Methods, kits and compositions for assessing and treating interstitial cystitis

Abstract

To provide methods and kits for identifying a companion animal having an increased risk of developing interstitial cystitis; along with compositions for treating the same.SOLUTION: A pet food composition comprises an effective amount of a fiber-bound polyphenol component, for use in increasing the level of a urinary cytokine selected from: Fms-like tyrosine kinase 3 (F1t3-L), a stem cell factor (SCF); and IL-12p40, in a companion animal.SELECTED DRAWING: Figure 1A

Description

[Background technology]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS This application claims the benefit of priority to U.S. Provisional Patent Application No. 63 / 129,088, filed December 22, 2020, the contents of which are incorporated by reference in their entirety herein.

[0002] Feline interstitial cystitis (FIC), also known as painful bladder syndrome (BPS), is a term used to describe a systemic condition in cats characterized by bladder pain, urinary frequency, urinary outlet obstruction (LUTD), and nocturia. FIC can be classified into two types: acute, which resolves spontaneously within 3-7 days regardless of treatment, and chronic, which persists or recurs frequently over weeks or months. FIC can also be seen at all ages, with felines between 2 and 7 years of age being at higher risk. Various scientists have proposed possible pathomechanisms that may lead to FIC, but the exact causal relationship of FIC is unclear.

[0003] Clinical diagnosis of FIC typically includes urinalysis, urine culture, and abdominal ultrasound. However, the diagnosis is situational. Typically, FIC is diagnosed by "ruling out" other possible causes, including stone formation, infection, neoplasia, or renal disease. Currently, there are no consistent or specific diagnostic molecular markers that can serve as reliable markers for evaluating FIC. Furthermore, all clinical trials rely heavily on subjective pet owner observation, interpretation, and quantification of clinical signs that may confirm FIC. This produces unreliable results due to inconsistent ability to reliably quantify owner urinary patterns and behavior.

[0004] Although several biomarker candidates have been identified and evaluated in humans, few have been evaluated in cats with FIC.Therefore, there is a need in the art for non-invasive urinary biomarkers for diagnosing FIC and food formulations that improve diagnostic biomarker levels and clinical symptoms to alleviate the condition.Particular embodiments of the present invention are designed to meet these and other objectives. Summary of the Invention

[0005] Some embodiments of the present invention provide a method for diagnosing or identifying a feline's tendency to develop interstitial cystitis. In certain embodiments, the diagnostic method comprises analyzing a cytokine biomarker. Biomarker levels may be analyzed from any bodily fluid, and in preferred embodiments, biomarker levels are obtained from urine. Additionally, preferred biomarkers tested include, but are not limited to, Fms-like tyrosine kinase 3 ligand (Flt3-L) and the cytokine stem cell factor (SCF).

[0006] Certain embodiments of the present invention are directed to the treatment of interstitial cystitis in felines comprising administering a pet food composition containing high levels of antioxidants.

[0007] A further embodiment of the present invention provides a kit for diagnosing or identifying the propensity of a feline to develop interstitial cystitis.

[0008] According to at least one embodiment, a method is provided for identifying a companion animal at increased risk for developing interstitial cystitis (IC). The method typically includes analyzing a biological sample obtained from the companion animal to determine a concentration of a urinary cytokine, analyzing a biological sample from a healthy companion animal to determine a concentration of a urinary cytokine, and comparing the urinary cytokine concentration of the companion animal to the urinary cytokine concentration of the healthy companion animal, where if the biological sample obtained from the companion animal contains a concentration of a urinary cytokine that is lower than the concentration of the urinary cytokine of the healthy companion animal, the companion animal is at increased risk for developing interstitial cystitis.

[0009] According to another embodiment, a method for identifying a companion animal that would benefit from a treatment to reduce the risk of developing interstitial cystitis is provided, the treatment comprising administering to the companion animal a composition comprising an effective amount of a fiber-bound polyphenol component. The method for identifying a companion animal that would benefit from a treatment to reduce the risk of developing interstitial cystitis comprises: analyzing a biological sample obtained from the companion animal to determine a concentration of urinary cytokines; analyzing a biological sample from a healthy companion animal to determine a concentration of urinary cytokines; and comparing the urinary cytokine concentration of the companion animal to the urinary cytokine concentration of the healthy companion animal, wherein a lower urinary cytokine concentration of the companion animal than the urinary cytokine concentration of the healthy companion animal indicates that the companion animal would benefit from a treatment to reduce the risk of developing interstitial cystitis.

