Methods and compositions for treating skeletal joints and uses thereof

Abstract

Disclosed is a joint treatment composition that, when administered to a mammal in a therapeutically effective amount, can reduce inflammation and even heal damaged or diseased skeletal joints.For example, the joint treatment composition is well suited for treating osteoarthritis and other skeletal diseases or injuries.The disclosed composition can dramatically reduce various biomarkers associated with skeletal health, including, for example, reducing tumor necrosis factor serum and protein in the body.

Description

Related Applications

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims priority to the benefit of U.S. Provisional Application No. 63 / 278,860, filed November 12, 2021, which is expressly incorporated by reference in its entirety herein. [Background technology]

[0002] Cartilage matrix is ​​composed of collagen, proteoglycans, and non-collagenous proteins and serves as a cushion and shock absorber within skeletal joints as it covers the ends of two bones that form a joint. Cartilage damage can be caused by several conditions, including joint injury, avascular necrosis, the effects of aging, osteoarthritis, autoimmune disorders, and rheumatoid arthritis. Damaged cartilage can cause pain and limit joint movement.

[0003] Inflammation is the response of tissues to injury and is characterized by increased blood flow to the tissue resulting in increased body temperature, redness, swelling, and pain. Inflammation can be classified as either acute or chronic. Acute inflammation is the body's initial response to a harmful stimulus and is accompanied by increased migration of plasma and white blood cells (particularly granulocytes) from the blood to the injured tissue. A series of biochemical events propagates and matures the inflammatory response involving various cells within the local vasculature, immune system, and injured tissue. Prolonged inflammation, known as chronic inflammation, results in progressive migration of cell types present at the site of inflammation and is characterized by the concomitant destruction and healing of tissue from the inflammatory process.

[0004] One joint health factor indicative of poor joint health due to injury or osteoarthritis is the level of tumor necrosis factor alpha (TNFα). TNFα is produced by monocytes, macrophages, and T lymphocytes. TNFα exists as both a soluble form, solTNF, which is thought to play an important role in inflammation, and a transmembrane form, tmTNF, which is involved in immune function.

[0005] TNFα exerts its primary effects on monocytes, synovial macrophages, fibroblasts, chondrocytes, and endothelial cells, stimulating proinflammatory cytokine and chemokine synthesis, which activates granulocytes and increases MHC class II expression, which promotes the secretion of matrix metalloproteinases (MMPs), leading to cartilage matrix degradation indicative of inflammation.

[0006] It is known to initiate the inflammatory cascade and is increased in the vicinity of inflamed or damaged tissue, and therefore TNFα inhibition is a target for pain and / or inflammation therapy and / or tissue destruction.

[0007] There are various methods for treating skeletal joints suffering from damage or disease, such as osteoarthritis. Most of these treatment methods are directed to controlling pain and inflammation. What is needed is a composition that can be administered to a mammal to not only treat the pain associated with poor joint health, but also repair damaged tissue to restore mobility. Summary of the Invention

[0008] The present disclosure is generally directed to a method for improving one or more of joint pain, joint health, joint mobility, or inflammation in a mammal. The method includes administering a nutritional supplement to the mammal, the supplement includes a type II collagen composition. In one aspect, the type II collagen composition includes non-denatured collagen, which has been found to be particularly effective. The type II collagen is administered to the mammal in an amount sufficient to combat the effects of skeletal joint disease or inflammation, such as osteoarthritis or other joint necrosis. It has been unexpectedly discovered that increasing the dose of type II collagen leads to greater joint health effectiveness.

[0009] In one embodiment, for example, the present disclosure is directed to a method of treating skeletal joint injury or disease in a mammal. The method includes providing a therapeutically effective amount of a joint healing composition to the mammal. The joint healing composition includes collagen and can be administered to the mammal in the form of a dose. More specifically, the joint healing composition is administered to the mammal in an amount greater than about 0.66 mg / kg body weight per day (which translates to a human equivalent dose of 40 mg).

[0010] In the past, it was widely believed that daily non-denatured (or natural) collagen doses (e.g., 40 mg collagen for some mammals) exceeding about 0.55 mg / kg body weight per day, for example, exceeding about 0.6 mg / kg body weight per day, would not provide additional beneficial effects to the user or mammal. However, it has been surprisingly and unexpectedly discovered that administering collagen, particularly non-denatured collagen, exceeding about 0.66 mg / kg body weight per day can have a dramatic beneficial effect on joint damage or disease. In this regard, the method of the present disclosure, in one aspect, can administer to a mammal a higher amount of collagen sufficient to reduce the severity of skeletal joint damage or disease, as shown through a change in at least one joint health marker, compared to administering a joint treatment composition of less than 0.3 mg / kg body weight per day, for example, compared to administering a joint treatment composition of less than 0.5 mg / kg body weight per day, for example, compared to administering a joint treatment composition of less than 0.6 mg / kg body weight per day.

[0011] For example, the joint treatment composition can be administered to a mammal such that the mammal is administered collagen, particularly undenatured type II collagen, in an amount of more than about 1 mg / kg body weight per day, for example more than about 2 mg / kg body weight per day. Collagen can be administered to a mammal, for example, in an amount of about 1.2 mg / kg body weight per day to about 3 mg / kg body weight per day.

[0012] The amount of collagen administered to mammals may depend on various factors, including type and amount of inflammation and type of mammals.In one embodiment, mammals, which may be human, are administered collagen in an amount of more than about 50mg per day, for example more than about 80mg per day, for example more than about 100mg per day, for example more than about 120mg per day, for example more than about 160mg per day, for example more than about 320mg per day, for example more than about 400mg per day, for example more than about 480mg per day, and generally less than about 3,000mg per day, for example less than about 1,000mg per day.

[0013] The joint treatment composition may be administered to a mammal in an amount and for a period of time sufficient to treat inflammation of skeletal joints and / or treat joint damage or disease, such as osteoarthritis. The effect of the composition of the present disclosure may be seen in improved mobility and / or by monitoring various biomarkers related to inflammation, joint metabolism, and / or joint health. For example, the joint treatment composition of the present disclosure may be administered to a mammal for a period of time sufficient to reduce serum tumor necrosis factor alpha by more than about 5%, such as more than about 15%, such as more than about 25%. Serum C-reactive protein may be reduced by more than about 10%, such as more than about 20%, such as more than about 30%, such as more than about 40%. Prostaglandin E2 may be reduced by more than about 8%, such as more than about 15%, such as more than about 25%, such as more than about 30%. Interleukin-1 serum levels may be reduced by more than about 10%, such as more than about 20%, such as more than about 30%, such as more than about 40%. Interleukin-6 serum may be reduced by more than about 10%, such as more than about 20%, such as more than about 30%, such as more than about 40%. Interleukin-1-B protein may be reduced by more than about 8%, such as more than about 10%, such as more than about 15%, such as more than about 20%. Interleukin-6 protein may be reduced by more than about 10%, such as more than about 15%, such as more than about 20%, such as more than about 30%, such as more than about 40%. Interleukin-10 protein may be reduced by more than about 10%, such as more than about 20%, such as more than about 30%, such as more than about 40%. Tumor necrosis factor alpha protein may be reduced by more than about 10%, such as more than about 20%, such as more than about 25%, such as more than about 35%. Cyclooxygenase 2 protein may be reduced by more than about 7%, such as more than about 10%, such as more than about 20%, such as more than about 30%. Nuclear factor kappa B protein may be decreased by more than about 10%, for example, more than about 20%, for example, more than about 30%, for example, more than about 35%. Matrix metallopeptidase 3 may be decreased by more than about 15%, for example, more than about 25%, for example, more than about 35%. Cartilage oligomeric matrix serum may be decreased by more than about 10%, for example, more than about 20%, for example, more than about 30%. Cartilage oligomeric matrix protein may be decreased by more than about 10%, for example, more than about 15%, for example, more than about 25%, for example, more than about 30%.On the other hand, transforming growth factor B may be increased by more than about 5%, such as more than about 10%, such as more than about 15%, such as more than about 20%.

[0014] The joint treatment composition of the present disclosure may be administered to a mammal for a period of time sufficient to change the mammal's Kellgren-Lawrence score by at least about 20%. The joint treatment composition of the present disclosure may also be administered to a mammal for a period of time sufficient to change the mammal's Mankin score by at least about 15%.

