Non-denatured type ii collagen in animal foods and treats

Abstract

Currently available non-denatured collagen products include powders and capsules that can optionally be incorporated into a final product that does not require heating or acidic conditions, or instead is consumed directly. This has been particularly problematic for administration to non-human mammals, such as pets and livestock, because when added as a dry powder to food or water, the supplements and capsules sink to the bottom of the bowl of food or liquid and remain unconsumed by the mammal, or in the case of capsules, can be difficult, if not impossible, to administer to the mammal.SOLUTION: The present disclosure relates to processed animal foods and / or treats comprising undenatured type II collagen. The undenatured type II collagen is introduced into the animal food and / or treat prior to processing and at least 30% or more of the undenatured collagen remains undenatured after processing of the animal food and / or treat.SELECTED DRAWING: None

Description

[Background technology]

[0001] In recent years, the use of collagen in the treatment of various conditions has become very popular. Collagen is a protein that can be found in muscles, bones, skin, blood vessels, and other parts of the body. There are various different types of collagen depending on its function and form. For example, type I collagen, the most abundant collagen, is made up of fibers found in tendons, ligaments, organs, and skin. Meanwhile, type II collagen helps build cartilage, the main structural entity located on the surface of bones that primarily form articulating joints. Type III collagen is a major component of the extracellular matrix that makes up organs and skin. Type III collagen also forms tissues within blood vessels and the heart. Type IV collagen is found in the skin primarily as a sheet-like structure in the basal lamina of the skin. Furthermore, collagen peptides are one or more portions of the alpha chains of any type of collagen formed by enzymatic hydrolysis of collagen. Collagen peptides are often used in beverages and food products because they are water-soluble and non-gelling.

[0002] For example, collagen production in the bodies of most mammals tends to slow down as they age. For example, many mammals, including domestic animals and livestock, suffer from age-related arthritis, as well as exercise-induced joint pain, muscle pain, cartilage loss, and bone loss. This can impair the animal's function in competitive or working environments, and can also impair the quality of life of domestic pets.

[0003] Collagen has also been found to effectively treat arthritis and other joint pain in mammals. For example, U.S. Patent No. 9,066,926 discloses a method for reducing exercise-induced joint pain in mammals by administering type II collagen to the mammal. This patent also discloses the mechanism of action by which this component acts: oral immune tolerance. This putative mechanism involves the stimulation of regulatory T cells (Tregs) located in gut-associated lymphoid tissue to specifically recognize antigenic determinants (epitopes) on native collagen proteins. Once induced, Tregs leave the intestinal region and migrate into the joint cavity, where they stimulate chondrocytes to build new type II collagen, thereby improving the structural integrity and flexibility of joint joints. One such example for which clinical data have been published is the knee. Patent No. 9,066,926 is incorporated herein by reference.

[0004] However, to date, undenatured collagen sources have only been available in "raw" form, meaning that undenatured collagen has not been incorporated into processed animal foods or treats because it is sensitive to high temperatures, mechanical processing, and pH changes. In particular, it is believed that to form a product containing undenatured collagen, it cannot be subjected to cooking, such as baking, frying, or other methods of heating, including incorporation into heated liquids or steam, which would denature the collagen. Furthermore, it was previously taught and believed that undenatured collagen would denature when exposed to acidic conditions, and therefore was included in compositions with a pH greater than 7. Furthermore, even mechanical processing, such as pressing and extrusion, was thought to adversely affect the amount of undenatured collagen in processed animal foods and treats.

[0005] Thus, currently available non-denatured collagen products include powders and capsules that can be optionally incorporated into final products that do not require heating or acidic conditions, or are instead consumed directly. This is because, when added to food or water as a dry powder, the supplements and capsules sink to the bottom of the food or liquid bowl and remain unconsumed by the mammal, or, in the case of capsules, cannot be administered to a mammal. Administration to non-human mammals, such as pets and livestock, has been particularly problematic because it can be difficult, if not impossible, to denatured collagen. Thus, many mammals, such as pets and livestock, face problems when administering undenatured collagen because they are reluctant to voluntarily ingest supplements or capsules.

[0006] Although collagen can provide various benefits when administered to mammals, there is a need for processed animal foods and / or treats containing non-denatured collagen. It would be an additional benefit to provide processed animal foods and / or treats that have a higher recovery rate of non-denatured collagen compared to pre-processed foods and / or treats. There is also a need for processed animal foods and / or treats containing non-denatured collagen to support healthy mammals. Furthermore, it would be beneficial to provide processed animal foods and / or treats containing non-denatured collagen to support trained mammals. Summary of the Invention

[0007] In general, the present disclosure relates to processed animal food and / or treat compositions comprising undenatured type II collagen after processing at a temperature of about 37° C. or higher. In one embodiment, the composition is a processed animal food composition. Additionally or alternatively, in some embodiments, the composition is a processed animal treat or chew product.

[0008] In one embodiment, undenatured type II collagen is incorporated into the animal food and / or treat composition as part of a collagen composition that includes one or more different types of collagen in addition to undenatured type II collagen. In some embodiments, the one or more different types of collagen include native type II collagen, collagen peptides, or mixtures thereof.

[0009] In a further embodiment, the amount of undenatured type II collagen incorporated into the composition before processing, and at least about 30% or more of the undenatured type II collagen, is recovered after processing. In one embodiment, 45% or more of the undenatured type II collagen is recovered after processing. Additionally or alternatively, in some embodiments, 60% or more of the undenatured type II collagen is recovered after processing. Furthermore, in one embodiment, 85% or more of the undenatured type II collagen is recovered after processing.

[0010] In another embodiment, the processed animal food and / or treat undergoes processing that includes withstanding temperatures of about 40° C. or greater. Additionally, in one embodiment, the processing lasts from 6 seconds to about 2 hours.

[0011] In one embodiment, the food and / or treat comprises one or more of a protein source, a grain, a flavoring, or a coloring.

[0012] The present disclosure also generally relates to a method of forming a processed animal food and / or treat, the method comprising combining undenatured type II collagen with at least one animal food and / or treat component and processing the undenatured type II collagen and the at least one animal food and / or treat component at a temperature of about 37° C. or greater, wherein at least about 30% or more of the undenatured type II collagen is recovered in the processed animal food and / or treat after processing compared to the amount of undenatured type II collagen before processing.

[0013] In one embodiment, the processing comprises exposing the undenatured type II collagen and at least one animal food and / or treat component to a temperature of about 40° C. for at least about 10 minutes. In a further embodiment, the processing comprises exposing the undenatured type II collagen and at least one animal food and / or treat component to a temperature of about 100° C. for at least about 1 minute. Furthermore, in one embodiment, the processing comprises exposing the undenatured type II collagen and at least one animal food and / or treat component to a temperature of about 100° C. The method comprises exposing the animal food and / or treat component to a temperature of at least about 120°C for at least about 1 minute. In one embodiment, the processing further comprises extruding the composition. Additionally, in one embodiment, the processing further comprises injection molding the composition. In a further embodiment, the processing comprises pelletizing the undenatured type II collagen and at least one animal food and / or treat component.