[0010] In yet another embodiment, a method of reducing the risk of developing interstitial cystitis in a feline subject includes the steps of analyzing a biological sample obtained from the companion animal to determine a urinary cytokine concentration; analyzing a biological sample from a healthy companion animal to determine a urinary cytokine concentration; comparing the urinary cytokine concentration of the companion animal to the urinary cytokine concentration of the healthy companion animal, where a lower urinary cytokine concentration of the companion animal than the healthy companion animal indicates that the companion animal would benefit from treatment; and administering to the companion animal a composition comprising an effective amount of a fiber-bound polyphenol component if the urinary cytokine concentration of the companion animal is lower than the urinary cytokine concentration of the healthy companion animal.

[0011] According to additional embodiments, kits are provided for identifying companion animals at higher risk of developing interstitial cystitis. The kits typically include a container for collecting a biological sample; a detection method selected from the following: ELISA, chromatographic analysis, aptamer-based quantitative assays, fluorescent tags or stains specific for a combination of two or more of their metabolites; a point-of-care test device with pre-loaded chromophores / antibodies specific for one or more of the urinary cytokines described herein; and instructions for use.

[0012] According to certain embodiments, a method is provided for treating, inhibiting or ameliorating symptoms associated with interstitial cystitis in a companion animal, the method comprising administering to a companion animal in need thereof a composition comprising an effective amount of fiber-bound polyphenols. [Brief description of the drawings]

[0013] [Figure 1A] FIG. 1A shows levels of specific urinary cytokines in healthy and diseased felines.

[0014] [Figure 1B] FIG. 1B shows urinary cytokine levels before and after interstitial cystitis diagnosis.

[0015] [Figure 1C] FIG. 1C shows urinary cytokine levels over several years.

[0016] [Figure 2A] Figures 2A and 2B show the relationship of various urinary cytokines in healthy and diseased felines. [Figure 2B] Same as above.

[0017] [Figure 3A] Figures 3A and 3B show the levels of two urinary cytokines in healthy and diseased felines. [Figure 3B] Same as above.

[0018] [Figure 4] FIG. 4 shows serum cytokine levels in healthy and diseased felines.

[0019] [Figure 5A] FIG. 5A shows the ROC curves for diseased and healthy felines generated from urinary cytokines.

[0020] [Figure 5B] FIG. 5B shows ROC curves for diseased and healthy felines generated from blood CBC, chemistry and cytokines.

[0021] [Figure 6A] Figures 6A and 6B show data on urinary cytokines, and blood CBC, chemistry and cytokines. [Figure 6B] Same as above. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0022] The following invention relates, in part, to diagnostic methods for assessing a feline's propensity to develop interstitial cystitis.

[0023] The following description of the preferred embodiments is merely exemplary in nature and is not intended to limit the invention in any respect. The description of the exemplary embodiments is intended to be read in conjunction with the accompanying drawings, which should be considered part of the entire written description. The following description of the preferred embodiments is merely exemplary in nature and is not intended to limit the invention in any respect. The description of the exemplary embodiments is intended to be read in conjunction with the accompanying drawings, which should be considered part of the entire written description.

[0024] As used herein and in the appended claims, the singular forms "a," "an," and "the" include plural references unless the context dictates otherwise. The singular form of any class of components refers not only to one chemical species within that class, but also to mixtures of those chemical species, for example, the singular term "protein" may refer to a mixture of compounds, each of which is also considered to be a protein. The terms "a" (or "an"), "one or more," and "at least one" may be used interchangeably herein. The terms "comprising," "including," and "having" may be used interchangeably. The term "include" should be interpreted as "including, but not limited to." The term "including" should be interpreted as "including, but not limited to."

[0025] As used throughout, "range" is used as a shorthand for describing any and all values ​​within the range. Any value within the range can be selected as the terminus of the range. In addition, all references cited herein are incorporated by reference in their entirety. In the event of a discrepancy between a definition in this disclosure and a definition in a cited reference, the present disclosure controls.

[0026] Unless otherwise defined, all technical and scientific terms and acronyms used herein have the same meaning as commonly understood by those skilled in the art.Furthermore, it is to be understood that the present invention is not limited to any particular method, protocol, or reagent described herein.The described methods, protocols, and reagents are merely examples and are for illustrative purposes only.

[0027] As used herein, the term "biological sample" may be used interchangeably with the terms "sample", "specimen", "biological material" and "biological material". A biological sample refers to any organic material obtained from a pet, including bodily fluids such as blood, saliva, urine, tissue samples from a biopsy or such as fur, and other clinical specimens such as breath condensate. Biological samples can be obtained in non-invasive and / or invasive manners. For example, a biological sample may be provided in a non-invasive manner, such as via a mouth swab, a collection of fur, or urine. In other examples, a biological sample may be provided in an invasive manner, such as by drawing blood with a needle or removing tissue via a biopsy.

[0028] In certain embodiments, the biological sample may further comprise one or more excipients. The excipients may be added to the biological sample at any time. For example, the excipients may be added to the biological sample during collection, transportation, preparation, and / or analysis of the sample.

