Animal food composition containing glycyrrhizin source

Abstract

The present disclosure relates to an animal food composition comprising a glycyrrhizin source for use in the prevention and / or treatment of allergic inflammatory skin diseases in animals, the animal food composition being formulated to supply glycyrrhizin to the animal in a daily dose in an amount of at least about 0.02 mg / kg body weight.

Description

Description of Related Applications

[0001] This application claims the benefit of priority to European Patent Application No. 22170352.3, filed on April 27, 2022, the content of which is incorporated herein by reference in its entirety.

Technical Field

[0002] The subject matter disclosed herein relates to the field of animal food compositions for preventing and / or treating allergic inflammatory skin diseases, particularly for preventing and / or treating atopic dermatitis.

Background Art

[0003] Reports strongly suggest that allergic diseases are increasing, particularly in Western societies (Non-Patent Document 1).

[0004] Allergic inflammatory skin diseases are chronic diseases characterized by clinical signs such as erythema, pain, redness, swelling, and the appearance of small blisters or papules on the skin. These diseases are the second most common allergic skin diseases in animals, particularly dogs, surpassed only by flea allergy. These allergic reactions can be caused by normally harmless substances such as grass, mold spores, house dust mites, and other environmental allergens.

[0005] The initial symptoms are usually associated with dermatitis and pruritus due to scratching and abrasion by affected animals, which results from the interaction between genetic and environmental factors. This allergic symptom appears as eczematous skin, and animals with atopic dermatitis (such as dogs) often suffer from pruritus, i.e., severe itching, hair loss, skin abrasion due to deep scratching, frequent licking of the paws, and excessive tear production. Secondary skin problems, including skin infections and excessive sebum excretion, are also common. In particular, atopic dermatitis is one of the most common allergic inflammatory skin diseases.

[0006] A common feature of such diseases across many symptoms is an immune response bias towards the expression of so-called T helper 2 cell (Th-2) cytokines, which is the expression of inflammatory mediators associated with the promotion of IgE and eosinophil responses. For example, increased interleukin IL-4 or IL-5 relative to Th-1 helper cell cytokines such as interferon-γ (IFN-γ) or Th-17 helper cell cytokines such as IL-17 seems to make the immune response more likely to result in an allergic phenotype. On the other hand, the Th-1 cytokine IFN-γ is known to inhibit IgE synthesis, Th-2 cell proliferation, and IL-4 receptor expression on T cells (Non-Patent Documents 2 and 3). Thus, bioactive substances that regulate the immune response to suppress Th-2 and even promote Th-1 cell proliferation and cytokine production can prove useful in the management of allergies and allergic diseases.

[0007] Some treatments for atopic dermatitis and pruritus, including the use of ultrafine palmitoylethanolamide (Non-Patent Document 4), antihistamines such as fexofenadine, or drugs such as cyclosporine, have already been described. Nevertheless, most of the available drugs are not suitable for long-term use due to significant side effects (and often exorbitant prices). Immunotherapy can improve symptoms if the allergen can be identified, but the success rate still varies. The ability to reduce drug dependence would be seen as a valuable contribution to the field and is the main purpose of nutritional interventions.

[0008] Indeed, there is now evidence that genetic factors can affect the immune system as well as the functional / structural characteristics of skin protectants. Known environmental factors are the allergen load and the microbial flora present on the skin. Drugs currently prescribed for allergic conditions typically target the individual characteristics of the "march" of the disease, which is typically immune-related, but only suppress the disease rather than provide long-term relief or cure. Nutritional management strategies are more likely to be successful if they target multiple characteristics simultaneously.

[0009] Numerous natural products derived from plant sources have shown potential for immunomodulation (Non-Patent Document 5, Non-Patent Document 6, and Non-Patent Document 7). Some of these phytochemical products are said to regulate immunological pathways related to allergic conditions. However, the level of scientific evidence for these beneficial effects varies, and in some cases, there is little evidence beyond what is recorded in traditional uses.

[0010] In particular, the roots of Spanish licorice (Glycyrrhiza glabra licorice), which have previously shown immunomodulatory properties, have attracted attention. Indeed, glycyrrhizin, a typical chemical component of licorice also known as glycyrrhizic acid, has been shown to affect the maturation and signaling activity of dendritic cells by suppressing specific Th-2 type responses and, concomitantly, increasing Th-1 responses (Non-Patent Document 8 and Non-Patent Document 9) (Non-Patent Document 10, Non-Patent Document 11, and Non-Patent Document 12). In addition to glycyrrhizin, other important components of licorice, namely isoliquiritigenin and naringenin, have been shown to affect the induction and function of regulatory T cells (Non-Patent Document 13).

[0011] Thus, in the past, glycyrrhizin was included in animal food compositions for the prevention and / or treatment of allergic inflammatory diseases: for example, Patent Document 1, the disclosure of which is hereby incorporated by reference in its entirety, describes the use of a composition for this purpose in animals, which combines (i) a glycyrrhizin source with (ii) one or more curcuminoid sources.

[0012] However, the amount of glycyrrhizin included in animal food compositions is usually very small. In fact, it has been previously shown that there is a relationship between the intake of glycyrrhizin in dogs and hypertension. Therefore, the administrative safety advice for glycyrrhizin is that, especially for dogs, the presumed safe intake (PSI) is 1 mg / kg of a complete feed giving a presumed 0.3 mg / day or 0.02 mg / kg body weight (Bwt) / day (EFSA Journal based on the study of Non-Patent Document 14, recommendation in 2015).

Prior Art Documents

Patent Documents

[0013]

Patent Document 1

Non-Patent Documents

[0014]

Non-Patent Document 1

Non-Patent Document 2

Non-Patent Document 3

Non-Patent Document 4

Non-Patent Document 5

Non-Patent Document 6

Non-Patent Document 7

Non-Patent Document 8

Non-Patent Document 9

Non-Patent Document 10

Non-Patent Document 11

Non-Patent Document 12

Non-Patent Document 13

Non-Patent Document 14

Summary of the Invention

Problems to be Solved by the Invention

[0015] Thus, the amount of the active compound contained in the composition such that the required anti-inflammatory properties are imparted is limited. Therefore, there is still a need for treatment options for allergic inflammatory skin diseases such as atopic dermatitis. Also, alternative natural products for treating or preventing such diseases are still needed. Therefore, there is still a need for a novel regulated diet designed to improve both immune function and barrier defense.

[0016] Furthermore, for allergic inflammatory skin diseases such as atopic dermatitis, there is still a need for treatment options, particularly novel regulated diets, that are not only particularly effective for the disease but also safe and have limited side effects for the individual to be treated.

Means for Solving the Problems

[0017] The objects and advantages of the disclosed subject matter are set forth in the following description and will be apparent from the description, and will be understood by practice of the disclosed subject matter.

[0018] A first aspect of the present disclosure is an animal food composition comprising a source of glycyrrhizin for use in a method of preventing and / or treating an inflammatory disease or disorder, an allergic inflammatory skin disease, the animal food composition being formulated to supply an animal with a daily amount of glycyrrhizin in an amount of at least about 0.02 mg / kg body weight.

[0019] In particular, the animal food composition can be formulated to supply an animal with a daily amount of glycyrrhizin in an amount from about 0.02 mg / kg body weight to about 2 mg / kg body weight.

[0020] In particular, the animal food composition can be formulated to supply an animal with a daily amount of glycyrrhizin in an amount from about 0.03 mg / kg body weight to about 2 mg / kg body weight, particularly from about 0.05 mg / kg body weight to about 1 mg / kg body weight, particularly from about 0.1 mg / kg body weight to about 0.8 mg / kg body weight, particularly from about 0.2 mg / kg body weight to about 0.6 mg / kg body weight.

[0021] The present disclosure also relates to an animal food composition comprising a glycyrrhizin source for use as a drug, the animal food composition being formulated to supply an animal with a daily dose of glycyrrhizin in an amount of at least about 0.02 mg / kg body weight.

[0022] In a particular embodiment, the animal food composition is selected from the group consisting of a nutritionally complete animal food composition, a dietary supplement, and a topping composition.

[0023] In a specific embodiment, the glycyrrhizin source of the animal food composition comprises licorice root extract.

[0024] In a particular embodiment, the animal food composition further comprises linoleic acid.

[0025] In a particular embodiment, the animal food composition further comprises EPA / DHA, taurine, lutein, vitamin E and / or a combination thereof.

[0026] In a particular embodiment, the allergic inflammatory skin disease is selected from the group consisting of atopic dermatitis, flea allergic dermatitis, urticaria, insect sting allergy, angioedema, inhalant allergy, inhalant allergic dermatitis, food allergic dermatitis, contact dermatitis, miliaria, eosinophilic granuloma, head and neck pruritus, and generalized pruritus.

[0027] A second aspect of the present disclosure is for use in a method of preventing and / or treating an inflammatory disease or disorder, an allergic inflammatory skin disease, (i) a first part comprising a glycyrrhizin source, and (ii) a second part comprising one or more additional components, A kit comprising, the kit being formulated to supply an animal with a daily dose of glycyrrhizin in an amount of at least about 0.02 mg / kg body weight.

[0028] A third aspect of the present disclosure relates to an animal food composition comprising a glycyrrhizin source, wherein the amount of glycyrrhizin is at least about 5 mg / kg based on the total mass of the dry matter of the animal food composition.

[0029] In particular, the amount of glycyrrhizin can be from about 5 mg / kg to about 100 mg / kg based on the total mass of the dry matter of the animal food composition.

[0030] In particular, the amount of glycyrrhizin can be from about 5 mg / kg to 100 mg / kg, particularly from about 10 mg / kg to about 75 mg / kg, particularly from about 15 mg / kg to about 50 mg / kg, and particularly from about 20 mg / kg to 45 mg / kg based on the total mass of the dry matter of the animal food composition.

[0031] In a particular embodiment, the animal food composition constitutes kibble and / or the animal food composition further comprises a protein source.

[0032] In particular, the animal food composition comprises a nutritionally complete food composition.

[0033] In particular, the animal food composition or kit is formulated for oral administration.

[0034] In particular, the glycyrrhizin source in the animal food composition or kit comprises licorice root extract.

[0035] In particular, the animal food composition may contain one or more additional ingredients.

[0036] In particular, one or more additional ingredients of the animal food composition or kit can be selected from linoleic acid, EPA / DHA, taurine, lutein, vitamin E and / or combinations thereof.

[0037] The foregoing has outlined the features and technical advantages of the present application in broad terms so that the following detailed description can be better understood.

[0038] Additional features and advantages of the present application that form the subject matter of the claims of the present application are described below. It will be recognized by those skilled in the art that the disclosed concepts and specific embodiments can be readily utilized as a basis for modifying or designing other structures for achieving the same objectives of the present application. Those skilled in the art should also be able to notice that such equivalent configurations do not depart from the spirit and scope of the present application as set forth in the appended claims. The novel features believed to be characteristic of the present application will be better understood from the following description, together with further objectives and features, with respect to both its construction and method of operation.

Brief Description of the Drawings

[0039]

Figure 1

Figure 2

Figure 3

DETAILED DESCRIPTION OF THE INVENTION

[0040] The presently disclosed subject matter relates to making available products for preventing and / or treating inflammatory diseases or disorders, allergic inflammatory skin diseases, particularly atopic dermatitis, i.e., canine atopic dermatitis, in animals, particularly dogs. In particular, this product may be useful for assisting in the management of canine atopic dermatitis.

[0041] The presently disclosed subject matter provides, herein, an animal food composition comprising a glycyrrhizin source in an amount greater than the recommended daily dose.

[0042] Surprisingly, according to the presently disclosed subject matter, the aqueous extract of licorice root can inhibit the release of interleukin-5 (IL-5) from activated T cells within a concentration range that is non-toxic and does not stimulate proliferation or the release of interleukin-2 (IL-2). Furthermore, the presently disclosed subject matter was able to show that even when consuming an amount of glycyrrhizin that is more than about 20 times the recommended daily dose for dogs, no significant blood pressure homeostasis is induced in adult dogs. Additionally, the presently disclosed subject matter also shows that even when consuming an amount of glycyrrhizin that is more than about 20 times the recommended daily dose for dogs, no changes are induced in potassium excretion or aldosterone secretion, and this result indicates that this high dose of glycyrrhizin does not affect renal function.

[0043] Therefore, the presently disclosed subject matter relates to an animal food composition comprising a source of glycyrrhizin that can be provided to animals, particularly canids such as dogs, in an amount greater than the normal recommended amount, while such a food composition remains safe, i.e., it does not cause an increase in blood pressure and does not affect renal function such as not increasing potassium excretion or aldosterone secretion. Such a composition is useful for preventing and / or treating inflammatory diseases or disorders, allergic inflammatory skin diseases in animals, more particularly in animals suffering from atopic dermatitis. In a particular embodiment, the animal is a canid. In a specific embodiment, the canid is a dog.

[0044] As used herein, inflammatory diseases or disorders include inflammation of the gastrointestinal tract.

[0045] Furthermore, the presently disclosed subject matter provides an animal food composition that exhibits good overall palatability and digestibility, particularly in dogs, and remains stable throughout its manufacturing process.

[0046] More particularly, the presently disclosed subject matter also provides animal food compositions, which may include nutraceuticals, topping compositions, and / or complete food compositions. The animal food compositions disclosed herein, consisting of either a nutraceutical or a complete food composition, contain a source of glycyrrhizin in the amounts specified herein. In some embodiments, the food composition is in the form of a wet food composition, such as a gravy composition. Alternatively, the food composition is in the form of a dry food composition, such as in the form of kibble.

[0047] For clarity, and not by way of limitation, the detailed description of the presently disclosed subject matter is divided into the following subsections: 1. Definitions; 2. Animal food compositions; 3. Methods of preparing animal food compositions; and 4. Therapeutic applications.

[0048] 1. Definitions The terms used herein generally have their ordinary meaning within the context of the present disclosure and in the particular context in which each term is used. Particular terms are discussed below or elsewhere in this specification to provide additional guidance to those skilled in the art in describing the compositions and methods of the present disclosure, and their methods of manufacture and use.

[0049] For the purposes of interpreting this specification, the following definitions apply, and terms used in the singular are to be construed to include the plural where appropriate, and vice versa. Any definition set forth below shall control over any conflicting definition in a document incorporated herein by reference.

[0050] As used herein, the use of a noun can mean "one" object when used in conjunction with the term "comprising" in the claims and / or the specification, but is also consistent with the meaning of "one or more", "at least one", and "one or a plurality of" objects.

[0051] The terms "about" or "approximately" in part mean within the allowable error range of a particular value determined by a person skilled in the art due to the limitations of the measurement system, i.e., how the value is measured or determined. For example, "about" can mean within 3 or more standard deviations according to the convention in the relevant technical field. Alternatively, "about" can mean in the range up to 20%, preferably up to 10%, more preferably up to 5%, and even more preferably up to 1% of a given value. Alternatively, especially with respect to a system or process, the term can mean within one order of magnitude, preferably within a factor of 5, and more preferably within a factor of 2 of the value.

[0052] The terms "comprise(s)", "include(s)", "having", "has", "can", "contain(s)", and variations thereof are intended, as used herein, to be transitional terms, terms, or words that do not exclude additional acts or structures and are without limitation. The present disclosure contemplates other embodiments that "comprise", "consist of", and "consist essentially of" the embodiments or elements presented herein, whether explicitly stated or not.