[0014] Nevertheless, the present disclosure also generally includes methods for improving one or more of joint health, muscle health, bone health, skin health, or fitness, comprising administering to a non-human mammal an effective amount of a processed animal food and / or treat according to this disclosure and / or any one or more of the embodiments discussed above.

[0015] definition As used herein, the terms "about," "approximately," or "generally," when used to modify a value, indicate that the value can be above or below 10% and still remain within the disclosed embodiments.

[0016] As used herein, the term "therapeutically effective amount" refers to a dosage or amount of a composition administered or delivered to a mammal in need of such treatment that results in a specific pharmacological or nutritional response. 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 an amount would be considered a "therapeutically effective amount" by those skilled in the art. Certain subjects may actually 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 achieved. It should be further understood that in certain cases, compositions or supplements may be measured as oral dosages or with respect to component levels that can be measured in blood. In other embodiments, dosages may be measured by the amount applied to the skin when the composition is included in a topical formulation.

[0017] 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.

[0018] As used herein, the terms "delivering" or "administering" refer to any route for providing a composition, product, or nutritionally 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.

[0019] 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, ovine, or porcine mammals. For purposes of this application, "mammal" does not include human subjects and may be used interchangeably with animal.

[0020] As used herein, "healthy" refers to the absence of disease or injury.

[0021] The term "physical activity" refers to activity that lasts for about 10 minutes or more, such as about 25 minutes or more, such as about 30 minutes or more, such as at least about 45 minutes or more, and that causes a mammal's heart rate to reach about 30% to about 85%, such as about 40% to about 80%, of its maximum heart rate, such as about 50% to about 75% of the mammal's maximum heart rate.

[0022] The term "intensive physical activity" means activity lasting for about 20 minutes or more, such as about 25 minutes or more, such as about 30 minutes or more, such as at least about 45 minutes or more, and which causes the mammal's heart rate to reach about 50% to about 99%, such as about 55% to about 95%, such as about 60% to about 90%, such as about 705 to about 85% of its maximum heart rate.

[0023] Unless otherwise stated, as used herein, "collagen" refers to all forms of collagen, whether denatured or not, 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).

[0024] Other features and aspects of the disclosure are discussed in more detail below. DETAILED DESCRIPTION OF THE INVENTION

[0025] 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.

[0026] In general, the present disclosure relates to processed animal foods and / or treats, where the processing includes a temperature of about 37° C. or greater, containing non-denatured collagen, e.g., in one embodiment, non-denatured type II collagen. In particular, the present disclosure has discovered that processed animal foods and / or treats can be formed using non-denatured collagen that has been carefully formulated to preserve epitopes on the non-denatured chains, even when the processed animal foods and / or treats require high temperature processing, high pressure processing, mechanical processing, high moisture content, and / or low pH levels for production and / or storage.

[0027] For example, non-denatured collagen according to the present disclosure can be maintained at temperatures above about 37°C, such as above about 40°C, for example above about 45°C, for example above about 50°C, for example above about 55°C, for example above about 60°C, for example above about 65°C, for example above about 70°C, for example above about 75°C, for example above about 80°C, for example above about 85°C, for example above about 90°C, for example above about 95°C, for example above about 100°C, for example above about 105°C, for example , may be contained in processed animal foods and / or treats that have been processed at temperatures including about 110°C or higher, for example, about 120°C or higher, for example, about 130°C or higher, for example, about 140°C or higher, for example, about 150°C or higher, for example, about 160°C or higher, for example, about 170°C or higher, for example, about 180°C or higher, for example, about 190°C or higher, for example, about 200°C or higher, for example, up to about 300°C or lower, for example, about 275°C or lower, for example, about 250°C or lower.

[0028] In one aspect, processed animal foods and / or treats processed according to one or more of the above temperatures may be processed for about 3 seconds or more, such as about 6 seconds or more, for example, about 1 minute or more, such as about 1.5 minutes or more, for example, about 2 minutes or more, such as about 5 minutes or more, for example, about 10 minutes or more, such as about 15 minutes or more, for example, about 20 minutes or more, for example, about 30 minutes or more, such as about 1 hour or more, for example, about 1.5 hours or more, such as about 2 hours or more, for example, in one aspect up to about 4 hours, such as about 5 hours or less, for example, about 4 hours or less, for example, about 3 hours or less, such as about 2 hours or less, for example, about 1 hour or less, for example, about 30 minutes or less, or any range or value therebetween.

[0029] Additionally or alternatively, the processed animal food and / or treats may be subjected to any one or more of the temperatures or times described above, and may be subjected to high temperature processing simultaneously or after high temperature processing. In such embodiments, the processed animal food and / or treat may be subjected to a high pressure process of about 50 psi or more, such as about 100 psi or more, for example, about 200 psi or more, for example, about 300 psi or more, such as about 400 psi or more, for example, about 500 psi or more, such as about 600 psi or more, for example, about 700 psi or more, for example, about 800 psi or more, such as about 900 psi or more, for example, about 1000 psi or more, such as about 1100 psi or more, for example, about 1200 psi or more, for example, about 1300 psi or more, such as about 1400 psi or more, for example, about 1500 psi or more, such as about 1600 psi or more, for example, about 1700 psi or more, such as about 1800 psi or more, for example, about 1900 psi or more, for example, about 2000 psi or more, up to about 300 psi.

[0030] For example, in one embodiment, the high pressure may additionally or alternatively be mechanical pressure such as extrusion, molding, pressing, punching, pulling, pelletizing, etc. In one embodiment, the mechanical pressure may be injection molding or pressure molding, or a combination thereof. Further, in one embodiment, the mechanical pressure may be extrusion. Nevertheless, in one embodiment, the mechanical pressure may be applied to any one or more of the other processing methods discussed herein.

[0031] In another embodiment, processing can be carried out at a pH of about 2.5 to about 7, e.g., about 3 to about 6, e.g., about 3.25 to about 5, e.g., about 3.5 to about 4.5. Of course, in one such embodiment, the low pH processing can be with respect to beverage processing; however, one or more processed food products may also undergo low pH processing.

[0032] Additionally or alternatively, in one embodiment, the processed animal food and / or treat can have a high moisture content either before processing, after processing, or both before and after processing, and still maintain a high level of non-denatured collagen. For example, in one embodiment, the processed animal food and / or treat can have a moisture content of about 5% or more, such as about 10% or more, such as about 15% or more, such as about 20% or more, such as about 25% or more, such as about 30% or more, or any range or value therebetween. In one embodiment, the animal food and / or treat can initially have a moisture content according to the above and can withstand being dried while maintaining good recovery of non-denatured collagen.

