WO2022091075A1

WO2022091075A1 - Combination compositions of probiotics with fermented wheat germ extract and uses thereof

Info

Publication number: WO2022091075A1
Application number: PCT/IL2020/051133
Authority: WO
Inventors: A. William Schwartz
Original assignee: Aili Life Sciences Ltd.; WEBB, Cynthia
Priority date: 2020-11-01
Filing date: 2020-11-01
Publication date: 2022-05-05

Abstract

The present invention provides advantageous combinations and compositions comprising fermented wheat germ extract, probiotics and optionally an organic compound. The combinations and compositions of the present invention may synergistically activate and enhance immune system function of subject. The invention further provides methods of using said combinations and compositions to prevent and/or treat medical conditions or to enhance immune system function.

Description

COMBINATION COMPOSITIONS OF PROBIOTICS WITH FERMENTED

WHEAT GERM EXTRACT AND USES THEREOF

FIELD OF THE INVENTION

The present invention relates to combinations or compositions comprising probiotics together with fermented wheat germ extract and optionally with an additional biologically active organic compound, the compositions being useful to prevent and/or treat medical conditions. In particular, the combination compositions provide specific strains of probiotics encapsulated with coated particles of fermented wheat germ extract.

BACKGROUND OF THE INVENTION

PROBIOTICS

Certain bacteria have been shown to benefit gastrointestinal function and overall physiological health. These helpful bacteria, known as “probiotics” supplement the natural flora of the gastrointestinal tract with additional bacteria. Supplementation with probiotic organisms has been shown to be effective in treating a broad spectrum of conditions including cancer, dermatitis, allergies, and upper respiratory infections. Specialized combination of probiotics and other compounds have the potential to ameliorate disease pathology by augmenting the body’s immune system and inflammatory response (Liu, Yuying et al. “Probiotics in Autoimmune and Inflammatory Disorders.” Nutrients Vol. 10, 10 1537. 18 Oct. 2018).

Examples of probiotic organisms include lactic acid producing bacteria such as Lactobacillus, Lactococcus and Bifidobacterium, and other bacteria such as Streptococcus. Probiotic organisms found in foods and nutritional supplements are not normally found in the gastrointestinal tract. Most of these foreign bacteria are unable to inhabit the intestinal environment and are flushed through and eliminated quickly from the body. As a result, these bacteria need to be ingested regularly in order to achieve desired health benefits. Despite their benefit, problems exist with the currently available probiotic formulations. Much of the bacteria found in supplements cannot survive the extreme, acidic conditions of the stomach.

Of the major groups of bacteria found in the intestine, at least Lactobacilli, Streptococci, and Bifidobacteria confer beneficial effects to mammals, particularly humans. Lactobacilli have been used for several hundred years for promoting human health. Lactobacilli found in the human intestinal tract include L. acidophilus, L. casei, L. fermentum, L. saliva roes, L. brevis, L. leichmannii, L. plantarum, and L. cellobiosus.

L. acidophilus has been shown to be useful in treating conditions such as antibiotic- induced imbalances in the gastrointestinal microflora, hypercholesterolemia, vaginal infections, E. coli infection, depressed immunity, cancerous tumors, chronic granulomatous disease, and lactose indigestion. (A. G. Shauss, Method of Action, Clinical Application, and Toxicity Data, 3 J. Advancement Med. 163 (1990)). Results of several studies suggest that dietary supplementation with L. acidophilus may reduce the risk of developing colon cancer.

Lactobacilli also produce organic acids that reduce intestinal pH thereby inhibiting the growth of acid-sensitive undesirable bacteria. In vitro studies have shown L. acidophilus to inhibit the growth of pathogenic bacteria such as Campylobacter pylori, Staphylococcus aureus, Pseudomonas aeruginosa, and Sarcina lutea. (K. M. Shahani et al., “Natural Antibiotic Activity of Lactobacillus Acidophilus and Bulgaricus.'' 11 Cultured Dairy Products J. 14(1976)).

Bifidobacteria are also known to benefit human health. These bacteria exert antimicrobial activity in the human intestine by producing short chain fatty acids (SCFAs) such as acetic, propionic, and butyric acids, as well as lactic and formic acids, as a result of carbohydrate metabolism. Both Lactobacilli and Bifidobacteria may produce other antimicrobial substances, such as bacteriocins, that also inhibit the growth and proliferation of certain harmful bacteria.

Additionally, SCFAs are believed to support normal gastrointestinal function by increasing colonic blood flow, stimulating pancreatic enzyme secretion, promoting sodium and water absorption and intestinal mucosal growth. Bifidobacteria are also believed to deconjugate bile salts to free bile acids, which are more inhibitory to susceptible bacteria than are the conjugated forms.

Without sufficient numbers of probiotics, intestinal ecology may be thrown off balance, which can potentially result in health problems. Probiotic supplements may be used to increase the number of probiotics in the intestinal tract. About Probiotics, supra. As mentioned in “Probiotics, Prebiotics & Synbiotics: Harnessing Enormous Potential,” Nutraceuticals World (September 2001), a trend regarding probiotics is finding novel delivery systems, particularly because acidity in the stomach is detrimental to many probiotics and may destroy as much as 90-95% of such probiotics during their passage through the stomach. To date, improving the protection of probiotics from stomach acid has included using enteric coated capsules and microencapsulation. Such types of microencapsulation include technology known as Probiocap™ by Institut Rosell. PREBIOTICS

“Prebiotics” are foods or nutrients that are used by specific bacteria that can be added to the diet to increase the chance of these probiotic bacteria growing and thriving in the intestine.

Known prebiotics include dietary fibers, such as polysaccharides and oligosaccharides, that have the ability to increase the number of probiotic bacteria, which leads to the benefit(s) conferred by the probiotic. A prebiotic may also provide one or more of the following benefits: (1) indirectly SCFAs that in turn have a trophic (nourishing) effect on the intestinal epithelium, supporting its integrity as a defense barrier against invading organisms; (2) indirectly produce immune stimulants, by the promotion of Bifidobacteria that excrete an end product inhibitory to pathogenic bacteria; (3) promote a host-mediated attack against tumor sites and promote certain strains of Lactobacilli that have immune-modulating activity, enhancing phagocyte activity in the blood; and (4) indirectly provide any of the benefits of an increased number of probiotic whose number increased due at least in part to the presence of the prebiotic. A prebiotic may also affect the production of certain bacteria enzymes, such as decreasing glucosidase that is associated with the absorption of intestinal cholesterol, associated with the formation of secondary bile acid that is considered a co-carcinogen. Examples of prebiotics include inulin, chicory, honey, Active Hexose Correlated Compound (AHCC).