[0053] As used herein, the words "about" or "approximately" in part mean within the allowable error range of a particular value determined by a person skilled in the art due to the limitations of the measurement system, i.e., how the value is measured or determined. For example, "about" can mean within 3 or more standard deviations according to the convention in the relevant technical field. Alternatively, "about" can mean in the range up to 20%, preferably up to 10%, more preferably up to 5%, and even more preferably up to 1% of a given value.

[0054] As used herein, the term "allergic" refers to an inability or uncontrolled hypersensitivity, particularly hypersensitivity, of the immune system to substances that are typically harmless, such as those present in the environment. As previously described herein, allergic inflammatory skin diseases are characterized by the occurrence of erythema, pain, redness, swelling, and small blisters or papules on the skin. Body areas frequently affected in atopic dermatitis include the head, wings, legs, abdomen, and axillae. Common allergens include, but are not limited to: airborne allergens such as pollen, animal dander, house dust mites, and mold; certain foods, particularly peanuts, nuts, wheat, soy, fish, shellfish, eggs, and milk; insect stings such as those from bees and wasps; drugs, particularly penicillin and penicillin-based antibiotics; latex and other substances that can cause allergic skin reactions; allergic inflammatory skin diseases include atopic dermatitis, scabies allergic dermatitis, urticaria, insect sting allergy, angioedema, inhalant allergy, inhalant allergic dermatitis, food allergic dermatitis, contact dermatitis, miliaria, eosinophilic granuloma, head and neck pruritus, and generalized pruritus. Therefore, it will be understood herein that the term "allergic" can be related to both "self-antigens" and exogenous antigens.

[0055] As used herein, the term "animal" or "animals" refers to ruminants, poultry, pigs, mammals, horses, mice, rats, rabbits, guinea pigs, hamsters, cows, cats, or canines. In a specific embodiment, the term animal relates to canines.For example, the animal can be, without limitation, a penguin, a falcon, an agouti, an American bittern, a snake, a bear, a condor, an ant, an impala, an antelope, an armadillo, an Australian brush turkey, a babirusa, a white-tailed eagle, a valmaine, a fox, a bat, a sambar deer, a bee, a beetle, a leopard cat, a binturong, a bird of paradise, a boa, a bonobo, a bontebok, a brown bear, a sea otter, a camel, a caracal, a chameleon, a cheetah, a chimpanzee, a chinchilla, a cichlid, a clouded leopard, a cobra, a cockatiel, a crocodile, a dolphin, an eel, a flamingo, a fossa, a frog, a common toad, a Galapagos tortoise, a gelada, a giant anteater, a giant panda, a gila monster, a giraffe, a goat, a sheep, a frog, a gorilla, a goldfinch, a Guam rail, a guanaco, a guenon, a hippopotamus, a kite, a crested ibis, an iguana, a jacana, a jaguar, a kagu, a kangaroo, a wallaby, a nightingale, a king snake, a kinkajou, a kiwi, a klipspringer, a koala, a koi, a Komodo dragon, a ladybug, a laughingthrush, a macaque, a leopard, a lion, a lizard, a tiger cat, a Congo African grey parrot, a crowned crane, a manatee, a mandrill, a mangabey, a marsupial, a meerkat, a metallic starling, a monkey, a serval, an okapi, an opossum, an orangutan, an oryx, an ostrich, a weasel, an owl, a painted dog, a parrot, a peacock, a pelican, a polar bear, a mountain gorilla, a prairie dog, a pronghorn, a Przewalski's horse, a raccoon, a ruddy shelduck, a red panda, a reindeer, a rhinoceros, a ringtail, a rock hyrax, a saiga, a scorpion, an asiatic seal, a secretary bird, a serval, a shaman, a snow leopard, a spider, a stork, a brown bear, a Suriname toad, a takin, a yak, a Tasmanian devil, a tenrec, a tiger, a great egret, a tuatara, a turtle, a vulture, an ibex, a waxbill, a whale, a Cape buffalo, a wolf, a wombat, or a zebra.

[0056] As used herein, the term "canid" encompasses animals including pet animals selected from the group including wolves, coyotes, jackals, dingoes, foxes, dogs, raccoon dogs, and domestic dogs. As used herein, "dog" encompasses wild dogs and domestic dogs. In a specific embodiment, the term "dog" means a domestic dog.

[0057] As used herein, the term "drug" refers to any substance or composition represented as having therapeutic, curative or prophylactic properties against animal diseases. Thus, pharmaceuticals include any substance or composition that can be used or administered to an animal for the purpose of performing a medical diagnosis or for the purpose of restoring, modifying or altering the physiological functions of an animal by exerting pharmacological, immunological or metabolic effects.

[0058] In the detailed description of the present specification, references to "embodiment", "an embodiment", "one embodiment", "various embodiments", etc. indicate that the described embodiments may include specific features, structures, or characteristics, but not all embodiments necessarily include those specific features, structures, or characteristics. Furthermore, such phrases do not necessarily refer to the same embodiment. Moreover, when a specific feature, structure, or characteristic is described in relation to one embodiment, it is considered within the knowledge of those skilled in the art to affect such feature, structure, or characteristic in relation to other embodiments, whether or not explicitly described. Reading this specification will make it clear to those skilled in the relevant art how to implement the present disclosure in alternative embodiments.

[0059] As used herein, the term "enantiomer" refers to a pair of stereoisomers that are non-superimposable mirror images of each other. A 1:1 mixture of a pair of enantiomers is a "racemic" mixture or racemic compound. This term is used, where appropriate, to designate a racemic mixture.

[0060] As used herein, the term "enantiopure" refers to a sample consisting of a single enantiomer within the limits of detection.

[0061] As used herein, the term "diastereoisomer" refers to stereoisomers that have at least two asymmetric atoms but are not mirror images of each other. Absolute stereochemistry is defined according to the Cahn-Ingold-Prelog R-S system. When a compound is a pure enantiomer, the stereochemistry of each chiral carbon can be designated as either R or S. A resolved compound of unknown absolute configuration can be designated as (+) or (-) according to the direction (dextrorotatory or levorotatory) in which it rotates plane-polarized light at the wavelength of the sodium D line. The compounds of the presently disclosed subject matter contain one or more asymmetric centers and can therefore give rise to enantiomers, diastereomers, and other stereoisomers that can be defined as (R)- or (S)- from the perspective of absolute stereochemistry. The presently disclosed subject matter is meant to include all such possible isomers, including racemic mixtures, optically pure forms, and intermediate mixtures. The optically active (R)- and (S)- isomers can be prepared using chiral synthons or chiral reagents or can be resolved using conventional techniques. When a compound contains a double bond, the substituents can be in the E or Z configuration. When a compound contains a disubstituted cycloalkyl, the cycloalkyl substituents can be in the cis or trans configuration. It is also intended that all tautomeric forms be included.

[0062] As used herein, the term "isomer" refers to different compounds that have the same molecular formula but differ in the arrangement and structure of their atoms. Also, as used herein, the term "stereoisomer" refers to any of the various stereoisomeric structures that may exist for a given compound of the presently disclosed subject matter and includes geometric isomers. Substituents are understood to be capable of bonding to the chiral centers of carbon atoms. Accordingly, the term stereoisomer includes enantiomers, diastereomers, or racemic compounds of a compound. Also, as used herein, the term "structural isomer" refers to different compounds that have the same number and types of atoms but differ in the connectivity of their atoms, including but not limited to tautomers, among others.

[0063] As used herein, the term "food composition" or "diet" encompasses all materials that may contain foodstuffs, diets, dietary supplements, or materials that may contain proteins, carbohydrates, and / or crude fat. Food may also include adjunct substances or additives such as minerals, vitamins, and flavorings (see Merriam-Webster's Collegiate Dictionary, 10th Edition, 1993). Such food compositions may or may not be nutritionally complete. In one embodiment, the animal food composition according to the presently disclosed subject matter is a nutritionally complete food composition. By way of example, the food compositions described herein may include, but are not limited to, proteins, crude fat, ash, crude fiber, starch, calcium, phosphorus, sodium, chloride, potassium, magnesium, iron, water, copper, manganese, zinc, selenium, vitamin A, vitamin D3, vitamin B1, vitamin B2, vitamin B6, vitamin B12, vitamin B7, vitamin B9, choline chloride, arachidonic acid, omega-3 fatty acid, or omega-6 fatty acid.

[0064] As used herein, "food supplement" refers to a concentrated source of nutrients, a substance for nutritional or physiological purposes, or a plant and plant preparation, which is intended to supplement deficiencies in the normal diet of animals. Food supplements are usually in the form of capsules, lozenges, tablets, pills, powder sachets, or liquids (ampoules, vials with droppers).

[0065] As used herein, the term "inflammation" refers to a local reaction to cellular injury characterized by capillary dilation, leukocyte infiltration, erythema, heat, and pain.

[0066] As used herein, "nutritionally complete" means that the composition provides complete and balanced nutritional requirements for an animal, in a particular embodiment, for a dog. Thus, a nutritionally sufficient feed is a feed that can be fed as the sole feed for the animal, e.g., a dog, and can sustain life without additional food (except water). The food composition can contain a carrier, diluent, or excipient. Depending on the intended use, the carrier, diluent, or excipient can be selected to be suitable for use in animals, e.g., for use in canines such as dogs. In a general aspect, a nutritionally complete composition includes at least one source of protein (or polypeptide) such as a protein extract; and at least one source of vitamins, minerals, trace elements, and fats.

[0067] As used herein, the term "prevent" can include reducing the likelihood of the occurrence or recurrence of a given health condition in an animal.

[0068] As used herein, the term "treating" or "treatment" means to cause, relieve, arrest the progression of, or improve a disorder, disease, or health condition to which such term applies, or one or more symptoms of such disorder, disease, or health condition.

[0069] As used herein, the term "wet food" or "wet food composition" generally refers to a food composition having a water content of about 30% by weight or more, generally about 40% by weight or more, based on the total weight of the food composition. Preferably, the wet food composition has a water content of less than about 90% by weight based on the total weight of the food composition. Generally, this is the final product of a process that includes a final step of sterilization (instead of a drying step). In a particular embodiment, the wet food includes chunks, more particularly chunks in gravy form. Preferably, the wet food includes chunks and gravy, jelly, loaf, mousse, terrine, bite-sized chunks. "Chunks and gravy" products are prepared by making a meat emulsion and passing this meat emulsion through a nozzle under pressure, and then include preformed cooked meat particles. Products such as cooked meat are cut into chunks, which are ultimately mixed with gravy or sauce. The two components are then filled into a container, usually a can or pouch, which is closed or sealed and sterilized. In contrast to ground loaf, chunks and gravy compositions have discrete chunks (i.e., pieces of ground meat and grains) that are physically separated during preparation. These discrete particles are present in the gravy-like liquid in the final container. When served, chunks and gravy products can flow out of the can and be easily mixed with other dry products. While chunks and gravy products can enhance the integrity of the individual components, the non-uniform distribution of chunks and gravy products may not be preferred by consumers. Wet food compositions are generally packaged in containers such as cans and are considered "wet" in appearance due to the moisture contained therein. In the art, generally two types of wet compositions are known. The first is known in the art as "ground loaf". Loaf products are typically prepared by contacting a mixture of ingredients under heat to produce an essentially homogeneous intracellular honeycomb-type mass or "ground loaf". The ground loaf mass is then packaged in a cylindrical container such as a can. During packaging, the ground loaf takes the shape of the container so that it must be cut when provided to a companion animal.The wet food composition is preferably packaged. By doing so, consumers can identify the ingredients contained in the food from the package and confirm that it is suitable for the specific pet in question. The packaging can be metal, plastic, paper, or cardboard.

[0070] As used herein, the term "dry food" or "dry food composition" generally refers to a food or composition having a moisture content of less than about 12% by weight, generally less than about 7% by weight, based on the total mass of the food composition. In particular embodiments, the dry food according to the presently disclosed subject matter has a moisture content of up to about 12% by mass. In some embodiments, the dry food has a moisture content of about 7% by weight or less, such as about 5% by weight or less. In particular embodiments, this dry food has a moisture content of more than about 3% by weight based on the total weight of the food composition. For example, in the examples provided herein, a dry food having a moisture content of about 9.5% by weight based on the total mass of the food composition is exemplified. In certain embodiments, the dry food consists of kibble. By way of non-limiting example, kibble includes microparticles; pellets; pieces of pet food, dehydrated meat, meat analogs, vegetables, and combinations thereof; and pet snacks such as meat and vegetable jerky, rawhide, and biscuits. The dry food composition can be manufactured by mixing the ingredients together and kneading to form a firm dough that can be cooked. Generally, the dry food composition can be the final product of a process that includes a drying step following an extrusion step.

[0071] The process of making dry food is typically carried out by baking and / or extrusion. The dough is typically fed into a machine called an expander or extruder that uses pressurized steam or hot water to cook the ingredients. While inside the extruder, the dough is subjected to extreme pressure and high temperatures. The dough is then forced through a die (holes of a specific size and shape) and cut off with a knife. The puffed dough pieces are passed through a dryer to become kibble, where the moisture is reduced to a predetermined target value and the food stability is maintained until ingestion. Lipids, oils, minerals, vitamins, and a cocktail of natural extracts can then be sprayed onto the kibble and, if desired, sealed in a package. It is preferred that the dry food composition be packaged. This allows the consumer to identify the ingredients contained in the food from the package and confirm that it is suitable for the particular pet in question, namely the dog. The package can be metal, plastic, paper, or cardboard.

[0072] As used herein, the term "semi-moist food" or "semi-moist food composition" generally refers to a food composition having a moisture content intermediate between about 12% and about 30% by weight based on the total weight of the food composition. Therefore, such semi-moist food compositions are generally the final product of a process that allows for a moisture content intermediate between dry food and wet food. In some embodiments, the process may include the step of adding a humectant. In some embodiments, the process includes an extrusion step and a subsequent treatment step with super-heated steam (SHS). In some embodiments, the semi-moist food according to the presently disclosed subject matter contains more than about 12% and up to about 30% moisture by weight based on the total weight of the food composition. By way of example, the semi-moist food composition has about 11% to about 20% moisture and / or a water activity of about 0.64 to about 0.75, preferably both, based on the total weight of the food composition.

[0073] As a non-limiting example, semi-moist foods can be obtained using a superheated steam (SHS) process, such as the processes and methods described in the published patent applications International Publication Nos. WO 2009 / 018990, WO 2009 / 018996, WO 2010 / 112097, WO 2014 / 122072, WO 2016 / 071372 and / or WO 2016 / 071367, the disclosures of which are hereby incorporated by reference in their entirety.

[0074] As disclosed herein, in certain embodiments, when the animal food composition is a semi-moist food, the composition consists of soft semi-moist kibble.

[0075] As disclosed herein, the "moisture content" of an animal food composition is (a) weighing a sample of the food composition to obtain a total mass, (b) heating the sample of step (a) in an oven at 135 ± 2 °C for 240 minutes until all moisture is removed to obtain a dried sample, (c) weighing the dried sample to obtain a dry mass, (d) calculating the difference between the total mass and the dry mass Water content = (total mass) - (dry mass) as and thereby obtaining the moisture content and can be determined by a loss on drying method including each step.

[0076] As used herein, the term "water activity" refers to the ratio of the vapor pressure of water in a food composition divided by the vapor pressure of water in the standard state. For the measurement of water activity, several methods known to those skilled in the art can be utilized, including the resistive electrolytic method, the capacitance method, or a dew point hygrometer. Most preferably, reference is made to the method specified in the international standard ISO 21087 for the measurement of water activity in animal foods and animal feeds.