[0033] Furthermore, regardless of the processing conditions selected, processing may, in one aspect, include baking, frying, steaming, boiling, autoclaving, or otherwise heating, cooking, or sterilizing the processed animal food and / or treat. Additionally or alternatively, processing may include mechanical processing such as emulsifying, shearing, gelling, homogenizing, or other mixing and / or incorporating processes known in the art.

[0034] In another embodiment, the present disclosure has found that the processed animal food and / or treat can also contain one or more additional ingredients, and the collagen remains undenatured due to the recovery rate described below. For example, in one embodiment, the processed animal food and / or treat can contain sweeteners, preservatives, spices, coloring agents, dyes, plant proteins, fruit components, flavorings, or other animal food and / or treat components known in the art, or combinations thereof. For example, in one embodiment, the processed animal food and / or treat can contain a sweetener such as sugar or an artificial sweetener, or can contain a sweetener syrup. Furthermore, in one embodiment, the processed animal food and / or treat can contain one or more fruit components, such as fruit juice or juice. In particular, it has been found that citrus components can be used, despite having a relatively low pH, and undenatured collagen can be recovered as discussed above. Of course, other animal food and / or treat components known in the art may be used, including gluten-free flours and components in addition to conventional flours and sweeteners.

[0035] Regardless of the processing conditions selected, undenatured type II collagen can be recovered from the animal food and / or treat by post-processing of the present disclosure, such that at least about 30% or more of the undenatured collagen in the processed animal food and / or treat is recovered after processing compared to the pre-processing amount of undenatured collagen, such that, compared to the amount of undenatured collagen contained in or added to the pre-processed animal food and / or treat, e.g., about 35% or more, such as about 40% or more, such as about 45% or more, such as about 50% or more, such as about 55% or more, such as about 60% or more, such as about 65% or more, such as about 70% or more, such as about 75% or more, such as about 80% or more, such as about 85% or more, such as about 90% or more, such as about 95% or more of the undenatured collagen remaining in the processed animal food and / or treat. Thus, in one aspect, an amount of undenatured collagen may be recovered from the processed animal food and / or treat after processing according to the percentages described above.

[0036] In one embodiment, the non-denatured collagen according to the present disclosure is incorporated into processed animal foods and / or treats as a collagen composition. The collagen composition may comprise one or more of any collagens defined above, and / or in one embodiment, may comprise 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 embodiment, the collagen composition contains type II collagen alone or in combination with one or more of type I collagen, type III collagen, type IV collagen, or collagen peptides. In one embodiment, the collagen composition may comprise a mixture of type II collagen (sometimes referred to as native type II collagen) and non-denatured type II collagen. Additionally or alternatively, the collagen composition may comprise a mixture of native type II collagen and non-denatured type II collagen, in addition to additional collagens such as type I, type III, type IV, or collagen peptides.

[0037] As mentioned above, in one aspect, the processed animal food and / or treat contains a collagen composition, particularly a type II collagen composition, such as a non-denatured type II collagen composition. The type II collagen for use in the present disclosure can be obtained from any suitable source. For example, the collagen can be derived from various mammalian sources, avian sources, or various fish species, or combinations thereof. For example, the collagen can be obtained from salmon, shark, poultry, pigs, eggshells, turkey cartilage, bovine cartilage, etc. 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 from Lonza Consumer Health Inc. under the UC-II® brand. The UC-II® brand is a natural ingredient containing glycosylated non-denatured type II collagen. The collagen composition can also include hydrolyzed collagen. The collagen composition can also include pure protein or active peptide fragments. In one embodiment, the collagen composition can be free of any bone or bone material. In other embodiments, the collagen composition can be free of any transforming growth factors (TGFs), bone morphogenetic proteins (BMPs), or both, hi yet another embodiment, the collagen composition comprises type II collagen and is completely free of type I collagen.

[0038] In preparing animal tissue for oral administration, in one embodiment, the type II collagen-containing tissue may first be dissected free of surrounding tissue and diced or otherwise ground into particles. The particulate or milled cartilage may be sterilized by a means that does not affect or denature the structure of the majority of the type II collagen in the tissue, such as low-temperature processing, and formed into a dose containing therapeutically effective levels of undenatured type II collagen. The dosage is generally at least about 0.01 grams, preferably about 0.02 to about 0.5 grams, of animal tissue per dose. Because it is a natural product, some sample-to-sample variation is expected. These variations can be minimized by blending after grinding. Blending can be assisted by analytical techniques that allow for the determination of the amount of undenatured type II collagen and other components.

[0039] 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.Through this sterilization process, non-denatured type II collagen also maintains its three-dimensional shape and preserves its biologically active 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.

[0040] In one embodiment, the collagen composition is present in a serving of processed animal food and / or treat in an amount of from about 1 mg to about 600 mg per gram of processed animal food and / or treat. For example, the collagen composition may be present in the processed animal food and / or treat in an amount of about 3 mg or more, such as about 5 mg or more, for example, about 7.5 mg or more, for example, about 10 mg or more, such as about 12.5 mg or more, for example, about 15 mg or more, such as about 25 mg or more, for example, about 50 mg or more, such as about 75 mg or more, for example, about 100 mg or more, about 125 mg or more, for example, about 150 mg or more, such as about 200 mg or more, for example, about 250 mg or more, such as about 300 mg or more, for example, about 350 mg or more, such as about 400 mg or more, for example, about 450 mg or more, for example, about 500 mg or more, such as about 550 mg or more, for example, about 600 mg or more, per gram of processed animal food and / or treat. The total amount of collagen composition present in 1 gram of processed animal food and / or treat is generally less than about 700 mg, e.g., less than about 600 mg, e.g., less than about 500 mg, e.g., less than about 400 mg, e.g., less than about 300 mg, e.g., less than about 250 mg, or any range or value therebetween. Additionally or alternatively, the collagen composition may be present in the processed animal food and / or treat in an amount of about 0.01 to about 10, e.g., about 0.1 to about 9, e.g., about 0.25 to about 8, e.g., about 0.5 to about 7.5, e.g., about 0.75 to about 5 weight percent of the processed animal food and / or treat composition, or any range or value therebetween. Furthermore, in one embodiment, the collagen composition may be a type II collagen composition, and it should be understood that substantially all of the collagen in the collagen composition is type II collagen.

[0041] In one embodiment, undenatured type II collagen may comprise all or substantially all of the total type II collagen in the collagen composition and thus may be present in the processed animal food and / or treat in the amounts discussed above. However, in one embodiment, undenatured type II collagen may comprise about 1% to about 95% of the total type II collagen and / or collagen composition, e.g., about 2.5% to about 75% of the total type II collagen or total collagen composition, e.g., about 5% to about 50%, e.g., about 10% to about 40%, or any range or value therebetween. Thus, in one embodiment, undenatured type II collagen may be present in the composition in an amount of about 0.1 mg to about 100 mg, e.g., about 0.5 mg to about 75 mg, e.g., about 0.75 mg to about 50 mg, e.g., about 1 mg to about 30 mg, or any range or value therebetween.

