WO2019203827A1

WO2019203827A1 - Composition and method for preventing or treating hyperuricemia or gout

Info

Publication number: WO2019203827A1
Application number: PCT/US2018/028252
Authority: WO
Inventors: Natarajan Ranganathan
Original assignee: Kibow Biotech Inc .
Priority date: 2018-04-19
Filing date: 2018-04-19
Publication date: 2019-10-24

Abstract

A composition comprising probiotic components and a curcuminoid is provided for use in reducing uric acid levels in blood and preventing or treating hyperuricemia or gout. In some embodiments, the probiotic components include a Lactobacillus acidophilus bacterium and a Lactobacillus rhamnosus bacterium and a curcuminoid.

Description

COMPOSITION AND METHOD FOR PREVENTING OR TREATING

HYPERURICEMIA OR GOUT

Background of the Invention

[0001] Hyperuricemia is a common condition, with some studies showing the prevalence as high as 21% in men and women. Hyperuricemia is especially common in patients with metabolic, cardiovascular and renal diseases. While the presence of hyperuricemia in the absence of gout has often been described as "asymptomatic", recent studies suggest hyperuricemia may have a contributory role in metabolic and cardiovascular diseases. Potential mechanisms involved in uric acid' s deleterious metabolic effects include the ability of soluble uric acid to increase oxidative stress, mitochondrial and endoplasmic reticulum dysfunction, endothelial dysfunction, and activation of the renin- angiotensin system, as well as to increase synthesis and secretion of proinflammatory factors. Depending on the cell type, uric acid may also cause increased cellular proliferation (i.e., vascular smooth muscle cells) or cellular apoptosis (renal tubular cells, endothelial cells ) .

[0002] Extrarenal excretion of uric acid via the intestine accounts for up to one-third of its total excretion. Uric acid is secreted into the gut where it is rapidly metabolized by bacterial flora. Moreover, it was recently shown that hyperuricemic patients with gout have a significantly different intestinal flora in comparison to normo-uricemic subjects. In addition, transporter-mediated intestinal secretion or uptake of uric acid is a significant step in the metabolism by gut flora. It has been found that endogenous uric acid is secreted from blood directly into the intestinal lumen at all intestinal segments. In humans, some polymorphic variants of the transporter BCRP (also known as DTR-binding cassette transporter, sub-family G, member 2, ABCG2), secretes uric acid into the intestine, resulting in decreased transporter activity, elevated serum uric acid, and an increased incidence of gout. Moreover, blockade of uric acid transport into intestine induced an increment in serum uric acid concentrations, suggesting again that the intestinal mechanism is essential to modulate uric acid systemic levels. Another transporter, SLC2A9, is also important in intestinal secretion, and knockdown of intestinal SLC2A9 can also result in hyperuricemia and features of metabolic syndrome .

[0003] A population that is highly dependant on gut excretion and metabolism are patients with end-stage renal disease who have not yet started dialysis and in whom hyperuricemia is extremely common. In these patients, uric acid excretion relies heavily on uricolysis by gut flora. Additionally, hyperuricemia has been associated with the risk of developing acute kidney injury and with the progression of chronic kidney disease. Moreover, the use of drugs aimed to reduce serum uric acid has shown promise in treating these diseases.

[0004] Current therapy for lowering serum uric acid levels includes inhibitors of xanthine oxidase (allopurinol, febuxostat) , recombinant uricase ( rasburicase ) and uricosuric agents (probenecid) . However, all these drugs may produce undesired health effects. Therefore, the development of alternative therapeutic strategies to reduce uric acid concentrations would be useful. Recent research has focused on the use of various microorganisms, known as probiotics, to affect removal of undesired waste products in vivo and in vitro. For example, it has been demonstrated that microencapsulated, genetically-engineered E. coli DH5 was effective in removing urea and ammonia in an in vitro system and in a uremic rat animal model. Further, it has been demonstrated that supplemental gum arabic fiber increases fecal nitrogen excretion and lowers urea nitrogen concentration in chronic renal failure patients consuming a low protein diet and that canine renal patients fed a diet containing a fermentable fiber blend improved clinical end- stage renal disease status. Moreover, US Patent No. 6,706,263 disclosed a microencapsulated and enteric coated composition that comprised a mixture of sorbents with specific adsorption affinities for uremic toxins wherein the sorbents claimed were oxystarch, locust bean gum and activated charcoal; and the bacteria claimed were ones that metabolize urea and ammonia.

[0005] A recent review describes the evidence supporting the clinical use of dietary supplements for treatment of gout (Andres, M. et al. 2014. Cochrane Data. Syst. Rev. 10: CD010156) . The dietary supplements reviewed included skim milk, lactose powder and vitamin C. The authors concluded that the available clinical evidence did not provide support for the utility of these treatments. The treatments examined were not treatments comprising probiotics administered either alone or in combination with other dietary supplements.

[0006] The effects of probiotics on blood uric acid levels have been studied to some extent. Asemi, Z. et al. (2014. Clin. Nutr. 33:198-203) reported on results of a clinical trial where the effects of a synbiotic composition on clinical endpoints in patients with diabetes was examined. One endpoint was blood uric acid levels. The synbiotic food consisted of a probiotic viable and heat-resistant Lactobacillus sporogenes (lxlO⁷ CFU) , 0.04 g inulin as a prebiotic with 0.38 g isomalt, 0.36 g sorbitol and 0.05 g stevia as sweetener (per gram) . Control food (the same substance without probiotic bacteria and prebiotic inulin) was packed in identical 9-gram packages. Patients were asked to consume the synbiotic and control foods three times a day for six weeks. The authors reported that uric acid levels were increased, not decreased, in patients taking the synbiotic food product, although markers of oxidative stress were reduced.

[0007] Bai et al. (2014. Renal Fall. 36:5, 790-794) studied the effect of a mutant probiotic organism to lower levels of uric acid in blood samples obtained from uremic patients. In this in vitro study, a new mutant bacterial strain, DUC3-17, from probiotic Lactobacillus bulgaricus was capable of decomposing uremic toxins. The authors suggested that this organism may be useful to assist in delaying the progression of chronic kidney disease.

[0008] In a recent clinical study examining the effect of a synbiotic formulation on azotemia in patients with chronic kidney disease (Dehghani, H. et al. 2016. IJKD 10:351-7), the authors reported that the intake of a synbiotic supplement could reduce blood urea nitrogen in patients with CKD in stages 3 and 4; however, it had no effect on the other markers of kidney function. The symbiotic known as FAMILACT (Zist Takhmir, Tehran, Iran) was used. This composition was stated to contain 500 mg of a mixture of seven different probiotic strains (Lactobacillus casei, Lactobacillus acidophilus, Lactobacillus bulgarigus, Lactobacillus rhamnosus, Bifidobacterium breve,

Bifidobacterium longum, Streptococcus thermophilus) , and a prebiotic which was a mixture of fructooligosaccharides .