[0029] The addition of excipients is well known in the art. Such excipients should be present in an amount that does not impair the purpose and effect provided by the present invention. Excipients may be included as stabilizers, preservatives, processing aids, pH buffers, bulking agents, diluents, color reagents, dyes. For example, ethylenediaminetetraacetic acid (EDTA) may be added to biological samples during collection to preserve the biological samples.

[0030] Examples of excipients may include boric acid and its derivatives, dimethylsulfoxide (DMSO), ethanol, polyethylene glycol, ethylenediaminetetraacetic acid (EDTA), formic acid and its derivatives, protease inhibitors, sodium salts such as sodium citrate and sodium metabisulfate, protease inhibitors.

[0031] Biological samples (e.g., blood, saliva, urine, exhaled breath condensate, tissue, etc.) may be used as diagnostic tools. For example, biological samples (e.g., biological fluids) may be used as diagnostic tools due to their ability to correlate to the health and / or condition of an animal (e.g., a human). The health condition may be related to a disease, disorder, or other condition. For example, biological fluids may be used to diagnose and / or determine the onset of disease, the progression of disease, and / or the progress of treatment for disease. Biological fluids may be used as diagnostic fluids in non-invasive and / or invasive manners. For example, in some examples, the biological fluid may be provided in a non-invasive manner, such as via a mouth swab, spitting from the mouth, or urine. In other examples, the biological fluid may be provided in an invasive manner, such as via drawing blood via an injection, or removing tissue. Detection (e.g., rapid detection) of antibodies in biological fluids (such as saliva) may be performed quickly and via compact tools (via toothbrushes, mouthguards, patches, etc.). Detecting a disease, disorder, or other condition via a biological fluid may enable point-of-care diagnosis of a disease, disorder, or other condition. While the present disclosure may provide numerous examples of biological fluids, it should be understood that such examples are for illustrative purposes only. Examples may extend to any biological sample, including, for example, biological fluids as well as biological tissues.

[0032] Detecting a disease, disorder, or other condition from a biological sample may enable point-of-care diagnosis of the disease, disorder, or other condition. Methods for quantifying one or more biomarkers in a biological sample are well known in the art (e.g., colorimetric reporting, nuclear magnetic resonance (NMR) spectroscopy, infrared (IR) spectroscopy, mass spectrometry, etc.). While the present disclosure may provide numerous examples relating to urine, it should be understood that such examples are for illustrative purposes only, and the examples may be extended to any biological sample, including bodily fluids or biological tissues.

[0033] As used herein, the term "biomarker" may be used interchangeably with the term "biological marker" and is used to refer to any measurable substance that can be used to determine organ function or any other biological state or condition. In certain embodiments, the biomarkers may include proteins such as immunoglobulins, polynucleotides such as DNA and RNA, and metabolites. In certain embodiments, the biomarkers may be cytokines, including but not limited to colony stimulating factors (CSFs), interferons (IFNs), interleukins (ILs), and tumor necrosis factors (TNFs). In a preferred embodiment, the biomarker is the FMS-related tyrosine kinase 3 ligand biomarker. In another embodiment, the biomarker is the cytokine stem cell factor.

[0034] Detection and quantification of biomarkers in a sample (such as urine) may be performed at any time after collection. For example, quantification of biomarkers in a biological sample may be performed within a short period of time (e.g., within 2 minutes, within 5 minutes, etc.) or after a longer storage, transport, preservation, or incubation step (e.g., within 6 hours, within 72 hours, etc.). In certain embodiments, the biological sample is preserved, transported, or stored at a temperature ranging from about -200°C to about 30°C, e.g., -80°C. As another example, tissue biopsies are preferably stored at -20°C for short-term storage and -80°C for long-term storage. Of course, these examples are for illustrative purposes only, and the appropriate temperature will depend on the type of biological sample used in the present invention. The biological sample should be stored at a temperature appropriate for the particular type of biological sample, as is generally understood by those of skill in the art.

[0035] One or more biomarkers in a sample may be used as a diagnostic tool for any type of disease, disorder, or other condition. For example, one or more biomarkers may indicate a tendency (e.g., likelihood of developing) that a cat has for a disease, disorder, and / or condition. In a preferred embodiment, one or more biomarkers are used to diagnose or identify a tendency for interstitial cystitis in a cat. In an even more preferred embodiment, FLT3L or the cytokine SCF are used to diagnose or identify a tendency for interstitial cystitis in a feline.