[0077] As used herein, "solvent" can refer to acetic acid, n-butanol, isopropanol, n-propanol, ethanol, methanol, formic acid, dimethylformamide, dimethyl sulfoxide, acetonitrile, acetone, dichloromethane, tetrahydrofuran, ethyl acetate, n-hexane, benzene, toluene, diethyl ether, chloroform, 1,4-dioxane, water, or a combination thereof, and preferably can refer to water. In a non-limiting embodiment, the solvent is a mixture of about 80% w / w water and about 20% w / w ethanol.

[0078] As used herein, the terms "mass percent" or "mass %" or "by mass" are meant to refer to either (i) the quantity by mass of a constituent / component in a composition as a percentage of the total mass of the composition, or (ii) the quantity by mass of a constituent / component in a material as a percentage of the mass of the final material or product. All mass percentages expressed herein are by mass of the total mass of the dry matter of the food composition, unless otherwise specified.

[0079] As used herein, the amount of a component expressed as mass / Mcal consists of the mass amount of that component per unit of metabolizable energy (ME) of the entire animal food composition. As used herein, the ME parameter is intended to represent the energy value of a food composition that is directly metabolized after ingestion. Within the scope of the presently disclosed subject matter, the ME value can be measured by any suitable method known in the art. By way of example, the ME value can be measured using a feeding trial. In practice, the gross energy (GE) of the food is determined in the laboratory and the amount of food eaten by the animal is recorded. The feces and urine of the animal are collected and the energy in each is determined and referred to as fecal energy (FE) and urinary energy (UE), respectively. Then, ME is calculated as follows: ME (kcal / kg) = [GE - (FE + UE)] / kg of food ingested.

[0080] Alternatively, the ME value can be measured by a mathematical method, particularly considering the percentages of crude fat (CF), crude protein (CP), and NFE (carbohydrates) in the composition. In practice, each percentage is multiplied by its respective at-water coefficient. Next, the sum of the results is multiplied by 10. This mathematical method can be represented by the following formula: ME (kcal / kg) = 10 × [(3.5 × CP) + (8.5 × CF) + (3.5 × NFE)].

[0081] Metabolizable energy can conventionally be determined according to standard methods, particularly according to the National Research Council.

[0082] As used herein, the amount of a component expressed as mass / kg of a food composition, e.g., a complete food, consists of the mass amount of the component per kilogram of mass of the food composition, e.g., the complete food. As used herein, a "complete food" can refer to a quantitative mixture of all feed ingredients formulated to meet the energy, protein, fat, vitamin, and mineral requirements of an animal. As used herein, the amount of a component expressed as mass / kg body weight (Bwt) / day is the daily amount of the component per kilogram of body weight of an animal, e.g., a dog. In certain embodiments, mass / kg body weight refers to the mass of the component fed to the animal per kilogram of the animal's body weight.

[0083] 2. Animal food compositions In certain embodiments, an animal food composition for preventing and / or treating an inflammatory disease or disorder can include glycyrrhizin, additional components, and additives. In certain embodiments, the additional components can include curcuminoids, linoleic acid, EPA / DHA, taurine, lutein, vitamin E, or combinations thereof. In certain embodiments, the additives can include proteins, crude fat, fiber, soluble nitrogen-free substances, carbohydrates, starch, and / or combinations thereof.

[0084] In certain embodiments, the presently disclosed subject matter can be formulated as an animal food composition. In a particular embodiment, the animal food composition can be fed to dogs according to feeding guidelines.

[0085] 2.1. Glycyrrhizin As used herein, the term "glycyrrhizin" refers to the root of Glycyrrhiza glabra, i.e., the components of licorice. Glycyrrhizin, the most prominent compound contained in licorice, is a triterpenoid glycoside. Licorice is a perennial plant that grows wild or is cultivated in Europe and Asia. Furthermore, glycyrrhizin is said to have numerous pharmacological effects such as anti-inflammatory, antiviral, antitumor, antioxidant, and hepatoprotective effects (Sato et al., 1996; Rahman et al., 2006).

[0086] The structure of glycyrrhizin contains a triterpene moiety (glycyrrhetinic acid) and two iduronic acid residues. Other names for glycyrrhizin include Glycyrrhizinic acid, Glycyrrhizic acid, glycyrrhetinic acid glycoside, and (3-beta,20-beta)-20-carboxy-11-oxo-30-norlan-12-en-3-yl-2-O-beta-D-glucopyranuronosyl-alpha-D-glucopyranuronate.

[0087] Glycyrrhizin can be extracted from various natural plants by several extraction techniques known to those skilled in the art, such as supercritical CO 2 2 extraction, aqueous two-phase extraction, solvent extraction, third-phase extraction, microwave-assisted extraction, or ultrasonic-assisted extraction.

[0088] Glycyrrhizin has the CAS registration number: 1405-86-3 and the following structure:

[0089]

Chemical formula

[0090] characterized by

[0091] According to one embodiment, glycyrrhizin can be one of its pharmaceutically acceptable salts and / or racemates, enantiomers, diastereoisomers or tautomers.

[0092] Non-limiting examples of pharmaceutically acceptable salts include ammonium salts such as ammonium glycyrrhizinate, and salts with acids or bases including alkali metal salts such as disodium glycyrrhizinate, trisodium glycyrrhizinate, and dipotassium glycyrrhizinate.

[0093] In one embodiment, glycyrrhizin can be found in or obtained chemically from some naturally occurring extracts and / or plants (Shabkhiz MA, Eikani MH, Bashiri Sadr Z, Golmohammad F. Superheated water extraction of glycyrrhizic acid from licorice root. Food Chem. 2016, 210: 396-401).

[0094] According to one embodiment, glycyrrhizin can be extracted from a glycyrrhizin source such as crude licorice. The glycyrrhizin source can be a glycyrrhizin-containing extract obtained from plant material. Preferably, the glycyrrhizin-containing extract is obtained from plant material containing glycyrrhizin such as crude licorice by at least the following steps: (a) providing a plant raw material containing glycyrrhizin; (b) pulverizing the plant material of step (a), preferably in a solvent, thereby obtaining a pulverized mixture; (c) extracting glycyrrhizin from the pulverized mixture, thereby obtaining a glycyrrhizin-containing extract; (d) Optionally, filtering the glycyrrhizin extract to obtain the filtered glycyrrhizin extract; (e) Optionally, concentrating the filtered glycyrrhizin extract; (f) Optionally, spray-drying the filtered glycyrrhizin extract to obtain a powder; and (g) Optionally, sieving the powder It can be obtained by an extraction method including.

[0095] As used herein, the "plant material containing glycyrrhizin" refers to a glycyrrhizin source, preferably licorice root, most preferably glycyrrhizin-containing licorice extract or glycyrrhizin-containing crude licorice.

[0096] According to some embodiments, the term "plants" or "plant" includes Glycyrrhiza. In a particular embodiment, Glycyrrhiza is selected from the group consisting of Glycyrrhiza acanthocarpa, Glycyrrhiza aspera, Glycyrrhiza astragalina, Glycyrrhiza bucharica, Glycyrrhiza echinata, Glycyrrhiza glabra, Glycyrrhiza glabra var. glandulifera, Glycyrrhiza glabra var. grandiflora, Glycyrrhiza glabra var. hirsuta, Glycyrrhiza glabra var. inflata, Glycyrrhiza glabra var. korshinskyi, Glycyrrhiza glabra var. lepidota, Glycyrrhiza glabra var. pallidiflora, Glycyrrhiza glabra var. scabra, Glycyrrhiza glabra var. triflora, Glycyrrhiza glabra var. uralensis, and Glycyrrhiza yunnanensis.

[0097] The term "licorice" or "liquorice" includes crude licorice, licorice root and Glycyrrhiza glabra root. In a non-limiting embodiment, the glycyrrhizin source consists of licorice root extract (LRE).

[0098] The amount of glycyrrhizin in a given plant source, extract, or food composition can be expressed as a mass per total mass of the dry matter of the given plant source, extract, or composition.

[0099] Licorice can contain from about 1% to about 30%, from about 1% to about 25%, from about 5% to about 20%, from about 6% to about 20%, or from about 6% to about 12% glycyrrhizin, based on the total mass of the dry matter of licorice. As used herein, from about 6% to about 12% glycyrrhizin includes about 6.0%, about 6.5%, about 7.0%, about 7.5%, about 8.0%, about 8.5%, about 9.0%, about 9.5%, about 10.0%, about 10.5%, about 11.0%, about 11.5%, about 12.0% glycyrrhizin, based on the total mass of the dry matter of licorice root.

[0100] Licorice can be provided from a commercial source such as Licorice Root PE 12% Glycyrrhizin formulation (Product number; ED161596) manufactured by Naturex.

[0101] In non-limiting embodiments, the glycyrrhizin source includes licorice, or an extract thereof, that contains glycyrrhizin. Any other glycyrrhizin source known to those skilled in the art, particularly any other glycyrrhizin source distinct from licorice, can also be used. According to some embodiments, the glycyrrhizin source can be pure glycyrrhizin. In non-limiting embodiments, the glycyrrhizin source includes a licorice root extract containing 12% glycyrrhizin prepared using a mixture of 80% water and 20% ethanol.

[0102] In certain embodiments, the presently disclosed subject matter relates to an animal food composition for use in a method of preventing and / or treating an inflammatory disease or disorder, an allergic inflammatory skin disease, the animal food composition being formulated to supply an animal with a daily amount of at least about 0.02 mg / kg body weight of glycyrrhizin.

[0103] In certain embodiments, the animal food composition is formulated to supply an animal with a daily amount of glycyrrhizin in an amount from about 0.02 mg / kg body weight to about 2 mg / kg body weight, from about 0.03 mg / kg body weight to about 2 mg / kg body weight, from about 0.05 mg / kg body weight to about 1 mg / kg body weight, from about 0.1 mg / kg body weight to about 0.8 mg / kg body weight, from about 0.2 mg / kg body weight to about 0.6 mg / kg body weight, from about 0.2 mg / kg body weight to about 1 mg / kg body weight, or from about 1 mg / kg body weight to about 2 mg / kg body weight.

[0104] As used herein, the daily amount of glycyrrhizin is an amount from about 0.02 mg / kg body weight to about 2 mg / kg body weight, which may include about 0.02 mg / kg body weight, about 0.03 mg / kg body weight, about 0.04 mg / kg body weight, about 0.05 mg / kg body weight, about 0.06 mg / kg body weight, about 0.07 mg / kg body weight, about 0.08 mg / kg body weight, about 0.09 mg / kg body weight, about 0.1 mg / kg body weight, about 0.2 mg / kg body weight, about 0.3 mg / kg body weight, about 0.4 mg / kg body weight, about 0.5 mg / kg body weight, about 0.6 mg / kg body weight, about 0.7 mg / kg body weight, about 0.8 mg / kg body weight, about 0.9 mg / kg body weight, about 1 mg / kg body weight, about 1.1 mg / kg body weight, about 1.2 mg / kg body weight, about 1.3 mg / kg body weight, about 1.4 mg / kg body weight, about 1.5 mg / kg body weight, about 1.6 mg / kg body weight, about 1.7 mg / kg body weight, about 1.8 mg / kg body weight, about 1.9 mg / kg body weight, and about 2 mg / kg body weight.

[0105] In another embodiment, the animal food composition is formulated to supply an animal with a daily amount of glycyrrhizin in an amount from about 0.03 mg / kg body weight to about 2 mg / kg body weight, from about 0.05 mg / kg body weight to about 1 mg / kg body weight, from about 0.1 mg / kg body weight to about 0.8 mg / kg body weight, or from about 0.2 mg / kg body weight to about 0.6 mg / kg body weight. In a particular embodiment, the animal food composition is formulated to supply an animal with a daily amount of glycyrrhizin in an amount of about 0.2 mg / kg body weight, about 0.4 mg / kg body weight, or about 0.6 mg / kg body weight.

[0106] In certain embodiments, an animal food composition comprising a glycyrrhizin source can be applied for use as a pharmaceutical, wherein the animal food composition is formulated to supply an animal with a daily dose of glycyrrhizin in an amount of at least about 0.02 mg / kg body weight. In certain embodiments, the pharmaceutical supplies an animal with a daily dose of glycyrrhizin in an amount from about 0.02 mg / kg body weight to about 2 mg / kg body weight, from about 0.02 mg / kg body weight to about 1 mg / kg body weight, or from about 1 mg / kg body weight to about 2 mg / kg body weight.

[0107] In certain embodiments, the animal food composition comprises a glycyrrhizin source comprising glycyrrhizin in an amount of at least about 5 mg / kg based on the total mass of the dry matter of the animal food composition. In certain embodiments, the amount of glycyrrhizin is from about 5 mg / kg to about 100 mg / kg based on the total mass of the dry matter of the animal food composition. In particular embodiments, the amount of glycyrrhizin is from about 5 mg / kg to about 50 mg / kg, from about 5 mg / kg to about 40 mg / kg, from about 10 mg / kg to about 75 mg / kg, from about 15 mg / kg to about 50 mg / kg, from about 20 mg / kg to about 45 mg / kg, or from about 50 mg / kg to about 100 mg / kg based on the total mass of the dry matter of the animal food composition.

[0108] In certain embodiments, the amount of glycyrrhizin is at least about 5 mg / kg based on the total mass of the dry matter of the animal food composition. As used herein, at least about 5 mg / kg based on the total mass of the dry matter of the animal food composition includes at least about 6 mg / kg, about 7 mg / kg, about 8 mg / kg, about 9 mg / kg, about 10 mg / kg, about 11 mg / kg, about 12 mg / kg, about 13 mg / kg, about 14 mg / kg, about 15 mg / kg, about 16 mg / kg, about 17 mg / kg, about 18 mg / kg, about 19 mg / kg, about 20 mg / kg, about 21 mg / kg, about 22 mg / kg, about 23 mg / kg, about 24 mg / kg, about 25 mg / kg, about 26 mg / kg, about 27 mg / kg, about 28 mg / kg, about 29 mg / kg, about 30 mg / kg, about 31 mg / kg, about 32 mg / kg, about 33 mg / kg, about 34 mg / kg, about 35 mg / kg, about 36 mg / kg, about 37 mg / kg, about 38 mg / kg, about 39 mg / kg, or about 40 mg / kg based on the total mass of the dry matter of the animal food composition.

[0109] In certain embodiments, the amount of glycyrrhizin is less than about 100 mg / kg based on the total mass of the dry matter of the animal food composition. As used herein, less than about 100 mg / kg based on the total mass of the dry matter of the animal food composition includes less than about 99 mg / kg, about 98 mg / kg, about 97 mg / kg, about 96 mg / kg, about 95 mg / kg, about 94 mg / kg, about 93 mg / kg, about 92 mg / kg, about 91 mg / kg, about 90 mg / kg, about 89 mg / kg, about 88 mg / kg, about 87 mg / kg, about 86 mg / kg, about 85 mg / kg, about 84 mg / kg, about 83 mg / kg, about 82 mg / kg, about 81 mg / kg, about 80 mg / kg, about 79 mg / kg, about 78 mg / kg, about 77 mg / kg, about 76 mg / kg, about 75 mg / kg, about 74 mg / kg, about 73 mg / kg, about 72 mg / kg, about 71 mg / kg, about 70 mg / kg, about 69 mg / kg, about 68 mg / kg, about 67 mg / kg, about 66 mg / kg, about 65 mg / kg, about 64 mg / kg, about 63 mg / kg, about 62 mg / kg, about 61 mg / kg, about 60 mg / kg, about 59 mg / kg, about 58 mg / kg, about 57 mg / kg, about 56 mg / kg, about 55 mg / kg, about 54 mg / kg, about 53 mg / kg, about 52 mg / kg, about 51 mg / kg, or about 50 mg / kg based on the total mass of the dry matter of the animal food composition.