[0009] Other probiotic compositions have been taught for uses other than to reduce uric acid levels or to affect markers of kidney function, or to treat gout. For example, US 5,756,088 teaches a prescription diet for the prevention and treatment of dog and cat dermatosis comprising a composition containing a poly-unsaturated fatty acid such as y-linolenic acid, g-linolenic acid and docosahexaenoic acid, and/or biotin, and an antiflatulent such as a lactic acid bacterium, a Bifidobacterium, a Lactobacillus, a butyric acid bacterium or a Bacillus, and optionally an oligosaccharide. US 7,993,903 teaches a composition for inhibiting cholesterol absorption in the intestinal tract, wherein the composition includes Bifidobacterium, and optionally a Lactobacillus bacterium and carbohydrate. US 2011/0171283 teaches a composition containing at least one nutrient, at least one disinfecting or decontaminating and/or at least one proteases inhibiting substance and/or complex of substances incorporated in an absorbent dressing for external care and/or treatment of wounds to a human or animal. In one embodiment, the protease inhibiting substance includes non-pathogenic acid producing micro organisms (e.g., bifidobacteria, lactococci, or lactobacilli ) and/or synbiotics (e.g., xylooligosaccharide) . US 2009/0252709 teaches a preventive or therapeutic agent for gastritis or ulcer, which includes as an active ingredient Bifidobacterium bifidum. This reference teaches that other microorganisms (e.g., Bifidobacterium or Lactobacillus bacteria) , as well as sugars such as xylooligosaccharide. WO 2007/140622 teaches a probiotic composition containing a mixture of a Propionibacterium, a Lactobacillus, a Bifidobacterium and a Streptococcus, wherein said composition can further include a prebiotic.

[0010] US Patent No. 6,706,287 discloses a pharmaceutical composition to alleviate the symptoms of uremia that comprised a probiotic, a prebiotic, a sorbent for inorganic phosphate, an adsorbent for specific uremic solutes other than urea, and a microorganism with high alkaline pH stability and high urease activity to aid in degrading urea. The composition was microencapsulated or enteric coated. The patent teaches that the prebiotic component ensured the viability of the probiotic component, while the microencapsulation or enteric coating protected the patient from infection by the organism. The probiotic species described included Sporosarcina ureae, Bacillus pasteurii, trained lactobacillus and Bacillus species and a novel Lactobacillus, known as KB-I.

[0011] US Patent No. 7,026,160 discloses use of a probiotic for affecting nitrogenous waste levels in patients with renal failure. The composition disclosed comprises Bacillus pasteurii , an organism which converts nitrogenous waste into non-toxic compounds in vivo. In a related patent, US Patent No. 7,998,470 disclosed compositions and methods for improving renal function with Streptococcus probiotic bacteria selected for converting nitrogenous waste into non-toxic compounds. By reducing the levels of nitrogenous wastes, the probiotic composition led to a reduced kidney burden. The Streptococcus thermophilus organisms taught were known as KB19, KB4 and KB25, and were selected because they were capable of reducing urea concentrations from 300 mg/dL to 20 mg/dL within 24 hours at a pH of 6.3.

[0012] US Patent No. 8,257,693 discloses a composition that contains both prebiotic and probiotic components that is used to reduce elevated levels of nitrogenous waste products and to promote a healthy bowel microenvironment. Specifically, the composition taught consists of Lactobacillus acidophilus, Streptococcus thermophilus, Bifidobacterium longum, psyllium husks and a pharmaceutical excipient, and also is enterically coated. In preferred embodiments, the composition organisms are viable organisms and are present in a ratio of about 1:1:8. The probiotic organisms are taught to make up about 50% by weight of the composition while psyllium husks make up about 45% of the composition. In a related patent (US Patent No. 8,481,025) the same composition of prebiotic and probiotic components is taught to be useful for removing nitrogenous waste products and for treating renal failure. The compositions are also taught to have a probiotic component that makes up from 20% to 70% of the composition, or to alternatively have a prebiotic component that makes up from 20% to 70% of the composition.

[0013] US Patent No. 9,655,932 discloses compositions and methods for reducing uric acid levels in the blood of patients, as well as methods for preventing or treating hyperuricemia or gout, by administering a composition containing a Lactobacillus bacterium, a Bifidobacterium bacterium, and a xylooligosaccharide . In specific embodiments, the composition consisted essentially of Lactobacillus acidophilus, Bifidobacterium longum, and a xylooligosaccharide .

Summary of the Invention

[0014] This invention provides a composition comprising at least two probiotic components and a curcuminoid. In certain embodiments, the at least two probiotic components include a Lactobacillus acidophilus bacterium and a Lactobacillus rhamnosus bacterium and the curcuminoid includes curcumin. In other embodiments, the composition is formulated as an extended release composition, and/or with a prebiotic component which comprises an xylooligosaccharide. Methods for reducing blood and urine levels of uric acid and preventing or treating hyperuricemia gout in a patient using the composition of invention are also provided.

Detailed Description of the Invention

[0015] One of the main functions of the normal, healthy kidney, besides its regulatory, endocrine, and metabolic functions, is the disposal of waste products. Any impairment of excretory function can lead to the accumulation of a variety of nitrogenous waste products including, urea, creatinine and uric acid. High concentrations of uric acid in the blood stream can lead to a condition known as hyperuricemia, which in turn can lead to development of symptoms consistent with a condition known as gout. Gout is a form of arthritis caused by excess uric acid in the bloodstream. The symptoms of gout are due to the formation of uric acid crystals in the joints and the body's response to them (swelling in joints is common) . Nitrogenous solutes in the circulating blood, such as uric acid, promote osmotic diffusion into the lumen because of the concentration gradient across the intestinal wall. This diffusion mechanism led to the concept of oral sorbents to augment gut-based clearance of nitrogenous waste products . Sorbents or microbes have demonstrated their ability to remove various compounds and nitrogenous wastes within the large bowel.

[0016] In addition to preventing or reducing hyperuricemia in blood as a method to prevent or treat gout, gout treatment can involve use of anti-inflammatory agents (Grosser, T. et aI. 2018. Pharmacotherapy of inflammation, fever, pain, and gout. In: Goodman and Gilman's The Pharmacological Basis of Therapeutics , 13^th edition. McGraw- Hill: New York, NY, chapter 38). Curcumin, an active ingredient in the turmeric plant, is a yellow-colored pigment that has been investigated for its anti inflammatory properties (e.g., Menon, V.P. and A.R. Sudheer. 2007. Adv. Exp. Med. Biol. 595:105-125; Jurenka, J.S. 2009. Altern. Med. Rev. 14 (2) : 141-153; Oliviera, F. et al. 2017. J. Sci . Food Agric. September 8 doi : 10.1002 /j sfa .8664 epub ahead of print). Curcumin has been studied in animals and in humans and has been shown to be safe and effective as an anti-inflammatory agent (Jurenka et al. 2009. Altern. Med. Rev. 14 (2) : 141-153) , although studies have not been done in humans to demonstrate the safe and effective doses for use to treat gout .

[0017] It has now been shown that combining probiotic components with a curcuminoid effectively reduces the blood concentration of nitrogenous waste products, specifically levels of uric acid in vivo in an animal model of the hyperuricemic state. Reducing the levels of uric acid, and relieving hyperuricemia, is known to be beneficial in gout patients (Grosser, T. et al. 2018. Pharmacotherapy of inflammation, fever, pain, and gout. In: Goodman and

Gilman's The Pharmacological Basis of Therapeutics, 13^th edition. McGraw-Hill: New York, NY, chapter 38). For the first time, the specific combination of two probiotic organisms selected for their ability to reduce nitrogenous waste products and an anti-inflammatory compound, curcumin, has been tested in vivo in a model of hyperuricemia and shown to be effective at reducing levels of uric acid and improving endpoints associated with hyperuricemia such as renal function, blood pressure, oxidative stress, and endothelial dysfunction. The results presented herein were unexpected given that curcumin has been shown to exhibit antibacterial activity against both Gram-positive bacteria ( e.g., Staphylococcus aureus and Enterococcus faecalis) and Gram-negative bacteria [e.g., Escherichia coli, Pseudomonas aeruginosa , Salmonella paratyphi) as well as Mycobacterium tuberculosis . See Schraufstatter & Bernt (1949) Nature 164 (4167) : 456; Tyangi, et al . (2015) PLoS One

10 (3) : e0121313; Teow, et al . (2016) J. Trop. Med.