[0015] The decrease and increase in the biomarkers may be compared to the initial levels in the mammal being treated. The effect on the biomarkers may occur over a period of time after administration of the joint treatment composition. The period may be, for example, about 10 days to about 60 days, such as about 20 days to about 40 days. In one embodiment, the effect is realized after a period of 28 days. The joint health composition may be administered in any suitable form. In one aspect, the joint health composition is a dietary supplement intended to be ingested. Such forms include solids, liquids, and gels, including capsules, tablets, and powders. The joint health composition may also be in lipid multiparticulate form (LMP), whether spray-dried, beadlet, or the like.

[0016] Other features and aspects of the disclosure are discussed in more detail below. [Brief description of the drawings]

[0017] A full and enabling disclosure of the present invention, including the best mode thereof, directed to one of ordinary skill in the art, is set forth more particularly in the remainder of the specification, which makes reference to the accompanying drawings, in which:

[0018] [Figure 1] 1 is a graphical representation of some of the results obtained in the examples below. [Diagram 2] 1 is a graphical representation of some of the results obtained in the examples below. [Diagram 3] 1 is a graphical representation of some of the results obtained in the examples below. [Figure 4] 1 is a graphical representation of some of the results obtained in the examples below. [Diagram 5] 1 is a graphical representation of some of the results obtained in the examples below. [Figure 6] 1 is a graphical representation of some of the results obtained in the examples below. [Figure 7] 1 is a graphical representation of some of the results obtained in the examples below. [Figure 8] 1 is a graphical representation of some of the results obtained in the examples below. [Figure 9] 1 is a graphical representation of some of the results obtained in the examples below. [Figure 10] 1 is a graphical representation of some of the results obtained in the examples below. [Figure 11] 1 is a graphical representation of some of the results obtained in the examples below. [Figure 12] 1 is a graphical representation of some of the results obtained in the examples below. [Figure 13] 1 is a graphical representation of some of the results obtained in the examples below. [Figure 14] 1 is a graphical representation of some of the results obtained in the examples below. [Figure 15] 1 is a graphical representation of some of the results obtained in the examples below. [Figure 16] 1 is a graphical representation of some of the results obtained in the examples below. [Figure 17] 1 is a graphical representation of some of the results obtained in the examples below. [Figure 18] 1 is a graphical representation of some of the results obtained in the examples below. [Figure 19] 1 is a graphical representation of some of the results obtained in the examples below. [Figure 20] 1 is a graphical representation of some of the results obtained in the examples below. [Figure 21] 1 is a graphical representation of some of the results obtained in the examples below. [Figure 22] 1 is a graphical representation of some of the results obtained in the examples below. [Diagram 23] 1 is a graphical representation of some of the results obtained in the examples below. [Figure 24] 1 is a graphical representation of some of the results obtained in the examples below. [Diagram 25] 1 is a graphical representation of some of the results obtained in the examples below. [Figure 26] 1 is a graphical representation of some of the results obtained in the examples below. [Figure 27] 1 is a graphical representation of some of the results obtained in the examples below. [Figure 28] 1 is a graphical representation of some of the results obtained in the examples below. [Figure 29] 1 is a graphical representation of some of the results obtained in the examples below. [Diagram 30] 1 is a graphical representation of some of the results obtained in the examples below. [Diagram 31] 1 is a graphical representation of some of the results obtained in the examples below. [Diagram 32] 1 is a graphical representation of some of the results obtained in the examples below. [Diagram 33] 1 is a graphical representation of some of the results obtained in the examples below. [Diagram 34] 1 is a graphical representation of some of the results obtained in the examples below. [Diagram 35] 1 is a graphical representation of some of the results obtained in the examples below. [Diagram 36] 1 is a graphical representation of some of the results obtained in the examples below. [Figure 37] 1 is a graphical representation of some of the results obtained in the examples below. [Figure 38] 1 is a graphical representation of some of the results obtained in the examples below. [Figure 39] 1 is a graphical representation of some of the results obtained in the examples below. [Diagram 40] 1 is a graphical representation of some of the results obtained in the examples below. [Diagram 41] 1 is a graphical representation of some of the results obtained in the examples below. [Diagram 42]1 is a graphical representation of some of the results obtained in the examples below. [Diagram 43] 1 is a graphical representation of some of the results obtained in the examples below. [Diagram 44] 1 is a graphical representation of some of the results obtained in the examples below.

[0019] Repeat use of reference characters in the present specification and drawings is intended to represent same or analogous features or elements of the invention.

[0020] [Definition] As used herein, the terms "about", "approximately", or "generally", when used to modify a value, indicate that the value may vary up or down by 10%, in one embodiment, such as 8%, for example 5%, such as 4%, for example 3%, for example 2%, for example 1% in one embodiment, and still remain within the disclosed embodiments.

[0021] As used herein, the term "therapeutically effective amount" shall mean a dosage or amount of a composition that results in a particular pharmacological or nutritional response to which the composition is administered or delivered to a mammal in need of such treatment. It is emphasized that in certain cases, a "therapeutically effective amount" administered to a particular subject is not necessarily effective to treat a disease or otherwise improve health as described herein, but such a dosage is considered a "therapeutically effective amount" by those skilled in the art. Certain subjects may in fact be "refractory" to a "therapeutically effective amount". For example, refractory subjects may have low bioavailability or genetic variation in certain receptors, metabolic pathways, or response capabilities such that clinical efficacy is not obtained. It is further understood that in certain cases, the composition or supplement may be measured as an oral dosage or in terms of component levels that may be measured in the blood. In other embodiments, the dosage may be measured in the amount applied to the skin when the composition is included with a topical formulation. In one aspect, a therapeutically effective amount refers to providing or ingesting an amount of collagen to maintain, repair, or improve joint health.

[0022] As used herein, "active" or "active ingredient" refers to a drug, medicament, pharmaceutical, therapeutic, dietary supplement, or other compound that may be desired to be administered to the body. An active ingredient may be a "small molecule" generally having a molecular weight of 2000 Daltons or less. An active ingredient may be a "bioactive." Bioactive ingredients include proteins, antibodies, antibody fragments, peptides, oligonucleotides, vaccines, and various derivatives of such materials. In one embodiment, the active ingredient is a small molecule. In another embodiment, the active ingredient is a bioactive. In yet another embodiment, the active ingredient is a mixture of a small molecule and a bioactive. Also, as used herein, the terms "active ingredient," "first active ingredient," "second active ingredient," and the like may be used to indicate active ingredients located at different locations within the particle, for example, one located within the core or one or more outer layers. However, the terms "first" or "second" do not necessarily indicate that the first active ingredient is different from the second active ingredient. For example, in certain embodiments, the active ingredient contained within the core can be the same as a second active ingredient contained within an outer layer disposed on the core. In certain other embodiments, the active ingredient contained within the core can be different from the second active ingredient contained within an outer layer disposed on the core.

[0023] The term "supplement" refers to a product that is added to the normal diet of a mammal, but may be combined with the normal food or beverage composition of a mammal. The supplement may be in any form, including but not limited to, a solid, liquid, gel, capsule, or powder. The supplement may also be administered simultaneously with or as a component of a food composition, which may include a food, beverage, pet food, snack, or treat. In one embodiment, the beverage may be an active beverage.

[0024] The term "nutraceutical" refers to any compound added to a nutritional source (e.g., a food, beverage, or dietary supplement) that provides a health or medical benefit in addition to its basic nutritional value.

[0025] As used herein, the term "delivering" or "administering" refers to any route for providing a composition, product, or nutritional functional food to a subject, as accepted as standard by the medical community. For example, the present disclosure contemplates routes of delivery or administration including oral ingestion and any other suitable delivery route, including transdermal, intravenous, intraperitoneal, intramuscular, topical, and subcutaneous.

[0026] As used herein, the term "mammal" includes any mammal that can benefit from improved joint health, resilience, and recovery, and may include, but is not limited to, canine, equine, feline, bovine, human, or porcine mammals.

[0027] As used herein, "healthy" refers to the absence of disease or injury.