[0110] 2.2. Additional ingredients In certain embodiments, an animal food composition for preventing and / or treating an inflammatory disease or disorder, an allergic inflammatory skin disease, further comprises linoleic acid.

[0111] In a particular embodiment, an animal food composition for preventing and / or treating an inflammatory disease or disorder, an allergic inflammatory skin disease, disclosed herein, further comprises one or more curcuminoids.

[0112] In certain embodiments, the animal food composition further comprises EPA / DHA, taurine, lutein, vitamin E and / or combinations thereof. The animal food composition may further comprise, in some embodiments, one or more of vitamin A, vitamin B 3 , vitamin C, zinc, vitamin D, one or more fatty acids, and / or combinations thereof.

[0113] 2.2.1. Curcuminoids As used herein, the term "curcuminoid" or "curcuminoids" refers to the phenols contained in turmeric, an Indian spice. Turmeric generally derives from the roots of the plant Curcuma longa. Curcuminoids have also been found in the roots of other species of the Zingiberaceae family of the Curcuma genus. In particular, turmeric may contain about 60% to about 80% by mass of curcumin, about 15% to about 30% by mass of demethoxycurcumin, and about 2% to about 6% by mass of bisdemethoxycurcumin, based on the total mass of the dry turmeric. The curcuminoids in the food compositions of the presently disclosed subject matter can be in any form, including powders or lipid extracts. In non-limiting embodiments, curcuminoids can be found in some naturally occurring extracts and / or plants, or can be obtained or synthesized chemically.

[0114] In non-limiting embodiments, the curcuminoid source comprises turmeric extract, i.e., Curcuma longa. In certain embodiments, the turmeric extract can be BCM-95® from Arjuna or turmeric extract from Naturex. Any other curcuminoid source known to those skilled in the art can be used.

[0115] Other available sources of curcuminoids can also be selected from liposomal curcumin, curcumin nanoparticles, curcumin phospholipid complexes (e.g., MERIVA procurement, "BCM-95" Arjuna having from about 20% to about 90% by mass of total curcuminoids from total curcuminoids), curcumin structural analogs (e.g., EF-24), demethoxycurcumin, bisdemethoxycurcumin, tetrahydrocurcumin, and any formulation designed to enhance the bioavailability of commercially available / DM, curcumin.

[0116] Generally, curcuminoids are found in other plants other than Curcuma longa, such as Curcuma xanthorrhiza and Curcuma zedoania. Curcuminoids in pure form have low solubility in water. Curcuminoids can be extracted from curcuminoid-containing plants, i.e., turmeric roots, using organic solvents such as ethanol and acetone.

[0117] In a particular embodiment, the curcuminoids according to the presently disclosed subject matter have the general formula (I):

[0118]

Chemical formula

[0119] can include compounds of, wherein R 1 and R 2 are independently selected from a hydrogen atom, a methoxy, methyl, hydroxy and ethoxy group, or a pharmaceutically acceptable salt and / or racemate, enantiomer, diastereoisomer or tautomer thereof.

[0120] In a non-limiting embodiment, R 1 and R 2 can be the same or different.

[0121] In a non-limiting embodiment, the curcuminoid is selected from the group consisting of curcumin, demethoxycurcumin, bis-methoxycurcumin, tetrahydrocurcumin, or combinations thereof.

[0122] For reference, curcumin (I), demethoxycurcumin (II), bis-methoxycurcumin (III) and tetrahydrocurcumin (IV) have the chemical structures:

[0123]

Chemical formula

[0124] which consist of.

[0125] In a particular embodiment, the curcuminoid is curcumin. In a special embodiment, the curcuminoid source (i.e., curcumin) can be turmeric.

[0126] In a particular embodiment, the animal food composition contains a curcuminoid source in an amount ranging from about 0.01 g / Mcal to about 0.1 g / Mcal, about 0.01 g / Mcal to about 2 g / Mcal, about 0.01 g / Mcal to about 5 g / Mcal, about 5 g / Mcal to about 10 g / Mcal, or about 0.01 g / Mcal to about 10 g / Mcal. In a particular embodiment, the animal food composition contains a curcuminoid source in an amount less than about 5 g / Mcal, less than about 2 g / Mcal, or less than about 0.5 g / Mcal.

[0127] In a special embodiment, the animal food composition contains a curcuminoid source in an amount ranging from about 0.01% by mass to about 0.1% by mass, about 0.01% by mass to about 5% by mass, about 5% by mass to about 10% by mass, about 0.1% by mass to about 10% by mass, or about 0.01% by mass to about 10% by mass based on the total mass of the dry matter of the food composition.

[0128] As used herein, from about 0.01% to about 10% by mass of the curcuminoid source may include about 0.01%, about 0.1%, about 1%, about 2%, about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, or about 10% by mass of the curcuminoid source, based on the total mass of the dry matter of the food composition.

[0129] As used herein, the curcuminoid source of from about 0.01% to about 0.1% can include from about 0.01% by weight, about 0.011% by weight, about 0.012% by weight, about 0.013% by weight, about 0.014% by weight, about 0.015% by weight, about 0.016% by weight, about 0.017% by weight, about 0.018% by weight, about 0.019% by weight, about 0.02% by weight, about 0.021% by weight, about 0.022% by weight, about 0.023% by weight, about 0.024% by weight, about 0.025% by weight, about 0.026% by weight, about 0.027% by weight, about 0.028% by weight, about 0.029% by weight, about 0.03% by weight, about 0.031% by weight, about 0.032% by weight, about 0.033% by weight, about 0.034% by weight, about 0.035% by weight, about 0.036% by weight, about 0.037% by weight, about 0.038% by weight, about 0.039% by weight, about 0.04% by weight, about 0.041% by weight, about 0.042% by weight, about 0.043% by weight, about 0.044% by weight, about 0.045% by weight, about 0.046% by weight, about 0.047% by weight, about 0.048% by weight, about 0.049% by weight, about 0.05% by weight, about 0.051% by weight, about 0.052% by weight, about 0.053% by weight, about 0.054% by weight, about 0.055% by weight, about 0.056% by weight, about 0.057% by weight, about 0.058% by weight, about 0.059% by weight, about 0.06% by weight, about 0.061% by weight, about 0.062% by weight, about 0.063% by weight, about 0.064% by weight, about 0.065% by weight, about 0.066% by weight, about 0.067% by weight, about 0.068% by weight, about 0.069% by weight, about 0.07% by weight, about 0.071% by weight, about 0.072% by weight, about 0.073% by weight, about 0.074% by weight, about 0.075% by weight, about 0.076% by weight, about 0.077% by weight, about 0.078% by weight, about 0.079% by weight, about 0.08% by weight, about 0.081% by weight, about 0.082% by weight, about 0.083% by weight, about 0.084% by weight, about 0.085% by weight, about 0.086% by weight, about 0.087% by weight, about 0.088% by weight, about 0.089% by weight, about 0.09% by weight, about 0.091% by weight, about 0.092% by weight, about 0.093% by weight, about 0.094% by weight, about 0.095% by weight, about 0.096% by weight, about 0.097% by weight, about 0.098% by weight, about 0.099% by weight to about 0.1% by weight of curcuminoid source, based on the total mass of the dry matter of the animal food composition.

[0130] In certain embodiments, the curcuminoid source can consist of pure curcuminoids.

[0131] By way of non-limiting example, an animal food composition suitable for small dogs (see the classification described herein), particularly in the form of dry food, can contain about 0.002% by weight of glycyrrhizin and about 0.0275% by weight of curcuminoids relative to the total mass of the composition. According to another example, an animal food composition suitable for medium or large dogs (see the classification described herein), particularly in the form of dry food, can contain about 0.003% by weight of glycyrrhizin and about 0.0275% by weight of curcuminoids relative to the total mass of the composition. The animal food compositions described herein can be formulated, particularly in the form of wet food, to supply an animal with a daily amount of about 77 mg / kg of curcuminoids.

[0132] 2.2.2. Linoleic acid As used herein, the term "linoleic acid" means a polyunsaturated omega-6 fatty acid, which is one of two essential fatty acids for animals.

[0133] To measure the content of linoleic acid in a food composition, one of ordinary skill in the art can refer to any well-known technique. As an example, a method based on the NF EN ISO 5508 / 5509 standard by gas chromatography can be used. Therefore, in some embodiments, the animal food compositions described herein contain one or more substances that are a source of linoleic acid.

[0134] Examples of linoleic acid sources, i.e., linoleic acid-containing substances, include vegetable oils, although animal oils or crude fats can also be used. Examples of linoleic acid sources include safflower oil, sunflower oil, soybean oil, sesame oil, canola oil, other vegetable oils or animal oils / crude fats, meat, or a combination of two or more thereof. In certain embodiments, the linoleic acid source is selected from the group consisting of safflower oil, sunflower oil, soybean oil, sesame oil, canola oil, meat, and combinations thereof.

[0135] In certain embodiments, the animal food composition comprises linoleic acid in an amount ranging from about 1 g / Mcal to about 20 g / Mcal, from about 1 g / Mcal to about 10 g / Mcal, from about 8 g / Mcal to about 12 g / Mcal, from about 10 g / Mcal to about 12 g / Mcal, or from about 10 g / Mcal to about 20 g / Mcal.

[0136] In certain embodiments, the animal food composition comprises linoleic acid in an amount ranging from about 2% to about 10%, from about 2% to about 5.5%, or from about 5.5% to about 10% by mass based on the total mass of the dry matter of the animal food composition.

[0137] 2.2.3. EPA / DHA As used herein, the term "EPA / DHA" refers to a fatty acid or mixture of fatty acids consisting of (i) eicosapentaenoic acid ethyl ester (EPA) only, (ii) docosahexaenoic acid ethyl ester (DHA) only, or (iii) a combination of eicosapentaenoic acid ethyl ester and docosahexaenoic acid ethyl ester (EPA+DHA). Therefore, as used herein, the amount of "EPA / DHA" means (i) the amount of EPA in the absence of DHA, (ii) the amount of DHA in the absence of EPA, or (iii) the amount of the combination of EPA and DHA.

[0138] In certain embodiments, the animal food composition comprises a combination of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in an EPA:DHA mass ratio ranging from about 0.0001:1000 to about 1000:0.0001. In certain embodiments, the EPA:DHA mass ratio is about 0.0001:100, about 1:1, or about 0.1:0.0001 based on the total mass of the dry matter of the animal food composition. In particular embodiments, the EPA:DHA mass ratio can be about 0.1:100, about 100:0.1, or from 0.1 to 100. In particular embodiments, the mass or energy ratio of EPA to DHA can range from 0.0001 to 1000 based on the total mass of the dry matter of the animal food composition.

[0139] According to some embodiments, the animal food composition comprises EPA / DHA in an amount of from about 0.1 g / Mcal to about 7 g / Mcal, from about 0.1 g / Mcal to about 3 g / Mcal, from about 1 g / Mcal to about 2 g / Mcal, from about 1.25 g / Mcal to about 1.75 g / Mcal, or from about 3 g / Mcal to about 7 g / Mcal. In certain embodiments, the animal food composition comprises EPA / DHA in an amount of from about 0.1 wt% to about 5 wt%, from about 0.1 wt% to about 2.5 wt%, or from about 2.5 wt% to about 5 wt% based on the total mass of the dry matter of the animal food composition.

[0140] 2.2.4. Taurine Taurine is a non-essential amino acid obtained from meat and fish. Taurine acts as a precursor molecule to stimulate the production of sphingolipids in the skin. Sphingolipids exhibit antibacterial properties.

[0141] Taurine according to the presently disclosed subject matter has the following structure:

[0142]

Chemical formula

[0143] Or may have one of its pharmaceutically acceptable salts and / or racemates, enantiomers, diastereoisomers or tautomers.

[0144] In certain embodiments, the animal food composition comprises taurine in an amount of from about 1 g / Mcal to about 2 g / Mcal, from about 1 g / Mcal to about 1.5 g / Mcal, or from about 1.5 g / Mcal to about 2 g / Mcal.

[0145] 2.2.5. Lutein As used herein, "lutein" refers to a xanthophyll, which is one of the naturally occurring carotenoids. Lutein is a lipophilic molecule and is generally insoluble in water.

[0146] Lutein is synthesized only in plants and is abundant in green leafy vegetables such as spinach, kale, and yellow carrots like other xanthophylls. Lutein is an isomer of zeaxanthin, differing only in the position of one double bond. The main natural stereoisomer of lutein is (3R,3’R,6’R)-β,ε-carotene-3,3’-diol. Lutein exists in plants as a fatty acid ester in which one or two fatty acids are bound to two hydroxyl groups.

[0147] Lutein according to the presently disclosed subject matter has the following structure:

[0148]

Chemical formula

[0149] Or may have one of its pharmaceutically acceptable salts and / or racemates, enantiomers, diastereoisomers or tautomers.

[0150] In certain embodiments, the animal food composition contains lutein in an amount from about 0.001 g / Mcal to about 0.002 g / Mcal, from about 0.001 g / Mcal to about 0.0015 g / Mcal, or from about 0.0015 g / Mcal to about 0.002 g / Mcal.

[0151] 2.2.6. Vitamin E Vitamin E is a general term for several biologically similar compounds, including those called tocopherols and tocotrienols, which share the same biological activity. The biologically active form of vitamin E in animal tissues (which is also the most active antioxidant) is α-tocopherol. Vitamin E cannot be synthesized in vivo. Vitamin E prevents the loss of cell membrane integrity that can negatively affect the function of cells and organs. Vitamin E can be in any form according to the aspects of the presently disclosed subject matter. It can be liquid, semi-solid or solid. Vitamin E can be a tocopherol or a tocotrienol. Vitamin E can be alpha tocopherol, (d-x or dl-oc) beta tocopherol (d-, B or dl-), gamma tocopherol (dot or dim), delta-tocopherol, alpha tocotrienol, beta-tocotrienol, gamma-tocotrienol or delta-tocotrienol. In a particular embodiment, vitamin E is alpha-tocopherol.

[0152] In certain embodiments, the source of vitamin E is not limited. In non-limiting embodiments, sources of vitamin E include vitamin E acetate, (e.g., tocopherol acetate), vitamin E acetate adsorbate, or spray dried vitamin E acetate. In certain embodiments, the source is synthetic, although natural sources may be used.

[0153] For reference, vitamin E according to the presently disclosed subject matter has the following structure:

[0154]

Chemical formula

[0155] Or it can have one of its pharmaceutically acceptable salts and / or racemates, enantiomers, diastereoisomers or tautomers.

[0156] In certain embodiments, the animal food composition contains vitamin E in an amount from about 0.2 g / Mcal to about 0.3 g / Mcal, from about 0.2 g / Mcal to about 0.25 g / Mcal, or from about 0.25 g / Mcal to about 0.3 g / Mcal.