2016:2853045; Moghadamtousi , et al . (2014) BioMed. Res.

International Article ID 186864.

[0018] The present invention provides a composition composed of at least two probiotic components and a curcuminoid and use of the same in the treatment of hyperuricemia in humans and as a preventative agent or a treatment for gout. Curcumin [ 1 , 7-bis ( 4 -hydroxy-3- methoxyphenyl ) -1, 6-heptadiene-3 , 5-dione] is a yellow pigment present found in plants of the genus Curcuma. In particular, curcumin is found in Curcuma longa (turmeric) and Curcuma amada (mango ginger) . As disclosed herein, the term "curcuminoid" refers to a mixture of curcumin, demethoxycurcumin and bisdide ethoxycurcumin, wherein curcumin is the major component of the curcuminoid. Ideally, the curcuminoid of this invention includes about 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% curcumin. Accordingly, in certain embodiments, the curcuminoid is isolated from any source that can supply a food grade isolated curcumin ingredient, e.g., from a species of the genus Curcuma. As used herein, "an isolated curcuminoid" refers to a curcuminoid that is separated from its natural environment [e.g., the rhizome of a Curcuma plant) and is substantially free from other components with which it is naturally associated.

[0019] The curcuminoid may be isolated by any suitable method including, e.g., solvent extraction. Briefly, rhizomes of turmeric are dried, powdered, and treated with a solvent to obtain an extract. Suitable solvents for this purpose include acetone, hexane, ethyl acetate, dicholoroethane, chloroform, etc. The extraction is conveniently carried out at moderate temperatures ( 40-55 °C) and the solvent is partially removed to yield a concentrate containing 30-60% solids. This solution is cooled (e.g. to 4°C) to obtain crystals of curcu inoid, which are isolated by any suitable method, e.g., filtration or centrifugation. See, e.g., US 2007/0148263.

[0020] A probiotic component of the present invention refers to a culture of live or freeze-dried microorganisms which, when applied to man or animal, beneficially affects the host by improving the properties of the indigenous microflora. In accordance with the composition and method of this invention, at least two probiotic components or microorganisms are used, in particular two Lactobacillus species that have been selected for their ability to reduce levels of nitrogenous wastes, such as uric acid. Ideally, the probiotic component of the present invention consists essentially of two Lactobacillus strains that have been selected for their ability to exert a beneficial effect on the host, survive transit through the intestinal tract, adhere to intestinal epithelial cell lining, and exhibit the ability to metabolize uric acid or other nitrogenous waste. Furthermore, a probiotic component should have a good shelf-life. In a preferred embodiment, the probiotic component is a mixture of two organisms, Lactobacillus acidophilus and Lactobacillus rhamnosus . In a more preferred embodiment, the strains KB27 and KB79 are used as they have been selected for their ability to reduce levels of nitrogenous waste products, such as uric acid.

[0021] Microorganisms also useful in the invention are those that have the ability, either through natural selection or by genetic manipulation, to catabolize various nitrogenous compounds (e.g. , urea, creatinine, uric acid and ammonia) by expressing or overexpressing one or more cognate catabolic enzymes. Exemplary microorganisms are those having an elevated level of urease or creatininase secretion .

[0022] A microorganism exhibiting elevated levels of catabolic enzyme secretion can be selected or trained by exposing a selected microorganism on increasing amounts of the metabolite of interest (e.g., urea, creatinine, uric acid and ammonia) . For example, a standard strain of Lactobacillus can be trained to express elevated levels of urease by sequential passage of the strain on increasing amounts of urea, e.g., a single colony growing on 0.5% urea is selected and applied to medium containing 1.0% urea, a single colony growing on 1.0% urea is selected and applied to medium containing 2.0% urea, etc. Using such a method, L. acidophilus and L. rhamnosus strains having the ability to grow on 5% urea are isolated. These strains proliferate in artificial intestinal fluid (AIF, US Pharmacopeia) in the pH range of 5.5 to 7.5, characteristic of the colon environment; use urea as a sole nitrogen source; and catabolize urea in the presence of other nitrogen sources. Moreover, these strains can survive 3 hours in acidic pH 3.0 with only a one-log loss in cfu and was able to pass through bile. Therefore, a specifically selected or trained bacterial isolate can be used as urea-targeted components in a probiotic product of the present invention. Accordingly, the Lactobacillus bacterium of the present invention, by selection or training, are selected as being capable of substantially reducing urea concentrations within 24 hours to about 50%, 40%, 30%, 20% or 10% of the starting amount of urea. [0023] Elevated levels of secretion can also be obtained by overexpressing the gene of interest (e.g. , via multiple copies or a promoter driving high levels of expression) in a prokaryotic microorganism, such as Lactobacillus. The gene of interest can be under the regulatory control of an inducible or constitutive promoter. Promoters for use in recombinant prokaryotic expression vectors are well- established in the art and can include the beta-lactamase (penicillinase) and lactose promoter systems (Chang et al . (1978) Nature 275:615; Goeddel et al . (1979), Nature 281:544), a tryptophan (trp) promoter system (Goeddel et al . (1980) Nucleic Acids Res. 8:4057; EPO App . Publ . No. 36, 776) and the tac promoter (De Boer et al . (1983) Proc. Natl. Acad. Sci. USA 80:21) . While these are commonly used promoters which are commercially available, one of skill in the art can appreciate that any other suitable microbial promoter can be used as well. Nucleic acid sequences encoding suitable prokaryotic promoters have been published thereby enabling one of skill in the art to readily isolate these promoters {e.g., by standard cloning or PCR methodologies) for cloning into plasmid or viral vectors (Siebenlist et al. (1980) Cell 20:269). The promoter and Shine-Dalgarno sequence (for prokaryotic host expression) are operably-linked to the DNA encoding the gene of interest, i.e., they are positioned so as to promote transcription of the messenger RNA from the DNA, and subsequently introduced into a suitable host cell.

[0024] As will be understood by those of skill in the art, expression vectors containing polynucleotides which encode a degradative enzyme of interest, e.g., a urease or creatininase , can be designed to contain signal sequences which direct secretion of enzyme of interest through a prokaryotic cell membrane. Such signal sequences are well- established in the art and can be taken from other enzymes/proteins known to be secreted into the extracellular environment.

[0025] Transforming the microorganisms as defined herein, describes a process by which exogenous DNA is introduced into a recipient cell and changes the recipient cell . It can occur under natural or artificial conditions using various methods well-known in the art. Transformation can rely on any known method for the insertion of foreign nucleic acid sequences into a prokaryotic host cell. The method is selected based on the type of host cell being transformed and can include, but is not limited to, viral infection, electroporation, heat shock, lipofection, and particle bombardment. Such "transformed" cells include stably-transformed cells in which the inserted DNA is capable of replication either as an autonomously replicating plasmid or as part of the host chromosome. This also includes cells which transiently express the inserted DNA or RNA for limited periods of time.