[0028] Unless otherwise stated, as used herein, "collagen" refers to all forms of collagen, with or without denaturation, with or without salts or stabilizers, as well as fibrillar and non-fibrillar collagens, including but not limited to fibril-associated collagens with interrupted triple helices (FACIT, IX, XII, XIV, XIX, XXI), including short chain collagens (generally types VII and X), basement membrane (type IV), Multiplexin (multiple triple helical domains with interruptions) (types XV, XVIII), and other types of collagens (types VI, VII). In one aspect, collagens can be maintained in triple helical structure and epitopes without chemical or enzymatic or heat or lower pH treatment.

[0029] As used herein, the term "biomarker" refers to a measurable substance in a mammal whose presence, concentration, or amount indicates some phenomenon, such as disease, infection, or inflammation, and may be a joint biomarker, an inflammatory biomarker, etc.

[0030] As used herein, the percent change of any one or more of the biomarkers discussed is based on the baseline value of the respective subject and / or marker and does not refer to the percent change from an intermediate or non-starting point. Thus, as used herein, "baseline" refers to day zero or starting point and can thus be used to determine the value of the marker measured at the starting point, which is then used to calculate the percent change of any of the biomarkers discussed herein from baseline.

[0031] Other features and aspects of the disclosure are discussed in more detail below. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0032] It should be understood by those skilled in the art that this discussion is merely a description of exemplary embodiments and is not intended to limit the broader aspects of the present disclosure.

[0033] Generally, the present disclosure is directed to a method for reducing inflammation, increasing mobility, and / or otherwise treating disease or injury in a skeletal joint in a mammal.The method comprises administering a therapeutically effective amount of a joint treatment composition to a mammal, such as an animal or human, affected as described above, for a period sufficient to improve the health of the mammal and the skeletal joint.The method of the present disclosure is particularly well suited for treating, for example, osteoarthritis and / or osteonecrosis.

[0034] The joint treatment composition of the present disclosure contains collagen, such as any suitable collagen in various forms. In one embodiment, the collagen contained in the joint treatment composition is undenatured type II collagen. Collagen has been used in the past as a supplement to improve joint health in healthy mammals. For example, PCT Application No. PCT / US2021 / 045853, filed August 13, 2021, and incorporated herein by reference, discloses supplements and methods for improving inflammation, joint health, joint mobility, and joint comfort in healthy mammals. In the past, it was believed that the benefits of administering collagen to a mammal were realized at lower doses, and that increasing the amount of collagen administered to the mammal had no additional benefits. Conversely, it has now been discovered that administering higher amounts of collagen to a mammal can unexpectedly and dramatically improve skeletal joint health in mammals suffering from diseases and disorders such as osteoarthritis and / or osteonecrosis. In fact, it has been discovered that the effectiveness of the supplement of the present disclosure continues to increase as the dose of undenatured collagen increases.

[0035] For example, in the past, it was widely believed that a daily collagen dose of more than about 0.55, for example more than about 0.6 mg / kg body weight per day (e.g., 40 mg collagen for some mammals, such as humans) would not provide additional beneficial effects to the user or mammal. However, it has been surprisingly and unexpectedly discovered that administering more than about 0.66 mg / kg body weight per day of collagen can have a dramatic beneficial effect on joint damage or disease. In this regard, the method of the present disclosure, in one aspect, can administer to a mammal a greater amount of collagen sufficient to reduce the severity of skeletal joint damage or disease as shown through a change in at least one biomarker, compared to administering a joint treatment composition of less than 0.3 mg / kg body weight per day, for example, compared to administering a joint treatment composition of less than 0.5 mg / kg body weight per day, for example, compared to administering a joint treatment composition of less than 0.6 mg / kg body weight per day.

[0036] When administered to a mammal suffering from skeletal joint disease or injury, the joint treatment composition of the present disclosure can dramatically reduce pain in addition to healing the skeletal joint. When administered to a mammal suffering from one or more skeletal joint injuries or illnesses, the joint treatment composition can dramatically improve mobility, which can be measured through Kellgren-Lawrence score and Mankin score, and improve joint and tissue health. For example, the joint treatment composition of the present disclosure can be administered to a mammal for a period of time sufficient to change the mammal's Kellgren-Lawrence score by at least about 20%. The joint treatment composition of the present disclosure can also be administered to a mammal for a period of time sufficient to change the mammal's Mankin score by at least about 15%.

[0037] It has also been discovered that administering a therapeutically effective amount of a joint treatment composition to a mammal can dramatically affect and improve various biomarkers associated with skeletal health and / or inflammation. For example, in a mammal suffering from a disease or injury of a skeletal joint, the joint treatment composition of the present disclosure can dramatically and unexpectedly reduce tumor necrosis factor alpha (TNFα), which is believed to play a key role in inflammation by initiating an inflammatory cascade. However, the method of the present disclosure can reduce serum TNFα in the mammal by more than about 5%, such as more than about 15%, such as more than about 25%, and generally less than about 120%, such as less than about 80%. Similarly, TNFα protein can be reduced by more than about 10%, such as more than about 20%, such as more than about 25%, such as more than about 35%, and generally less than about 80% in the mammal. The reduction can occur after a period of administering the joint treatment composition at least once every three days, such as at least every day. The above results may, for example, be shown after 20 weeks of treatment, such as after 15 weeks of treatment, for example after 12 weeks of treatment, such as after 10 weeks of treatment, for example after 8 weeks of treatment, such as after 6 weeks of treatment, for example after 4 weeks of treatment.

[0038] The joint treatment composition of the present disclosure can also reduce serum interleukin-1 levels in mammals suffering from osteoarthritis or other injuries or diseases of skeletal joints. Serum interleukin levels can be reduced by more than about 10%, for example more than about 20%, for example more than about 30%, for example more than about 40%, and generally less than about 100%. The above results can occur after treatment with the joint treatment composition for a period of 20 weeks of treatment, for example after 15 weeks of treatment, for example after 12 weeks of treatment, for example after 10 weeks of treatment, for example after 8 weeks of treatment, for example after 6 weeks of treatment, for example after 4 weeks of treatment.

[0039] The joint treatment composition of the present disclosure can also reduce serum interleukin-6 levels in mammals suffering from osteoarthritis or other injuries or diseases of skeletal joints. Serum interleukin levels can be reduced by more than about 10%, for example more than about 20%, for example more than about 30%, for example more than about 40%, and generally less than about 120%. The above results can occur after treatment with the joint treatment composition for a period of 20 weeks of treatment, for example after 15 weeks of treatment, for example after 12 weeks of treatment, for example after 10 weeks of treatment, for example after 8 weeks of treatment, for example after 6 weeks of treatment, for example after 4 weeks of treatment.

[0040] The joint treatment composition of the present disclosure can also reduce serum C-reactive protein levels in mammals suffering from osteoarthritis or other injuries or diseases of skeletal joints. Serum C-reactive protein levels can be reduced by more than about 10%, for example more than about 20%, for example more than about 30%, for example more than about 40%, and generally less than about 90%. The above results can occur after treatment with the joint treatment composition for a period of 20 weeks of treatment, for example after 15 weeks of treatment, for example after 12 weeks of treatment, for example after 10 weeks of treatment, for example after 8 weeks of treatment, for example after 6 weeks of treatment, for example after 4 weeks of treatment.

[0041] The joint treatment composition of the present disclosure can also reduce prostaglandin E2 levels in mammals suffering from osteoarthritis or other injuries or diseases of skeletal joints.Prostaglandin E2 levels can be reduced by more than about 8%, for example more than about 15%, for example more than about 25%, for example more than about 30%, and generally less than about 80%.The above results can occur after treatment with the joint treatment composition for a period of 20 weeks of treatment, for example after 15 weeks of treatment, for example after 12 weeks of treatment, for example after 10 weeks of treatment, for example after 8 weeks of treatment, for example after 6 weeks of treatment, for example after 4 weeks of treatment.

[0042] The joint treatment composition of the present disclosure may also reduce interleukin-1-B protein levels in mammals suffering from osteoarthritis or other injuries or diseases of skeletal joints. Interleukin-1-B protein levels may be reduced by more than about 8%, for example more than about 10%, for example more than about 15%, for example more than about 20%, and generally less than about 60%. The above results may occur after treatment with the joint treatment composition for a period of 20 weeks of treatment, for example after 15 weeks of treatment, for example after 12 weeks of treatment, for example after 10 weeks of treatment, for example after 8 weeks of treatment, for example after 6 weeks of treatment, for example after 4 weeks of treatment.