While known prebiotics break down to provide carbohydrates for probiotics, some probiotics also require amino acids for nourishment.

W02015/160314 and WO2018/209139 to the inventor of the present invention describe compositions comprising prebiotic and probiotic components.

FERMENTED WHEAT GERM EXTRACT

Fermented wheat germ extract (FWGE) is a clinically tested composition attributed with cancer fighting properties. A multi center study of 170 patients with colorectal cancer who received 9 g of Avemar (a commercially available fermented wheat germ extract) daily demonstrated that the co-administration of Avemar with other treatments significantly inhibited disease progression, incidence of metastasis, and survival rate. Fermented wheat germ extract has also been implicated in the modulation of cellular messengers and growth factors to inhibit tumor growth (Ulmer, Christoph, et al. "Immunologic and biochemical effects of the fermented wheat germ extract Avemar." Experimental Biology and Medicine 230.2 (2005): 144-149). US20120121612 describes compositions comprising the component of fermented wheat germ extract (“FWGE") active in reducing, inhibiting or preventing the proliferation of cancer cells and/or tumors, and methods of making and using such compositions. Gyula Bencze et al., (Scientific Reports volume 10, Article number: 14174, 2020) showed that a highly concentrated form of FWGE is an effective agent that increases normal mitochondrial functionality. CREATINE

Creatine is an organic compound found in mammalian skeletal muscle, brain, and other organs. Creatine is used by the body during times of increased energy demands to rapidly resynthesize ATP from ADP through the anaerobic conversion of phosphorylated creatine (phosphocreatine) to creatine in a reversible reaction by the enzyme creatine kinase. Creatine has been shown to be a critical molecule, buffering ATP levels in cancer-targeting CD8 T cells through maintaining a readily available high- energy phosphate reservoir (Wyss, M. et al. “Creatine and creatinine metabolism.” Physiological reviews 80.3 (2000): 1107-1213.) Creatine uptake deficiency severely impaired CD8 T cell responses to tumor challenge in vivo and to antigen stimulation in vitro, while supplementation of creatine through either direct administration or dietary supplement significantly suppressed tumor growth in multiple mouse tumor models. (Di Biase, S. et al. “Creatine uptake regulates CD8 T cell antitumor immunity.” Journal of Experimental Medicine (2019): jem-20182044).

Fermented wheat germ extract (FWGE) is a nutrient supplement with medical value as demonstrated in a wide range of potential disease targets, including anti-tumor efficacy against many tumor types in vitro and in vivo. 2-methoxy benzoquinone and 2,6- dimethoxybenzene, the two major components of FWGE, are suggested to exert main biological properties of FWGE although additional compounds present therapeutic potential. (Tai, Cheng-Jeng, et al. "Fermented wheat germ extract induced cell death and enhanced cytotoxicity of cisplatin and 5 -fluorouracil on human hepatocellular carcinoma cells." Evidence-Based Complementary and Alternative Medicine (2013)).

Fermented wheat germ polypeptides (FWGP) isolated from FWGE contain a variety of polypeptides that find use in the reduction, inhibition and/or prevention of cancer cell proliferation and/or tumor growth. The active fraction of FWGE, the FWGP, comprises polypeptides and peptides substantially isolated or purified from (i.e., separated from) non-proteinaceous components of FWGE, e.g., substantially purified or isolated from sugars, lipids, nucleic acids and insoluble components of FWGE. The active polypeptides in FWGP includes but is not limited to those taught by US9480725.

Despite a vast abundance of different medications, nutraceuticals and food supplements, there is a need for developing safe, convenient and inexpensive natural products that allow enhancement of immune system function and treatment of patients dealing with cancer.

SUMMARY OF THE INVENTION

The present invention relates to specialized and advantageous combinations comprising a botanical extract, at least one probiotic component, and optionally an additional biologically active component which is an organic compound. The invention further relates to the uses of said combinations as supportive and/or preventative and/or therapeutic treatments in pathologies relating to the immune system, metabolism, and cancer. The combinations of the present invention interact synergistically to enhance the immune system.

According to one aspect, the present invention provides a combination comprising a probiotic component and a fermented wheat germ extract (FWGE), wherein the combination enhances immune system function.

In some embodiments of the combination, the probiotic component is selected from the group consisting of Lactobacillus rhamnosus, Lactobacillus reuteri, Bifidobacterium animalis subsp. lactis (BB12)), Bifidobacterium bifidum, and Bifidobacterium longum.

In one embodiment, the probiotic component of the combination comprises Lactobacillus rhamnosus and Lactobacillus reuteri. In another embodiment, the probiotic component of the combination comprises Lactobacillus rhamnosus. In another embodiment, the probiotic component of the combination comprises Lactobacillus rhamnosus GG. In another embodiment the probiotic component consists essentially of L. rhamnosus GG. In a further embodiment, the probiotic component of the combination consists essentially of Lactobacillus reuteri.

In a particular embodiment the probiotic component of the combination comprises Lactobacillus rhamnosus GG and the prebiotic component comprises fermented wheat germ extract.

In one particular embodiment, the prebiotic component comprises metabolites and/or probiotic cell wall fractions of probiotic, e.g. derived from cultures of Lactobacillus rhamnosus and/or Lactobacillus reuteri. In some embodiments of the combination, the combination further comprises at least one organic compound selected from creatine and Vitamin D. In certain embodiments the organic compound is creatine. In other embodiments the organic compound is Vitamin D.

In one embodiment of the method, the botanical extract and a probiotic component in the combination may be administered simultaneously or sequentially.

In another variation of the combination, the FWGE, the probiotic component, and the organic compound are formulated for oral delivery. The oral delivery formulation may include the FWGE, the probiotic component, and the organic compound together in a single unit dosage form. In such dosage form, the FWGE and the probiotic component are separated by a barrier preventing their interactions and disintegration of the FWGE.