[0157] 2.3. Additives In some embodiments, the animal food compositions described herein are nutritionally complete and may also contain additives such as proteins, crude fat, crude fiber, soluble nitrogen-free extract, ash, minerals, vitamins, or seasonings.

[0158] As will be readily understood, every embodiment of the animal food compositions encompassed by the presently disclosed subject matter contains various components, each present in a predetermined mass percentage in the composition as compared to the total mass of the dry matter of the animal food composition.

[0159] The animal food compositions disclosed herein contain proteins, crude fat, ash, fiber, NFE, and, optionally, one or more additional additives such as vitamins, minerals, etc., and the sum of the masses of the components contained therein corresponds to 100% by mass based on the total mass of the dry matter of the animal food composition.

[0160] 2.3.1. Protein In certain embodiments, the animal food compositions according to the presently disclosed subject matter may further contain a protein source. Thus, the protein level should be high enough to ensure maintenance of the fat-free body mass. The animal food composition may contain one or more distinct protein sources.

[0161] In a particular embodiment, the animal food compositions described herein may contain multiple proteins contained in the protein source used in the manufacturing process. In certain embodiments, the animal food compositions described herein may contain multiple proteins from multiple protein sources.

[0162] In certain embodiments, the protein is not hydrolyzed. In some other embodiments, the protein may be present in at least a partially hydrolyzed form or may even be completely hydrolyzed. The animal food compositions according to the presently disclosed subject matter can incorporate proteins in the form of meat or animal-derived materials such as beef, chicken, turkey meat, lamb meat, fish, plasma, bone marrow, or combinations thereof. In certain embodiments, the animal food compositions described herein can be meat-free. In non-limiting embodiments, meat-free animal food compositions include meat alternative protein sources such as soybeans, corn gluten, or any other protein-containing soy product to provide a protein source. The animal food compositions disclosed herein can include additional protein sources such as soy protein concentrate, milk protein, gluten, and the like.

[0163] 2.3.2. Crude fat The animal food compositions according to the present disclosure may further contain a nutritionally appropriate amount of crude fat. As used herein, the expression "crude fat" includes any crude fat and / or oil that is food acceptable, regardless of their consistency at room temperature, i.e., regardless of whether the "crude fat" is present in an essentially fluid form or an essentially solid form. The animal food compositions according to the presently disclosed subject matter can include crude fat derived from animals and / or plants. The crude fat can be supplied by any of a variety of sources known to those of skill in the art. Non-limiting examples of plant-based crude fat sources include wheat, sunflower, safflower, rapeseed, olive, borage, flaxseed, peanut, blackcurrant seed, cottonseed, wheat germ, corn germ, and oils derived from these and other plant-based crude fat sources. Non-limiting examples of animal sources include, for example, chicken crude fat, turkey meat crude fat, beef crude fat, duck crude fat, pork crude fat, lamb meat crude fat, fish oil, or meat, meat by-products, seafood, dairy products, eggs, and the like. The crude fat content of the food can be measured by any of a number of methods known to those of skill in the art.

[0164] 2.3.3. Fiber Fibers are optionally included in the animal food compositions disclosed herein. The expression "fiber" is similar to "dietary fiber" and for the purposes of the presently disclosed subject matter is to be construed as total fiber including both soluble and insoluble fibers. Soluble fiber can be defined as non-starch polysaccharides that are resistant to digestion and absorption in the small intestine and resistant to fermentation in the large intestine, as opposed to insoluble fiber which can be defined as being resistant to digestion and absorption in the small intestine and undergoing complete or partial fermentation in the large intestine. Soluble fiber is thought to have a prebiotic effect by supplying short-chain fatty acids as an energy source to colonocytes. Insoluble fiber is thought to be useful for transit and ballast effects. Non-limiting examples of fibers include beet pulp, guar gum, chicory root, psyllium, pectin, blueberry, cranberry, pumpkin, apple, oatmeal, beans, citrus, barley, or lentils, and a second group including cellulose, whole grain wheat products, wheat oatmeal, corn bran, flax seeds, grapes, celery, green beans, cauliflower, potato skins, fruit skins, vegetable skins, peanut hulls, and soy fiber.

[0165] 2.3.4. Soluble nitrogen-free extract (NFE) As used herein, and as conventionally understood in the art, soluble nitrogen-free extract (NFE) consists of the soluble carbohydrate fraction that may be included, as necessary, in the animal food compositions disclosed herein. NFE, when present in the animal food composition, may include soluble polysaccharides, starch, gums, mucilage, and pectin. Thus, as conventionally known in the art, NFE does not include the insoluble carbohydrate fraction in the crude fiber material that may be present in the food composition in certain embodiments. Usually, the content of soluble nitrogen-free extract in the food composition is determined by subtracting the contents of other components (protein, crude fat, crude fiber, ash) from the total dry matter of the food composition. In embodiments where the qualitative and quantitative characteristics of the food composition are expressed as energy density (e.g., metabolizable energy density in units of g / Mcal), the NFE content is determined by subtracting the energy value of each of the other components (protein, crude fat, crude fiber, ash) from the energy value of the entire food composition. In embodiments where the qualitative and quantitative characteristics of the food composition are expressed as mass percentage (e.g., mass percentage based on the total mass of the dry matter of the composition), the NFE content is determined by subtracting the mass percentages of the other components (protein, crude fat, crude fiber, ash) from the total mass of the food composition.

[0166] 2.3.5. Carbohydrates As used herein, the term "carbohydrate" refers to a mixture of polysaccharides and sugars that are metabolized as energy when hydrolyzed in the body. The carbohydrate content of a food can be measured by several methods known to those skilled in the art. Carbohydrates can be provided from a variety of carbohydrate sources known to those skilled in the art, including starch (e.g., any type, corn, wheat, barley), beet pulp (which contains a small amount of sugars), and psyllium.

[0167] 2.3.6. Starch An important source of soluble nitrogen-free substances optionally contained in the animal food composition disclosed in this specification contains starch. The term "starch" used herein refers to a polysaccharide consisting of amylose and amylopectin. Starch is usually present as granules in many plant tissues, typically between 1 μm and 100 μm in diameter, depending on the plant source. Chemically, starch is a polysaccharide composed of α-D-glucopyranosyl units linked by α-D(1-4) and / or α-D(1-6) bonds, consisting of two molecular types: amylose, a linear polyglucan composed of approximately 1000 α-D(1-4) linked glucoses, and amylopectin, a branched glucan composed of approximately 4000 glucose units with branching occurring as α-D(1-6) bonds. As used herein, starch encompasses various crystalline structures of A-type, B-type, and C-type starches, which contain amylopectin in different proportions. A-type starch is mainly contained in grains, B-type starch is mainly contained in tubers and starches rich in amylose, and C-type starch consists of a mixture of both A-type and B-type and is mainly contained in leguminous plants. Generally, digestible starch is degraded (hydrolyzed) in the small intestine by the enzymes α-amylase, glucoamylase, and sucrose-iso-maltase to produce free glucose, which is then absorbed. The starch contained in the animal food disclosed in this specification can be any starch suitable for dietary purposes. In fact, the natural starch contained in the starting materials used in the preparation of the animal food composition described herein is liable to change during the manufacturing process. The amount of starch in the animal food composition includes the amount of starch contained in the total amount of the raw materials of the animal food composition. However, the amount of starch contained in the animal food composition disclosed in this specification is equal to the total amount of starch contained in the starting materials used in the preparation of the animal food composition. If not yet known, the starch content of the starting materials used in the preparation of the animal food composition described herein can be determined according to known prior art techniques in the art, particularly according to known polarization methods such as those in accordance with NF EN ISO 10520.In some embodiments where the starting material may contain modified starch or alpha starch, the starch content can be determined according to NF EN ISO 15914.

[0168] 2.3.7. Ash The ash content is a measure of the total amount of minerals contained therein when specified as a result of the analytical determination of the animal food composition described herein. The mineral content is a measure of the amount of specific inorganic components contained therein, and the inorganic components include calcium (Ca), sodium (Na), potassium (K) and chlorine (Cl).

[0169] 2.4. Formulation of animal food compositions The presently disclosed subject matter further relates to an animal food composition comprising a glycyrrhizin source, wherein the amount of glycyrrhizin is more than 5 mg / kg based on the total mass of the dry matter of the animal food composition. The animal food composition described herein can be as described above.

[0170] According to one embodiment, the amount of glycyrrhizin is from about 5 mg / kg to about 100 mg / kg based on the total mass of the dry matter of the animal food composition.

[0171] In certain embodiments, the amount of glycyrrhizin can be from about 5 mg / kg to about 100 mg / kg, from about 10 mg / kg to about 75 mg / kg, from about 15 mg / kg to about 50 mg / kg, or from about 20 mg / kg to about 45 mg / kg based on the total mass of the dry matter of the animal food composition.

[0172] In certain embodiments, the glycyrrhizin source can contain at least one plant of the genus Glycyrrhiza, preferably licorice root, i.e., licorice. In certain embodiments, the glycyrrhizin source is licorice. In a particular embodiment, the glycyrrhizin source in the animal food composition comprises licorice root extract.

[0173] In a particular embodiment, the glycyrrhizin source is present in an amount ranging from about 0.01 g / Mcal to about 0.1 g / Mcal, based on the total mass of the dry matter of the animal food composition. In a particular embodiment, the glycyrrhizin source is present in an amount ranging from about 0.01 wt% to about 10 wt%, based on the total mass of the dry matter of the animal food composition.

[0174] In certain embodiments, the animal food composition is a nutritionally complete animal food composition, a nutritional supplement, or a topping composition. In some embodiments, the animal food composition consists of kibble. In a particular embodiment, the animal food composition includes a nutritionally complete animal food composition. In a particular embodiment, the animal food composition is formulated for oral administration. In certain embodiments, the animal food composition includes a nutritional supplement and / or a drug.

[0175] In a particular embodiment, the animal food composition described herein further includes linoleic acid. In certain embodiments, the linoleic acid source is selected from the group consisting of safflower oil, sunflower oil, soybean oil, sesame oil, canola oil, meat, and combinations thereof.

[0176] In certain embodiments, the animal food composition further includes curcuminoids, linoleic acid, EPA / DHA, taurine, lutein, vitamin E, and / or combinations thereof. In certain embodiments, the animal food composition further includes EPA / DHA, taurine, lutein, vitamin E, and / or combinations thereof. In some embodiments, the animal food composition further includes a protein source.

[0177] In certain embodiments, the animal food composition comprises a glycyrrhizin source. In certain embodiments, the animal food composition comprises a glycyrrhizin source and a curcuminoid. In certain embodiments, the animal food composition comprises a glycyrrhizin source and linoleic acid. In certain embodiments, the animal food composition comprises a glycyrrhizin source and EPA / DHA. In certain embodiments, the animal food composition comprises a glycyrrhizin source and taurine. In certain embodiments, the animal food composition comprises a glycyrrhizin source and lutein. In certain embodiments, the animal food composition comprises a glycyrrhizin source and vitamin E. In certain embodiments, the animal food composition comprises a glycyrrhizin source, a curcuminoid, and linoleic acid. In certain embodiments, the animal food composition comprises a glycyrrhizin source, a curcuminoid, and EPA / DHA. In certain embodiments, the animal food composition comprises a glycyrrhizin source, a curcuminoid, and taurine. In certain embodiments, the animal food composition comprises a glycyrrhizin source, a curcuminoid, and lutein. In certain embodiments, the animal food composition comprises a glycyrrhizin source, a curcuminoid, and vitamin E. In certain embodiments, the animal food composition comprises a glycyrrhizin source, linoleic acid, and EPA / DHA. In certain embodiments, the animal food composition comprises a glycyrrhizin source, linoleic acid, and taurine. In certain embodiments, the animal food composition comprises a glycyrrhizin source, linoleic acid, and lutein. In certain embodiments, the animal food composition comprises a glycyrrhizin source, linoleic acid, and vitamin E. In certain embodiments, the animal food composition comprises a glycyrrhizin source, EPA / DHA, and taurine. In certain embodiments, the animal food composition comprises a glycyrrhizin source, EPA / DHA, and lutein. In certain embodiments, the animal food composition comprises a glycyrrhizin source, EPA / DHA, and vitamin E. In certain embodiments, the animal food composition comprises a glycyrrhizin source, taurine, and lutein.In certain embodiments, the animal food composition comprises a glycyrrhizin source, taurine, and vitamin E. In certain embodiments, the animal food composition comprises a glycyrrhizin source, lutein, and vitamin E. In certain embodiments, the animal food composition comprises a glycyrrhizin source, curcuminoid, linoleic acid, and EPA / DHA. In certain embodiments, the animal food composition comprises a glycyrrhizin source, curcuminoid, linoleic acid, and taurine. In certain embodiments, the animal food composition comprises a glycyrrhizin source, curcuminoid, linoleic acid, and lutein. In certain embodiments, the animal food composition comprises a glycyrrhizin source, curcuminoid, linoleic acid, and vitamin E. In certain embodiments, the animal food composition comprises a glycyrrhizin source, curcuminoid, EPA / DHA, taurine, and lutein. In certain embodiments, the animal food composition comprises a glycyrrhizin source, curcuminoid, EPA / DHA, taurine, and vitamin E. In certain embodiments, the animal food composition comprises a glycyrrhizin source, EPA / DHA, taurine, lutein, and vitamin E. In certain embodiments, the animal food composition comprises a glycyrrhizin source, curcuminoid, linoleic acid, EPA / DHA, taurine, and lutein. In certain embodiments, the animal food composition comprises a glycyrrhizin source, curcuminoid, linoleic acid, EPA / DHA, taurine, and vitamin E. In certain embodiments, the animal food composition comprises a glycyrrhizin source, linoleic acid, EPA / DHA, taurine, lutein, and vitamin E. In certain embodiments, the animal food composition comprises a glycyrrhizin source, curcuminoid, linoleic acid, EPA / DHA, taurine, lutein, and vitamin E.

[0178] In a particular embodiment, the animal food composition of the presently disclosed subject matter comprises licorice in an amount that provides a daily amount of glycyrrhizin from about 0.02 mg / kg body weight to about 2 mg / kg body weight. In certain embodiments, the animal food composition further comprises curcuminoids in an amount from about 0.01 g / Mcal to about 10 g / Mcal, linoleic acid in an amount from about 1 g / Mcal to about 20 g / Mcal, EPA / DHA in an amount from about 0.1 g / Mcal to about 7 g / Mcal, taurine in an amount from about 1 g / Mcal to about 2 g / Mcal, lutein in an amount from about 0.001 g / Mcal to about 0.002 g / Mcal, vitamin E in an amount from about 0.2 g / Mcal to about 0.3 g / Mcal, or a combination thereof.

[0179] In a particular embodiment, the animal food composition of the presently disclosed subject matter comprises licorice in an amount that provides a daily amount of glycyrrhizin from about 0.02 mg / kg body weight to about 1 mg / kg body weight. In certain embodiments, the animal food composition further comprises curcuminoids in an amount from about 0.01 g / Mcal to about 5 g / Mcal, linoleic acid in an amount from about 10 g / Mcal to about 20 g / Mcal, EPA / DHA in an amount from about 0.1 g / Mcal to about 3 g / Mcal, taurine in an amount from about 1 g / Mcal to about 1.5 g / Mcal, lutein in an amount from about 0.001 g / Mcal to about 0.0015 g / Mcal, vitamin E in an amount from about 0.2 g / Mcal to about 0.25 g / Mcal, or a combination thereof.