[0036] As described herein, electronic assays may be used to quantify biomarkers in biological fluids (e.g., blood, saliva, urine, etc.). Biomarkers may be associated with diseases, disorders, and / or conditions. For example, biomarkers may indicate the presence of a disease, disorder, and / or condition. Biomarkers may indicate risk (e.g., risk of developing) a disease, disorder, and / or condition. Electronic assays may use impedance, such as impedance cytometry, to quantify biomarkers in biological fluids. The biomarkers may include proteins. The biomarkers may include immunoglobulin G (IgG) and / or immunoglobulin A (IgA). Quantification of the biomarkers in the biological fluid may be performed in a short period of time (e.g., within 5 minutes, within 2 minutes, etc.) and via compact, lightweight, and / or inexpensive equipment. In embodiments, machine learning techniques (such as supervised machine learning techniques) may be used to determine (e.g., aid in determining) the quantification of the biomarkers (such as immunoglobulins) in the biological fluid.

[0037] In some embodiments, the invention is a pet food composition used to treat any of the pathologies or conditions described herein. In a preferred embodiment, the invention is a method of treating interstitial cystitis in a feline comprising administering the pet food composition. Preferably, the pet food composition comprises a high level of antioxidants. In a more preferred embodiment, the pet food composition comprises a high level of antioxidants and is fed daily for at least 28 days.

[0038] The pet food composition may be in the form of a kibble. In other embodiments, the pet food composition is in the form of a multi-layered kibble and / or a multi-layered kibble including a coating. Furthermore, the coating may include a taste enhancer. The term "taste" as used herein encompasses all of the various characteristics of food sensed by an animal, such as texture, taste, and smell. For example, the coating may include a taste enhancer to increase the taste of the pet food composition. In certain embodiments, the composition has a taste equivalent to that of a control composition.

[0039] In certain embodiments, the kibble is shaped by extrusion. In other embodiments, the composition is in a form selected from a loaf, a stew, a "meat and gravy" form, a congee, shreds having a moisture content of greater than 50%, and a product that can be extruded from a syringe. In another embodiment, the present invention comprises between 6% and about 12% moisture by weight.

[0040] In some embodiments, the kibble may include a binder. In certain embodiments, binders include monosaccharides such as glucose, fructose, mannose, arabinose, disaccharides and trisaccharides such as sucrose, lactose, maltose, trehalose, lactulose, corn and rice syrup solids, dextrins such as corn, wheat, rice and tapioca dextrins, maltodextrins, starches such as rice, wheat, corn, potato, tapioca starch or these starches modified by chemical modification, alginates, chitosan, gums such as carrageen and gum arabic, glycerol, sorbitol, malic acid, sorbitol ... The preferred saccharides and / or denatured proteins include, but are not limited to, polyols such as glycerol, tartrate, xylitol, erythritol; esters of polyols such as sucrose esters, polyglycol esters, glycerol esters, polyglycerol esters, sorbitan esters; sorbitol; molasses; honey; gelatin; peptides; proteins and denatured proteins such as whey liquid, whey powder, concentrated whey, whey isolate, whey protein isolate, high lactose whey by-product, meat broth solids such as chicken broth, chicken broth solids, soy protein, and egg whites, or combinations thereof.

[0041] In certain embodiments, the binder includes, but is not limited to, lipid and / or lipid derivative.Lipid can be used in combination with water and / or other binder components.Lipid can include vegetable fats such as soybean oil, corn oil, rapeseed oil, olive oil, safflower oil, palm oil, coconut oil, palm kernel oil and their partially and fully hydrogenated derivatives, animal fats and their partially and fully hydrogenated derivatives, and wax.

[0042] In certain embodiments, the present invention may include additional ingredients including, but not limited to, additives, minerals, vitamins, carbohydrate sources, fats, proteins, additional fiber, amino acids, carotenoids, antioxidants, fatty acids, glucose mimetics, probiotics, prebiotics, and the like.

[0043] The pet food composition may contain additives known in the art. Such additives should be present in amounts that do not impair the purpose and effect provided by the present invention. Examples of additives include substances that have a stabilizing effect, sensory stimulants, processing aids, and substances that provide nutritional benefits.

[0044] Stabilizing substances may extend the shelf life of the composition.Suitable examples may include preservatives, antioxidants, synergists and scavengers, packaging gases, stabilizers, emulsifiers, thickeners, gelling agents, and wetting agents.Examples of emulsifiers and / or thickeners include gelatin, cellulose ethers, starches, starch esters, starch ethers, and modified starches.

[0045] Additives for coloring, flavoring, and nutritional purposes can include coloring agents, salts (including but not limited to sodium chloride, potassium citrate, potassium chloride, and other edible salts), vitamins, minerals, and flavorings. The amount of such additives in the composition is typically up to about 5% by weight (based on the dry matter of the composition). In some instances, the pet food composition includes additives in an amount of up to about 4% by weight, up to about 3.5% by weight, up to about 3% by weight, up to about 2.5% by weight, up to about 2% by weight, up to about 1.5% by weight, or up to about 1% by weight, based on the total weight of the pet food composition on a dry matter basis. Additionally or alternatively, the pet food composition can include up to about 1% by weight, up to about 0.5% by weight, or up to about 0.1% by weight of additives, based on the total weight of the pet food composition on a dry matter basis. Other additives can include antioxidants, omega-3 fatty acids, omega-6 fatty acids, glucosamine, chondroitin sulfate, plant extracts, herbal extracts, and the like.