In some embodiments, FWGE is referred as a prebiotic.

According to another aspect, the present invention provides a composition comprising a probiotic component and a fermented wheat germ extract (FWGE), wherein the probiotic component and the FWGE are separated by a barrier. According to some embodiments, the probiotic component comprises Lactobacillus rhamnosus. According to specific embodiments the combination composition comprises FWGE-SC and L. rhamnosus GG. According to certain embodiments, the composition of FWGE and L. rhamnosus further comprises a biologically active component which is an organic compound. According to some embodiments, the organic compound is selected from creatine and Vitamin D.

According to another aspect, the combination or the composition of the present invention is for use in treating and/or preventing cancer.

In one aspect the combination or the composition presented in the invention is for use in enhancing immune system function of a subject. In one embodiment, the combination or the composition presented in the invention is for use in enhancing and supporting immune system function of a subject during radiation and chemotherapy.

In some embodiments the combination or the composition presented in the invention is for use in enhancing and supporting immune system function in patients having a viral infection or at risk of viral infection. In some embodiments the viral risk may be COVID-19 infection.

In one embodiment, the combination presented in the invention is disclosed for use in supporting anti-cancer activities of CD8 T cells. In one embodiment, the combination presented in the invention is disclosed for use in supporting anti-cancer activities by increasing the concentration of NK cells.

According to some embodiment, the present invention provides a method for reducing viability of cancer cells. The method of the present invention comprises administering to the cancer cells a combination comprising effective amounts of a FWGE, a probiotic component, and optionally an organic compound, wherein the effective amounts are sufficient to synergistically reduce the viability of the cancer cells.

According to some embodiments, the combination or the composition of the present invention is for modulating mitochondria energy metabolism.

In one embodiment of the method, the immune cell activation includes supporting anti-tumor activities via modulating mitochondria energy metabolism. In another embodiment, the immune cell activation includes activation of one or more of NK cells, monocytes and macrophages.

In some embodiments of the composition, the prebiotic component promotes a growth of the probiotic component.

In some embodiments of the composition, the at least one probiotic component is selected from the group consisting of Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus paracasei, Leuconostoc mesenteroides, Lactobacillus bulgaricus, Lactobacillus sasei, Lactobacillus salivarius, Pediococcus pentosaceus, Streptococcus thermophiles, Bacillus subtilis, Bacillus coagulans, Enteroccous faecium, Bifidobacterium bifidum, Bifidobacterium lactis, Bifidobacterium longum, Lactobacillus rhamno sus, Lactobacillus reuteri, and Bifidobacterium inf antis.

In an alternative embodiment, a kit is disclosed for activating and/or enhancing immune cell function. The kit comprises: an effective amount of a botanical extract such as fermented wheat germ extract, an effective amount of at least one probiotic, optionally an effective amount of at least one organic compound selected from creatine and Vitamin D and instructions for using the combination to enhance or stimulate immune system function. According to some embodiments, the kit is useful to treat or prevent a condition for which activating immune cell function may be therapeutic or prophylactic; wherein the amounts of the fermented wheat germ extract and the at least one probiotic are effective in combination to synergistically activate immune cell function.

In another variation to the kit, the synergistic activation of immune cell function comprises one or more of the following: upregulating expression of CD69 on lymphocytes and monocytes; activation of one or more of NK cells, monocytes and macrophages; and synergistic reduction cancer cell viability.

In some embodiments, a method for enhancement of immune system function in a subject is provided. In some embodiments, the method comprises identifying the subject, administering a therapeutically effective amount of at least one botanical extract component and a therapeutically effective amount of at least one probiotic component, and improving at least one indication of enhancement of immune system function. In some embodiments of the method, the subject is a mammal. In some embodiments of the method, the subject is a human. In some embodiments of the method, the botanical extract component is FWGE. In some embodiments of the method, the at least one probiotic component is selected from the group consisting of Lactobacillus plantarum, Lactobacillus acidophilus, Lactobacillus paracasei, Leuconostoc mesenteroides, Lactobacillus bulgaricus, Lactobacillus sasei, Lactobacillus salivarius, Pediococcus pentosaceus, Streptococcus thermophiles, Bacillus subtilis, Bacillus coagulans, Enteroccous faecium, Bifidobacterium bifidum, Bifidobacterium lactis, Bifidobacterium longum, Lactobacillus rhamnosus, Lactobacillus reuteri, and Bifidobacterium inf antis. In some embodiments of the method, the at least one indication of enhancement of immune system function is selected from the group consisting of increase in red blood cell count, increase in hemoglobin content, increase in hematocrit, improved antigen presentation and improved immunoglobulin secretion, macrophage antigen presentation activity and inhibition of tumor-derived immune suppressive factors, enhance macrophage proliferation and activation, promote differentiation of Thl cells; increase macrophage production of IL-12, increase NK activity; promote apoptosis of cancer cells.

In some embodiments of the method, the therapeutically effective amount of the at least one botanical extract component is about 0.1 mg to about 15 g. In some embodiments of the method, the therapeutically effective amount of the at least one probiotic component is about 0.5 million CFU to about 100 billion CFU.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to combinations and compositions of matter including a botanical extract, a probiotic compound, and optionally an organic compound. The combinations and compositions of the present invention are useful for enhancing the immune system, for mediating mitochondrial metabolism in a subject and for enhancing the immune response to cancer or during cancer treatment in a subject. According to one aspect, the present invention provides a combination comprising a probiotic component, a botanical extract and optionally a carrier. According to some embodiments, the botanical extract is a wheat germ extract. According to some embodiments, the wheat germ extract is a fermented wheat germ extract (FWGE). The combinations of the present invention enhances immune system function. Thus according to some embodiments, the present invention provides a combination comprising a probiotic component and a fermented wheat germ extract (FWGE), wherein the combination enhances immune system function. In some embodiments of the invention FWGE is referred as prebiotic.

The term "probiotic" and "probiotic component" are used interchangeably and refer to bacteria that may form or supplement the at least a part of the transient or endogenous flora and thereby exhibit a beneficial prophylactic and/or therapeutic effect on the host organism. According to some embodiments, the probiotic component comprises bacteria selected from Lactobacillus rhamnosus, Lactobacillus reuteri, Bifidobacterium animalis subsp. lactis (BB12)), Bifidobacterium bifidum, Bifidobacterium longum and any combination thereof.