[0026] As will be appreciated by the skill artisan, a microorganism can also be exposed to a mutagen to cause changes in the genetic structure of the organism so that it expresses elevated levels of a catabolic enzyme of interest .

[0027] Transformed or mutagenized strains are subsequently selected for the ability to grow in the presence of the metabolite which is degraded by the catabolic enzyme of interest. By way of example, a strain transformed with nucleic acid sequences encoding a urease is selected for high levels of urease secretion by growing said strain on high levels of urea. Levels of urease secretion can also be detected using standard enzymatic assays. As disclosed herein, the strain can be sequentially subcultured on increasing levels of urea to further enhance urease secretion. In one embodiment, a urease-secreting strain is a Lactobacillus acidophilus or a Lactobacillus rhamnosus bacterium that is capable of substantially reducing urea concentrations within 24 hours to about 50%, 40%, 30%, 20% or 10% of the starting amount of urea.

[0028] The probiotics according to the invention can be obtained by fermentation and can be stored after fermentation and before addition to the composition of the present invention for a time and at a temperature that prevents substantial loss of probiotic cfu. For example, the probiotic component can be fermented until a final concentration of 10⁶ to 10¹², or 10⁷ to 10¹², or 10⁸ to 10¹¹ cfu per mL of fermented medium is achieved.

[0029] When the probiotic component is composed of at least two microorganisms, each microorganism can be 10, 15, 20, 30, 40, 50, 60, 70, 80, or 90% of the probiotic component, wherein the total of all microorganisms is 100%. An exemplary probiotic component is composed of about 30-70% Lactobacillus acidophilus bacterium and about 30-70% Lactobacillus rhamnosus bacterium. In particular embodiments, the probiotic component is composed of about 50% L. acidophilus bacterium and about 50% L. rhamnosus bacterium.

[ 0030 ] As used herein, the probiotic component is one component or additive to a food product or to an enterically coated tablet, capsule, powder, soft gel, gelcap, or liquid. Accordingly, the probiotic component is included at a concentration of 10⁸ cfu/mL, 10⁹ cfu/mL, 10¹⁰ cfu/mL, 10¹¹ cfu/mL, or 10¹² cfu/mL when added as a liquid or 10⁸ cfu/g, 10⁹ cfu/g, 10¹⁰ cfu/g, 10¹¹ cfu/g, or 10¹² cfu/g when added as a freeze-dried powder. In one embodiment, the probiotic component is about 20% to about 70% of the total finished product weight. In particular embodiments, the probiotic component is about 50% of the total finished product weight.

[0031] The curcuminoid component of the present invention is present at an amount of from 100 to 300 mg, and makes up 30 to 50% by weight of the finished product. In particular embodiments, the curcuminoid is present in the composition of this invention at an amount of from 100-200 mg. Notably, studies in human beings have indicated no dose-limiting toxicities for oral curcumin at doses as high as 8 g/day (Kanai, et al . (2011) Cancer Chemother. Pharmacol. 68 (1) : 157-64) .

[0032] In another embodiment, the composition of the present invention can be formulated to contain a prebiotic component, in addition to the probiotic component and the curcumin component. A prebiotic of the present invention refers to a non-digestive food that beneficially affects the host by selectively stimulating the growth and/or activity of one or more non-pathogenic bacteria in the colon. Prebiotic components of the present invention are considered to have anti-carcinogenic, anti-microbial, hypolipidemic and glucose modulatory activities. They can also improve mineral absorption and balance. Furthermore, bacteria belonging to the Lactobacillus family are stimulated by the presence of the prebiotic component and proliferate. Pharmacokinetically, the prebiotic components reach the colon largely intact. An exemplary prebiotic component includes, but is not limited to, an oligosaccharide such as xylooligosaccharide . In addition to a xylooligosaccharide, the prebiotic component of the invention can also include fructo-oligosaccharide, inulin, isomaltose oligosaccharide, trans-galacto-oligosaccharide, or soy-oligosaccharide; a pyrodextrin such as arabinogalactan, lactilol, lactosucrose, or lactulose; or a fiber source such as oat gum, pea fiber, apple fiber, pectin, guar gum, psyllium husks, glucomannan or guar gum hydrolysate (BENEFIBER® (dietary fiber supplement; Novartis AG)). In a preferred embodiment, the prebiotic component comprises a xylooligosaccharide .

[0033] The amount of prebiotic component added to the probiotic component is 100 milligrams per serving, 500 milligrams per serving, 1 gram per serving, 5 grams per serving or 10 grams per serving. In one embodiment, the prebiotic component is not less than 100 milligrams and not more than 10 grams per serving. In one embodiment, the prebiotic component is about 20% to about 70% of the total product weight. In particular embodiments, the prebiotic component is about 35% of the total product weight.

[0034] A composition of the present invention is a single oral dosage form that combines the beneficial properties of the probiotic component with the curcuminoid. The finished product could include a food product, dietary supplement, comestible medical food or pharmaceutical product. The ingestion of the composition of the present invention reduces the blood concentration of nitrogenous waste products that accumulate in the circulating blood stream, specifically uric acid. Furthermore, repeated ingestion of the composition of the present invention will have a highly beneficial effect upon the intestinal microflora by localization and colonization in the large intestine of microbes known to promote a healthy intestinal microenvironment. In particular embodiments of this invention, the probiotic component consists essentially of a Lactobacillus acidophilus bacterium and a Lactobacillus rhamnosus bacterium, in combination with a curcuminoid. In certain embodiments, the composition of the invention consists essentially of a Lactobacillus acidophilus bacterium, a Lactobacillus rhamnosus bacterium, and a curcuminoid. In other embodiments, the composition consists essentially of a Lactobacillus acidophilus bacterium, a

Lactobacillus rhamnosus bacterium, a curcuminoid and a prebiotic. In further embodiments, the composition consists essentially of a Lactobacillus acidophilus bacterium, a

Lactobacillus rhamnosus bacterium, a curcuminoid and a xylooligosaccharide . Other non-essential ingredients such as fillers, additives, excipients, flavors, sweetening agents, binders or bulking agents may also be included. In specific embodiments, the composition of the present invention consists of Lactobacillus acidophilus strain KB27, Lactobacillus rhamnosus strain KB79, and a curcuminoid. In an alternative embodiment, the composition of the present invention consists of Lactobacillus acidophilus strain KB27, Lactobacillus rhamnosus strain KB79, a curcuminoid and a xylooligosaccharide.

[0035] As indicated herein, the composition of the present invention can take the form of a food product including, but is not limited to, a health bar, health drink, yogurt, dahi, or sachet or an enterically coated tablet, capsule, powder, soft gel, gelcap, or liquid. In addition to containing the probiotic component, curcuminoid and optional prebiotic, the composition of the present invention can further contain various fillers or additives .

[0036] Optional additives of the present composition include, without limitation, pharmaceutical excipients such as magnesium stearate, talc, starch, sugars, fats, antioxidants, amino acids, proteins, nucleic acids, electrolytes, vitamins, derivatives thereof or combinations thereof. In one embodiment, an additive of the synbiotic product is carob flour, for example, locust bean gum. In particular embodiments, a gel cap contains fillers such as magnesium stearate, talc and starch.

[0037] Further, to increase the palatability of a food product containing the composition of the present invention, it may be desirable to add flavors, sweetening agents, binders or bulking agents.