[0046] In certain embodiments, the pet food composition contains vitamins and minerals in the amounts required to avoid deficiencies and maintain health. These amounts are readily available in the art. The Association of American Feed Control Officials (AAFCO) provides recommended amounts of these ingredients for dogs and cats (see Association of American Feed Control Officials. Official Publication, pp. 126-140 (2003)). Minerals may be specifically maintained at optimal levels known to those skilled in the art to reduce the incidence of stone formation.

[0047] Vitamins that may be mentioned include, by way of example, Vitamin A, Vitamin B1 (thiamine or related sources such as thiamine nitrate), Vitamin B2 (riboflavin), Vitamin B3 (niacin), Vitamin B5 (pantothenic acid or related sources such as calcium pantothenate), Vitamin B6 (pyridoxine or related sources such as pyridoxine hydrochloride), Vitamin B8 (folic acid), Vitamin B12, Vitamin C (ascorbic acid), Vitamin D (such as a Vitamin D3 supplement), Vitamin E, Vitamin H (biotin), Vitamin K, acetate, choline-related sources such as choline and choline chloride, and inositol.

[0048] Minerals and trace elements can include, for example, calcium, phosphorus, sodium, potassium, magnesium, copper, zinc, choline, and iron salts. Mineral sources can include, for example, sodium selenite, monosodium phosphate, calcium carbonate, potassium chloride, ferrous sulfate, zinc oxide, manganese sulfate, copper sulfate, manganese oxide, potassium iodide, and / or cobalt carbonate.

[0049] The term "carbohydrate" as used herein includes polysaccharides (e.g., starch, dextrin) and sugars (e.g., sucrose, lactose, maltose, glucose, fructose) that are metabolized for energy when hydrolyzed. Examples of high carbohydrate ingredients suitable for inclusion in the compositions disclosed herein include, but are not limited to, corn, grain sorghum, wheat, barley, and rice.

[0050] In certain embodiments, the carbohydrate component comprises a mixture of one or more carbohydrate sources. Examples of carbohydrates or carbohydrate components may include cereals, grains, corn, wheat, rice, oats, corn grits, sorghum, grain sorghum / milo, wheat bran, oat bran, amaranth, durum, and / or semolina.

[0051] By properly balancing the carbohydrate sources, one skilled in the art can manipulate the texture of the final product, for example, short chain polysaccharides tend to be sticky or starchy and long chain polysaccharides tend to be less sticky or starchy than short chains, and the desired texture of the hybrid food product is achieved by using long chain polysaccharides and modified starches (such as native or modified starches, cellulose, and the like).

[0052] The carbohydrate mixture may additionally include optional ingredients such as added salt, spices, seasonings, vitamins, minerals, flavorings, colorants, and the like. The amounts of the optional additives will depend at least in part on the nutritional requirements of the animal at different life stages.

[0053] In some embodiments, the present invention may include from about 5% to about 25% by weight fat. For example, the pet food composition may include fat in an amount of from about 5% to about 25% by weight, from about 5% to about 20% by weight, from about 5% to about 15% by weight, from about 5% to about 10% by weight, from about 10% to about 25% by weight, from about 10% to about 20% by weight, from about 10% to about 15% by weight, from about 15% to about 25% by weight, from about 15% to about 20% by weight, or from about 20% to about 25% by weight, based on the total weight of the pet food composition. Fats or fat component sources may include chicken fat, chicken fat, turkey fat, lard, tallow, beef tallow, vegetable oil, corn oil, soybean oil, cottonseed oil, palm oil, palm kernel oil, linseed oil, canola oil, rapeseed oil, fish oil, menhaden oil, anchovy oil, and / or olestra.

[0054] In some embodiments, the present invention may comprise about 5% to about 30% by weight of protein. For example, the pet food composition may comprise about 5% to about 30% by weight, about 5% to about 25% by weight, about 5% to about 20% by weight, about 5% to about 15% by weight, about 5% to about 10% by weight, about 10% to about 30% by weight, about 10% to about 25% by weight, about 10% to about 20% by weight, about 10% to about 15% by weight, about 15% to about 30% by weight, about 15% to about 25% by weight, about 15% to about 20% by weight, about 20% to about 30% by weight, or about 20% to about 25% by weight, based on the total weight of the pet food composition. The term "protein" refers to a polypeptide, or peptide, or polymer of amino acids. The term encompasses naturally occurring and non-naturally occurring (synthetic) polymers, as well as polymers in which artificial chemical mimetics have been substituted for one or more amino acids. The term also encompasses fragments, variants, and homologs that have the same or substantially the same properties and perform the same or substantially the same functions as the original sequence. The term encompasses polymers of any length, including those containing about 2-1000, 4-800, 6-600, 8-400 amino acids. The term encompasses amino acid polymers that are synthesized, as well as those that are isolated and purified from natural sources. Under some embodiments, the terms "polypeptide," "peptide," or "protein" are used interchangeably.