Thus, according to some embodiments, the present invention provides a combination of FWGE and a probiotic component comprising bacteria selected from Lactobacillus rhamnosus, Lactobacillus reuteri, Bifidobacterium animalis subsp. lactis (BB12)), Bifidobacterium bifidum, Bifidobacterium longum and any combination thereof. According to one embodiment, the combination comprises FWGE and Lactobacillus rhamnosus. According to another embodiment, the combination comprises FWGE and Lactobacillus reuteri. According to yet another embodiment, the combination comprises FWGE, Lactobacillus rhamnosus and Lactobacillus reuteri. According to one embodiment, the combination comprises FWGE and BB12. According to another embodiment, the combination comprises FWGE and Bifidobacterium bifidum. According to yet another embodiment, the combination comprises FWGE and Bifidobacterium longum. According to any one of the embodiments and aspects of the present invention, the Lactobacillus rhamnosus is Lactobacillus rhamnosus GG. The term "Lactobacillus rhamnosus GG" as used herein refers to American Type Culture Collection (ATCC) accession number 53103 strain of Lactobacillus rhamnosus as known in art.

The terms "fermented wheat germ extract" and "FWGE" are used herein interchangeably and refer to concentrated extract of wheat germ derived from the wheat germ that were fermented using yeasts. The polypeptides in the FWGE typically have molecular weights in the range of about 5 kD to about 100 kD. In some embodiments, the polypeptides in the FWGE have molecular weights in the range of about 10-90 kD, 10- 80 kD, 10-70 kD, 10-60 kD, 12-50 kD, 5-60 kD, 5-50 kD, 5-40 kD, 5-30 kD or 5-20 kD. In some embodiments, the polypeptides in the FWGE have molecular weights in the range of 5 to 20 kD.

The term "FWGP", "FWGE-super concentrate" and "FWGE-SC" are used interchangeably and refers to FWGE isolated or purified from certain peptides or polypeptides. FWGP is the concentration active fraction of FWGE. Example of FWGP is METRATOL® sold by American BioSciences, Inc. In some embodiments, the FWGP is substantially isolated or purified from peptides or polypeptides with molecular weights that are larger or smaller than the stated range of molecular weight. For example, in some embodiments, the FWGP is substantially isolated or purified from peptides or polypeptides that are larger than 100 kD and smaller than 5 kD, for example, peptides or polypeptides that are larger than 90 kD and smaller than 10 kD, for example, peptides or polypeptides that are larger than 80 kD and smaller than 10 kD, for example, peptides or polypeptides that are larger than 70 kD and smaller than 10 kD, for example, peptides or polypeptides that are larger than 60 kD and smaller than 10 kD, for example, peptides or polypeptides that are larger than 50 kD and smaller than 12 kD, for example, peptides or polypeptides that are larger than 60 kD and smaller than 5 kD, for example, peptides or polypeptides that are larger than 50 kD and smaller than 5 kD, for example, peptides or polypeptides that are larger than 40 kD and smaller than 5 kD, for example, peptides or polypeptides that are larger than 30 kD and smaller than 5 kD, for example, peptides or polypeptides that are larger than 20 kD and smaller than 5 kD. According to some embodiments, the active component from FWGE comprises polypeptides having a molecular weight in the range of about 5-100 kiloDalton (kD), for example, a molecular weight in the range of about 12-50 kD.

According to some embodiments, the FWGP comprises peptides or polypeptides having molecular weight above 5 kD and below 100 kD. According to some embodiments, the FWGP comprises components having molecular weight from 5 to 100 kD, from 10 to 95 kD, from 15 to 90 kD, from 20 to 80kD, from 5 to 80 kD or from 10 to 60 kD.

According to any one of the embodiments of the present invention, the term FWGE may be replaced by the term FWGP. Any embodiments referring to fermented wheat germ extract encompass also the meaning of FWGP. According to any one of the above embodiments, the combination further comprises an additional biologically active agent being an organic molecule. According to some embodiments, the organic molecule participates in cell energy metabolism. According to some embodiments the organic molecule is Vitamin D. According to some embodiments, the organic molecule is creatine. According to another embodiment, the organic molecule is phosphocreatine. Thus, in some embodiments, the present invention provides a combination of FWGE, a probiotic component, and creatine. According to some embodiments, the probiotic component comprises Lactobacillus rhamnosus, Lactobacillus reuteri, Bifidobacterium animalis subsp. lactis (BB12)), Bifidobacterium bifidum, Bifidobacterium longum and combination thereof. According to one embodiment, the probiotic component comprises Lactobacillus rhamnosus, Lactobacillus rhamnosus GG, Lactobacillus reuteri, and a combination thereof.

According to any one of the above embodiments, the combination provides a synergistic enhancement of immune system function.

The term “enhancing” as used herein refers to promoting, improving, augmenting, typically increasing the parameters, and being measured relatively to a control sample.

The terms “inducing an immune response”, "enhancing an immune system", "enhancing an immune function", "enhancing an immune system function" and “enhancing an immune response” are used interchangeably and refer to the stimulation of an immune response (i.e., either passive or adaptive) to antigens.

The components of the combination may be co-administered in a regimen selected from a single combined composition, separate individual compositions administered substantially at the same time, and separate individual compositions administered under separate schedules and include treatment regimens in which the agents are not necessarily administered by the same route of administration or at the same time.

The term “co-administration” encompasses administration of a first and second and optionally the third agents in an essentially simultaneous manner, such as in a single dosage form, e.g., a capsule or tablet having a fixed ratio of first, second and third compounds, or in multiple dosage forms for each. The agents can be administered in a sequential manner in either order. When co-administration involves the separate administration of each agent, the agents are administered sufficiently close in time to have the desired effect (e.g., complex formation). The term “sequential manner” refers to an administration of two or more compounds at a different times, and optionally in different modes of administration. The agents can be administered in a sequential manner in either order.

The terms “substantially simultaneous manner” refers to administration of two or more compounds with only a short time interval between them. In some embodiments, the time interval is in the range of from 0.5 to 60 minutes.