[0043] The joint treatment composition of the present disclosure can also reduce interleukin-6 protein levels in mammals suffering from osteoarthritis or other injuries or diseases of skeletal joints. Interleukin-6 protein levels can be reduced by more than about 10%, for example more than about 15%, for example more than about 20%, for example more than about 30%, for example more than about 40%, and generally less than about 80%. The above results can occur after treatment with the joint treatment composition for a period of 20 weeks of treatment, for example after 15 weeks of treatment, for example after 12 weeks of treatment, for example after 10 weeks of treatment, for example after 8 weeks of treatment, for example after 6 weeks of treatment, for example after 4 weeks of treatment.

[0044] The joint treatment composition of the present disclosure can also reduce interleukin-10 protein levels in mammals suffering from osteoarthritis or other injuries or diseases of skeletal joints. Interleukin-10 protein levels can be reduced by more than about 10%, for example more than about 20%, for example more than about 30%, for example more than about 40%, and generally less than about 100%. The above results can occur after treatment with the joint treatment composition for a period of 20 weeks of treatment, for example after 15 weeks of treatment, for example after 12 weeks of treatment, for example after 10 weeks of treatment, for example after 8 weeks of treatment, for example after 6 weeks of treatment, for example after 4 weeks of treatment.

[0045] The joint treatment composition of the present disclosure may also reduce cyclooxygenase 2 protein levels in mammals suffering from osteoarthritis or other injuries or diseases of skeletal joints. Cyclooxygenase 2 protein levels may be reduced by more than about 7%, for example more than about 10%, for example more than about 20%, for example more than about 30%, and generally less than about 90%. The above results may occur after treatment with the joint treatment composition for a period of 20 weeks of treatment, for example after 15 weeks of treatment, for example after 12 weeks of treatment, for example after 10 weeks of treatment, for example after 8 weeks of treatment, for example after 6 weeks of treatment, for example after 4 weeks of treatment.

[0046] The joint treatment composition of the present disclosure can also reduce nuclear factor kappa B protein levels in mammals suffering from osteoarthritis or other injuries or diseases of skeletal joints. Nuclear factor kappa B protein levels can be reduced by more than about 10%, for example more than about 20%, for example more than about 30%, for example more than about 35%, and generally less than about 80%. The above results can occur after treatment with the joint treatment composition for a period of 20 weeks of treatment, for example after 15 weeks of treatment, for example after 12 weeks of treatment, for example after 10 weeks of treatment, for example after 8 weeks of treatment, for example after 6 weeks of treatment, for example after 4 weeks of treatment.

[0047] The joint treatment composition of the present disclosure can also reduce matrix metallopeptidase 3 levels in mammals suffering from osteoarthritis or other injuries or diseases of skeletal joints. Matrix metallopeptidase 3 levels can be reduced by more than about 15%, for example more than about 25%, for example more than about 35%, and generally less than about 75%. The above results can occur after treatment with the joint treatment composition for a period of 20 weeks of treatment, for example after 15 weeks of treatment, for example after 12 weeks of treatment, for example after 10 weeks of treatment, for example after 8 weeks of treatment, for example after 6 weeks of treatment, for example after 4 weeks of treatment.

[0048] The joint treatment composition of the present disclosure can also reduce the serum level of cartilage oligomer matrix in mammals suffering from osteoarthritis or other damage or disease of skeletal joints.The serum level of cartilage oligomer matrix can be reduced by more than about 10%, for example more than about 20%, for example more than about 30%, and generally less than about 80%.The above results can occur after treatment with the joint treatment composition for a period of 20 weeks of treatment, for example after 15 weeks of treatment, for example after 12 weeks of treatment, for example after 10 weeks of treatment, for example after 8 weeks of treatment, for example after 6 weeks of treatment, for example after 4 weeks of treatment.

[0049] The joint treatment composition of the present disclosure can also reduce cartilage oligomeric matrix protein levels in mammals suffering from osteoarthritis or other injuries or diseases of skeletal joints. Cartilage oligomeric matrix protein levels can be reduced by more than about 10%, for example more than about 15%, for example more than about 25%, for example more than about 30%, and generally less than about 80%. The above results can occur after treatment with the joint treatment composition for a period of 20 weeks of treatment, for example after 15 weeks of treatment, for example after 12 weeks of treatment, for example after 10 weeks of treatment, for example after 8 weeks of treatment, for example after 6 weeks of treatment, for example after 4 weeks of treatment.

[0050] The joint treatment composition of the present disclosure may also increase transforming growth factor beta levels in mammals suffering from osteoarthritis or other injuries or diseases of skeletal joints. Transforming growth factor B protein levels may increase by more than about 5%, for example more than about 10%, for example more than about 15%, for example more than about 20%, and generally less than about 65%. The above results may occur after treatment with the joint treatment composition for a period of 20 weeks of treatment, for example after 15 weeks of treatment, for example after 12 weeks of treatment, for example after 10 weeks of treatment, for example after 8 weeks of treatment, for example after 6 weeks of treatment, for example after 4 weeks of treatment.

[0051] As shown above, the effects on biomarkers in mammals can be dramatic and far exceed previously recorded biomarker effects when mammals are administered collagen.

[0052] As mentioned above, the joint treatment composition of the present disclosure contains collagen, either alone or in combination with various other additives and ingredients. The joint treatment composition may include one or more of any collagens defined above, and / or in one aspect, may include one or more of type I collagen, type II collagen, type III collagen, type IV collagen, or collagen peptides, or a mixture thereof. In one aspect, the joint treatment composition contains type II collagen, either alone or in combination with one or more of type I collagen, type III collagen, type IV collagen, or collagen peptides. In one aspect, the joint treatment composition may include a mixture of type II collagen (sometimes referred to as native type II collagen) and undenatured type II collagen. Additionally or alternatively, the joint treatment composition may include a mixture of native type II collagen and undenatured type II collagen, in addition to additional collagens such as type I, type III, type IV, or collagen peptides. Furthermore, in one aspect, the joint treatment composition includes whole collagen protein, biologically active peptide fragments of collagen, or a combination thereof.

[0053] In one aspect, one or more types of collagen in the joint treatment composition may be heat sterilized, such as by autoclaving, and / or may also include alkalizing or acid, and / or salts, such as organic or inorganic salts. Thus, in one aspect, a portion of the collagen may be at least partially hydrolyzed. In one aspect, collagen is hydrolyzed by any process or compound, including by acid-base agents, enzymes, heat or other extreme temperatures, chemicals, UV, salts, or combinations thereof. In one aspect, hydrolyzed collagen is non-denatured collagen that has been at least partially hydrolyzed by any method, and in one aspect, may include non-denatured collagen with any portion being denatured. However, as discussed, in one aspect, at least a portion of the joint treatment composition is non-denatured.

[0054] As mentioned above, in one aspect, the supplement contains a joint treatment composition, particularly a type II collagen composition, such as a non-denatured type II collagen composition, an at least partially hydrolyzed type II collagen composition, or a combination thereof. Type II collagen for use in the present disclosure can be obtained from any suitable source. For example, collagen can be derived from various mammalian sources, avian sources, or can be derived from various fish species or combinations thereof. For example, collagen can be obtained from salmon, shark, poultry, pig, eggshell, turkey cartilage, bovine cartilage, and the like. In one embodiment, for example, type II collagen can be obtained as disclosed in U.S. Patent No. 7,083,820 to Schilling, which is incorporated by reference. For example, non-denatured type II collagen is commercially available as UC-II® brand from InterHealth Nutraceuticals. UC-II® brand is a natural ingredient that contains glycosylated non-denatured type II collagen. The joint treatment composition can also include hydrolyzed collagen. The joint treatment composition can also include pure protein or active peptide fragments. In one embodiment, the joint treatment composition may be free of any bone or bone material. In another embodiment, the joint treatment composition may be free of any transforming growth factor (TGF), bone morphogenetic protein (BMP), or both. In yet another embodiment, the joint treatment composition includes type II collagen and is completely free of type I collagen.