[0038] Flavors which can optionally be added to the present compositions are those well-known in the pharmaceutical art. Examples include, but are not limited to, synthetic flavor oils, and/or oils from plants leaves, flowers, fruits and so forth, and combinations thereof are useful. Examples of flavor oils include, but are not limited to, spearmint oil, peppermint oil, cinnamon oil, and oil of wintergreen (methylsalicylate ) . Also useful are artificial, natural or synthetic fruit flavors such as citrus oils including lemon, orange, grape, lime, and grapefruit, and fruit essences including apple, strawberry, cherry, pineapple and so forth.

[0039] Sweetening agents can be selected from a wide range of materials such as water-soluble sweetening agents, water-soluble artificial sweeteners, and dipeptide-based sweeteners, including salts thereof and mixtures thereof, without limitation.

[0040] Binders can be selected from a wide range of materials such as hydroxypropylmethylcellulose, ethylcellulose, or other suitable cellulose derivatives, povidone, acrylic and methacrylic acid co-polymers, pharmaceutical glaze, gums (e.g., gum tragacanth) , milk derivatives (e.g. , whey), starches (e.g. , corn starch) or gelatin, and derivatives, as well as other conventional binders well-known to persons skilled in the art. Examples of bulking substances include, but are not limited to, sugar, lactose, gelatin, starch, and silicon dioxide. [0041] When the above-mentioned additives are included in the composition of the present invention, they are generally less than 15% of the total composition weight. In particular embodiments, they are less than 1 to 5% of the total composition weight.

[0042] To facilitate targeting of the composition of the present invention to the gastrointestinal tract, controlled release formulations, also known as extended release formulations, are useful, preferably for oral administration. These include, but are not limited to, osmotic pressure-controlled gastrointestinal delivery systems; hydrodynamic pressure-controlled gastrointestinal delivery systems; membrane permeation-controlled gastrointestinal delivery systems, which include microporous membrane permeation-controlled gastrointestinal delivery devices; gastric fluid-resistant intestine targeted controlled-release gastrointestinal delivery devices; gel diffusion-controlled gastrointestinal delivery systems; and ion-exchange-controlled gastrointestinal delivery systems, which include cationic and anionic drugs. Additional information regarding controlled release drug delivery systems can be found in Yie W. Chien, Novel Drug Delivery Systems, 1992 (Marcel Dekker, Inc.) .

[0043] For example, enteric coatings can be applied to tablets or capsules such that at low pH the coatings remain insoluble. However, as the pH increases in the gastrointestinal tract, the acidic functional groups are capable of ionization, and the polymer swells or becomes soluble in the intestinal fluid. Enteric coatings promote the compounds remaining physically incorporated in the dosage form for a specified period when exposed to gastric juice. Yet the enteric coatings are designed to disintegrate in intestinal fluid. Materials used for enteric coatings include shellac {e.g., esters of aleurtic acid), cellulose acetate phthalate (CAP), poly (methacrylic acid-co-methyl methacrylate, cellulose acetate trimellitate (CAT), poly (vinyl acetate phthalate) (PVAP) , hydroxypropyl methylcellulose phthalate (HPMCP) , fatty acids, waxes, plastics, plant fibers (e.g., amylose starch) and plant proteins (e.g., zein or AQUA-ZEIN™) , or dextrins . Various enteric coating materials can be selected on the basis of testing to achieve an enteric-coated dosage form designed ab initio to have a preferable combination of dissolution time, coating thicknesses and diametral crushing strength. (Porter et al . (1970) J. Pharm. Pharmacol. 22:42p). It is contemplated that the enteric coating can be either food grade or pharmaceutical material which is generally used in the production of various drug or dietary supplements.

[ 0044 ] Colon-specific dosage forms protect the active ingredient from release in duodenum and jejunum, and promote eventual release of the active ingredient into ileum or colon. Several approaches are used for site- specific drug delivery including, but not limited to pH- sensitive polymer drug delivery; time-controlled release delivery (see, e.g., Gazzaniga, et al. (1994) Int. J. Pharm. 108:77-83; Fukui, et al. (2000) Int. J. Pharm. 204 (1-2) : 7-15) ; microbially triggered drug delivery (see, e.g., Peters, et al. (1993) Int. J. Pharm. 94:125-134; Swift (1992) Proc. ACS Div. Poly. Mat. Sci . Eng. 66:403- 404; Huang, et al . (1979) J. Appl . Polym. Sci. 23:429-437); polysaccharide-based drug delivery systems such as chitosan, chitosan derivatives, amidated pectin, and chondroitin sulfate; pressure-controlled drug delivery (see, e.g., Muraoka, et al . (1998) J. Control. Release

52 (1-2 ): 119-29) ; novel colon targeted delivery (see, e.g. , US 6,368,629; Katsuma, et al . (2004) J. Pharm. Sci. 93 (5) : 1287-1299) ; and osmotic controlled drug delivery (OROS-CT; Alza Corporation) .

[0045] Depending on whether the composition is to be consumed by an adult human, child or animal (e.g., companion animal or livestock) , it can be produced in various sizes and with various ingredients suitable for the intended recipient. For example, while a gel cap size of 0 or 1 may be suitable for humans, a gel cap size of 2, 3, 4, or 5 may be more suitable for a companion animal.

[0046] Further, because the probiotic and curcuminoid components of the present invention are generally recognized as safe (GRAS) status, they can be consumed one, two or three times daily or more.

[0047] The present invention also relates to a method for reducing uric acid levels in the blood of a subject. The method of this invention involves administering an effective amount of a composition of the present invention so that the levels of uric acid in the blood are decreased or reduced, desirably to a normal range. Accordingly, in the context of the present invention, an "effective amount" is defined as an amount capable of producing statistically significant changes in blood uric acid levels in vivo. For example, normal uric acid levels in males and females is in the range of 2.5 to 7.0 mg/dL and 1.5 to 6.0 mg/dL, respectively (Jin, et al . (2012) Front. Biosci. 17:656-69). Generally, hyperuricemia in adults is defined as a blood uric acid concentration greater than 7.0 mg/dL in men and 6.0 mg/dL in women. Accordingly, a reduction in the level of uric acid in the blood of a subject having or at risk of having hyperuricemia or gout preferably reduces the level of uric acid to an amount within the normal range. As one of skill in the art can appreciate, means for determining the levels of uric acid are well-known to the skilled laboratory clinician.

[0048] As a composition of the present invention reduces the levels of uric acid in the blood, this composition is useful in a method for preventing or treating hyperuricemia or gout. Such a method involves administering a composition of the present invention to a subject having or at risk of having hyperuricemia or gout. Subjects having or at risk of having hyperuricemia or gout are those in the upper range of normal uric acid levels (e.g., 6 to 7 mg/dL for men and 5 to 6 mg/dL for women) or above the normal uric acid levels (e.g., more than 7 mg/dL for men and 6 mg/dL for women) . Desirably, an effective amount of a composition for preventing or treating hyperuricemia or gout is an amount sufficient to effect beneficial or desired results, including clinical results, and, as such, an effective amount of a composition is one which results in the alleviation or amelioration of one or more symptoms associated with hyperuricemia or gout (e.g., crystal deposits in joints or tendons), delay or slowing of disease progression (i.e., a delay of crystal deposits in joints or tendons), or preventing crystal deposits in joints or tendons of subject with a history of hyperuricemia or gout.

[0049] The invention will be further described in the following examples, which do not limit the scope of the invention described in the claims.