According to any one of the above embodiments, the combination may be formulated as a single formulation, wherein the probiotic component and the FWGE are separated by a barrier.

According to another aspect, the present invention provides a composition comprising a probiotic component and botanical extract. According to some embodiments, the probiotic component and the extract are separated by a barrier. Thus, in some embodiments, the present invention provides a composition comprising a probiotic component and botanical extract, wherein, the probiotic component and the extract are separated by a barrier. According to some embodiments, the barrier is a water impervious or water resistant coating. According to some embodiments, the extract is a fermented wheat germ extract (FWGE). According to another embodiment, the extract is FWGP. The composition of the present invention enhances immune system function. Thus, according to some embodiments, the present invention provides compositions comprising a probiotic component and a fermented wheat germ extract (FWGE), wherein the composition enhances immune system function.

According to some embodiments, the probiotic component comprises bacteria selected from Lactobacillus rhamnosus, Lactobacillus reuteri, Bifidobacterium animalis subsp. lactis (BB12), Bifidobacterium bifidum, Bifidobacterium longum and any combination thereof. Thus, according to some embodiments, the present invention provides a composition comprising FWGE and a probiotic component comprising bacteria selected from Lactobacillus rhamnosus, Lactobacillus reuteri, Bifidobacterium animalis subsp. lactis (BB12), Bifidobacterium bifidum, Bifidobacterium longum and any combination thereof. According to one embodiment, the composition comprises FWGE and Lactobacillus rhamnosus. According to another embodiment, the composition comprises FWGE and Lactobacillus rhamnosus GG. According to another embodiment, the composition comprises FWGE and Lactobacillus reuteri. According to yet another embodiment, the composition comprises FWGE, Lactobacillus rhamnosus and Lactobacillus reuteri. According to one embodiment, the composition comprises FWGE and BB12. According to another embodiment, the composition comprises FWGE and Bifidobacterium bifidum. According to any one of the above embodiments, the FGWE is FGWP.

According to any one of the above embodiments, the composition further comprises an additional biologically active organic molecule. According to some embodiments, the organic molecule participates in cell energy metabolism. According to some embodiments, the organic molecule is creatine. According to another embodiment, the organic molecule is phosphocreatine. According to some embodiments the organic molecule is Vitamin D. Thus, in some embodiments, the present invention provides a composition comprising FWGE, a probiotic component, and creatine. According to some embodiments, the probiotic component comprises Lactobacillus rhamnosus, Lactobacillus reuteri, Bifidobacterium animalis subsp. lactis (BB12)), Bifidobacterium bifidum, Bifidobacterium longum and combination thereof. According to one embodiment, the probiotic component comprises Lactobacillus rhamnosus, Lactobacillus rhamnosus GG, Lactobacillus reuteri, and a combination thereof. According to any one of the above embodiments, the FGWE is FGWP.

According to some embodiments, the barrier is configured to prevent from penetration of FWGE to the probiotic compound and vice versa. According to some embodiments, the barrier is a sub-coating separating FWGE and the probiotic compound. As such, according to some embodiments, the composition comprises at least two compartments or layers. According to some embodiments, one compartment comprises FWGE and the second compartment comprises the probiotic compound, wherein the compartments are separated by the barrier. According to another embodiments, the composition comprises an inner shell comprising FWGE and the outer shell comprising the probiotic compound, having a barrier sub-coating separating the two shells. According to yet another embodiments, the composition comprises an inner shell comprising the probiotic compound and the outer shell comprising FWGE, having a barrier sub-coating separating the two shells. According to some embodiments, the composition comprises an organic compound having biological activity. In some embodiments the organic compound is selected from Vitamin D and creatine. According to some embodiment, the organic compound, such as Vitamin D and/or creatine, is admixed with FWGE, with the probiotic compound or with both FWGE and the probiotic compound. According to yet another embodiment, the creatine is located in yet another compartment or layer, which may be located in any possible configuration. According to some embodiments, the barrier is a layer, a coating, a membrane and any other possible obstacle preventing contact of FWGE and the probiotic component. According to some embodiments, the barrier, e.g. the coating, maintains the probiotic at a very low moisture content. According to some embodiments, the water content in the probiotic compound is less than 0.5%, less than 0.2%, less than 0.1%, less than 0.08% or less than 0.05%. According to some embodiments, the water activity of the probiotic compound in the composition is less than 1, less than 0.9, less than 0.8, less than 0.7, less than 0.6 or less than 0.5. According to some embodiments, the water activity of the probiotic compound in the composition is from about 0.01 to about 0.9, from about 0.05 to about 0.8, from about 0.1 to about 0.6. The term “water activity” as used herein refers to the partial vapor pressure of water in a substance divided by the standard state partial vapor pressure of water or alternatively by partial vapor pressure of pure water at the same temperature. Thus, the coating of the present invention prevents or inhibits penetration of water to the probiotic compound in the composition from e.g. FWGE.

According to some embodiments, the barrier is a moisture barrier. According to another embodiment, the barrier is a water impermeable barrier. According to yet another embodiment, the barrier is enteric coating as known in art. According to some embodiments, the barrier is BONULAC® or silicone release coating. According to other embodiments, the barrier may comprises at least one of CAP (Cellulose Acetate Phthalate), CAT (Cellulose Acetate Trimellitate), PVAP (Polyvinyl Acetate Phthalate), and HPMCP (Hypromellose Phthalate), fatty acids, waxes, shellac, plastics and plant fibers. According to some embodiments, the barrier is a natural oil. According to other embodiments, the barrier is a vegetable oil. According to some embodiments, the vegetable oil is selected from coconut oil, palm oil, palmist oil, soy oil, rapeseed oil, sunflower oil, cottonseed oil, peanut oil, olive oil, canola oil, castor oil, corn oil, flaxseed oil, safflower oil, and sesame oil. According to some embodiment, the vegetable oil is a non-hydrogenated vegetable oil. According to one embodiment, the vegetable oil is a refined oil. According to some embodiments, the barrier is a non-hydrogenated, refined palm oil. Examples of such barrier or coating is a commercial product REVEL®-A sold by Bunge Loders Croklaan B.V.