[0042] Furthermore, in one embodiment, the collagen composition may further comprise a preservative 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, a preservative salt, or a combination thereof. In such an embodiment, Total type II collagen, including native and undenatured type II collagen, can account for about 1% to about 99% of the collagen composition, e.g., about 2.5% to about 90%, e.g., about 5% to about 80%, e.g., about 7.5% to about 70%, e.g., about 10% to about 60%, e.g., about 15% to about 50%, e.g., about 20% to about 35%, or any range or value therebetween. Thus, in one embodiment, the total amount of type II collagen, including native and undenatured type II collagen, in the collagen composition can be about 1 mg to about 1000 mg, e.g., about 2.5 mg to about 500 mg, e.g., about 5 mg to about 250 mg, e.g., about 7.5 mg to about 100 mg, e.g., about 10 mg to about 40 mg, or any range or value therebetween. Of course, in one embodiment, preservative salts are not used.

[0043] Furthermore, in one embodiment, when the type II collagen contains undenatured type II collagen, the undenatured type II collagen may have a high oxygen radical absorbance capacity (ORAC) as measured according to ORAC 6.0. Specifically, 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 (ROS), peroxyl radicals, hydroxyl radicals, superoxide anions, and peroxynitrite. Specifically, 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. Therefore, if the assay being tested contains an antioxidant, the antioxidant absorbs 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 test material.

[0044] For example, the ORAC assay for peroxyl radicals measures the antioxidant capacity of a sample to protect a fluorescent protein (fluorescein) from damage caused by peroxyl radicals generated from 2,2'-azobis(2-amidinopropane) dihydrochloride (AAPH). The ORAC assay for hydroxyl radicals measures the antioxidant capacity of a sample to protect a fluorescent protein (fluorescein) from damage caused 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 caused 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 caused by superoxide generated from xanthine oxidase. The ORAC assay for singlet oxygen measures the antioxidant capacity of a sample to protect hydroethidine from damage caused 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.

[0045] Thus, in one aspect, a collagen composition having undenatured type II collagen according to the present disclosure can have a total ORAC of about 200 μmol TE / g or more, such as about 250 μmol TE / g or more, for example, about 300 μmol TE / g or more, for example, about 350 μmol TE / g or more, for example, about 400 μmol TE / g or more, for example, about 450 μmol TE / g or more, for example, about 500 μmol TE / g or more, for example, about 550 μmol TE / g or more, for example, about 600 μmol TE / g or more, for example, about 700 μmol TE / g or more, for example, about 750 μmol TE / g or more, for example, about 800 μmol TE / g or more, for example, about 825 μmol TE / g or more, up to about 1000 μmol TE / g, or any range or value therebetween.

[0046] Further, in one aspect, a collagen composition comprising 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, for example, about 5 μmol TE / g or more, for example, about 7.5 μmol Te / g or more, for example, about 10 μmol TE / g or more, for example, up to about 10.5 μmol TE / g or more, up to about 50 μmol TE / g, or any range or value therebetween.

[0047] Similarly, in one aspect, a collagen composition having undenatured type II collagen according to the present disclosure can have an ORAC to hydroxyl radicals of about 10 μmol TE / g or more, e.g., about 15 μmol TE / g or more, e.g., about 20 μmol TE / g or more, e.g., about 25 μmol TE / g or more, e.g., about 27.5 μmol TE / g or more, e.g., about 30 μmol TE / g or more, up to about 40 μmol TE / g, or any range or value therebetween.

[0048] 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, for example, about 1 μmol TE / g or more, for example, about 1.5 μmol TE / g or more, for example, about 2 μmol TE / g or more, for example, about 2.25 μmol TE / g or more, up to about 5 μmol TE / g, or any range or value therebetween.

[0049] 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, for example about 600 μmol TE / g or more, such as about 650 μmol TE / g or more, for example about 700 μmol TE / g or more, for example about 725 μmol TE / g or more, up to about 1000 μmol TE / g, or any range or value therebetween.

[0050] Furthermore, in one aspect, a collagen composition having undenatured type II collagen according to the present disclosure can have an ORAC to hypochlorite of about 25 μmol TE / g or more, such as about 30 μmol TE / g or more, for example, about 35 μmol TE / g or more, for example, about 40 μmol TE / g or more, for example, about 45 μmol TE / g or more, for example, up to about 50 μmol TE / g or more, up to about 75 μmol TE / g, or any range or value therebetween.

[0051] Furthermore, in one embodiment, when the type II collagen comprises undenatured type II collagen, the undenatured type II collagen has a molecular weight of about 10,000 daltons or more, for example, 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, for example, 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. 000 daltons or more, such as about 60,000 daltons or more, for example about 65,000 daltons or more, for example about 70,000 daltons or more, for example about 75,000 daltons or more, for example about 80,000 daltons or more, for example about 85,000 daltons or more, such as about 90,000 daltons or more, for example about 95,000 daltons or more, for example about 100,000 or more, up to about 350,000 daltons or less, or any range or value therebetween.

[0052] Furthermore, it should be understood that, up to now, it has been contemplated that undenatured collagen may undergo processing along with processed animal foods and / or treats. However, in one aspect, undenatured collagen may be incorporated into animal foods and / or treats both before and after processing. Thus, in one aspect, undenatured collagen may be incorporated into animal foods and / or treats in the amounts discussed above. It may be included in the animal food and / or treat before processing, and the amount of collagen composition may be added or incorporated into the animal food and / or treat after processing according to the amounts discussed above with respect to the collagen composition. The amounts selected for pre-processing and post-processing may be the same or different and may be based on the total unmodified amount desired to be present in the final composition.

[0053] Various aspects and advantages have been discussed, and in one aspect, collagen composition is incorporated into a suitable delivery form before being incorporated into a dosage form as discussed below.In one aspect, the composition of the present disclosure can be included as 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 (for example, type II collagen), and the composition can be incorporated into aqueous applications using emulsion.

[0054] Nevertheless, 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 containing arabinose and galactose. Gum arabic is generally soluble in water and edible. In some embodiments, gum arabic may include 100% modified gum arabic, such as Ticamulsion® A-2010 gum arabic powder. In certain embodiments, gum arabic may be a mixture or blend of gum arabic and modified gum arabic. For example, in certain embodiments, gum arabic may include Ticamulsion® 3020.

[0055] In certain aspects, the oil-in-water emulsion contains about 10% to about 30% gum arabic by weight. In some embodiments, the oil-in-water emulsion contains about 15% to about 25% gum arabic by weight. In some embodiments, the oil-in-water emulsion contains less than about 20% gum arabic by weight, e.g., less than 15%, e.g., less than 10%, e.g., less than 5%.