[0180] In a particular embodiment, the animal food composition of the presently disclosed subject matter comprises licorice in an amount that provides at least about 0.02 mg / kg body weight of glycyrrhizin per day. In certain embodiments, the animal food composition further comprises linoleic acid in an amount from about 8 g / Mcal to about 12 g / Mcal, EPA / DHA in an amount from about 1 g / Mcal to about 2 g / Mcal, taurine in an amount from about 1 g / Mcal to about 2 g / Mcal, lutein in an amount from about 0.001 g / Mcal to about 0.002 g / Mcal, vitamin E in an amount from about 0.2 g / Mcal to about 0.3 g / Mcal, or a combination thereof.

[0181] In a particular embodiment, the animal food composition of the presently disclosed subject matter comprises licorice in an amount that provides a daily amount of glycyrrhizin of at least about 0.02 mg / kg body weight. In certain embodiments, the animal food composition further comprises linoleic acid in an amount from about 10 g / Mcal to about 12 g / Mcal, EPA / DHA in an amount from about 1.25 g / Mcal to about 1.75 g / Mcal, taurine in an amount from about 1 g / Mcal to about 1.5 g / Mcal, lutein in an amount from about 0.001 g / Mcal to about 0.0015 g / Mcal, vitamin E in an amount from about 0.2 g / Mcal to about 0.25 g / Mcal, or combinations thereof.

[0182] In a particular embodiment, the animal food composition of the presently disclosed subject matter further comprises licorice in an amount that provides a daily amount of glycyrrhizin of at least about 0.02 mg / kg body weight, linoleic acid in an amount of about 10.5 g / Mcal, EPA / DHA in an amount of about 1.5 g / Mcal, taurine in an amount of about 1.2 g / Mcal, lutein in an amount of about 0.0013 g / Mcal, and vitamin E in an amount of about 0.244 g / Mcal.

[0183] In certain embodiments, the animal food composition further comprises a source of protein, crude fat, fiber, NFE, carbohydrate, starch, or combinations thereof.

[0184] In certain embodiments, the animal is a canid. In a particular embodiment, the canid is a dog. In a particular embodiment, the animal food composition comprises or consists of dry, wet, and / or semi-moist food compositions.

[0185] The presently disclosed subject matter further provides a kit as described herein. The kit is for use in a method of preventing and / or treating an inflammatory disease or disorder, an allergic inflammatory skin disease: (i) a first part comprising a source of glycyrrhizin, and (ii) a second part comprising one or more additional components, and the kit is formulated to provide a daily dose of glycyrrhizin in an amount of at least about 0.02 mg / kg body weight. In certain embodiments, the kit comprises glycyrrhizin as disclosed herein and one or more additional components.

[0186] In certain embodiments, the glycyrrhizin source may comprise at least one plant of the genus Glycyrrhiza, preferably licorice root, i.e., licorice. In certain embodiments, the glycyrrhizin source is licorice. In particular embodiments, the glycyrrhizin source in the kit comprises licorice root extract.

[0187] In particular embodiments, the kit contains dry, wet and / or semi-moist food compositions. In certain embodiments, the kit contains a nutritionally complete animal food composition, a dietary supplement, or a topping composition. In some embodiments, the kit contains kibble. In one embodiment, the kit is formulated for oral administration. In one embodiment, the kit contains a dietary supplement and / or a drug.

[0188] 2.5. Dog breeds and feeding guidelines The present disclosure relates in particular to a method for evaluating the health and well-being of animals. The characteristics of companion animals vary by size, sex, breed, and species. Considering that domesticated animals receive most of the veterinary care, the examined trends and individual animal characteristics of the most common domesticated animals (e.g., dogs) can generally provide an indication of the effectiveness of the examined methods when applied to other animals.

[0189] The present disclosure relates in particular to a method for determining the oral biometrics of animals of the genus Canis. The genus Canis includes domestic dogs (Canis lupus familiaris), wolves, coyotes, foxes, jackals, dingos, and the present disclosure can be used for all of these animals. In some embodiments, the subject is a domestic dog, referred to herein simply as a dog. This genus Canis is part of the Canidae family, which includes a number of extant species.

[0190] As used herein, the expression "size category" refers to a definition of an animal (e.g., dog, cat, etc.) in terms of the average body weight of a particular animal species. Animals of the same breed (e.g., dog, cat, etc.) may have relatively uniform physical characteristics such as size, coat color, physiology, and behavior patterns compared to animals of different breeds. Although the following discussion focuses on dogs, it should be noted that other companion animals and wild animals are also intended to be included within the scope of the present disclosure and the present disclosure is not intended to be limited to dogs.

[0191] The dog can be of any breed, including toy dog breeds, small dog breeds, medium dog breeds, large dog breeds, or giant dog breeds. Non-limiting examples of toy dog breeds include Affenpinscher, Australian Silky Terrier, Bichon Frise, Bolognese, Cavalier King Charles Spaniel, Chihuahua, Chinese Crested Dog, Coton de Tulear, English Toy Terrier, Brussels Griffon, Havanese, Italian Greyhound, Pekingese, King Charles Spaniel, Löwchen (Little Lion Dog), Maltese, Miniature Pinscher, Papillon, Pekingese, Pomeranian, Pug, Russian Toy Terrier, and Yorkshire Terrier. Examples of small dog breeds include, but are not limited to, French Bulldog, Beagle, Dachshund, Welsh Corgi Pembroke, Miniature Schnauzer, Cavalier King Charles Spaniel, Shih Tzu, and Boston Terrier. Examples of medium dog breeds include, but are not limited to, Bulldog, Cocker Spaniel, Shetland Sheepdog, Border Collie, Basset Hound, Siberian Husky, Dalmatian, Doberman, and Staffordshire Bull Terrier. Examples of large dog breeds include, but are not limited to, Bearded Collie, Great Dane, Neapolitan Mastiff, Scottish Deerhound, Bordeaux Mastiff, Newfoundland, English Mastiff, Saint Bernard, Leonberger, and Irish Wolfhound. Crossbreeds are generally classified as toy, small, medium, and large dog breeds according to their weight. In certain embodiments, the dog is a toy dog breed. In certain embodiments, the dog is a medium, large, or giant dog breed. In some embodiments, the dog is a mix of two or more dog breeds. Even in such cases, the mixed-breed dog can be classified by size according to its weight and may exhibit traits (e.g., behavioral traits, genetic traits, etc.) associated with each of the two or more dog breeds found in the dog.

[0192] The Fédération Cynologique Internationale currently recognizes 346 purebred dog breeds. Dog breeds can be identified, for example, by observing physical characteristics or through genetic analysis. A pedigree dog is the offspring of two dogs of the same breed and is eligible for registration with an approved club or association that maintains the registration of dogs of that breed. There are several pedigree registration systems, but the Kennel Club is the most well-known.

[0193]

Table 1

[0194] In certain embodiments, the dog size categories are selected according to the 2017 Salt et al. reference (Table 1) (Salt et al., Growth standard charts for monitoring bodyweight in dogs of different sizes PLoS ONE 12: e0182064.). In other embodiments, the dog size categories are selected according to alternative nomenclature. Small dog breeds may correspond to animals with an average weight of about 6.5 kilograms to about 9 kilograms. Medium dog breeds may correspond to animals with an average weight of about 9 kilograms to about 30 kilograms. Large dog breeds may correspond to animals with an average weight of about 30 kilograms to about 40 kilograms. Giant dog breeds may correspond to animals with an average weight exceeding about 40 kilograms.

[0195] In other embodiments, the dog size categories are selected according to alternative nomenclature. Small dog breeds may correspond to animals with an average weight of up to about 10 kg. Medium dog breeds may correspond to animals with an average weight of about 10 kg to about 25 kg. Large dog breeds may correspond to animals with an average weight of about 25 kg to about 45 kg. Extra-large dog breeds may correspond to animals with an average weight of at least about 45 kg.

[0196] It is well known that the daily amount of food to be fed to an animal can depend on the weight of that animal. This is particularly true for dogs. The recommended daily amount of food is as shown in Table 2.

[0197]

Table 2

[0198] In a particular embodiment, the amount of glycyrrhizin is from about 5 mg / kg body weight to about 100 mg / kg body weight, from about 10 mg / kg body weight to about 75 mg / kg body weight, from about 15 mg / kg body weight to about 50 mg / kg body weight, or from about 20 mg / kg body weight to about 45 mg / kg body weight, based on the total mass of the dry matter of the animal food composition.

[0199] As a result, one of ordinary skill in the art can readily determine the amount, or range of amounts, of glycyrrhizin that should be comprised in the mass of the food composition such that animals of the Canidae family, particularly animals such as dogs, are provided with a preferred daily amount of glycyrrhizin as disclosed herein, e.g., an amount expressed as the daily amount of glycyrrhizin per kg of body weight.

[0200] As an example, in a small dog with a body weight of 9 kg, for which, e.g., a complete food composition in the form of kibble is supplied daily at 180 g, in order to prevent and / or treat allergic skin diseases, e.g., atopic dermatitis, the amount of glycyrrhizin contained therein must be more than about 0.27 (0.03 × 9) mg. Therefore, the complete food shall contain an amount of glycyrrhizin greater than about 0.15 mg per 100 g of complete food, or greater than about 0.015 mg per kg of complete food.

[0201] As already stated herein, the animal food composition containing glycyrrhizin can include a nutritional supplement composition (i.e., an incomplete food composition) or a complete food composition.

[0202] 3. Method for preparing an animal food composition The presently disclosed subject matter also encompasses a method for manufacturing an animal food composition as defined herein. The process for manufacturing the described animal food composition may be carried out according to any method known in the art.

[0203] Animal food compositions can be manufactured by mixing the ingredients and kneading them to form a firm dough or meat emulsion that can be cooked, i.e., by kneading to make a mixture. This also applies to liquids in which the ingredients are mixed and homogenized before the cooking step in the package. The process of making dry food embodiments is typically done by baking and / or extrusion. The dough is usually fed into a machine called an expander and / or extruder, which uses pressurized steam or water to cook the ingredients. While in the extruder, the dough is subjected to extreme pressure and high temperatures. The dough is then forced through a die (holes of a specific size and shape) and then cut off with a knife. The puffed dough is passed through a dryer to form kibble so that the moisture is reduced to a predetermined target value and the food stability is maintained until ingestion. The kibble can then be sprayed with fat, oil, minerals, vitamins, a cocktail of natural extracts, flavoring agents, and sealed in a package if desired.

[0204] The composition can be a dry food, wet food, or semi-moist food as described in the presently disclosed subject matter.

[0205] Therefore, the presently disclosed subject matter is a method for manufacturing an animal food composition comprising a glycyrrhizin source, a) preparing a glycyrrhizin source, b) optionally, mixing one or more additional ingredients and / or additives to thereby provide a mixture, and c) optionally, heating the mixture, and provides a method comprising.

[0206] In non-limiting embodiments, the animal food composition produced in the manufacturing method comprises kibble and / or a protein source.

[0207] In certain embodiments, the glycyrrhizin source is mixed, in step b), with a component selected from the group consisting of a curcuminoid source, linoleic acid, EPA / DHA, taurine, lutein, vitamin E, and combinations thereof.

[0208] In certain embodiments, the glycyrrhizin source is mixed, in step b), with a component selected from the group consisting of a linoleic acid source, EPA / DHA, taurine, lutein, vitamin E, and combinations thereof.

[0209] In embodiments, the glycyrrhizin source is mixed, in step b), with a component selected from the group consisting of a linoleic acid source, EPA / DHA, taurine, lutein, vitamin E, protein, crude fat, fiber, or combinations thereof.

[0210] 4. Therapeutic Applications The presently disclosed subject matter provides for the use of the animal food compositions described herein for the prevention and / or treatment of inflammatory diseases or disorders, allergic inflammatory skin diseases, preferably atopic dermatitis, including atopic dermatitis in canines, particularly dogs.

[0211] As described herein, allergic inflammatory skin diseases can include at least one of atopic dermatitis, flea allergic dermatitis, urticaria, insect sting allergy, angioedema, inhalant allergy, inhalant allergic dermatitis, food allergic dermatitis, contact dermatitis, miliary dermatitis, eosinophilic granuloma, head and neck pruritus, and generalized pruritus, preferably atopic dermatitis, including atopic dermatitis in canines, particularly dogs.

[0212] In certain embodiments, the animal food composition is used in a method for preventing and / or treating an animal suffering from an inflammatory disease or disorder, an allergic inflammatory skin disease, more particularly, atopic dermatitis, i.e., canine atopic dermatitis.

[0213] In a particular embodiment, the animal food composition is used for preventing and / or treating atopic dermatitis. According to a further embodiment, the animal food composition is used for preventing and / or treating canine atopic dermatitis. According to yet a further embodiment, the animal food composition is used for reducing the dosage and frequency of drugs necessary for managing an inflammatory disease or disorder, or a state of an allergic inflammatory skin disease.

[0214] The presently disclosed subject matter also includes a method for preventing and / or treating an inflammatory disease or disorder of an animal, an allergic inflammatory skin disease, comprising at least the step of feeding an animal with the animal food composition according to the presently disclosed subject matter.

[0215] In a specific embodiment, the animal is suffering from an allergic inflammatory skin disease, preferably, the animal is suffering from atopic dermatitis, more preferably, the animal is a dog suffering from canine atopic dermatitis.

[0216] The animal food composition described herein enables the prevention or treatment of an inflammatory disease or disorder, an allergic inflammatory skin disease, preferably atopic dermatitis. In a particular embodiment, the animal is fed only the food composition described herein. In a particular embodiment, the animal is partially fed the food composition described herein and another food composition. In a particular embodiment, the food composition is fed to the animal as a nutritional supplement or a drug. Beneficial effects are obtained in any of these disclosed dietary therapies.

[0217] In a particular embodiment, "partially" means that the animal food composition occupies at least about 30% of the animal diet, preferably at least about 30% of the animal diet, more preferably at least about 70% of the animal diet, still more preferably at least about 80% of the animal diet, and even more preferably at least about 90% of the animal diet.

[0218] Generally, the foodstuff is supplied daily to animals, particularly dogs. In certain embodiments, the animal food composition as disclosed herein can be supplied to the animal to be treated as the sole nutritionally complete food during the treatment period. According to these embodiments, the nutritionally complete animal food composition is supplied daily to the animal during the treatment period.

[0219] In certain embodiments, the animal food composition described herein can be supplied to the animal to be treated alternately with another animal food composition, preferably another nutritionally complete animal food composition, and this other nutritionally complete food composition can be selected from among known animal food compositions, including a variety of commercially available animal food compositions, particularly a variety of commercially available dog food compositions.

[0220] In a particular embodiment, the animal food composition is supplied alternately daily and thus according to a two-day schedule. In certain embodiments, the animal food composition is supplied according to a schedule of every 3, 4, 5, 6, 7 or more days.

[0221] In the daily practice of feeding animals, particularly dogs, it should be understood that the animal owner cannot always feed the animal with the animal food composition as if it were every day. In certain embodiments, the beneficial effect of preventing or treating an inflammatory disease or disorder, an allergic inflammatory skin disease, particularly preventing or treating canine allergic dermatitis, is sufficiently provided when the animal food composition described herein is fed to the animal alternately every other day. In a particular embodiment, if the animal is fed every 3, 4, 5, 6 or 7 days, the beneficial effect will decrease and a longer treatment period may be required.