[0055] In preparing animal tissue for oral administration, in one embodiment, type II collagen-containing tissue can be first cut free of surrounding tissue, and then diced or otherwise ground into particles. Particulate cartilage or milled cartilage can be sterilized by means that do not affect or denature the structure of the major portion of type II collagen in the tissue, such as low-temperature processing, and formed into a dose that contains therapeutically effective levels of undenatured type II collagen. Being a natural product, some variation from sample to sample is expected. These variations can be minimized by blending after grinding. Blending can be assisted by analytical techniques that allow the amount of undenatured type II collagen and other components to be measured.

[0056] Nevertheless, the present disclosure has found that by carefully forming particles and sterilizing type II collagen as discussed above, non-denatured type II collagen can be resistant to gastric acid and digestive enzymes in the stomach.This sterilization process also allows non-denatured type II collagen to retain its three-dimensional shape and preserve the bioactive epitope region.Without wishing to be bound by theory, it is believed that the epitope region contains the ability to induce oral immune tolerance as discussed above.In particular, the epitope region allows non-denatured collagen to bind to Peyer's patches, which have the ability to induce oral immune tolerance process.

[0057] The amount of the joint treatment composition administered to the mammal, the number of administrations per unit time, and the length of treatment can all vary depending on various factors, including the type of mammal being treated, the injury or disease being treated, etc. In one embodiment, for example, the joint treatment composition is administered to the mammal such that the mammal is administered collagen in an amount of at least 0.66 mg / kg body weight per day. For example, a therapeutically effective amount may include administering to the mammal an amount of collagen that is greater than about 1 mg / kg body weight per day, such as greater than about 1.2 mg / kg body weight per day, such as greater than about 1.4 mg / kg body weight per day, such as greater than about 1.6 mg / kg body weight per day, such as greater than about 1.8 mg / kg body weight per day, such as greater than about 2 mg / kg body weight per day, such as greater than about 2.2 mg / kg body weight per day, such as greater than about 2.4 mg / kg body weight per day, such as greater than about 2.6 mg / kg body weight per day, and generally less than about 5 mg / kg body weight per day, such as less than about 4 mg / kg body weight per day.

[0058] In one particular embodiment, collagen is administered to the mammal in an amount between about 1.2 mg / kg body weight per day and about 3 mg / kg body weight per day.

[0059] When collagen is administered to larger mammals, including humans, according to the present disclosure, collagen can be administered to the mammal in an amount of more than about 50 mg per day, such as more than about 80 mg per day, such as more than about 120 mg per day, such as more than about 160 mg per day, such as more than about 320 mg per day, such as more than about 480 mg per day, such as more than about 640 mg per day, such as more than about 800 mg per day. The amount of collagen administered to the mammal is generally less than about 5,000 mg per day, such as less than about 3,000 mg per day, such as less than about 2,000 mg per day, such as less than about 1,000 mg per day, such as less than about 500 mg per day.

[0060] The joint treatment composition may be orally administered to a mammal and may be in the form of a dietary supplement. The supplement may be in the form of individual dosage containers that can be swallowed, chewed, drunk, etc. The dosage containers may be capsules or tablets. In one embodiment, the amount of collagen in each dosage container may be from about 40 mg to about 200 mg. In one aspect, each of the dosage containers may be administered to a mammal at least twice a day, such as at least three times a day, such as at least four times a day, and generally less than eight times a day, such as less than six times a day, such as less than five times a day.

[0061] In one embodiment, the undenatured type II collagen may form all or substantially all of the total type II collagen in the collagen composition and thus may be present in the supplement in the amounts discussed above. However, in one embodiment, the undenatured type II collagen may comprise about 0.5% to about 95% of the total type II collagen and / or collagen composition, such as about 1% to about 75% of the total type II collagen or total collagen composition, such as about 1.5% to about 50%, such as about 2% to about 40%, such as about 2.5% to about 15%, or any range or value therebetween.

[0062] In one embodiment, the collagen composition may further comprise a preserving salt, such as potassium chloride. Thus, in one embodiment, the total amount of the collagen composition discussed above may comprise type II collagen and / or undenatured type II collagen, alone or in combination with additional collagen, preserving salt, or combinations thereof. In such an embodiment, the total type II collagen, including native and undenatured type II collagen, may comprise about 1% to about 99% of the collagen composition, such as about 2.5% to about 90%, such as about 5% to about 80%, such as about 7.5% to about 70%, such as about 10% to about 60%, such as about 15% to about 50%, such as about 20% to about 35%, or any range or value therebetween.

[0063] When the type II collagen comprises undenatured type II collagen, the undenatured type II collagen may have a large oxygen radical absorbance capacity (ORAC) as measured according to ORAC 6.0. In particular, the ORAC test measures antioxidant scavenging activity against oxygen radicals known to be involved in aging and the pathogenesis of common diseases, and consists of six ORAC assays that evaluate the antioxidant capacity of materials against primary reactive oxygen species, peroxyl radicals, hydroxyl radicals, superoxide anions, and peroxynitrite. In particular, the ORAC assay involves introducing a reactive oxygen species (ROS) introducer into the assay system, which causes the release of specific ROS that degrade the probe and change its emission wavelength or intensity. Thus, when the assay being tested contains an antioxidant, the antioxidant absorbs the ROS and protects the probe from degradation. The degree of probe preservation indicates the antioxidant capacity of the material, and the results are expressed as μmol Trolox equivalents (TE) / g of the test material.

[0064] For example, the ORAC assay for peroxyl radicals measures the antioxidant capacity of a sample to protect a fluorescent protein (fluorescein) from damage by peroxyl radicals generated from 2,2'azobis(2amidinopropane) dihydrochloride (AAPH). The ORAC assay for hydroxyl radicals measures the antioxidant capacity of a sample to protect a fluorescent protein (fluorescein) from damage by hydroxyl radicals generated from the reaction of cobalt with hydrogen peroxide. The ORAC assay for peroxin nitrite measures the antioxidant capacity of a sample to protect dihydrorhodamine-123 from damage by peroxin nitrite radicals generated from 3-morpholinosyndnonimine hydrochloride. The ORAC assay for superoxide measures the antioxidant capacity of a sample to protect hydroethidine from damage by superoxide generated from xanthine oxidase. The ORAC assay for singlet oxygen measures the antioxidant capacity of a sample to protect hydroethidine from damage by singlet oxygen generated from the reaction of lithium molybdate with hydrogen peroxide. Finally, the ORAC assay for hypochlorite measures the antioxidant capacity of a sample to protect the fluorescent protein fluorescein from damage by hypochlorous acid radicals generated from sodium hypochlorite.

[0065] Thus, in one aspect, a collagen composition having undenatured type II collagen according to the present disclosure may have a total ORAC of about 200 μmol TE / g or more, such as about 250 μmol TE / g or more, such as about 300 μmol TE / g or more, such as about 350 μmol TE / g or more, such as about 400 μmol TE / g or more, such as about 450 μmol TE / g or more, such as about 500 μmol TE / g or more, such as about 550 μmol TE / g or more, such as about 600 μmol TE / g or more, such as about 700 μmol TE / g or more, such as about 750 μmol TE / g or more, such as about 800 μmol TE / g or more, such as about 825 μmol TE / g or more, up to about 1000 μmol TE / g, or any range or value therebetween.

[0066] Further, in one aspect, a collagen composition having undenatured type II collagen according to the present disclosure may have an ORAC to peroxyl radicals of about 1 μmol TE / g or more, such as about 2.5 μmol TE / g or more, such as about 5 μmol TE / g or more, such as about 7.5 μmol Te / g or more, such as about 10 μmol TE / g or more, such as up to about 10.5 μmol TE / g or more, up to about 50 μmol TE / g, or any range or value therebetween.

[0067] Similarly, in one aspect, a collagen composition having undenatured type II collagen according to the present disclosure may have an ORAC to hydroxyl radicals of about 10 μmol TE / g or more, such as about 15 μmol TE / g or more, such as about 20 μmol TE / g or more, such as about 25 μmol TE / g or more, such as about 27.5 μmol TE / g or more, such as about 30 μmol TE / g or more, up to about 40 μmol TE / g, or any range or value therebetween.

[0068] Additionally or alternatively, in one aspect, a collagen composition having undenatured type II collagen according to the present disclosure may have an ORAC to peroxynitrite of about 0.5 μmol TE / g or more, such as about 1 μmol TE / g or more, such as about 1.5 μmol TE / g or more, such as about 2 μmol TE / g or more, such as about 2.25 μmol TE / g or more, up to about 5 μmol TE / g, or any range or value therebetween.

