least one probiotic or enzyme, the at least one probiotic or enzyme being selected from the group consisting of: (i) a probiotic having β-glycosidase activity, or a β-glycosidase; (ii) (iii) a probiotic having both β-glycosidase activity and esterase activity, (iv) a first probiotic having β-glycosidase activity and a second probiotic having esterase activity, (v) a probiotic having β-glycosidase activity and an esterase, (vi) a probiotic having β-glycosidase and esterase activity, and (vii) a β-glycosidase and an esterase. [Embodiment 20] 20. The method of embodiment 19, wherein the synovitis is associated with a condition selected from the group consisting of lupus, gout, rheumatoid arthritis, osteoarthritis, osteochondritis disease, osteoarthritis, and combinations thereof. [Embodiment 21] A method for preventing or treating cartilage destruction in an individual, the method comprising orally administering to the individual an effective amount of a composition comprising oleuropein and at least one probiotic or enzyme, wherein the at least one probiotic or enzyme is selected from the group consisting of: (i) a probiotic having β-glycosidase activity, or a β-glycosidase; (ii) a probiotic having esterase activity, or an esterase; (iii) a probiotic having both β-glycosidase activity and esterase activity; (iv) a first probiotic having β-glycosidase activity and a second probiotic having esterase activity; (v) a probiotic having β-glycosidase activity and an esterase; (vi) a probiotic having β-glycosidase and esterase activity; and (vii) a β-glycosidase and an esterase. [Embodiment 22] The method of any one of embodiments 17-21, wherein the individual is an elderly individual. [Embodiment 23] 23. The method of any one of embodiments 15 to 22, wherein the at least one probiotic comprises a probiotic having β-glucosidase activity. [Embodiment 24] The method according to any one of embodiments 15 to 23, wherein the at least one probiotic having β-glycosidase activity is selected from the group consisting of probiotics without esterase activity, probiotics with esterase activity, and mixtures thereof. [Embodiment 25] The method of any one of embodiments 15 to 24, wherein the at least one probiotic forms an oleuropein metabolite selected from the group consisting of oleuropein aglycone, hydroxytyrosol acetate, hydroxytyrosol, elenolic acid, and mixtures thereof. [Embodiment 26] A composition comprising a combination of oleuropein and at least one probiotic or enzyme, the at least one probiotic or enzyme being selected from the group consisting of (i) a probiotic having β-glycosidase activity or a β-glycosidase, (ii) a probiotic having esterase activity or an esterase, (iii) a probiotic having both β-glycosidase activity and esterase activity, (iv) a first probiotic having β-glycosidase activity and a second probiotic having esterase activity, (v) a β-glycosidase activity and a second probiotic having esterase activity, (vi) a β-glycosidase activity and a second probiotic having esterase activity, (vii) a β-glycosidase activity and a second probiotic having esterase activity, (viii) a probiotic having β-glycosidase activity and a second probiotic having esterase activity, (viiii) a probiotic having both β-glycosidase activity and an ... (vi) a probiotic with β-glycosidase and esterase activity, and (vii) a β-glycosidase and esterase, wherein the composition comprises the combination in an amount effective to obtain an effect selected from the group consisting of (i) maintaining or restoring balance in bone or cartilage metabolism, (ii) maintaining or improving bone or cartilage health, (iii) maintaining or improving mobility in the elderly, (iv) treating or preventing synovitis, and (v) combinations thereof. [Embodiment 27] 27. The composition of embodiment 26, wherein the composition is a food product comprising components selected from the group consisting of proteins, carbohydrates, fats, and combinations thereof. [Embodiment 28] 1. A method for making a composition for obtaining an effect selected from the group consisting of (i) maintaining or restoring balance to bone or cartilage metabolism, (ii) maintaining or improving bone or cartilage health, (iii) maintaining or improving mobility in the elderly, (iv) treating or preventing synovitis, and (v) a combination thereof, the method comprising adding an effective amount of a combination of oleuropein and at least one probiotic or enzyme to at least one component selected from the group consisting of protein, carbohydrate, and fat, the at least one probiotic or enzyme having (i) β-glycosidase activity. (ii) a probiotic with esterase activity or an esterase, (iii) a probiotic with both β-glycosidase activity and esterase activity, (iv) a first probiotic with β-glycosidase activity and a second probiotic with esterase activity, (v) a probiotic with β-glycosidase activity and an esterase, (vi) a probiotic with β-glycosidase and esterase activity, and (vii) a β-glycosidase and an esterase. [Embodiment 29] 29. The method of embodiment 28, further comprising adding a food additive selected from the group consisting of acidulants, thickening agents, pH adjusting buffers or agents, chelating agents, colorants, emulsifiers, excipients, flavorings, minerals, osmotic agents, pharma- ceutically acceptable carriers, preservatives, stabilizers, sugars, sweeteners, texturizing agents, vitamins, minerals, and combinations thereof to the at least one ingredient. [Embodiment 30] 1. A method for enhancing the therapeutic and / or prophylactic effect of oleuropein in an individual, the method comprising orally administering to the individual at least one probiotic or enzyme selected from the group consisting of: (i) a probiotic having β-glycosidase activity, or a β-glycosidase; (ii) a probiotic having esterase activity, or an esterase; (iii) a probiotic having both β-glycosidase activity and esterase activity; (iv) a first probiotic having β-glycosidase activity and a second probiotic having esterase activity; (v) a probiotic having β-glycosidase activity and an esterase; (vi) a probiotic having β-glycosidase and esterase activity; and (vii) a β-glycosidase and an esterase. [Embodiment 31] The method of embodiment 30, wherein the at least one probiotic is administered to the individual in the same composition as the oleuropein, such that the at least one probiotic and the oleuropein are administered simultaneously. [Embodiment 32] The method of embodiment 30, wherein the at least one probiotic is administered to the individual in a composition separate from the oleuropein. [Embodiment 33] The method of embodiment 30, wherein the at least one probiotic forms an oleuropein metabolite selected from the group consisting of oleuropein aglycone, hydroxytyrosol acetate, hydroxytyrosol, elenolic acid, and mixtures thereof, and the oleuropein and the oleuropein metabolites provide the therapeutic effect and / or the prophylactic effect for a longer duration than would be obtained by administering the oleuropein metabolite itself.

Claims

[Claim 1] A composition for treating and preventing a condition in an individual, the condition being selected from the group consisting of (i) loss of balance in bone or cartilage metabolism, (ii) loss of bone or cartilage health, (iii) reduced mobility, (iv) synovitis, and (v) combinations thereof, the composition comprising oleuropein and at least one probiotic or enzyme, the at least one probiotic or enzyme being (i) a probiotic having β-glycosidase activity or a β-glycosidase, (ii) a probiotic having esterase activity or an esterase, (iii) a probiotic having β-glycosidase activity and an esterase, (iv) a first probiotic having β-glycosidase activity and a second probiotic having esterase activity; (v) a probiotic having β-glycosidase activity and an esterase; (vi) a probiotic having β-glycosidase and esterase activity; and (vii) a β-glycosidase and an esterase, wherein the composition comprises at least one of the probiotics or enzymes in an amount effective to achieve the effect.

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