Example 1 : Efficacy of Composition

[0050] To demonstrate the efficacy of the present invention to prevent and treat hyperuricemia and gout, experiments were performed in a well-established rat model of hyperuricemia (Cristobal-Garcia, M. et al. 2015. Oxidat. Med. Cell. Longev. Volume 2015, Article ID 535686, 8 pages) . The presence of hyperuricemia in the absence of gout has often been described as an asymptomatic condition. Yet, recent studies suggest hyperuricemia may have a contributory role to metabolic and cardiovascular diseases, and potential mechanisms involved in uric acid' s deleterious metabolic effects include the ability of soluble uric acid to increase oxidative stress, mitochondrial and endoplasmic reticulum dysfunction, endothelial dysfunction, to activate the renin-angiotensin system, and to increase synthesis and secretion of proinflammatory factors. In the present invention, the hyperuricemic rat model was used to demonstrate that the combined probiotic-curcumin composition had activity to reduce uric acid levels, thus treating hyperuricemia. Then, such beneficial activity would be indicative of the potential of a composition to prevent and/or treat gout. It is noted that allopurinol, a human therapeutic drug used to treat gout has been shown to have activity to reduce uric acid levels in this rat model of hyperuricemia, as well as to have activity to improve other endpoints of hyperuricemia in the animals (Cristdbal-Garcia, M. et al. 2015. Oxidat . Med. Cell. Longev. Volume 2015, Article ID 535686, 8 pages). Therefore, a hyperuricemia model in rats was employed to demonstrate the efficacy of the composition of the present invention.

[0051] Thirty male Wistar rats were ordered from Envigo Mexico (Mexico City, Mexico) , and given five days to acclimate to the housing facility prior to be being trained for baseline systolic blood pressure (SBP) measurement. Baseline urine was collected by placing rats in metabolic cages (Tecniplast, Varese, Italy) for 16 hours with food and water ad libitum. During the study itself, SBP was measured by tail cuff manometry in conscious animals previously accustomed to this procedure (NIBP System IN125/R, ADInstruments Inc. Dunedin, New Zealand). Three consecutive measurements were recorded, and the mean reported. After SBP measurement, a sample of blood was taken from the tail vein (600 pL) of the animal, and plasma was stored at -20°C until further processing.

[0052] Five groups of six rats each were studied, with groups matched by body weight and SBP levels. Oxonic acid, potassium salt, was administered daily by gavage (using a flexible polyethylene tube attached to a syringe) in the morning hours (750 mg/kg BW, Sigma-Aldrich Chemical, St Louis, MO), for five weeks. Two probiotic formulations containing a specific combination of bacteria were tested. Both product formulations contained two probiotic strains, L. acidophilus KB27 (5.0 B CFU/day) and L. rhamnosus KB79 (5.0B CFU/day) . The FI formulation contained only the probiotics while the F2 formulation was supplemented through the addition of the anti-inflammatory ingredient, curcumin (25.0 mgs per day). The Lactobacillus strains were mixed with food, and five-gram portions, in the form of balls, were prepared and stored at -20°C. A normal regular diet, and a diet containing "placebo" was prepared and stored in the same way. To guarantee a uniform feeding pattern, each rat received two balls of food in the morning, three hours after oxonic acid dosing, followed by another two balls of food at night. All animals consumed all food during the day. Tap water was provided ad libitum to all groups.

[0053] The following groups were included in the study: 1) control group (C) which were normal, healthy rats receiving a normal diet, and no oxonic acid; 2) HU-ND group which consisted of oxonic acid-induced hyperuricemic rats that received the normal, regular diet; 3) HU-P group which consisted of oxonic acid-induced hyperuricemic rats that received the placebo-containing diet; 4) HU-F1 group which consisted of oxonic acid-induced hyperuricemic rats that received the Fl-containing diet; and 5) HU-F2 group which consisted of oxonic acid-induced hyperuricemic rats that received the F2-containing diet.

[0054] SBP measurements and blood samples were obtained at the end of week 3 and week 5. At the end of the study (after 5 weeks on treatment) , rats were euthanized by deep anesthesia with inhaled isoflurane and exsanguination via abdominal aorta. Immediately, kidneys were washed by perfusion with cold phosphate-buffered saline (PBS) and the right kidney was excised, separated into cortex and medulla, and stored in liquid nitrogen until further processing. The left kidney was fixed by perfusion for histological analysis. Samples of the small intestine (ileum) were also taken and stored in liquid nitrogen.

[0055] The primary endpoint monitored in all animals was plasma uric acid concentration. The secondary endpoints that were monitored in the rats included SBP, urine uric acid levels, urine oxidative stress (TBARS) , creatinine clearance, urine and plasma nitrites and nitrates, urine N- acetyl-beta-D-glucosaminidase (NAG) activity (marker of tubular damage) , renal histology, and markers of oxidative stress (protein oxidation and lipid peroxidation) and uric acid concentration in renal cortex homogenates. Also, the expression of the urate transporter, ABCG2 , in ileum was assessed by western blot.

[0056] Uric acid levels in plasma and urine were measured using a commercial enzymatic kit (Sekisui Diagnostics, Charlottetown, PE. Canada) . Week 5 plasma samples were also tested using a colorimetric assay (Quantichrom Uric Acid Assay, BioAssay Systems, San Francisco, CA) . Urine TBARS and products of nitric oxide metabolism in plasma and urine (nitrates and nitrites) were measured with commercial kits (Cayman Chemical, Ann Arbor, MI) . Plasma and urine creatinine were measured with a commercial kit (SpinReact, Girona, Spain) , and creatinine clearance was calculated. Urine NAG activity was determined using 4-nitrophenyl-N- acetyl-beta-D-glucosaminide as substrate. One unit of enzymatic activity (U) represents the amount of enzyme, which hydrolyzes one micromole of substrate per minute at 37°C. The results were expressed as U/24 hours.

[0057] Renal cortex oxidative stress was measured in renal cortex homogenates. The determination of carbonyl groups in the proteins of renal cortex homogenates was measured using the reaction with 2 , 4-Dinitrophenylhydrazine (DNPH) as previously described (Tapia, E. et al . 2016. Free Radio.

Res. 50 ( 7 ) : 781-92 ) . For the 4-HNE assay, a 50 mg sample of kidney cortex was homogenized in ice-cold PBS, and a colorimetric assay was performed as previously described (Goicoechea, M. et al . 2015. Am. J. Kidney Dis . 65(4) :543-

9) . The results were expressed as nmol of 4-HNE/mg protein.

[0058] Renal cortex uric acid content was measured after extracting uric acid from renal cortex according to known methods (see, e.g., Tapia, et al . (2013) AJP Renal Physiol.

304(6) :F727-F736) . Uric acid concentration was measured with a commercial enzymatic kit (Sekisui, Diagnostics, Charlottetown, PE, Canada) and corrected by protein concentration (Bradford method) .