[0069] In one aspect, a collagen composition having undenatured type II collagen according to the present disclosure may have an ORAC for singlet oxygen of about 500 μmol TE / g or more, such as about 550 μmol TE / g or more, such as about 600 μmol TE / g or more, such as about 650 μmol TE / g or more, such as about 700 μmol TE / g or more, such as about 725 μmol TE / g or more, up to about 1000 μmol TE / g, or any range or value therebetween.

[0070] Further, in one aspect, a collagen composition having undenatured type II collagen according to the present disclosure may have an ORAC to hypochlorite of about 25 μmol TE / g or more, such as about 30 μmol TE / g or more, such as about 35 μmol TE / g or more, such as about 40 μmol TE / g or more, such as about 45 μmol TE / g or more, such as up to about 50 μmol TE / g or more, up to about 75 μmol TE / g, or any range or value therebetween.

[0071] Further, in one embodiment, when the type II collagen comprises undenatured type II collagen, the undenatured type II collagen has an average molecular weight of about 10,000 daltons or more, such as about 15,000 daltons or more, for example, about 20,000 daltons or more, for example, about 25,000 daltons or more, for example, about 30,000 daltons or more, for example, about 35,000 daltons or more, such as about 40,000 daltons or more, for example, about 45,000 daltons or more, for example, about 50,000 daltons or more, for example, about 55, 000 daltons or more, such as about 60,000 daltons or more, such as about 65,000 daltons or more, such as about 70,000 daltons or more, such as about 75,000 daltons or more, such as about 80,000 daltons or more, such as about 85,000 daltons or more, such as about 90,000 daltons or more, such as about 95,000 daltons or more, such as about 100,000 or more, up to about 350,000 daltons or less, or any range or value therebetween.

[0072] Various aspects and advantages have been discussed, and in one aspect, the collagen composition is incorporated into a suitable delivery form prior to incorporation into a dosage form as discussed below. In one aspect, the composition of the present disclosure may be included as a capsule or tablet.

[0073] Alternatively, the composition can be an oil-in-water emulsion as a delivery form.In particular, in one aspect, such a configuration can allow one or more oil-soluble and / or one or more water-soluble active components to be contained in the same delivery form.Alternatively, only oil-soluble components can be used (e.g., type II collagen), and emulsion can be used to incorporate the composition into aqueous applications.However, it should be understood that in one aspect, water-soluble type II collagen can be used in supplements.

[0074] The oil-in-water emulsion may also contain at least one functional gum, such as gum arabic. Generally, gum arabic is a complex mixture of glycoproteins and polysaccharides that contain arabinose and galactose. Gum arabic is generally soluble in water and is edible.

[0075] The oil-in-water emulsion may also contain water, hi certain embodiments, the oil-in-water emulsion may contain about 5% to 35% water by weight.

[0076] In some embodiments, the oil-in-water emulsion may contain one or more stabilizers or suspension promoters. For example, in certain embodiments, the oil-in-water emulsion may contain one or more gums, such as gellan gum or xantham gum. When included, the gellan gum or xantham gum may be present in an amount of less than about 3.5% by weight of the oil-in-water emulsion, such as less than about 2.5% by weight, such as less than about 1.5% by weight, such as less than about 1.0% by weight, such as less than about 1.0% by weight. An example of a stabilizer is:

[0077] Silica may be added in an amount of less than about 2% by weight, such as less than about 1.5% by weight, such as less than about 1% by weight, such as less than about 0.5% by weight.

[0078] In some aspects, the oil-in-water emulsions disclosed herein may be used in any suitable dosage form, such as tablets, gummy chewables, edible films, lozenges, liquid suspensions, syrups, lipid micelles, spray dried dispersions, nanoparticles, and the like, which may also be incorporated into additional supplements.

[0079] Alternatively, the oil-in-water emulsions may be contained in nutritional products such as supplements, foods, or beverages. For example, in certain aspects, the oil-in-water emulsions may be incorporated into liquid nutritional products such as dietary supplements or infant formula for consumption by a mammal. Additionally, the oil-in-water emulsions provided herein may be added to any liquid nutritional product designed to provide nutritional supplementation to a mammal.

[0080] The nutritional product may include any suitable composition for consumption by a mammal. Such compositions include complete foods or beverages intended to supply the necessary dietary requirements for a mammal or food supplements such as sweets and snacks. Food compositions may include pellets, beverages, bars, canned prepared foods, milkshake drinks, juices, dairy products, or any other functional food composition. Food compositions may also include supplements in any form such as pills, soft gels, gummy figurines, wafers, powders, etc.

[0081] The supplement according to the present disclosure can be administered to mammals, including by oral, enteral or inhalation administration of whole collagen protein or biologically active peptide fragments of collagen.For example, in one aspect, it is believed that whole collagen protein or biologically active peptide fragments of collagen enhance the content of trans-L-hydroxyproline based on the total weight of amino acids contained in collagen type II, and enable the efficient production of trans-L-hydroxyproline for enhancing efficacy.

[0082] The supplement composition of the present disclosure may further comprise one or more excipients as additional additives in the composition. Exemplary, but non-limiting, excipients and / or additives include anti-adherents such as magnesium stearate; binders such as sugars, sugar alcohols, gelatin, and synthetic polymers; coatings such as cellulose ethers, hydroxypropylmethylcellulose (HPMC), shellac, corn protein zein, gelatin, fatty acids, fats, oils, and / or waxes; colorants such as titanium dioxide and azo dyes; disintegrants such as modified starch sodium starch glycolate and cross-linked polymers including polyvinylpyrrolidone and carboxymethylcellulose; fillers such as maltodextrin; mint, lyco, and the like. flavorings such as scallion, anise, vanilla, and fruit flavors including peach, banana, grape, strawberry, blueberry, raspberry, and mixed berry; glidants such as fumed silica, talc, and magnesium carbonate; lubricants such as talc, silica, and fats including vegetable stearin, magnesium stearate, and stearic acid; preservatives such as antioxidants, vitamins, retinyl palmitate, selenium, the amino acids cysteine ​​and methionine, citric acid, sodium citrate, and parabens; adsorbents; sweeteners such as sucrose and sucralose; and vehicles such as petrolatum and mineral oil.

[0083] In one aspect, the supplement composition of the present disclosure may be combined with various additives and components that can improve one or more properties of the composition. For example, in one embodiment, the additive composition may be combined with a stabilizer package that can help stabilize at least one property of the composition. In a particular embodiment, for example, the stabilizer package may be added to the composition in an amount sufficient to reduce the hydrophobic properties of the composition and / or prevent the composition from absorbing moisture. The stabilizer package may also be combined with the composition to improve the handling properties of the composition. For example, the stabilizer package may enable the composition to have better flow properties, especially when granular.

[0084] In one aspect, the supplement composition may be combined with a polymer binder in conjunction with a stabilizer package.In addition, a coating material may be applied to the composition after the composition is combined with the polymer binder and stabilizer package.The coating material may, for example, contain at least one fat.According to the present disclosure, the above components may be added to any suitable pharmaceutical composition in addition to the composition of the present disclosure.For example, the above components may be added to any pharmaceutical composition that contains carnitine or amino acid.

[0085] The polymer binder and stabilizer package may be combined with the supplement composition in a manner that homogenously incorporates the stabilizer package into the product. In one embodiment, for example, the composition of the present disclosure is first combined with the polymer binder, such as by a spray drying process, and then combined with the stabilizer package. The polymer binder may include any suitable pharma- ceutically acceptable polymer, such as a film-forming polymer and / or a polysaccharide. Specific examples of polymer binders that may be used according to the present disclosure include starch, maltodextrin, gum arabic, arabinogalactan, gelatin, and mixtures thereof. In one embodiment, the polymer binder is added to the pharmaceutical composition in an amount of at least about 5% by weight, such as at least about 8% by weight, such as at least about 10% by weight, such as at least about 15% by weight. One or more polymer binders are present in the composition in an amount of less than about 50% by weight, such as less than about 45% by weight, such as less than about 40% by weight, such as less than about 35% by weight, such as less than about 30% by weight.