[0056] The oil-in-water emulsion may also contain water. In certain embodiments, the oil-in-water emulsion contains deionized water. Nevertheless, in certain embodiments, the oil-in-water emulsion may contain any water suitable for mammalian consumption and incorporation into dietary supplements designed for mammalian consumption.

[0057] The amount of water incorporated into the oil-in-water emulsion can vary depending on the desired hygroscopic and water-soluble components incorporated into the oil-in-water emulsion. In certain aspects, the oil-in-water emulsion can contain about 5% to 35% water by weight. In some embodiments, the oil-in-water emulsion can contain about 10% to about 30% water by weight. In some embodiments, the oil-in-water emulsion can contain about 15% to about 20% water by weight. In some embodiments, the oil-in-water emulsion can contain less than about 20% water by weight, for example, less than about 15% water by weight, for example, less than about 10% water by weight.

[0058] In some embodiments, the oil-in-water emulsion may contain one or more stabilizers or suspension-promoting agents. 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.

[0059] In other embodiments, the oil-in-water emulsion may contain one or more stabilizers, such as silica. When included, silica may be present in an amount 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.

[0060] In addition, in one embodiment, oil-in-water emulsion can also contain one or more fat-soluble components or nutrients.In certain embodiments, one or more fat-soluble components or nutrients can be incorporated into the oil phase of oil-in-water emulsion.Suitable fat-soluble components include but are not limited to retinol, vitamin E derived from mixed tocopherols, beta-carotene, ubiquinone, lecithin, sunflower lecithin, vitamin D, cannabinoids, hemp extract, vitamin K, phosphatidylcholine, and combinations thereof.

[0061] In certain embodiments, at least one or more lipid-soluble components may be incorporated into the oil-in-water emulsion in an amount of from about 0% to about 50% by weight. For example, in some embodiments, the oil-in-water emulsion contains less than about 50% by weight of one or more lipid-soluble components, such as less than about 40% by weight, such as less than about 30% by weight, such as less than about 20% by weight, such as less than about 10% by weight, or such as less than about 5% by weight.

[0062] Additionally, in one embodiment, the oil-in-water emulsion may contain one or more additional antioxidants in one or more of the water-soluble phase or the oil / fat-soluble phase.

[0063] In some embodiments, the oil-in-water emulsions disclosed herein may be used in any suitable dosage form, such as gummy chewables, edible films, lozenges, liquid suspensions, syrups, lipid micelles, spray-dried dispersions, nanoparticles, etc., which may also be incorporated into further processed animal foods and / or treats. Regardless of the dosage form, it should be clear that the dosage form, animal food, and / or treat is processed at a temperature of at least 37°C as discussed herein. Accordingly, it should be clear that dosage forms, animal foods, and / or treats that do not include a processing step as defined herein are not encompassed by the above definition. Accordingly, in one embodiment, the dosage form, animal food, and / or treat does not include tablets or capsules.

[0064] The animal food or treat composition may comprise any suitable composition for consumption by mammals. Such compositions include complete foods or food supplements, such as treats and snacks, intended to provide the necessary dietary requirements of mammals. The food composition may comprise pellets, treats, bars, canned prepared foods, or any other functional food composition.

[0065] The animal food and / or treat compositions 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, and 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, lycopersicon esculentum, and the like. Flavorings such as squirrel, anise, vanilla, and peach, banana, grape, strawberry, blueberry, raspberry, and mixed berry; glidants such as fumed silica, talc, and magnesium carbonate; lubricants such as talc, silica, and vegetable stearin, magnesium stearate, and fats containing stearic acid; antioxidants; vitamins; retinyl palmitate; selenium; the amino acids cysteine ​​and methionine; preservatives such as citric acid, sodium citrate, and parabens; adsorbents; sweeteners such as sucrose and sucralose; and flavorings such as petrolatum and mineral oil. Vehicles include:

[0066] In one aspect, the processed animal food and / or treat 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 in a granular form.

[0067] In one embodiment, the animal food and / or snack 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 the stabilizer package. The coating material may, for example, contain at least one fat. According to the present disclosure, the above-mentioned components may be added to any suitable pharmaceutical composition in addition to the composition of the present disclosure. For example, the above-mentioned components may be added to any pharmaceutical composition containing carnitine or amino acids.

[0068] The polymer binder and stabilizer package may be combined with the processed animal food and / or treat composition in a manner that the stabilizer package is homogeneously incorporated 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 pharmaceutically acceptable polymer, such as a film-forming polymer and / or a polysaccharide. Specific examples of polymer binders that can 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, for example, at least about 8% by weight, for example, at least about 10% by weight, for example, at least about 15% by weight. The one or more polymeric binders are present in the composition in an amount less than about 50% by weight, such as less than about 45% by weight, for example, less than about 40% by weight, such as less than about 35% by weight, for example, less than about 30% by weight.

[0069] In one embodiment, the polymer binder can include a starch, such as a modified starch. The starch can be derived from, for example, corn or waxy corn. In one embodiment, the starch can include HI-CAP100 starch sold by National Starch and Chemical Company.

[0070] In another embodiment, the polymer binder may comprise arabinogalactan.Arabinogalactan is a soluble polysaccharide that not only functions as a polymer binder but also can provide other benefits.For example, arabinogalactan can enhance adaptive immune response in some situations.Arabinogalactan is described, for example, in U.S. Patent No. 8,784,844, which is incorporated herein by reference.

[0071] In one embodiment, arabinogalactan can be used as a polymer binder. Larch arabinogalactan is a highly branched polysaccharide composed of galactose and arabinose units in an approximately 6:1 ratio. It is extracted from large trees. The polysaccharide has a galactan backbone with galactose and arabinose side chains. Arabinogalactan is commercially available from Lonza Ltd.

[0072] 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 carboxylic acid salt. In a 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. The combination of oxide particles and a carboxylic acid salt has been found to provide numerous advantages and benefits when combined with the composition. For example, the stabilizer package has been found to stabilize the composition and reduce its hydrophobicity. This composition also produces a granular, flowable product that is easy to handle.

[0073] The oxide particles that may be added to the animal food and / or treat composition may comprise silica. For example, the oxide particles may comprise precipitated silica particles. The silica particles may have a particle size (d50, laser diffraction after ISO test 13320) of less than about 55 microns, for example, less than about 40 microns, for example, less than about 30 microns, for example, less than about 25 microns, for example, less than about 20 microns, for example, less than about 15 microns, for example, less than about 12 microns, for example, less than about 10 microns, for example, less than about 8 microns, for example, less than about 6 microns, for example, less than about 4 microns, for example, less than about 2 microns, for example, less than about 1 micron. The particle size is typically greater than about 0.5 microns, for example, greater than about 1 micron. The particles may have a specific surface area (ISO test 9277) of greater than about 120 m / g, for example, greater than about 130 m / g, for example, greater than about 150 m / g, for example, greater than about 170 m / g, for example, greater than about 200 m / g, for example, greater than about 220 m / g. The specific surface area is generally less than about 500 m / g. Oxide particles such as silica particles can be present in the pharmaceutical composition in an amount greater than about 0.01 wt%, for example, greater than about 0.05 wt%, for example, greater than about 0.1 wt%. Oxide particles are generally present in an amount less than 5 wt%, for example, less than about 2 wt%, for example, less than about 1.5 wt%, for example, less than 0.5 wt%.