According to another embodiments, the barrier has a cut-off size of 5 kD. According to some embodiments, the barrier has a cut-off size of from 0.1 to 5 kD. According to some embodiments, the barrier has a cut-off size of from 0.2 to 4 kD, from 0.3 to 3 kD, from 0.4 to 2 kD, from 0.5 to 1 kD, from 0.6 to 0.8 kD, or from 0.5 to 3 kD. According to some embodiments, the barrier has a cut-off size equal or below 3 kD, 2 kD, or 1 kD.

According to some embodiments, the barrier is a coating on FWGE, probiotic compound or both. According to some embodiments, the composition comprises a mixture of FWGE coated particles and/or probiotic compound coated particles. According to one embodiment, the composition comprises a mixture of (i) FWGE coated particles and probiotic compound particles; (ii) FWGE particles and probiotic compound coated particles or (iii) FWGE coated particles and probiotic compound coated particles. According to some embodiments, the FWGE is microencapsulated. According to some embodiments, the FWGE is FWGE-SC.

According to some embodiments, the composition comprises FWGE particles coated with a non-hydrogenated, refined palm oil and particles of the probiotic compound. According to some embodiments, the composition comprises FWGP particles coated with a non-hydrogenated, refined palm oil and particles of the probiotic compound. According to some embodiments, the probiotic compound is Lactobacillus rhamnosus GG.

According to some embodiments, the composition further comprises at least one organic compound selected from creatine and Vitamin D. According to some embodiments, the organic compound is present as separate particles or admixed with FWGE, with probiotic compound or both. According to some embodiments, the coating is a barrier as described above. According to another embodiment, the coating is an enteric coating as known in the art. According to some embodiments, the particles are pellets or granules. According to any one of the above embodiments, the composition is further enteric coated. According to some embodiments, the particles are further enclosed in a capsule. According to some embodiments, the capsules are enteric coated capsules.

According to any one of the above embodiments the composition comprises a carrier. The term “carrier” as used herein includes as a class any compound or composition useful in facilitating storage, stability, administration, cell targeting and/or delivery of the topical composition, including, without limitation, suitable vehicles, diluents, emollients, solvents, excipients, pH modifiers, salts, colorants, rheology modifiers, thickeners, lubricants, humectants, antifoaming agents, erodible polymers, hydrogels, surfactants, emulsifiers, emulsion stabilizers, adjuvants, surfactants, preservatives, chelating agents etc.

According to some embodiments, the carrier is an edible carrier. The term “edible carrier” refers to compounds, materials, compositions, and/or dosage forms which are suitable for use in contact with the tissues of a subject. Each carrier must also be “acceptable” in the sense of being compatible with the other ingredients of the formulation. The term "edible carrier" as used herein means an organic material that can be consumed, digested or passed through the digestive system of an animal or human without any toxic effect. These edible carrier materials can exist as either a solid or liquid at room temperature.

According to some embodiments, the carrier is a pharmaceutically acceptable carrier.

In some embodiments, the prebiotic ingredient in the combination may be an extract of a probiotic microbe, such as an isolated cell wall fraction or an isolated metabolite produced by a probiotic culture.

According to some embodiments, the composition further comprises microcrystalline Cellulose M112D and magnesium stearate.

In some embodiments, the prebiotic, such as FWGE, promotes growth of bacteria in the gut, increases adhesion of probiotic bacteria in the gut, displaces pathogens, or provides a fermentable dose of carbohydrate to probiotic bacteria (symbiotic) or selected commensal bacteria and increases the levels of those microbial populations in the gastrointestinal tract. Non-limiting examples of microbial populations include lactobacilli and bifidobacteria.

In some embodiments, the amount of prebiotic in the composition ranges from about 1 mg to about 0.5 g. In some embodiments, the amount of prebiotic in the composition ranges from about 0.5 g to about 5 g. In some embodiments, the amount of prebiotic in the composition ranges from about 1 g to about 3 g. In some embodiments, a therapeutically effective amount of prebiotic is about 0.1 mg to about 5 g.

In some embodiments, the amount of creatine in the composition ranges from about 0.5 mg to about 5 g. In some embodiments, the dose of creatine ranges from about 0.5 mg/day to about 5 g/day. In some embodiments, the dose of creatine is about 1800 mg/day. In some embodiments, the dose of creatine ranges from about 0.1 mg/kg to about 150 mg/kg based on an average human weight of about 60 kg. In some embodiments, the amount of FWGE in the composition ranges from about 0.5 mg to about 15 g. In other embodiments, the amount of FWGE in the composition ranges from about 5 mg to about 12 g. In some embodiments, the amount of FWGE in the composition ranges from about 10 mg to about 10 g, from 20 mg to 8 g, from 40 mg to 6 g. In some embodiments, the amount of FWGE in the composition ranges from about 0.05 g to about 2 g, or from 0.1 g to 1.2 g. In some embodiments, the dose of FWGE ranges from about 0.5 mg/day to about 15 g/day. In some embodiments, the dose of FWGE is about 1800 mg/day. In some embodiments, the dose of FWGE ranges from about 0.1 mg/kg to about 150 mg/kg based on an average human weight of about 60 kg. In some embodiments, the amount of FWGP in the composition ranges from about 0.5 mg to about 500 mg. In other embodiments, the amount of FWGP in the composition ranges from about 1 mg to about 450 mg, from 5 to 450, from 10 to 400 mg, from 20 to 300 mg, from 30 to 200 mg, from 35 to 150 mg, from 40 to 120 mg, from 50 to 100 mg, from 60 to 80 mg, from 20 to 80 mg or from 30 to 60 mg of FWGP. In some embodiments, the amount of FWGP in the composition ranges from about 1 to 100 mg. In some embodiments, the amount of FWGP in the composition ranges from about 60 to 150 mg. In some embodiments, the amount of FWGP in the composition ranges from about 80 to 120 mg. In some embodiments, the amount of FWGP in the composition ranges from about 90 to 115 mg. from 2 to 60 mg, from 3 to 50 mg, from 5 to 45 mg, from 3 to 20 mg or from 10 to 60 mg.