[0086] In one embodiment, the polymeric binder may include a starch, such as a modified starch. The starch may be derived, for example, from corn or waxy corn. In one embodiment, the starch may include HI-CAP100 starch sold by National Starch and Chemical Company. In an alternative embodiment, the polymeric binder may include arabinogalactan.

[0087] Once the polymer binder is combined with the composition, such as by a spray drying process, the resulting mixture can then be combined with a stabilizer package. In one embodiment, the stabilizer package includes oxide particles combined with a salt of a carboxylic acid. In one particular embodiment, the stabilizer package may include a dry product, such as a powder or granular product, combined with the composition and the polymer binder. It has been found that the combination of oxide particles and salt of a carboxylic acid provides numerous advantages and benefits when combined with the composition. For example, it has been found that the stabilizer package stabilizes the composition and reduces the hydrophobicity of the composition. This composition also produces a product that is easy to handle, granular, and flowable.

[0088] The oxide particles that may be added to the supplement composition may include silica.For example, the oxide particles may include precipitated silica particles.The silica particles may have a particle size (d50, laser diffraction after ISO test 13320) of less than about 55 microns, such as less than about 40 microns, such as less than about 30 microns, such as less than about 25 microns, such as less than about 20 microns, such as less than about 15 microns, such as less than about 12 microns, such as less than about 10 microns, such as less than about 8 microns, such as less than about 6 microns, such as less than about 4 microns, such as less than about 2 microns, such as less than about 1 micron.The particle size is typically greater than about 0.5 microns, such as greater than about 1 micron. The particles may have a specific surface area (ISO test 9277) of more than about 120 m2 / g, for example more than about 130 m2 / g, for example more than about 150 m2 / g, for example more than about 170 m2 / g, for example more than about 200 m2 / g, for example more than about 220 m2 / g. The specific surface area is generally less than about 500 m2 / g. The oxide particles, such as silica particles, can be present in the pharmaceutical composition in an amount of more than about 0.01% by weight, for example more than about 0.05% by weight, for example more than about 0.1% by weight. The oxide particles are generally present in an amount of less than 5% by weight, for example less than about 2% by weight, for example less than about 1.5% by weight, for example less than 0.5% by weight.

[0089] In addition to the oxide particles, the stabilizer package may also include a salt of a carboxylic acid. The salt of a carboxylic acid may include a salt of a fatty acid. The fatty acid may have, for example, a carbon chain length of about 6 carbon atoms to about 40 carbon atoms, for example, about 12 carbon atoms to about 28 carbon atoms. In one embodiment, the salt of a carboxylic acid may include a stearate. Stearates that may be used include calcium stearate, sodium stearate, magnesium stearate, mixtures thereof, and the like. In one embodiment, the salt of a carboxylic acid may include both hydrophilic and hydrophobic groups. The salt of a carboxylic acid may be present in the composition in an amount greater than about 0.5% by weight, for example, greater than about 1% by weight, for example, greater than about 1.5% by weight. The salt of a carboxylic acid is generally present in an amount less than about 5% by weight, for example, less than about 4% by weight, for example, less than about 3% by weight.

[0090] In addition to the polymer binder and stabilizer package, the composition may include various other components and ingredients. In one embodiment, for example, the composition may include a citric acid ester, such as a mono-citric acid ester and / or a diglyceride of a fatty acid. The composition may also include a lecithin, such as lecithin obtained from rapeseed, sunflower, etc. The above components may be present in the composition in relatively small amounts, such as less than about 2% by weight, for example less than about 1.5% by weight, for example less than about 1% by weight. The above components are generally present in an amount greater than about 0.05% by weight, for example greater than about 0.1% by weight.

[0091] Further, in one embodiment, the supplement may be formulated into a food and / or supplement for athletic, sports, or daily nutritional purposes. In such an embodiment, the supplement may further include at least one vitamin, such as at least one of vitamin B, vitamin C, and vitamin E. The vitamin may be contained in the supplement in an amount of about 50 μg / g to about 5000 μg / g of the supplement, such as about 100 μg / g to about 4500, such as about 250 μg / g to about 4000 μg / g, such as about 400 μg / g to about 3500 μg / g, or any range or value therebetween. The above ranges may be for any one vitamin alone or the total amount of all vitamins. In one embodiment, vitamin E is present in the supplement in an amount of about 100 μg / g to about 1000 μg / g, such as about 250 μg / g to about 750 μg / g, such as about 400 μg / g to about 600 μg / g, or any range or value therebetween. In another embodiment, vitamin C is present in the supplement in an amount of about 1000 μg / g to about 5000 μg / g, such as about 2000 μg / g to about 4000 μg / g, such as about 3000 μg / g to about 3750 μg / g, or any range or value therebetween.

[0092] Further, in one embodiment, the supplement contains at least one mineral, such as at least one of potassium, magnesium, zinc, or calcium. The mineral may be contained in the supplement in an amount of about 1 mg / g to about 50 mg / g, for example, about 2.5 mg / g to about 45 mg / g, for example, about 5 mg / g to about 40 mg / g, or any range or value therebetween. The above range may be the total amount of any one mineral or one mineral. In one embodiment, the supplement contains potassium in an amount of about 9.5 mg / g to about 12 mg / g, for example, about 9.75 mg / g to about 11.5 mg / g, for example, about 10 mg / g to about 11 mg / g, or any range or value therebetween. Similarly, in one embodiment, the supplement contains magnesium in an amount of about 1 mg / g to about 10 mg / g, for example, about 2.5 mg / g to about 7.5 mg / g, for example, about 4 mg / g to about 6 mg / g, or any range or value therebetween. Further, in one embodiment, the supplement contains calcium in an amount of about 1 mg / g to about 50 mg / g, e.g., about 2.5 mg / g to about 47.5 mg / g, e.g., about 5 mg / g to about 45 mg / g, e.g., about 10 mg / g to 40 mg / g, e.g., about 20 mg / g to about 37.5 mg / g, e.g., about 30 mg / g to about 35 mg / g, or any range or value therebetween.

[0093] Thus, in one aspect, the supplement according to the present disclosure may include, in addition to undenatured collagen, one or more additional joint supplements, such as hydroxycitric acid, glucosamine, chondroitin, methylsulfonylmethane, eggshell membrane, mussel, or combinations thereof, and / or a collagen absorption enhancer, such as vitamin C.

[0094] Furthermore, the supplement may be suitable for administration to any mammal. For example, the mammal may be a human or a dog. The composition may be provided to a mammal of any age, from birth to adulthood in the mammal. In various embodiments, the mammal may be a human, a dog, a cat, a horse, a pig, a sheep, or a cow. In many embodiments, the mammal may be in early to late adulthood. EXAMPLES

[0095] Nonetheless, certain embodiments of the present disclosure may be better understood in accordance with the following examples, which are intended to be non-limiting and exemplary in nature.

[0096] Example 1 The following example demonstrates some of the benefits and advantages of the present disclosure. In this example, Wistar rats were collected and divided into seven groups. Older rats aged 18-20 months were tested separately from younger rats aged 8 weeks. Except for the old and young rat control groups, the rats were injected intra-articularly with monosodium iodoacetate ("MIA") to simulate joint diseases such as osteoarthritis.

[0097] Except for the control groups of old and young rats that received only MIA, the remaining rat groups were administered the joint treatment composition according to the present disclosure with varying amounts of collagen for each group. The collagen used in the experiment was undenatured type II collagen, designated "UCII". More specifically, the following rat groups were tested: [Table 1]

[0098] As above, each group consisted of 7 rats. The rats were tested after 3, 7, 14, and 28 days.

[0099] The following tests were carried out: [Table 2]

[0100] The measurements of paw area, stride length, and step width in MIA-induced osteoarthritis rats on days 3, 7, 14, and 28 are shown below. [Table 3] [Table 4]

[0101] The results obtained in the above table are also shown in Figures 5-6. As shown, the joint treatment composition of the present disclosure dramatically and unexpectedly improved mobility in MIA-induced rats.

[0102] Rats were also tested for Kellgren-Lawrence scores and knee diameters after 28 days, and the results are shown in Figures 7-10.

[0103] Groups of rats were also tested for Mankin score, which is aimed at reducing the grade of lesion score in knee diameter, and the results are shown in Figures 11-12.