[0074] 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 a carbon chain length of, for example, about 6 carbon atoms to about 40 carbon atoms, e.g., 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 wt %, e.g., greater than about 1 wt %, e.g., greater than about 1.5 wt %. The salt of a carboxylic acid is generally present in an amount less than about 5 wt %, e.g., less than about 4 wt %, e.g., less than about 3 wt %.

[0075] In addition to the polymer binder and stabilizer package, the composition may contain various other components and ingredients. In one embodiment, for example, the composition may contain a citric acid ester, such as a mono-citric acid ester and / or a diglyceride of a fatty acid. The composition may also contain 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 amounts greater than about 0.05% by weight, for example, greater than about 0.1% by weight.

[0076] Furthermore, in one embodiment, the processed animal food and / or treats can be formulated into foods and / or treats for sports or daily nutritional purposes. In such an embodiment, the processed animal food and / or treats can further include at least one vitamin, such as at least one of vitamin B, vitamin C, and vitamin E. The vitamins can be present in amounts of about 50 ... Vitamin E may be present in processed animal foods and / or treats in amounts ranging from about 0 μg / g to about 5000 μg / g, e.g., about 100 μg / g to about 4500 μg / g, e.g., about 250 μg / g to about 4000 μg / g, e.g., 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 processed animal foods and / or treats in amounts ranging from about 100 μg / g to about 1000 μg / g, e.g., about 250 μg / g to about 750 μg / g, e.g., about 400 μg / g to about 600 μg / g, or any range or value therebetween. In another embodiment, vitamin C is present in the processed animal food and / or treat in an amount from about 1000 μg / g to about 5000 μg / g, e.g., from about 2000 μg / g to about 4000 μg / g, e.g., from about 3000 μg / g to about 3750 μg / g, or any range or value therebetween.

[0077] Furthermore, in some embodiments, the processed animal food and / or treat contains at least one mineral, such as at least one of potassium, magnesium, zinc, or calcium. The mineral may be present in the processed animal food and / or treat in an amount of about 1 mg / g to about 50 mg / g, e.g., about 2.5 mg / g to about 45 mg / g, e.g., about 5 mg / g to about 40 mg / g, or any range or value therebetween. The above range may be for any one mineral or the total amount of one mineral. In one embodiment, the processed animal food and / or treat contains potassium in an amount of about 9.5 mg / g to about 12 mg / g, e.g., about 9.75 mg / g to about 11.5 mg / g, e.g., about 10 mg / g to about 11 mg / g, or any range or value therebetween. Similarly, in one embodiment, the processed animal food and / or treat contains magnesium in an amount of about 1 mg / g to about 10 mg / g, e.g., about 2.5 mg / g to about 7.5 mg / g, e.g., about 4 mg / g to about 6 mg / g, or any range or value therebetween. Furthermore, in one embodiment, the processed animal food and / or treat 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.

[0078] Furthermore, the processed animal food and / or treat may further include at least one additive that improves sports performance or contributes to reducing oxidative stress. For example, in one embodiment, the additive may be one or more of curcumin, spirulina, astaxanthin, or other carotenoids. Furthermore, in one embodiment, the present disclosure may include one or more microalgae with high superoxide dismutase (SOD) and / or ORAC levels. In particular, such microalgae may help further reduce oxidative stress and may contribute additional anti-inflammatory properties and protection against infections, including improved immune health. Furthermore, in one embodiment, the additive may include one or more probiotics.

[0079] Additionally, in one embodiment, the processed animal food and / or treat may be formulated to include other components for daily nutrition suitable for consumption and dietary support of a non-human mammal, such as additional protein sources, one or more grains, dietary fiber, starches, fruits, vegetables, or combinations thereof. In one embodiment, the additional protein sources may be plant or animal-based, or a combination thereof.

[0080] Nevertheless, in one embodiment, for example, the processed animal foods and / or treats of the present disclosure are specifically formulated to improve joint health, muscle health, cartilage health, bone health, or a combination thereof. For example, the processed animal foods and / or treats may be used to treat non-arthritic joint pain, joint discomfort in healthy mammals, lack of joint flexibility in healthy mammals, muscle pain in healthy mammals, or poor adaptation in healthy mammals. The processed animal foods and / or treats of the present disclosure may be used to improve immune health, bone health, or brain health, and may also improve triglyceride and / or cholesterol levels in healthy mammals and / or mammals that regularly engage in physical activity and / or vigorous physical activity. Furthermore, the processed animal foods and / or treats of the present disclosure may also improve joint health, bone health, muscle health and pain, and cartilage health due to age-related decline. Thus, in one embodiment, the processed animal foods and / or treats may also include, in addition to undenatured collagen, one or more additional joint supplements, such as hydroxycitric acid, glucosamine, chondroitin, or a combination thereof, and / or a collagen absorption enhancer, such as vitamin C.

[0081] However, it should be understood that in one embodiment, additional joint supplements are not required in processed food and / or treats.For example, in one embodiment, processed animal food and / or treats can comprise the ingredients for the daily nutrition of non-human mammals, and comprise high levels of crude protein, fat, and dietary fiber.In one embodiment, crude protein can comprise one or more protein meals, such as dietary protein products formed from one or more animal protein sources, which can comprise mammalian bone, organ, cartilage, and skin.

[0082] For example, in one embodiment, a non-human processed food and / or treat may contain about 18% to about 40% crude protein by weight, about 4% to about 30% fat by weight, and about 2% to about 20% total dietary fiber by weight. In another embodiment, the animal processed food and / or treat may be a low-fat diet to promote weight loss. A typical low-fat diet may contain about 18% to about 22% protein by weight, about 8% to about 10% fat by weight, and about 1% to about 3% crude fiber by weight. In particular, it should be clear to those skilled in the art that appropriate levels for non-human mammals do not correspond to levels required or recommended for human nutrition.