In some embodiments, the amount of probiotic in the composition ranges from about 0.5 million CFU to about 100 billion CFU. In some embodiments, the amount of probiotic in the composition ranges from about 1 billion CFU to about 50 billion CFU. In some embodiments, the amount of probiotic in the composition ranges from about 50 billion CFU to about 100 billion CFU. In some embodiments, the amount of probiotic in the composition ranges from about 10 billion CFU to about 80 billion CFU, from 20 billion to 60 billion or from 30 billion to 50 billion CFU.

In some embodiments, the dose of probiotic ranges from about 100 million CFU to about 100 billion CFU. In some embodiments, the dose of probiotic ranges from about 200 million CFU to about 10 billion CFU. In some embodiments, the dose of probiotic ranges from about 1 billion CFU to about 50 billion CFU. In some embodiments, the dose of probiotic ranges from about 50 billion CFU to about 100 billion CFU.

In some embodiments, the dose of probiotic ranges from about 10,000 CFU/kg to about 10 million CFU/kg. In some embodiments, the dose of probiotic ranges from about 100,000 CFU/kg to about 1 million CFU/kg based on an average human weight of about 60 kg. In some embodiments, the dose of probiotic ranges from about 1 million CFU/kg to about 10 million CFU/kg based on an average human weight of about 60 kg. In some embodiments, a therapeutically effective amount of probiotic is about 0.5 million CFU to about 100 billion CFU based on an average human weight of about 60 kg.

In some embodiments, the amount of Lactobacillus rhamnosus in the composition ranges from about 0.5 million CFU to about 100 billion CFU. In other embodiments, the amount of Lactobacillus rhamnosus GG in the composition ranges from about 0.5 million CFU to about 100 billion CFU. In some embodiments, the dose of Lactobacillus rhamnosus ranges from about 100 million CFU/day to about 100 billion CFU/day. In some embodiments, the dose of Lactobacillus rhamnosus is about 12 billion CFU/day. In some embodiments, the dose of Lactobacillus rhamnosus ranges from about 10,000 CFU/kg to about 10 million CFU/kg based on an average human weight of about 60 kg. In some embodiments, the amount of Lactobacillus rhamnosus in the composition ranges from about 10 billion CFU to about 80 billion CFU, from 20 to 60 or from 30 to 50 billion CFU.

In some embodiments, the amount of Lactobacillus reuteri in the composition ranges from about 0.5 million CFU to about 100 billion CFU. In some embodiments, the dose of Lactobacillus reuteri ranges from about 0.5 million CFU/day to about 10 billion CFU/day. In some embodiments, the dose of Lactobacillus reuteri is about 12 billion CFU/day. In some embodiments, the dose of Lactobacillus reuteri ranges from about 10,000 CFU/kg to about 10 million CFU/kg based on an average human weight of about 60 kg. In some embodiments, the amount of Lactobacillus reuteri in the composition ranges from about 10 billion CFU to about 80 billion CFU, from 20 to 60 or from 30 to 50 billion CFU.

According to some embodiments, the composition comprises from 1 to 30 mcg, from 2 to 25 mcg, from 5 to 20 mcg of Vitamin D. Alternatively, the composition comprises from 10 to 1200 IU, from 20 to 1100 IU, from 30 to 1000 IU, from 50 to 900 UI, from 75 to 800 IU, from 100 to 600 IU, from 200 to 400 IU or from 300 to 1000 IU. Any known form of Vitamin D, e.g. Di, D2, D3, D4 and D5 is encompassed. According to some embodiments, Vitamin D id D2 or D3.

According to some embodiments, the composition comprises from 1 to 400 mg of FWGP and from 100 million CFU to about 100 billion CFU of probiotic. According to some embodiments, the composition comprises from 5 to 350 mg of FWGP and from 10 million CFU to about 80 billion CFU of probiotic. According to some embodiments, the composition comprises from about 10 to about 200mg, from about 20 to about 180, from about 40 to about 150mg, from about 60 to about 130mg, from about 80 to about 120 mg, from about 10 to 100 mg, from 2 to 60 mg, from 3 to 50 mg, from 5 to 45 mg, from 3 to 20 mg or from 10 to 60 mg of FWGP and from about 1 billion CFU to about 100 billion CFU, from about 10 to about 80 billion CFU, from about 15 billion CFU to about 80 billion CFU, from 20 billion to 60 billion or from 30 billion to 50 billion CFU of probiotic. According to some embodiments, the probiotic is Lactobacillus rhamnosus. According to other embodiments, the probiotic is Lactobacillus rhamnosus GG. According to yet another embodiment, the probiotic is Lactobacillus reuteri. According to some embodiments, the composition further comprises creatine. According to some embodiments, the composition comprises from about 80 to about 120 mg of FWGP and from about 20 billion CFU to about 60 billion CFU of the probiotic. According to other embodiments, the composition comprises from about 80 to about 120 mg of FWGP and from about 20 billion CFU to about 60 billion CFU of the Lactobacillus rhamnosus GG According to some embodiments, the composition comprises from about 0.5 mg to about 2 g or creatine. According to other embodiments, the composition comprises Vitamin D. According to some embodiments, the composition comprises from about 80 to about 120 mg of FWGE-SC and from about 20 billion CFU to about 60 billion CFU of the probiotic. According to other embodiments, the composition comprises from about 80 to about 120 mg of FWGE-SC and from about 20 billion CFU to about 60 billion CFU of the Lactobacillus rhamnosus GG. According to some embodiments, the composition comprises from about 0.5 mg to about 2 g of creatine. According to other embodiments, the composition comprises from about 1 mcg to about 20 mcg of Vitamin D.

According to some embodiments, the composition comprises from 1 to 15 g of FWGE and from 0.5 million CFU to about 100 billion CFU of probiotic. According to some embodiments, the composition comprises from 2 to 12 mg of FWGE and from 10 million CFU to about 80 billion CFU of probiotic. According to some embodiments, the composition comprises from about 3 to 10 g, from 4 to 8 g, or from 5 to 6 g of FWGE and from about 1 billion CFU to about 100 billion CFU, from about 10 to about 80 billion CFU, from about 15 billion CFU to about 80 billion CFU, from 20 billion to 60 billion or from 30 billion to 50 billion CFU of probiotic. According to some embodiments, the probiotic is Lactobacillus rhamnosus. According to other embodiments, the probiotic is Lactobacillus rhamnosus GG. According to yet another embodiment, the probiotic is Lactobacillus reuteri. According to some embodiments, the composition further comprises creatine. According to some embodiments, the composition comprises from about 0.5 mg to about 2 g or creatine. According to some embodiments, the composition further comprises Vitamin D, e.g. 1 to 30 mcg of Vitamin D.