[0104] Below is a summary of the changes in Kellgren-Lawrence and Mankin scores. [Table 5] [Table 6]

[0105] After 28 days, various biomarkers were tested, including serum and protein levels, and the results are shown in Figures 13-44.

[0106] A variety of different safety markers were tested and showed no significant differences between the treatments. These results are as follows: [Table 7] [Table 8]

[0107] As shown in the attached drawings, rats administered the joint treatment composition of the present disclosure showed a dramatic improvement in mobility. There was a significant dose response in old rats at 14 days and in young rats at 7 days. In addition, anti-inflammatory properties were observed. Anti-inflammatory properties were only observed after 3 days in younger rats. It was found that higher doses of collagen produced significant effects in terms of anti-inflammatory properties.

[0108] These and other modifications and variations to the present invention may be implemented by those of ordinary skill in the art without departing from the spirit and scope of the present invention, which is particularly set forth in the appended claims. In addition, it should be understood that aspects of the various embodiments may be interchanged in whole or in part. Moreover, those skilled in the art will appreciate that the foregoing description is by way of example only, and is not intended to limit the invention as further set forth in such appended claims.

Claims

1. 1. A nutritional supplement for use in treating skeletal joint injuries or diseases in a mammal, comprising:

1. The nutritional supplement comprising a therapeutically effective amount of a joint treatment composition, the joint treatment composition comprising collagen, the joint treatment composition being for administration in a dose, the therapeutically effective amount being an amount of at least about 0.66 mg / kg body weight per day, sufficient to reduce the severity of the skeletal joint injury or disease as indicated by a change in at least one marker of joint health compared to administering the joint treatment composition at less than 0.3 mg / kg body weight per day.

2. 10. The nutritional supplement of claim 1, wherein the therapeutically effective amount of the joint treatment composition administered to the mammal comprises at least 1 mg / kg body weight of collagen per day.

3. 10. The nutritional supplement of claim 1, wherein the therapeutically effective amount of the joint treatment composition administered to the mammal comprises at least 2 mg / kg body weight of collagen per day.

4. 10. The nutritional supplement of claim 1, wherein the therapeutically effective amount of the joint treatment composition administered to the mammal comprises at least 1.2 mg / kg body weight to about 3 mg / kg body weight of collagen per day.

5. The nutritional supplement of any one of claims 1 to 4, wherein the collagen comprises undenatured type II collagen.

6. 5. The nutritional supplement of any one of claims 1 to 4, wherein the joint treatment composition is for administration to the mammal in a therapeutically effective amount sufficient to reduce serum tumor necrosis factor by at least about 5%, such as at least about 15%, such as at least about 25%, and generally less than about 80%, after 28 days of treatment.

7. 5. The nutritional supplement of any one of claims 1 to 4, wherein the joint treatment composition is for administration to the mammal in a therapeutically effective amount sufficient to reduce serum C-reactive protein by at least about 10%, such as at least about 20%, such as at least about 30%, such as at least about 40%, and generally less than about 90%, after 28 days of treatment.

8. 5. The nutritional supplement of any one of claims 1 to 4, wherein the joint treatment composition is for administration to the mammal in a therapeutically effective amount sufficient to reduce prostaglandin E2 by at least about 8%, such as at least about 15%, such as at least about 25%, such as at least about 30%, and generally less than about 80%, after 28 days of treatment.

9. 5. The nutritional supplement of any one of claims 1 to 4, wherein the joint treatment composition is for administration to the mammal in a therapeutically effective amount sufficient to reduce interleukin-10 protein by at least about 10%, such as at least about 20%, such as at least about 30%, such as at least about 40%, and generally less than about 100%, after 28 days of treatment.

10. 5. The nutritional supplement of any one of claims 1 to 4, wherein the joint treatment composition is for administration to the mammal in a therapeutically effective amount sufficient to reduce tumor necrosis factor by at least about 10%, such as at least about 20%, such as at least about 25%, such as at least about 35%, and generally less than about 80% after 28 days of treatment.

11. 5. The nutritional supplement of any one of claims 1 to 4, wherein the joint treatment composition is for administration to the mammal in a therapeutically effective amount sufficient to reduce cyclooxygenase 2 protein by at least about 7%, such as at least about 10%, such as at least about 20%, such as at least about 30%, and generally less than about 90%, after 28 days of treatment.

12. 5. The nutritional supplement of any one of claims 1 to 4, wherein the joint treatment composition is for administration to the mammal in a therapeutically effective amount sufficient to reduce nuclear factor kappa B by at least about 10%, such as at least about 20%, such as at least about 30%, such as at least about 35%, and generally less than about 80%, after 28 days of treatment.

13. 5. The nutritional supplement of any one of claims 1 to 4, wherein the joint treatment composition is for administration to the mammal in a therapeutically effective amount sufficient to reduce matrix metallopeptidase 3 by at least about 15%, such as at least about 25%, such as at least about 35%, and generally less than about 75% after 28 days of treatment.

14. 5. The dietary supplement of any one of claims 1 to 4, wherein the joint treatment composition is for administration to the mammal in a therapeutically effective amount sufficient to increase transforming growth factor B protein by at least about 5%, such as at least about 10%, for example at least about 15%, such as at least about 20%, for example at least about 25%, such as at least about 30%, for example at least about 35%, such as at least about 40%, for example at least about 45%, such as at least about 50%, for example at least about 55%, such as at least about 60%, for example at least about 65%, and generally less than about 80%, after 28 days of treatment.

15. 5. The nutritional supplement of any one of claims 1 to 4, wherein the joint treatment composition is for administration to the mammal in a therapeutically effective amount sufficient to reduce cartilage oligomeric matrix serum by at least about 10%, such as at least about 20%, such as at least about 30%, and generally less than about 80% after 28 days of treatment.

16. 5. The nutritional supplement of any one of claims 1 to 4, wherein the joint treatment composition is for administration to the mammal in a therapeutically effective amount sufficient to reduce cartilage oligomeric matrix protein by at least about 10%, such as at least about 15%, such as at least about 25%, such as at least about 30%, and generally less than about 80%, after 28 days of treatment.

17. 5. The nutritional supplement of any one of claims 1 to 4, wherein the joint treatment composition is for administration to the mammal in a therapeutically effective amount sufficient to reduce interleukin-1 serum by at least about 10%, such as at least about 20%, such as at least about 30%, such as at least about 40%, and generally less than about 100% after 28 days of treatment.

18. 5. The nutritional supplement of any one of claims 1 to 4, wherein the joint treatment composition is for administration to the mammal in a therapeutically effective amount sufficient to reduce serum interleukin-6 by at least about 10%, such as at least about 20%, such as at least about 30%, such as at least about 40%, and generally less than about 120% after 28 days of treatment.

19. 5. The nutritional supplement of any one of claims 1 to 4, wherein the joint treatment composition is for administration to the mammal in a therapeutically effective amount sufficient to reduce interleukin-1-B protein by at least about 8%, such as at least about 10%, such as at least about 15%, such as at least about 20%, and generally less than about 60%, after 28 days of treatment.

20. 5. The nutritional supplement of any one of claims 1 to 4, wherein the joint treatment composition is for administration to the mammal in a therapeutically effective amount sufficient to reduce interleukin-6 protein by at least about 10%, such as at least about 15%, for example at least about 20%, such as at least about 30%, for example at least about 40%, and generally less than about 80%, after 28 days of treatment.

21. 5. The nutritional supplement of any one of claims 1 to 4, wherein the joint treatment composition is for administration to a mammal in a therapeutically effective amount sufficient to alter the mammal's Kellgren-Lawrence score by at least about 20%.

22. 5. The nutritional supplement of any one of claims 1 to 4, wherein the joint treatment composition is for administration to a mammal in a therapeutically effective amount sufficient to cause a change in the mammal's Mankin score by at least about 15%.

23. 5. A dietary supplement according to any one of claims 1 to 4, wherein the collagen is for administration to the mammal in an amount of more than 50 mg per day, such as more than 80 mg per day, for example more than 80 mg per day, such as more than 100 mg per day, for example more than 120 mg per day, such as more than 480 mg per day, for example more than 320 mg per day, such as more than 480 mg per day, and generally less than about 800 mg per day.

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