[0083] Furthermore, the processed animal food and / or treats can be suitable for administration to any non-human mammal. For example, the mammal can be a dog, cow, cat, or horse. The composition can be fed to non-human mammals of any age, from birth to adulthood in the mammal. In various embodiments, the mammal can be a dog, cat, horse, pig, sheep, or cow. In many embodiments, the mammal can be in early to late adulthood. For example, an active mammal can have an age that is at least 10%, such as at least 15%, such as at least 20%, such as at least 25%, such as at least 30%, such as at least 35%, such as at least 40%, such as at least 45%, such as at least 50%, such as at least 55%, such as at least 60%, such as at least 65%, such as at least 70%, such as at least 75%, such as at least 85%, such as at least 90%, such as at least 95% of its average lifespan. The mammal may have an age that is less than about 95% of its average lifespan, such as less than about 90%, for example, less than about 85%, for example, less than about 80%, for example, less than about 75%, for example, less than about 70%, for example, less than about 65%, for example, less than about 60%, for example, less than about 55%, for example, less than about 50%, for example, less than about 45%, for example, less than about 40%, for example, less than about 35%, for example, less than about 30%, for example, less than about 25%, for example, less than about 20%, for example, less than about 15%, for example, less than about 10%. Determination of lifespan may be based on actuarial tables, calculations, etc.

[0084] Nevertheless, 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.

[0085] Example 1 Animal and plant-based dry food Batches of animal-based dry food were formed by mixing chicken breast, dried egg, whole wheat flour, wheat flour, oatmeal, cornmeal, oat bran, canola oil, and water. Batches of vegetable-based dry food were formed by mixing Natural Balance plant-based dry dog ​​formula with dried egg, whole wheat flour, wheat flour, oatmeal, cornmeal, oat bran, canola oil, and water. Both plant- and animal-based dry foods were formulated with UC-II® brand undenatured type II collagen, as shown in Table 1 below, and cooked in a 120°C oven and dried to 10% moisture. After cooking and drying, samples were tested for undenatured collagen retained after processing and cooking, and are presented in Table 1 as % recovery. Animal-based foods are designated as "A" samples, and plant-based foods are designated as "P" samples. [Table 1]

[0086] As shown in Table I, both animal and plant-based foods demonstrated excellent recovery of undenatured type II collagen after formulation and cooking, even at moisture levels typical of pet foods. The greater than 100% recovery is at least partially due to the endogenous undenatured type II collagen contained in the chicken meat used in the animal-based foods, but may also be partially due to the moisture content of the foods. While not wishing to be bound by theory, it is believed that the increased recovery of undenatured type II collagen may be at least partially due to interactions with other nutrients contained in the foods. Furthermore, due to the low variability and potential of laboratory assays, for plant-based samples where greater than 100% recovery is reported, this value may be considered essentially 100% recovery.

[0087] Example 2 Extruded Pet Food Animal-based pet foods were formed according to Example 1, except that the pet food was extruded into the desired shape before cooking. Prior to injection molding, UC-II® brand undenatured type II collagen was added to the extruded powder mixture, as shown in Table 2 below. [Table 2]

[0088] Example 3 Extruded Dental Stick Potato flour, coconut glycerin, natural chicken flavor, pea protein, canola oil, powdered cellulose, coconut oil, dried cultured nonfat milk, citric acid (preservative), sodium hexametaphosphate, vanilla, natural mint flavor, zinc propionate, mixed tocopherols (preservative), green tea extract, and rosemary extract were combined to form a dental stick mixture. The dental stick mixture was then extruded and baked as discussed above for forming pet food, except that the extruded product was formed into a dental stick. The dental stick contained UC-II® brand undenatured type II collagen, as shown in Table 3 below. A control dental stick was formed in the same manner as the example, except that UC-II® brand undenatured type II collagen was not added. Both the example and the control were subjected to recovery testing, the results of which are shown in Table 3. [Table 3]

[0089] Example 4 Brown rice flour containing undenatured type II collagen Brown rice flour was blended with various levels of UC-II® brand undenatured type II collagen. The blended brown rice flour was found to be stable (e.g., maintained the level of undenatured type II collagen) and suitable for use in preparing processed foods such as pet food and / or treats. Brown rice was blended with UC-II® brand undenatured type II collagen in ratios of 80:20, 50:50, and 20:80 UC-II® brand undenatured type II collagen to brown rice flour. After blending and incorporation (e.g., no separation), samples were allowed to dry and analyzed to determine the level of undenatured type II collagen remaining in the blended brown rice, and the results are shown in Table 4. [Table 4]

[0090] Example 5 Pet food and / or treats containing undenatured collagen for inflammation and joint pain in exercised Labrador retrievers Objective: The purpose of this study was to evaluate the effectiveness of undenatured type II collagen in mediating inflammation and joint pain in Labrador retrievers during and after endurance running exercise. UC-II® brand undenatured type II collagen is believed to reduce inflammation and joint pain when administered orally. This will be evidenced by changes in inflammatory and general health indicators, including: 1) improvement or significant maintenance of gait parameters across pressure walkways; 2) improvement or significant maintenance of joint health and inflammatory biomarkers (CK, COMP, IL-6, and N:L ratio); 3) improvement or significant maintenance of objective lameness scores; and 4) improvement in activity during running exercise.

[0091] Animals and Housing: Forty healthy Labrador retrievers (20 males and 20 females) will be used during the study. Body weights ranged from 22 to 38 kg, with an average of 30 kg. All dogs were fit at the start of the study, with a body composition score of 3 to 6. The dogs' ages ranged from 5 to 11 years, with an average of 7.5 years. All dogs were housed individually overnight and were socially grouped in an outdoor yard for 6 hours per day, depending on the weather and study conditions. All dogs received their assigned food and treatment once daily in the morning and always had free access to an automatic water dispenser. All dogs were vaccinated on schedule and received monthly preventative vaccinations. Receive heartworm and parasite prevention.

[0092] Running Exercise: All dogs begin an outdoor endurance running program after a two-week loading period. The running regimen is prescribed as follows: Weeks 3–5: 2 miles twice per week; Weeks 6–8: 4 miles twice per week; Weeks 9–11: 5 miles twice per week; Week 12: 2 miles twice per week; and Week 13: 10 miles once per week. During the runs, all dogs wear Article® accelerometer collars to quantify activity intensity and Garmin® GPS collars to quantify distance and speed. All dogs run in groups with an all-terrain vehicle and are free to run, stop, swim, play, and more. Every effort is made to keep dogs active during each run, but dogs that refuse are allowed to return to their kennels.

[0093] Gait analysis: Gait analysis was performed using a pressure walkway (Gait4Dogs) on all days: baseline, 24 hours before the first 3-mile run, 24 hours and 48 hours after the first 3-mile run, 24 hours before the 10-mile run, and 24 hours and 48 hours after the 10-mile run. Each dog passed through the pressure walkway 6-12 times at each time point, obtaining at least 3-4 valid walkway samples for analysis. For each valid walk, various temporal, pressure, and spatial parameters were collected for each limb. Calculations were also performed to provide average symmetry ratios for all limbs, forelimbs, and hindlimbs, as well as for left and right forelimbs and hindlimbs for each parameter.