[0055] Proteins may be provided by any of a variety of sources known to those skilled in the art, including plant sources, animal sources, microbial sources, or combinations thereof. For example, animal sources may include, for example, meat, meat by-products, seafood, dairy products, eggs, and the like. Meat may include, for example, animal flesh, such as chicken, fish, mammals (including cows, pigs, sheep, goats, and the like). Meat by-products may include, for example, lungs, kidneys, brains, livers, stomachs, intestines. Plant proteins may include, for example, soybeans, cottonseeds, and peanuts. Microbial sources may be used to synthesize amino acids (e.g., lysine, threonine, tryptophan, methionine) or intact proteins (such as proteins from the sources listed below).

[0056] Examples of proteins, or protein ingredients, may include chicken meal, chicken, chicken by-product meal, lamb, lamb meal, turkey, turkey meal, beef, beef by-products, offal, fish meal, intestines, kangaroo, white fish, venison, soy meal, soy protein isolate, soy protein concentrate, corn gluten meal, corn protein concentrate, distillers dried grains and / or distillers dried grains solution, and single cell proteins such as, for example, yeast, algae, and / or bacterial cultures.

[0057] Protein can be intact, completely hydrolyzed, or partially hydrolyzed. The protein content of food can be determined by several methods known to those skilled in the art, such as the 988.05 method published by the Association of Official Analytical Chemists in Official Methods of Analysis ("OMA"). The amount of protein in the compositions disclosed herein can be determined based on the amount of nitrogen in the composition according to methods known to those skilled in the art.

[0058] Exemplary amino acids may include 1-tryptophan, taurine, histidine, carnosine, alanine, cysteine, arginine, methionine, tryptophan, lysine, asparagine, aspartic acid, phenylalanine, valine, threonine, isoleucine, histidine, leucine, glycine, glutamine, taurine, tyrosine, homocysteine, ornithine, citrulline, glutamic acid, proline, and / or serine. Sources of carotenoids may include lutein, astaxanthin, zeaxanthin, bixin, lycopene, and / or beta-carotene. Sources of antioxidant components may include tocopherols (vitamin E), vitamin C, vitamin A, plant-derived materials, carotenoids (discussed above), selenium, and / or CoQ10 (coenzyme Q10). In one preferred embodiment, the pet food composition contains high levels of arginine and its derivatives, and / or low levels of tryptophan and its derivatives. In another preferred embodiment, the pet food composition contains high levels of polyunsaturated fatty acids (e.g., alpha-linolenic acid, arachidonic acid, EPA, DHA).

[0059] Exemplary fatty acid components may include arachidonic acid, alpha-linoleic acid, gamma-linolenic acid, linoleic acid, eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA), and / or fish oil (as a source of EPA and / or DHA). Sources of glucose mimetics may include glucose antimetabolites (including 2-deoxy-D-glucose, 5-thio-D-glucose, 3-O-methylglucose), anhydrosugars (including 1,5-anhydro-D-glucitol, 2,5-anhydro-D-glucitol, 2,5-anhydro-D-mannitol), mannoheptulose, and / or avocado extracts containing mannoheptulose.

[0060] Further ingredients include beef broth, brewer's dried yeast, eggs, egg products, ground flax, DL-methionine, amino acids, leucine, lysine, arginine, cysteine, cystine, aspartic acid, polyphosphates, sodium pyrophosphate, sodium tripolyphosphate; zinc chloride, copper gluconate, stannous chloride, stannous fluoride, sodium fluoride, triclosan, glucosamine hydrochloride, chondroitin sulfate, green mussels, blue lipped mussels. mussel), methylsulfonylmethane (MSM), boron, boric acid, phytoestrogens, phytoandrogens, genistein, daidzein, L-carnitine, chromium picolinate, chromium tripicolinate, chromium nicotinate, acid / base modifiers, potassium citrate, potassium chloride, calcium carbonate, calcium chloride, sodium bisulfate; eucalyptus, lavender, peppermint, plasticizers, colorants, flavors, sweeteners, buffers, slip aids, carriers, pH adjusters, natural ingredients, stabilizers, biological additives such as enzymes (including proteases and lipases), chemical additives, coolants, chili powders, etc. The composition may include, for example, surfactants, denaturants, drug astringents, emulsifiers, topical soothing agents, fragrance compounds, humectants, opacifying agents (such as zinc oxide and titanium dioxide), antifoaming agents (such as silicones), preservatives (butylated hydroxytoluene (BHT) and butylated hydroxyanisole (BHA), propyl gallate, benzalkonium chloride, EDTA, benzyl alcohol, potassium sorbate, parabens and mixtures thereof), reducing agents, solvents, hydrotropes, solubilizing agents, suspending agents (non-surfactant), solvents, viscosity increasing agents (aqueous and non-aqueous), sequestering agents, and / or keratolytic agents.