[0222] While not intended to be bound by any particular theory, in certain embodiments, to efficiently prevent or treat an inflammatory disease or disorder, an allergic inflammatory skin disease, for example, to completely and efficiently prevent or treat canine allergic dermatitis, it is necessary to administer the animal food composition described herein to an animal, particularly a dog, at least alternately, most preferably daily.

[0223] The period of feeding the animal food composition described herein can range from several weeks to several years, particularly depending on the severity of the allergic inflammatory skin disease, i.e., atopic dermatitis.

[0224] As already defined in the present disclosure, treatment for a short period, i.e., 1 month, may be sufficient to reduce pruritus.

[0225] In certain embodiments, the animal food composition is supplied to the animal to be treated for a longer period, such as 9 months or more, for example, 12 months or more, according to a feeding schedule that includes (i) feeding only the animal food composition described herein to an animal, particularly a dog, or (ii) alternating the supply of the animal food composition described herein with another animal food composition.

[0226] The present disclosure also relates to a method for preventing and / or treating an inflammatory disease or disorder, an allergic inflammatory skin disease in an animal suffering from an allergic inflammatory skin disease, the method comprising at least the step of providing feeding advice comprising the animal food composition as defined by the presently disclosed subject matter.

[0227] The present disclosure further relates to the use of a glycyrrhizin source for preparing a composition for the treatment of allergic inflammatory skin diseases.

[0228] In certain embodiments, the animal food composition inhibits the release of one or more interleukin targets from activated T cells. As provided herein, overexpression of interleukin-4 (IL-4) and / or interleukin-5 (IL-5) can indicate a predisposition to an allergic phenotype in an immune response. In certain embodiments, the animal food composition inhibits the release of IL-5 from activated T cells. In certain embodiments, the animal food composition inhibits the release of IL-4 from activated T cells. In certain embodiments, the animal food composition inhibits the release of both IL-5 and IL-4 from activated T cells. In alternative embodiments, the animal food composition does not stimulate the proliferation or release of interleukin-2 (IL-2).

[0229] In certain embodiments, the animal food composition can be used to inhibit the release of IL-4 or IL-5 from activated T cells. In certain embodiments, the animal food composition can be used to prevent and / or treat a disease characterized by an increase in the release of IL-4 or IL-5 by activated T cells. In a particular embodiment, the disease characterized by an increase in the release of IL-4 or IL-5 by activated T cells is selected from the group consisting of allergic inflammatory skin diseases, and in particular, atopic dermatitis, flea allergic dermatitis, urticaria, insect sting allergy, angioedema, inhalant allergy, inhalant allergic dermatitis, food allergic dermatitis, contact dermatitis, miliaria, eosinophilic granuloma, head and neck pruritus, and generalized pruritus. In a particular embodiment, the disease characterized by an increase in the release of IL-4 or IL-5 by activated T cells is atopic dermatitis.

[0230] The presently disclosed subject matter also includes a kit comprising a source of glycyrrhizin for use in preventing and / or treating an inflammatory disease or disorder, an allergic inflammatory skin disease, preferably atopic dermatitis, i.e., canine atopic dermatitis.

[0231] The present disclosure further relates to the use of a kit comprising a first part comprising a source of glycyrrhizin and a second part comprising one or more additional components for preparing a composition for the treatment of allergic inflammatory skin diseases.

[0232] In certain embodiments, the presently disclosed subject matter is a kit comprising a source of glycyrrhizin for use in a method of preventing and / or treating an inflammatory disease or disorder, an allergic inflammatory skin disease, the kit being formulated to supply an animal with a daily amount of glycyrrhizin in an amount of at least about 0.02 mg / kg body weight.

[0233] In certain embodiments, the kits described herein can be supplied to an animal to be treated alternately with another animal food composition or kit, preferably another nutritionally complete animal food composition or kit, and this other nutritionally complete food composition can be selected from among known animal food compositions, including a wide variety of commercially available animal food compositions, particularly a wide variety of commercially available dog food compositions.

[0234] In a particular embodiment, the kit is supplied alternately daily, and thus according to a two-day schedule. In certain embodiments, the kit is supplied according to a schedule of every 3, 4, 5, 6, or 7 days.

[0235] The kits described herein are used in a method of preventing or treating an inflammatory disease or disorder, an allergic inflammatory skin disease, preferably atopic dermatitis. In certain embodiments, an animal is fed only the kits described herein. In certain embodiments, an animal is partially fed the kits described herein and another food composition or kit. In certain embodiments, the animal is fed the kit as a nutritional supplement or a drug. Beneficial effects are obtained in any of these disclosed dietary therapies.

Examples

[0236] For purposes of understanding rather than limitation, the presently disclosed subject matter will be better understood by reference to the following examples, which are provided by way of illustration of the presently disclosed subject matter and not by way of limitation.

[0237] Example 1: Safety test of glycyrrhizin Materials and methods The purpose of this clinical trial was to demonstrate the safety of three doses of glycyrrhizin (Gly) (much higher than the doses recommended for animal intake, especially canine intake) administered to dogs in the form of licorice root extract for 28 days.

[0238] Dogs: Sixteen healthy adult dogs (male and female) owned by individuals participated in the study. Specifically, those dogs were beagles, Brittany spaniels, and dachshunds weighing between 5 kg and 21 kg. Twelve were female and four were male. More females were selected in this study because past studies in other animal species had shown that blood pressure was more likely to increase with exposure to glycyrrhizin.

[0239] Glycyrrhizin source: The glycyrrhizin source was provided by a licorice root extract containing 12% glycyrrhizin formulated in gelatin capsules. The licorice root extract was obtained commercially from Naturex (Avignon, France) with product reference number DAB40178. This product was accompanied by an analytical certificate showing that it contained 12.6% w / w glycyrrhizin by HPLC method.

[0240] Commercially available pharmaceutical-grade gelatin capsules (Capsuline, located in Dania Beach, Florida, USA) were filled with a mixture of accurately weighed licorice powder and 100% pure maltodextrin powder (Bulk Powders, located in Chester, UK). The weighed combination of licorice powder and maltodextrin powder was prepared such that when mixed together and put into capsules, glycyrrhizin was administered to dogs daily at doses following a schedule of 0.2 mg / kg Bwt / day, then 0.4 mg / kg Bwt / day, and then 0.6 mg / kg Bwt / day.

[0241] Dosage: The dosage to be administered to each dog was calculated based on five different body weight groups. Separate capsules were weighed for each group. Licorice root extract was mixed with maltodextrin to deliver the same volume of all powder materials to each dog. The capsules were placed in the meat meal of wet pet food and fed to the dogs.

[0242] Over a period of 28 days, dogs were fed a total of three different dosages of glycyrrhizin. These dosages were 0.2 mg / kg Bwt / day, 0.4 mg / kg Bwt / day, and 0.6 mg / kg Bwt / day.

[0243] Each dog received the following protocol cycle, which was repeated for each test concentration. The first cycle administered 0.2 mg of glycyrrhizin (Gly) / kg Bwt / day. The second cycle was administered at 0.4 mg Gly / kg Bwt / day. The third cycle was administered at 0.6 mg Gly / kg Bwt / day.

[0244] Protocol cycle: Day 1: Baseline measurements were taken: blood pressure, and the concentrations of potassium, sodium, calcium, creatinine, and aldosterone in blood and urine.

[0245] Day 0: Administration of licorice was initiated (the concentration of glycyrrhizin was varied according to the first, second, or third cycle of the protocol as indicated previously). It was administered once a day to each dog along with the morning diet.

[0246] Days 1 - 27: It was administered once a day to each dog along with the morning diet.

[0247] Days 2, 4, 7, 14, 21, and 28: Blood pressure and the concentrations of potassium, sodium, calcium, creatinine, and aldosterone in blood and urine were measured.

[0248] Day 28: Washout was initiated and the dogs were returned to maintenance diet only, i.e., licorice was not fed to the dogs. The washout period was continued for 2 weeks. On days 34 and 41, blood pressure and the concentrations of potassium, sodium, calcium, creatinine, and aldosterone in blood and urine were measured again.

[0249] If there was a significant change in the measured values of the dogs at the end of the cycle compared to the baseline measured values, the test was terminated. If there was no significant change in the measured values of the dogs at the end of the cycle compared to the baseline measured values, the dog was advanced to the next cycle of the protocol.

[0250] Details of the protocol: Licorice root extract was administered successively for each cycle of the protocol according to the following predetermined glycyrrhizin (Gly) concentrations: 0.2 mg / kg Bwt / day, 0.4 mg / kg Bwt / day, and 0.6 mg / kg Bwt / day. The second and third cycles of the protocol were initiated only after it was shown that there were no significant changes in blood pressure (BP) or blood and urine parameters in the previous cycle.

[0251] Blood pressure was measured multiple times for each animal, and the first measurement value was discarded. Blood pressure was measured by the oscillometric method. For each measurement, if it was shown that the mean blood pressure of the dog had risen to a potentially harmful level (usually above 160 mmHg), the dog could be excluded from the test (temporarily or permanently) at the discretion of the veterinarian. All dogs were acclimated to the oscillometric device and the blood pressure measurement process before the start of the test. If signs of stress were observed in a dog before or during blood pressure measurement, the measurement of that dog was aborted and resumed when the dog had calmed down. If necessary, the dog was temporarily given intensive acclimation. If the problem was not solved by these measures, the dog was permanently excluded from the test and replaced with a strictly conforming alternative animal. The typical blood pressure ranges of the three dog breeds used were known and were considered when judging the normality of the blood pressure measured during the test. The normal reference ranges of blood pressure for the three dog breeds used were as follows: Beagle: Systolic 140±15, Diastolic 79±13 Dachshund: Systolic 142±10, Diastolic 85±15 Brittany Spaniel: Systolic 131±16, Diastolic 74±14

[0252] The conditions under which blood pressure measurement was performed were as follows.

[0253] The environment was isolated and quiet. The subject was in a sedated state and was acclimated to the measurement room for 5 to 10 minutes before measurement. Thereafter, the animal was gently restrained in a comfortable position, ideally in the ventral recumbent or lateral recumbent position, to limit the vertical distance from the base of the heart to the blood pressure cuff. The blood pressure cuff width was approximately 30% to 40% of the outer circumference of the blood pressure cuff site. The size of the blood pressure cuff was tracked and the same size was used each time. The blood pressure cuff was attached to the right forelimb that had been pre-shortened where the probe was applied. This reduced the stress and time constraints on the collection day. The right forelimb was used for all animals for each measurement. Whenever possible, the same person performed the blood pressure measurement on the same dog each time.

[0254] Measurements were taken only when the subject was calm and not moving. The first measurement value was discarded. A total of five consecutive and consistent values were recorded. In some subjects, blood pressure tended to decrease as the measurement was being taken. In such animals, the measurement was continued until the decrease leveled off, and then five consecutive consistent values were recorded.

[0255] Blood pressure measurements were repeated as necessary while changing the position of the measurement band in order to obtain consistent values. The blood pressure value was based on the average of the five recorded measurement values.

[0256] To measure the concentrations of potassium, sodium, calcium, creatinine, and aldosterone in the blood, a small blood sample (4 ml) was collected on the days indicated in the above protocol description. To measure the concentrations of potassium, sodium, and calcium in urine, free-catch urine (minimum 3 ml) was collected on each of the days indicated above. Urine creatinine concentration was also measured so that the fractional excretion rate (FER) of sodium, potassium, and calcium could be calculated.

[0257] FER is calculated according to the formula: (urinary potassium × serum creatinine × 100) / (serum potassium × urinary creatinine) as follows.

[0258] Since licorice root extract (LRE) / Gly was evaluated as a single component, it was most effective to administer it directly using the measured amount of licorice in the capsule. Therefore, an appropriate dose of LRE was weighed into gelatin capsules and then administered in parallel with the dog's normal feeding schedule.

[0259] Statistics: To evaluate the effect of the control diet (glycyrrhizin dosage), a linear mixed model was used. Animals were modeled as a random term. The Turkey HSD method was used for alpha correction of multiple comparisons. The significance level was set at 5% for two-sided tests.

[0260] Two methods of modeling blood pressure were evaluated. In the first method, the blood pressure difference between baseline and after 4 weeks was calculated for each dog and each sequence and used as the dependent variable. In the second method, the blood pressure after 4 weeks was used as the dependent variable, and the blood pressure at baseline was used as a covariate in the model.

[0261] Results Blood pressure: The overall mean of systolic blood pressure (mmHg) over time at each test dose of glycyrrhizin (Gly) is shown in Table 3 below:

[0262]

Table 3

[0263] These results indicate that there was no significant difference in blood pressure between D0 and any subsequent time point at any of the test doses.

[0264] Fractional excretion of potassium: The overall mean of the fractional excretion of potassium over time at each test dose of glycyrrhizin (Gly) is shown in Table 4 below. The results are expressed as the percentage of potassium excreted in urine compared to the amount of potassium filtered and retained in the kidney.

[0265]

Table 4

[0266] These results indicate that there was no significant difference in the fractional excretion of potassium between D0 and any subsequent time point at any of the test doses.

[0267] Plasma aldosterone concentration: The overall mean of plasma aldosterone concentration over time at each test dose of glycyrrhizin (Gly), expressed in picomoles / L, is shown in Table 5 below:

[0268]

Table 5

[0269] These results indicate that there was no significant difference in plasma aldosterone concentration between D0 and any subsequent time points at any of the tested doses.

[0270] In conclusion, the results of this study suggest that glycyrrhizin did not have a significant effect on blood pressure homeostasis, fractional potassium excretion, and plasma aldosterone concentration in adult dogs at the tested doses and duration.

[0271] Example 2: Immunomodulatory potential of glycyrrhizin Materials and methods Preparation of licorice root extract for bioassay. Licorice root extract (LRE) was obtained from William Ransom & Son PLC (UK). For preparation, 20 g of root powder was suspended in 100 ml of 20% ethanol solution in distilled water. Next, this suspension was heated at 60 °C for 2 hours with moderate stirring, and then the obtained extract was filtered through Whatman No. 4 filter paper. This extraction process was repeated once more with the filtered solids. Thereafter, the collected filtrate was concentrated overnight at 40 °C using a rotary evaporator to obtain a paste-like extract. The fresh phytochemical extract was stored at -20 °C under a nitrogen atmosphere. According to the supplier's specifications, the glycyrrhizin concentration of the extract was 18% w / w. The concentration of LRE is based on the wet mass of the extract. All assays were performed on the same extract.

[0272] The licorice root extract was weighed and suspended in dimethyl sulfoxide (DMSO - Sigma, UK) at 100 mg / mL, and further diluted to 4 mg / mL with complete tissue culture medium (CTCM; RPMI-1640 medium (Sigma) supplemented with 5% fetal bovine serum (Sigma) adjusted to 5%, 2 mM L-glutamine (Sigma), 100 U / mL penicillin, and 100 μg / mL streptomycin (Sigma)). These stocks were further diluted with CTCM and placed in assay wells to achieve final concentrations between 0.3 μg / mL and 1000 μg / mL.

[0273] Isolation of human peripheral blood mononuclear cells (HuPBMC). HuPBMC were isolated using the method of Telford et al. (Telford et al., International Immunopharmacology. 4 (2004) 1455-1466). Briefly, heparinized blood was obtained from healthy human volunteers aged 20 - 50 years after obtaining consent (Nottingham University Medical School Research Ethics Committee, BT / 04 / 2005), diluted with an equal volume of RPMI-1640 medium (Sigma), and layered on Histopaque 1077 (Sigma). After centrifugation at 710×g for 20 minutes, the buffy coat was collected and washed twice with RPMI-1640. The cells were suspended at 106 / mL in CTCM. The mononuclear cell concentration was determined by cytology to be 85%.