[0059] Results showed that all treatment groups had similar linear growth trends during the study (5 weeks) . Rats on the HU-F2 diet tended to gain less body weight as compared to the other groups, but the results were not statistically significant . [0060] The primary efficacy endpoint in the study was the effect on uric acid levels in the oxonic acid-treated rats. Results showed that ingestion of the compositions of the present invention, Fi and F2, prevented the development of hyperuricemia. At baseline, all treatment groups had comparable values of plasma uric acid. Rats fed the control diet and receiving no oxonic acid (no treatment) exhibited no changes in uric acid levels during the 5 weeks of the study. In rats treated with oxonic acid and on a normal diet and those that received the placebo diet, exhibited a small, but significant, increase in uric acid levels in blood after 3 weeks, an effect that was further accentuated after 5 weeks. Administration of oxonic acid with either the Fl or F2 diet supplement in rats prevented the rise of blood uric acid at both 3 and 5 weeks; there was no apparent difference in the effects between Fl and F2 treatments (compositions with or without curcumin added) . At the end of the study, however, rats in the HU-Fl and HU- F2 groups did exhibit a higher blood uric acid level as compared to their respective baselines; the levels achieved were significantly lower than blood uric acid levels achieved in the groups not administered the compositions of the present invention, with the differences greater at 5 weeks as compared to 3 weeks of exposure.

[0061] Urinary excretion of uric acid was another endpoint monitored in the study. Results showed that there was a significant increase in urinary excretion of uric acid after 5 weeks of treatment in the rats that received normal diet and placebo diet as compared to untreated control rats (no oxonic acid) . Administration of the compositions of the present invention, Fl and F2 groups, prevented the rise in uric acid urinary excretion that occurred with oxonic acid- induced hyperuricemia. Intrarenal uric acid levels also were monitored, and oxonic acid administration was shown to induce accumulation of intrarenal uric acid in rats receiving a normal diet or a placebo diet. In contrast, rats administered the compositions of the present invention, groups FI and F2, exhibited statistically significant decreases in the level of uric acid accumulation in renal tissue; these treatments did not completely abolish the accumulation of uric acid in the kidney tissue.

[0062] In addition, the administration of the probiotic compositions of the present invention prevented renal and extra-renal physiological changes that are typically induced by oxonic acid in this model of hyperuricemia. For example, there were no changes in SBP among the groups when baseline levels were examined (Table 1) . Then after 3 and 5 weeks of oxonic acid treatment, rats receiving either the normal diet or the placebo diet exhibited a small increase in SBP (Table 1) . At 5 weeks, treatment with either the Fl or F2 compositions prevented the oxonic acid-induced increases in SBP. Rats in groups receiving Fl and F2 treatments tended to have lower SBP levels after 5 weeks as compared to their baseline SBP levels, although the effects were not statistically significant.

TABLE 1

n=6 per group; *P<0.05 versus baseline ; ¹ p<0.05 versus no treatment; ² p, 0.05 versus HU-ND; ³p<0.05 versus HU-P.

[0063] Changes in creatinine clearance, a measure of kidney function, were monitored. Results showed that there were no changes in creatinine clearance among the groups at baseline, as well as after 3 weeks of follow-up. After 5 weeks of oxonic acid exposure, rats receiving normal diet (HU-ND) and placebo diet (HU-P) exhibited significant decreases in kidney function as measured by creatinine clearance (-30 to -40%) . This effect was prevented by administration of the F2 composition, and partially prevented in rats receiving the Fl composition (no curcumin) .

[0064] NAG urinary activity is a marker of tubular injury, and was another endpoint examined in the rats. Oxonic acid- induced hyperuricemia produced significant increases in the urinary activity of this enzyme in groups that received normal diet or placebo. Administration of the compositions of the present invention, Fl and F2, partially prevented the rise of NAG activity by the end of the study.

[0065] Renal histologic analysis was performed at the end of the study (after 5 weeks of oxonic acid exposure) . Results showed that afferent arteriolopathy was produced in oxonic acid-treated rats with hyperuricemia (defined as an increase in arteriolar wall area) ; tubulointerstitial inflammation and fibrosis were also observed with oxonic acid treatment, and were commonly seen in HU-P rats, while in the normal diet hyperuricemic group, the fibrosis was not as commonly observed. Administration of either of the compositions of the present invention (Fl and F2) prevented the renal histopathologic changes as compared to changes observed in the kidneys of rats in the HU-P group.

[0066] Administration of the compositions of the present invention also prevented oxidative stress and endothelial dysfunction induced by hyperuricemia. TBARS urinary excretion is a measure of systemic lipid peroxidation secondary to oxidative stress. Hyperuricemic rats receiving normal diet or placebo exhibited statistically significant increases in the excretion of urinary TBARS, an effect that was partially blocked by the exposure to either the Fl or F2 composition. At the end of 5 weeks of treatment, oxonic acid-induced hyperuricemia was associated with a significant increase in lipid peroxidation in the renal cortex. Again, this effect was partially prevented by exposure to the compositions of the present invention (Fl and F2 ) , although addition of curcumin to the treatment resulted in a statistically significant difference as compared to use of the Fl treatment. Hyperuricemia induced a marked reduction in plasma and urinary N02/N03 at the end of 5 weeks; reduction in levels of these nitric oxide byproducts is linked to oxidative stress. Again, treatment of rats with the compositions of the present invention resulted in a partial rescue of urinary levels of the nitric oxide byproducts, with the addition of curcumin again linked to a more favorable result . Strong correlations also were noted between systemic and renal markers of oxidative stress and endothelial dysfunction, where exposure to the compositions of the present invention resulted in decreased expression of the ABCG2 transporter in the ileum. Oxonic acid-induced hyperuricemia was associated with a statistically significant increase in expression of this transporter.

[0067] Considered together, these results in a well- established model of hyperuricemia, one that has been used to study drugs used to treat gout, clearly demonstrated that the compositions of the present invention provide important protection against the adverse effects linked to hyperuricemia, as well as preventing or treating hyperuricemia in the rats. The addition of curcumin to the composition containing a probiotic component, two Lactobacillus strains, appeared to provide additional benefits .

Example 2 : Yogurt Food Product

[0068] A yogurt food product can be prepared from one gallon of commercially available whole, homogenized, pasteurized milk which is heated to boiling and quickly allowed to cool to approximately 45°C. To this is added approximately one ounce of yogurt starter culture containing lactic acid bacteria of the genus Lactobacillus . The mixture is mixed well and allowed to ferment at 37 °C for 10 to 12 hours. Curcumin is subsequently added along with one or more whole fruit additives, flavorings, sweetening agents, binders, or other additives to obtain a product of desired consistency or to suit the palette of the prospective consumer. In one embodiment of the present invention, a food product comprises components to meet the special dietary needs of individuals with renal insufficiency .

Example 3 : Health Bar

[0069] Health bars are prepared by combining various excipients, such as binders, additives, flavorings, colorants and the like, along with the probiotic component (i.e., Lactobacillus acidophilus and Lactobacillus rhamnosus) and curcumin, and mixing to a plastic mass consistency. The mass is then either extruded or molded to form "candy bar" shapes that are then dried or allowed to solidify to form the final product.

Example 4 : Medical Food

[0070] A medical food can be prepared by combining rolled oats, dehydrated apples, honey, inulin, carob flour, cinnamon, sugar, vanilla extract, and lyophilized cultures of L. acidophilus and L. rhamnosus (10⁸ - 10¹⁰ cfu each) .

These ingredients are mixed in appropriate proportions with curcumin and formed into a rectangular bar approximately 12.5 to 15 centimeters in length, 3 to 4 centimeters in width and 1 centimeter in height and placed into a sterile vacuum oven for 12 to 24 hours to obtain an edible food product of the desired consistency.