[0061] The probiotic component may comprise any suitable bacteria, yeast, microorganism, and / or any mixture thereof. A variety of probiotic microorganisms are known in the art. In certain embodiments, the probiotic component may comprise bacteria of the order Lactobacillales, bacteria of the genera Bacillus, Bacteroides, and / or Bifidobacterium, yeasts of the order Saccharomycetales, including the family Saccharomycetes and the genera Candida, and / or any mixture thereof. Probiotics may or may not form spores.

[0062] In certain embodiments, the pet food composition may include polyphenols. In some embodiments, the polyphenol source includes a phenolic compound selected from ellagic acid, gallic acid, protocatechuic acid, p-hydroxybenzoic acid, catechin, and combinations of two or more thereof. In some embodiments, the polyphenol source includes pecan shells, or any other components of pecan nuts. Examples of additional sources of polyphenols may include tea extract, rosemary extract, rosmarinic acid, coffee extract, pecan shells, caffeic acid, turmeric extract, blueberry extract, grape extract, grape seed extract, and / or soybean extract.

[0063] The pet food composition preferably comprises an effective amount of a fiber-bound polyphenol component. The amount of fiber-bound polyphenol in the pet food composition can optionally be from about 0.1% to about 60% by weight. For example, in various embodiments, the pet food composition can contain from about 0.1% to about 60% by weight, from about 0.1% to about 50% by weight, from about 0.1% to about 40% by weight, from about 0.1% to about 30% by weight, from about 0.1% to about 25% by weight, from about 0.1% to about 20% by weight, from about 0.1% to about 17% by weight, from about 0.1% to about 14% by weight, from about 0.1% to about 11% by weight, from about 0.1% to about 9% by weight, from about 0.1% to about 7% by weight, from about 0.1% to about 5% by weight, or from about 0.1% to about 10% by weight, based on the total weight of the pet food composition. Amount% to about 3% by weight, about 1% to about 60% by weight, about 1% to about 50% by weight, about 1% to about 40% by weight, about 1% to about 30% by weight, about 1% to about 25% by weight, about 1% to about 20% by weight, about 1% to about 17% by weight, about 1% to about 14% by weight, About 1% to about 11% by weight, about 1% to about 9% by weight, about 1% to about 7% by weight, about 1% to about 5% by weight, about 1% to about 3% by weight, about 5% to about 60% by weight, about 5% to about 50% by weight, about 5% to about 40% by weight, about 5% to about 30% by weight, About 5% to about 25% by weight, about 5% to about 20% by weight, about 5% to about 17% by weight, about 5% to about 14% by weight, about 5% to about 11% by weight, about 5% to about 9% by weight, about 5% to about 7% by weight, about 10% to about 60% by weight, about 10% to about 50% by weight Weight%, about 10% to about 40% by weight, about 10% to about 30% by weight, about 10% to about 25% by weight, about 10% to about 20% by weight, about 10% to about 17% by weight, about 10% to about 14% by weight, about 15% to about 60% by weight, about 15% to about 50% by weight The fiber-bound polyphenol component may be present in an amount of about 15% by weight to about 40% by weight, about 15% by weight to about 30% by weight, about 15% by weight to about 25% by weight, about 15% by weight to about 20% by weight, about 20% by weight to about 60% by weight, about 20% by weight to about 50% by weight, about 20% by weight to about 40% by weight, about 20% by weight to about 30% by weight, about 20% by weight to about 25% by weight, about 30% by weight to about 60% by weight, about 30% by weight to about 50% by weight, about 30% by weight to about 40% by weight, about 40% by weight to about 60% by weight, or about 40% by weight to about 50% by weight.

[0064] The pet food composition may be determined by any of a variety of methods for feed analysis known to those skilled in the art. Feed analysis may be performed to measure any of the nutritional contents listed herein, including moisture, protein, fiber, carbohydrate, energy, vitamins, minerals, energy, fat, and ash content.

[0065] Protein content may be measured and reported in any of a variety of ways known to those skilled in the art. Protein may be reported as crude protein (CP) to measure both true protein content and non-protein nitrogen. Crude protein content may be further differentiated between degradable intake protein (DIP), undegradable intake protein (UIP), and metabolizable protein (MP). In certain embodiments, protein content may be differentiated to include heat damaged or insoluble crude protein (ICP), adjusted crude protein (ACP), digestible protein (DP).