[0274] Cytotoxicity of LRE against Jurkat E6.1 T cell line and HuPBMC. Cytotoxicity was evaluated using the human T lymphoblastic leukemia Jurkat E6.1 cell line (Walsh et al., 2008). In 96-well U-bottom plates (Costar), triplicates of pre-diluted LRE extract with final concentrations ranging from 0.3 to 1000 μg / mL in 200 μL of CTCM containing 10 5 Jurkat E6.1 cells per well were tested. After culturing at 37°C in 5% CO 2 for 24 hours, the cell viability was evaluated using the dye exclusion method (20 μL of 0.4% trypan blue (Sigma, UK) in physiological saline) by counting 10 random areas of the reticle of the objective lens under the microscope. The viability was determined by comparison with the control culture of cells without LRE addition.

[0275] The same cytotoxicity assay was performed using normal and fresh human peripheral blood mononuclear cells. The same pre-diluted extract was tested in triplicates in the range of 0.1 - 1000 μg / ml and incubated with 10 5 HuPBMC in 5% CO 2They were cultured at 37°C for 24 hours. The cell viability was evaluated as described above using the trypan blue dye exclusion method. The viability was determined by comparison with cells cultured without LRE.

[0276] Proliferation and cytokine release assays of HuPBMC. Anti-CD3 antibody (BD Pharmingen, UK) and anti-CD28 antibody (BD Pharmingen, UK) were used in combination to stimulate T cell proliferation and cytokine expression of HuPBMC, and the regulatory effect of glycyrrhiza extract was evaluated. Cell proliferation was determined by the incorporation of [3H]-thymidine. All in vitro assays were performed as six replicates of the assay.

[0277] The pre-diluted licorice root extract (LRE) was tested at final concentrations ranging from 0.3 to 1000 μg / mL in 200 μL of CTCM containing 10 5 HuPCM per well. In this assay, sub-optimal levels of anti-CD3 and anti-CD28 (final assay concentrations of 100 ng / mL each) were used. By using sub-optimal stimulation, potential stimulatory and inhibitory effects can be determined. Dexamethasone alone was also tested in triplicate at a final concentration of 0.5 μM as a positive control (maximum immunosuppression). DMSO alone was tested as a vehicle control.

[0278] In the interleukin-2 assay, after culturing at 37°C for 24 hours in 5% CO 2 50 μL of the medium was removed from each well and stored at -20°C until the IL-2 concentration was analyzed. At this time, the cells were pulsed with 0.25 μCi of [3H] thymidine (GE Healthcare, UK) per well and further cultured for 24 hours, and then harvested onto GF / c filter plates (Packard, UK) using a Packard Filtermate harvester. The radioactivity retained on the filter was measured using a Packard Topcount NXT scintillation counter, and EC 50(Effective concentration that reduces proliferation by 50%) was determined by non-linear regression using Prism 3.0 software. For the remaining cytokine assays, after culturing at 37°C in 5% CO 2 for 48 hours, 150 μL of medium was removed from each well (immediately before cell harvesting) and stored at -20°C prior to IL-2, IL-5, IL-17, and IFN-γ analysis.

[0279] Cytokine analysis. The concentrations of IL-2, IL-5, IL-17, and IFN-γ in the conditioned medium were determined by ELISA according to the following protocol: 96-well assay plates (Nunc, located in the UK) were coated with 50 μL of 1 μg / mL of the relevant capture antibody (BD Pharmingen, located in the UK; αIL-2, 555051; αIL-5, 554393; αIFN-γ, 551221; αIL-17, 560486) in carbonate / bicarbonate buffer at pH 9.6 and incubated overnight at 4°C. The wells were then incubated with 200 μL of 1% bovine serum albumin (BSA, Sigma, located in the UK) in phosphate-buffered saline (PBS) for 1 hour at room temperature and then washed three times with PBS + 0.05% Tween 20 (Sigma, located in the UK). Subsequently, 50 μL of the cell culture supernatant samples were diluted with an equal volume of PBS / Tween + 1% BSA. The prepared samples and cytokine standards (Human, BD Pharmingen, located in the UK: IL-2, 555602; IL-5, 554606; IFN-γ, 554616; IL-17 Sigma, H7791) were added to the relevant wells. After incubation overnight at 4°C, the wells were washed three times with PBS / Tween, and 50 μL per well of 0.5 μg / mL of the relevant biotinylated detection antibody (Human, BD Pharmingen, located in the UK: αIL-2, 555040; αIL-5, 554491; αIFN-γ, 554550; αIL-17, 555067) in PBS / Tween + 1% BSA was added and incubated for 1 hour at room temperature. The washed plates were incubated with 50 μL / well of 1:1000 streptavidin / peroxidase (BD Pharmingen, located in the UK) in PBS / Tween + 1% BSA for 30 minutes at room temperature, and after the final wash with PBS / Tween, they were incubated with 100 μL per well of 0.1 mg / mL of tetramethylbenzidine substrate (Sigma, located in the UK) in 0.1 M acetate buffer at pH 6. 2 SO 4 was added to stop the reaction, and the plates were read at 450 nm using a Dynex MRX plate reader.

[0280] Statistical analysis. In the cytokine assay, the LRE concentration was compared to the control of anti-CD3 / CD28 stimulation only. Following one-way ANOVA, Dunnett's post hoc test was performed to determine the significant suppression of cell survival, proliferation, and cytokine secretion using JMP14. In Dunnett's post hoc comparison, the reference data was provided by culturing the positive control of stimulation only.

[0281] Results Cytotoxicity of licorice root extract against Jurkat E6.1T cell line and human PBMC. Jurkat E6.1 cells have been previously used to evaluate the potential immunotoxicity of compounds (Walsh et al., Marine Drugs. 6 (2008) 291-307). Here, it provides a useful method to determine whether LRE causes cell stress / cell death in monocytes within the concentration range used in subsequent immunomodulatory assays. Licorice root extract did not show evidence of toxicity against the Jurkat cell line at concentrations up to 100 μg / ml (Figure 1). At 300 μg / ml, cytotoxicity was observed, showing approximately 30% cell death (p < 0.01), and increased to 40% with the 1 mg / ml extract (p < 0.01). It was concluded that the maximum safe concentration of the 100 μg / ml extract is acceptable for the interpretation of immunological cell assays.

[0282] When a similar toxicity assay was performed using HuPBMC cells, equivalent results were obtained (Figure 1). The cell survival rate also decreased in this case at an extract concentration of 300 μ / ml (p < 0.01), but did not decrease further at 1 mg / ml, further confirming that the maximum extract concentration interpretable in the cell assay is 100 μg / ml.

[0283] Anti-CD3 / CD28 stimulated the proliferation and interleukin-2 release by HuPBMC. The proliferation of the HuPBMC population was determined in the presence of LRE. In parallel with CD3 / CD28 stimulation, the effect of LRE was evaluated in the range of 0.3 - 1000 μg / ml (Figure 2). In the range of 0.3 - 30 μg / ml of LRE, no effect was observed. The proliferation decreased by approximately 40% at 100 μg / ml. Therefore, at concentrations lower than the cytotoxic threshold, the proliferation is affected by LRE.

[0284] Interleukin-2 (IL-2) is an important cytokine produced by activated T cells in that it plays a role in controlling the development and proliferation of immune cells. Therefore, IL-2 can be used as an indicator of the activation state of the mononuclear cell population. When evaluated in a T cell activation model, there was no evidence that interleukin-2 release was affected by LRE over the concentration range of 0.3 - 100 μg / ml (Figure 2). However, at 300 μg / ml of LRE, the secretion of IL-2 decreased by approximately 30%, and at 1000 μg / ml of LRE, it further decreased by 10%.

[0285] Summarizing these findings, the cytotoxic effect of LRE was observed at concentrations of 300 μg / ml and above. However, the proliferation response was more sensitive to the extract and showed a significant decrease at a concentration of 100 μg / ml (p < 0.05). Interleukin-2 secretion did not seem to be as sensitive to LRE as cell proliferation, and 300 μg / ml was the first concentration to cause a significant decrease (p < 0.05). However, the effect at high concentrations was not so dramatic.

[0286] Two possible interpretations of the findings at high LRE concentrations were that the anti-proliferative and anti-IL-2 effects were simply associated with cellular stress / death and could be indicators of early cellular stress detectable before the onset of cell death. Alternatively, the anti-proliferative and anti-IL-2 effects might be different from cell death.

[0287] Expression of interleukin-5, interleukin-17, and interferon-γ by stimulated HuPBMCs. To further elucidate the potential immunomodulatory effects of LRE, particularly its effects on different types of T-helper (Th) cells, titrated concentrations of the extract were cultured with anti-CD3 / CD28-activated HuPBMCs. The activity of Th1 cells was inferred by quantifying the amount of interferon-γ secreted into the medium.

[0288] Similarly, Th2 activity was detected by quantifying the proxy interleukin-5. Finally, Th17 activity was inferred by detection of interleukin-17 (IL-17) (Figure 3). The immunomodulatory effects were evaluated by comparing the cytokine secretion levels observed in the presence of only the activator (anti-CD3 / CD28 in the case of humans) and calibrating them to 100% activity by doing so.

[0289] In the HuPBMC assay, no significant decrease in the secretion of interferon-γ (IFNγ) and IL-17 was seen between LRE concentrations of 0.1 - 10 μg / ml. However, the secretion of interleukin-5 decreased significantly at 3 μg / ml and 10 μg / ml, showing decreases of 30% (P < 0.05) and 40% (P < 0.005), respectively, compared to the positive control. At an LRE concentration of 30 μg / ml, a greater decrease was shown, with a decrease of more than 50% (P < 0.0001) compared to the positive control, and at 100 μg / ml, a 75% decrease was shown (P < 0.0001). At LRE concentrations above 100 μg / ml, the expression of all three cytokines was significantly suppressed compared to the positive control (P < 0.0001). The effects observed at these high concentrations were probably related to the cytotoxicity of LRE. Therefore, the differential effects on cytokines were interpreted at concentrations below 100 μg / ml. At a concentration of 100 μg / ml, an effect on proliferation was observed, so it was a bit more ambiguous. At concentrations above 100 μg / ml, all findings seemed to be explainable by cytotoxicity. A summary of the differential effects of LRE on the three cytokines is shown in the results presented in the figure mentioned earlier.

[0290] Conclusion In summary, the inventors have provided evidence that the aqueous extract of licorice root can inhibit the release of IL-5 from activated T cells within a concentration range that is non-cytotoxic and does not stimulate proliferation or IL-2 release. Further, within this concentration range, it does not significantly affect the Th-1 cytokine IFN-γ or the Th-17 cytokine IL-17.

[0291] While the presently disclosed subject matter and its advantages have been described in detail, it is to be understood that various changes, substitutions, and modifications can be made herein without departing from the spirit and scope of the disclosed subject matter. Further, the scope of this application is not intended to be limited to the processes, and to the specific embodiments of the compositions of matter, methods, and acts described herein. As will be readily understood by those skilled in the art, processes, compositions of matter, methods, or acts that presently exist or are later developed that perform substantially the same function or achieve substantially the same result as the corresponding embodiments described herein may be utilized in accordance with the presently disclosed subject matter. Accordingly, the appended claims are intended to include such processes, machines, manufactures, compositions of matter, methods, or acts within their scope.

[0292] Various patents, patent applications, publications, product descriptions, protocols, and sequence accession numbers are cited throughout this application, and those inventions are hereby incorporated by reference in their entirety for all purposes.

Claims

1. An animal food composition containing a glycyrrhizin source for use in a method for preventing and / or treating an inflammatory disease or disorder, an allergic inflammatory skin disease, the animal food composition being formulated to supply an animal with a daily dose of glycyrrhizin in an amount of at least about 0.02 mg / kg body weight.

2. The animal food composition according to claim 1, wherein the animal food composition is formulated to supply an animal with a daily dose of glycyrrhizin in an amount from about 0.02 mg / kg body weight to about 2 mg / kg body weight.

3. The animal food composition according to claim 1 or 2, wherein the animal food composition is formulated to supply an animal with a daily dose of glycyrrhizin in an amount from about 0.03 mg / kg body weight to about 2 mg / kg body weight, particularly from about 0.05 mg / kg body weight to about 1 mg / kg body weight, particularly from about 0.1 mg / kg body weight to about 0.8 mg / kg body weight, particularly from about 0.2 mg / kg body weight to about 0.6 mg / kg body weight.

4. The animal food composition according to any one of claims 1 to 3, wherein the animal food composition is selected from the group consisting of a nutritionally complete animal food composition, a dietary supplement, and a topping composition.

5. The animal food composition according to any one of claims 1 to 4, wherein the glycyrrhizin source contains licorice root extract.

6. The animal food composition according to any one of claims 1 to 5, wherein the composition further contains linoleic acid.

7. The animal food composition according to any one of claims 1 to 6, wherein the composition further contains EPA / DHA, taurine, lutein, vitamin E and / or a combination thereof.

8. The animal food composition according to any one of claims 1 to 7, wherein the allergic inflammatory skin disease is selected from the group consisting of atopic dermatitis, flea allergic dermatitis, urticaria, insect sting allergy, angioedema, inhalant allergy, inhalant allergic dermatitis, food allergic dermatitis, contact dermatitis, miliaria, eosinophilic granuloma, head and neck pruritus, and generalized pruritus.

9. For use in a method for preventing and / or treating an allergic inflammatory skin disease, (i) a first part containing a glycyrrhizin source, and (ii) a second part containing one or more additional components, A kit comprising, formulated for supplying a daily amount of glycyrrhizin to an animal in an amount of at least about 0.02 mg / kg body weight.

10. An animal food composition comprising a glycyrrhizin source, wherein the amount of glycyrrhizin is at least about 5 mg / kg based on the total mass of the dry matter of the animal food composition.

11. The animal food composition according to claim 10, wherein the amount of the glycyrrhizin is from about 5 mg / kg to about 100 mg / kg based on the total mass of the dry matter of the animal food composition.

12. The animal food composition according to claim 10 or 11, wherein the amount of the glycyrrhizin is from about 5 mg / kg to 100 mg / kg, particularly from about 10 mg / kg to about 75 mg / kg, particularly from about 15 mg / kg to about 50 mg / kg, and particularly from about 20 mg / kg to 45 mg / kg, based on the total mass of the dry matter of the animal food composition.

13. The animal food composition according to any one of claims 10 to 12, wherein the animal food composition constitutes kibble and / or the animal food composition further comprises a protein source.

14. The animal food composition according to any one of claims 10 to 13, wherein the animal food composition comprises a nutritionally complete food composition.

15. The animal food composition according to any one of claims 10 to 14 or the kit according to claim 9, formulated for oral administration.

16. The animal food composition according to any one of claims 10 to 15 or the kit according to claim 9 or 15, wherein the glycyrrhizin source consists of licorice root extract.

17. The animal food composition according to any one of claims 10 to 16, further comprising one or more additional ingredients.

18. The animal food composition according to claim 17 or the kit according to claim 9, 15 or 16, wherein the one or more additional ingredients are selected from linoleic acid, EPA / DHA, taurine, lutein, vitamin E and / or combinations thereof.

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