[0094] Biological Sampling: For biomarker and hematological purposes, blood samples will be collected at baseline, 24 hours before the first 3-mile run, 24 hours after the first 3-mile run, 24 hours before the 10-mile run, and 24 hours after the 10-mile run. Biomarkers creatine kinase (CK), interleukin-6 (IL-6), and cartilage oligomatrix protein (COMP) will be assessed using commercially available ELISA kits. Blood tests will be analyzed using an automated hematology machine (Abaxis HM5).

[0095] Pain assessment: Pain is assessed by subjective analysis using the LOAD questionnaire. Three trained technicians observe the dogs in the kennel and outdoors and score them based on the questionnaire items. Data from all three technicians is collected and analyzed.

[0096] Using the statistical method JMP 14.0, mixed models were created to compare the differences and changes in intake, body weight, blood biomarkers, blood tests, running activity, running locomotion speed, and gait analysis between treatments. Gender was analyzed as a fixed effect. A p-value of 0.05 or less was considered significant.

[0097] Experimental Design: To evaluate the effectiveness of undenatured type II collagen in mediating inflammation and joint pain during and after exercise, 40 healthy Labrador retrievers (20 males and 20 females) were used in the study. The dogs were divided into two equalized treatment groups based on age, weight, and genetics. The treatment group received a 40 mg capsule of the test supplement UC-II® brand undenatured type II collagen daily, while the placebo group received a capsule of maltodextrin daily. After a two-week acclimation / loading period, each dog began an outdoor endurance running regimen with gradually increasing distances over 11 weeks. Each dog underwent blood draws, gait analysis, and pain assessments at baseline before the start of the loading period, before and after the first 3-mile run, and before and after the 10-mile run.

[0098] Example 6 Effect of undenatured type II collagen (UTIIC) on sodium monoiodoacetate-induced osteoarthritis (OA) in rats Male Wistar rats were divided into three groups: (i) control, (ii) vehicle-treated MIA-induced rats, and (iii) UC-II® brand undenatured type II collagen-treated MIA-induced rats (4 mg / kg). OA was induced in male Wistar rats by intra-articular injection of sodium monoiodoacetate (MIA: 1 mg). Treatment began one week before MIA injection and continued for 30 days. Biomarker testing was performed 24 days after MIA. The results of metabolic marker testing are shown in Table 5, and inflammatory markers are shown in Table 6. [Table 5] [Table 6]

[0099] As shown in Table 6, undenatured type II collagen was shown to reduce MIA-induced Kellgren-Lawrence scores (53.3%) in OA by reducing articular cartilage damage in rats (P<0.05). Pro-inflammatory cytokines [IL-1β (7.8%), IL-6 (18.0%), TNF-α (25.9%), COMP (16.4%)] were also significantly reduced. Suppression of the production of IL-1β (32.4%) and CRP (32.4%) was also found after treatment with undenatured type II collagen (P<0.0001). Undenatured type II collagen was also found to inhibit the production of PGE2 (19.6%) and the expression of IL-1β, IL-6, TNF-α, COX-2, MCP-1, NF-κB, MMP-3, and RANKL (P<0.001). Col-1 and OPG levels were increased in MIA-induced OA rats (P<0.001). Furthermore, MIA+UCII mice were found to have higher superoxide dismutase levels (44.19±2.15 U / mL) compared with MIA mice (37.98±3.19). Thus, it is further evident that undenatured type II collagen in its undenatured form is beneficial to mammals in terms of, for example, joint health, cartilage health, bone health, and muscle health, to name a few.

[0100] These and other modifications and variations to the present invention may be practiced by those of ordinary skill in the art without departing from the spirit and scope of the 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. Furthermore, 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 processed animal food and / or treat composition comprising: Contains undenatured type II collagen, A composition wherein the processed animal food and / or treat is processed at a temperature of about 37°C or higher.

2. The composition of claim 1 , wherein the composition comprises a processed animal food.

3. The composition of claim 1 , wherein the composition comprises a processed animal treat.

4. 4. The composition of any one of claims 1 to 3, wherein the undenatured type II collagen is incorporated into the processed animal food and / or treat composition as part of a collagen composition comprising one or more different types of collagen in addition to the undenatured type II collagen.

5. The composition of any one of claims 1 to 4, wherein the one or more different types of collagen comprise native type II collagen, collagen peptides, or a mixture thereof.

6. 6. The composition of claim 1, wherein an amount of undenatured type II collagen is incorporated into the composition prior to processing, and at least about 30% or more of the undenatured type II collagen is recovered after processing.

7. 7. The composition of claim 6, wherein 60% or more of the undenatured type II collagen is recovered after processing.

8. 8. The composition of any one of claims 1 to 7, wherein the processed animal food and / or treat undergoes processing that includes withstanding temperatures of about 40°C or higher.

9. The composition of any one of claims 1 to 8, wherein the processing lasts from 6 seconds to about 2 hours.

10. 10. The composition of any one of claims 1 to 9, wherein the animal food and / or treat comprises one or more of a protein source, a grain, a flavoring, or a coloring agent.

11. 11. The composition of any one of claims 1 to 10, wherein the processed animal food and / or treat comprises from about 15% to about 40% by weight crude protein, from about 5% to about 30% by weight fat, and from about 2% to about 20% by weight dietary fiber.

12. The composition of any one of claims 1 to 11, wherein the processed animal food and / or treat comprises processed animal meal.

13. 1. A method of forming a processed animal food and / or treat, comprising: combining undenatured type II collagen with at least one animal food and / or treat component; processing the undenatured type II collagen and at least one animal food and / or treat component at a temperature of about 37°C or greater; The method, wherein at least about 30% or more of the undenatured type II collagen is recovered in the processed animal food and / or treat after processing compared to the amount of undenatured type II collagen before processing.

14. The processing comprises combining the undenatured type II collagen and at least one animal food product and 14. The method of claim 13, further comprising exposing the snack component to a temperature of about 40°C.

15. 15. The method of claim 13 or 14, wherein said processing comprises exposing said undenatured type II collagen and at least one animal food and / or treat component to a temperature of about 37°C for at least about 10 minutes.

16. 14. The method of claim 13, wherein said processing comprises exposing said undenatured type II collagen and at least one animal food and / or treat component to a temperature of about 100°C for at least about 1 minute.

17. The method of any one of claims 13 to 16, wherein the processing comprises a pelletizing process.

18. The method of any one of claims 13 to 17, wherein the processing further comprises extruding the composition.

19. The method of any one of claims 13 to 17, wherein the processing further comprises injection molding the composition.

20. 20. A method for improving one or more of joint health, cartilage health, muscle health, bone health, skin health, inflammatory markers, or fitness, comprising administering to a non-human mammal an effective amount of a processed animal food and / or treat according to any one of claims 1 to 19.

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