[0066] Fiber content may be measured and reported by any of a variety of methods known to those skilled in the art. Fiber content may be reported as total dietary fiber (TDF, a combination of soluble and insoluble fiber), crude fiber (CF), neutral detergent fiber (NDF), acid detergent fiber (ADF) and / or acid detergent lignin (ADL). Crude fiber is generally known to estimate the indigestible portion of the plant material found in a pet food composition. ADF measures cellulose and lignin, which are components of plant cell walls. NDF measures the total material found in the plant cell wall and also includes the hemicellulose in addition to the fiber content measured as ADF. ADL measures only the lignin portion of the plant cell wall.

[0067] Energy content may be measured and reported in any of a variety of ways known to those skilled in the art and may be reported as digestible energy (DE), metabolizable energy (ME), net energy (NE), total digestible nutrients (TDN), ether extractable (EE), relative feed value (RFV), relative feed quality (RFQ).

[0068] Embodiments of the invention will now be further described by the following non-limiting examples. EXAMPLES

[0069] A retrospective study was performed on serum and urine samples from 44 cats. Of the 44 cats, 6 cats had a clinical diagnosis of FIC, 3 cats had a clinical diagnosis of feline lower urinary tract disease (FLUTD), 13 cats had a clinical diagnosis of cystitis (UB), and 23 cats were used as healthy controls.

[0070] Cytokine analysis was performed on serum and urine samples measuring 19 cytokines (sFas, TNFα, IL-12p40, SCF, PDGF-BB, IL-13, IL-18, IL-6, IL-4, IL-2, GM-CSF, KC, RANTES, SDF-1, FLT-3L, IL-1β, IFNγ, MCP-1 and IL-8). Cytokine analysis demonstrated that the urinary cytokine Flt3-L was significantly decreased in FIC cats compared to healthy cats (Table 1 below). Shaded cells indicate clinically significant differences (p<0.05) between groups. [Table 1]

[0071] Univariate analysis was performed that identified urinary Flt3-L and the cytokine SCF as having a diagnostic predictive potential of 76% and 72%, respectively (Table 2 below). Shaded cells indicate clinically significant differences (p≦0.5) between groups. [Table 2]

[0072] After analysis, two cats clinically diagnosed with FIC were fed either "Feline Metabolic AOX upgrade dry" or "Feline k / d Chick Canned" cat food for 28 days. Urine samples were taken on day 28 (when the experiment ended) and several years after diagnosis. Cytokine analysis demonstrated that urinary Flt3-L levels in the FIC group were close to the mean levels of urinary Flt3-L levels in healthy cats.

[0073] The present invention has been described with reference to several embodiments that have been described in considerable detail for the purpose of providing a complete disclosure of the invention, but such embodiments are merely exemplary and are not intended to limit or represent an exhaustive enumeration of all aspects of the invention. The scope of the invention should be determined from the claims appended hereto. Moreover, it will be apparent to one skilled in the art that numerous changes can be made in these details without departing from the spirit and principles of the invention.

Claims

1. A pet food composition containing an effective amount of fiber-bound polyphenol component for use in increasing the levels of urinary cytokines selected from Fms-like tyrosine kinase 3 (Flt3-L), stem cell factor (SCF), and IL-12p40 in companion animals.

2. The pet food composition according to claim 1, wherein the fiber-bound polyphenol component comprises materials selected from oat fiber, psyllium husk, tomato pomace, flaxseed, beet pulp, carrot powder, brewed rice, and two or more combinations thereof.

3. The pet food composition according to claim 1 or claim 2, wherein the fiber-bound polyphenol component comprises a material selected from brewed rice, oat fiber, plantain husk, and two or more combinations thereof.

4. The pet food composition according to any one of claims 1 to 3, wherein the fiber-bound polyphenol component comprises a material selected from tomato pomace, flaxseed, beet pulp, carrot powder, and two or more combinations thereof.

5. The pet food composition according to any one of claims 1 to 4, wherein the pet food composition increases the level of urinary cytokines after about 14 days.

6. The pet food composition according to any one of claims 1 to 5, wherein the pet food composition increases the level of urinary cytokines after about 28 days.

7. The pet food composition according to any one of claims 1 to 6, wherein the pet food composition increases the level of urinary cytokines after about 21 days.

8. The pet food composition according to any one of claims 1 to 7, wherein the urinary cytokine is Flt3-L.

9. The pet food composition according to any one of claims 1 to 8, wherein the companion animal is a feline.

10. The pet food composition according to any one of claims 1 to 9, wherein the pet food composition is in the form of kibble.

11. The pet food composition according to any one of claims 1 to 10, wherein the fiber-bound polyphenol component is present in the pet food composition in an amount of about 0.1% to about 60% by weight.

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