Example 5 : Dietary Supplement

A dietary supplement of the present invention can be prepared by combining the two bacteria, Lactobacillus acidophilus (20 to 40%) and Lactobacillus rhamnosus (20 to 40%) , aseptically freeze-drying the bacteria and combining the processed bulk bacteria (e.g., about 50% of the total synbiotic product weight) with the curcumin component, wherein the final curcumin component is about 35% of the total product weight. Fillers such as magnesium stearate, talc and starch (e.g., about 1 to 5% of the total product weight) are added to the composition and enterically coated gel caps are produced according to the method of Kim et al ((1988) J. Lndust. Microbiol. 3:253-257). Approximately 10 to 40 billion CFU of the freeze-dried microorganism is contained in each capsule (i.e., approximately 30 to 120 billion CFU microorganisms per gram) that is enterically coated with hydroxy-propylmethyl cellulose phthalate by spraying over a fluidized bed of capsules. The resulting dietary supplement has a low surface area, is relatively non-porous and can protect the contents therein from the low pH that is found in the gastric environment for several hours and will release the contents into the bowel wherein the pH is relatively neutral or slightly alkaline. Advantageously, approximately 90-95% of the microorganisms can survive to be released into the gastric environment.

Example 6 : Pharmaceutical Product

[0071] A pharmaceutical product for persons suffering from gout or hyperuricemia can be prepared by aseptically freeze-drying a Lactobacillus acidophilus and a Lactobacillus rhamnosus, combining the processed bulk microorganisms with a curcuminoid compound, and preparing the finished product as enterically coated capsules according to the method of Kim et al. ((1988) supra) or tablets, powders, soft gels, gelcaps, or liquids according to standard methods. For example, the ingredients in each capsule are enterically coated with hydroxy-propylmethyl cellulose phthalate by spraying over a fluidized bed of capsules. The resulting pharmaceutical product has a low surface area, is relatively non-porous and can protect the contents therein from the low pH that is found in the gastric environment for several hours and will release the contents into the bowel, wherein the pH is relatively neutral or slightly alkaline.

Example 7 : Reduction in Uric Acid Levels in Patients with Hyperuricemia and Gout

[0072] A clinical study will be performed under the approval of an Institutional Review Board (IRB) consistent with Good Clinical Practice. The study will be designed as a double-blind, placebo-controlled, randomized crossover clinical trial. The hypothesis to be tested is that adjunct therapy which targets the mechanistic actions of probiotics and prebiotics in patients with hyperuricemia, and signs and symptoms of gout, will lead to modulation of the gut microbiome, reduction in uric acid levels, a decrease in inflammation and an improved quality of life (QoL) . The composition to be tested against placebo treatment is a mixture consisting essentially of Lactobacillus acidophilus, Lactobacillus rhamnosus , xylooligosaccharide and curcumin.

[0073] The product composed of Lactobacillus acidophilus, Lactobacillus rhamnosus , xylooligosaccharide and curcumin will be orally administered to gout patients in a cross over experiment. The primary aim is to study the mechanisms by which the formulation provides benefits to patients with hyperuricemic gouty arthritis. Secondary study aims will focus on 1) the impact of the composition on microbial shift in gout patients; 2) the impact of the composition on uric acid levels (reductions are sought); and 3) the impact of the formulation on QoL. In addition, biomarkers of disease will be monitored such as urinary N-acetyl beta D glucosamine (NAG) , thiobarbituric acid reactive species (TBARS ) , and ABCG2 transporters. The study will be a double-blind, placebo-controlled randomized trial with a one to one arm. Patients will be switched from one treatment to the next halfway through the study after a four-week washout period. Inclusion criteria for patients will be: 1) age of 30 to 75 years old; 2) baseline serum uric acid levels above 8 mg/dL; and 3) completed informed consent. Exclusion criteria will include: 1) pregnant or nursing women; 2) HIV/AIDS/liver disease; 3) refusal to sign informed consent; 4) active dependency on drugs or alcohol; 5) any medical, debilitating disease , disorder or social condition that in the judgement of the investigator would interfere with, or serve as a contraindication to, adherence to the study protocol, or the ability to give informed consent or affect overall prognosis of the patients; 6) current use of anti-coagulant therapy or long term antibiotics; and 7) refusal to participate.

[0074] All patients will be seen by the investigator at baseline, 5 weeks, 9 weeks and 14 weeks. Clinical data (history, physical exam, laboratory data and management including prescribed prescriptions, OTC medications and supplements) will be reviewed by the investigator ( s ) and clinical research team member. Participants will switch from one group to the other at the midway point (week 9) , after a 4-week washout period. The composition to be administered will be formulated into acid-resistant gel caps containing 30 billion CFUs per capsule of L. acidophilus, L. rhamnosus, xylooligosaccharide and curcumin . All subjects will be administered two capsules per day of the composition or the placebo. The placebo will be formulated to be identical in appearance to the test composition but will not contain the mixture of probiotic organisms with curcumin. Placebo will be a probiotic product from the market having no uricase activity. Doses to be administered to the patients were chosen based on extrapolating from the doses found to be effective in rats. Given that the probiotic organisms being used are ones with a long history of safe use, the doses administered to humans are typically in the same range. In the case of curcumin doses in the composition, they were also chosen based on extrapolation from the doses shown to be effective in rats and based on the fact that curcumin also is an ingredient commonly found in the diet and known to be safe for human consumption.

[0075] It is expected that the composition of the invention will reduce blood levels of uric acid to less than 6 mg/dL, with hyperuricemia typically defined as a blood level above 7.0 mg/dL. Other endpoints monitored will include clinical signs and symptoms linked to gout such as changes in the swelling of the joints in the fingers and toes, a common finding in patients with gout. Both self-reported gout symptoms and measurements of joint swelling before and after treatment in the study are also reported. The results of this analysis will demonstrate that the composition of the invention measurably reduces swelling of the joints in subjects exhibiting symptoms of gout.

[ 0076 ] At least 50 patients with hyperuricemia and gout are included in the study to allow for a statistical analysis to be performed that could identify statistically significant changes in blood uric acid levels as compared to baselines in each patient.

Claims

What is Claimed is :

1. A composition comprising at least two probiotic components and a curcuminoid.

2. The composition of claim 1, wherein the at least two probiotic components include a Lactobacillus acidophilus bacterium and a Lactobacillus rhamnosus bacterium.

3. The composition of claim 1, wherein the curcuminoid comprises curcumin.

4. The composition of claim 1, wherein the composition further comprises a xylooligosaccharide .

5. The composition of claim 1, wherein the composition is formulated as an extended release composition.

6. A method for reducing uric acid levels in the blood comprising administering an effective amount of a composition comprising at least two probiotic components and a curcuminoid to a subject in need of treatment thereby reducing the subject's uric acid levels.

7. The method of claim 6, wherein the at least two probiotic components include a Lactobacillus acidophilus bacterium and a Lactobacillus rhamnosus bacterium.

8. The method of claims 6, wherein the curcuminoid comprises curcumin.

9. The method of claim 6, wherein the composition further comprises a xylooligosaccharide .

10. A method for preventing or treating hyperuricemia or gout comprising administering an effective amount of a composition comprising at least two probiotic components and a curcuminoid to a subject in need of treatment thereby preventing or treating the subject's hyperuricemia or gout.

11. The method of claim 10, wherein the at least two probiotic components include a Lactobacillus acidophilus bacterium and a Lactobacillus rhamnosus bacterium.

12. The method of claims 10, wherein the curcuminoid comprises curcumin.

13. The method of claim 10, wherein the composition further comprises a xylooligosaccharide.