Various proportions of FWGE and probiotic are contemplated in the compositions according to the present disclosure. For example, in some embodiments, compositions can comprise from about 0.5% to about 99.5% FWGE, about 0.5% to about 99.5% probiotic, and about 0.5 to about 99.5% organic compound. The proportions of FWGE, probiotic, and organic compound in the compositions can be adjusted based on need by one of ordinary skill in the art. For example, in some embodiments, compositions can comprise about 2.5% FWGE, about 9.5% probiotic, and about 2.5% organic compound. In some embodiments, compositions can comprise about 5% FWGE, about 90% probiotic, and about 5% organic compound. In some embodiments, compositions can comprise about 7.5% FWGE, about 85% probiotic, and about 7.5% organic compound. In some embodiments, compositions can comprise about 10% FWGE, about 80% probiotic, and about 10% organic compound. In some embodiments, compositions can comprise about 12.5% FWGE, about 75% probiotic, and about 12.5% organic compound. In some embodiments, compositions can comprise about 15% FWGE, about 70% probiotic, and about 15% organic compound. In some embodiments, compositions can comprise about 17.5% FWGE, about 65% probiotic, and about 17.5% organic compound. In some embodiments, compositions can comprise about 20% FWGE, about 60% probiotic, and about 20% organic compound. In some embodiments, compositions can comprise about 22.5% FWGE, about 55% probiotic, and about 22.5% organic compound. In some embodiments, compositions can comprise about 25% FWGE, about 50% probiotic, and about 25% organic compound. In some embodiments, compositions can comprise about 27.5% FWGE, about 45% probiotic, and about 27.5% organic compound In some embodiments, compositions can comprise about 5% FWGE, about 90% probiotic, and about 5% organic compound. In some embodiments, compositions can comprise about 7.5% prebiotic, about 85% probiotic, and about 7.5% organic compound. In some embodiments, compositions can comprise about 10% FWGE, about 80% probiotic, and about 10% organic compound. In some embodiments, compositions can comprise about 12.5% FWGE, about 85% probiotic, and about 12.5% organic compound. In some embodiments, compositions can comprise about 15% FWGE, about 70% probiotic, and about 15% organic compound. In some embodiments, compositions can comprise about 30% FWGE and about 40% probiotic. In some embodiments, compositions can comprise about 25% FWGE and about 75% probiotic. In some embodiments, compositions can comprise about 30% FWGE and about 70% probiotic. In some embodiments, compositions can comprise about 35% FWGE and about 65% probiotic. In some embodiments, compositions can comprise about 40% FWGE and about 55% probiotic. In some embodiments, compositions can comprise about 50% FWGE and about 50% probiotic. In some embodiments, compositions can comprise about 55% FWGE and about 45% probiotic. In some embodiments, compositions can comprise about 60% FWGE and about 40% probiotic. In some embodiments, compositions can comprise about 65% FWGE and about 35% probiotic. In some embodiments, compositions can comprise about 70% FWGE and about 30% probiotic. In some embodiments, compositions can comprise about 75% FWGE and about 25% probiotic. In some embodiments, compositions can comprise about 80% FWGE and about 20% probiotic. In some embodiments, compositions can comprise about 85% FWGE and about 15% probiotic. In some embodiments, compositions can comprise about 90% FWGE and about 10% probiotic. In some embodiments, compositions can comprise about 95% pre FWGE and about 5% probiotic. According to some embodiments, the FWGE is FWGP.

Various proportions of FWGE and Lactobacillus rhamnosus are contemplated in the compositions according to the present disclosure. For example, in some embodiments, compositions can comprise from about 0.5% to about 99.5% FWGE and about 0.5% to about 99.5% Lactobacillus rhamnosus. The proportions of FWGE and Lactobacillus rhamnosus in the compositions can be adjusted based on need by one of ordinary skill in the art. For example, in some embodiments, compositions can comprise about 5% FWGE and about 95% Lactobacillus rhamnosus. In some embodiments, compositions can comprise about 10% FWGE and about 90% Lactobacillus rhamnosus. In some embodiments, compositions can comprise about 15% FWGE and about 85% Lactobacillus rhamnosus. In some embodiments, compositions can comprise about 20% FWGE and about 80% Lactobacillus rhamnosus. In some embodiments, compositions can comprise about 25% FWGE and about 75% Lactobacillus rhamnosus. In some embodiments, compositions can comprise about 30% FWGE and about 70% Lactobacillus rhamnosus. In some embodiments, compositions can comprise about 35% FWGE and about 65% Lactobacillus rhamnosus. In some embodiments, compositions can comprise about 40% FWGE and about 55% Lactobacillus rhamnosus. In some embodiments, compositions can comprise about 50% FWGE and about 50% Lactobacillus rhamnosus. In some embodiments, compositions can comprise about 55% FWGE and about 45% Lactobacillus rhamnosus. In some embodiments, compositions can comprise about 60% FWGE and about 40% Lactobacillus rhamnosus. In some embodiments, compositions can comprise about 65% FWGE and about 35% Lactobacillus rhamnosus. In some embodiments, compositions can comprise about 70% FWGE and about 30% Lactobacillus rhamnosus. In some embodiments, compositions can comprise about 75% FWGE and about 25% Lactobacillus rhamnosus. In some embodiments, compositions can comprise about 80% FWGE and about 20% Lactobacillus rhamnosus. In some embodiments, compositions can comprise about 85% FWGE and about 15% Lactobacillus rhamnosus. In some embodiments, compositions can comprise about 90% FWGE and about 10% Lactobacillus rhamnosus. In some embodiments, compositions can comprise about 95% FWGE and about 5% Lactobacillus rhamnosus. According to some embodiments, the FWGE is FWGP. According to some embodiments, the Lactobacillus rhamnosus is Lactobacillus rhamnosus GG.

In some embodiments, the composition of the present invention is a pharmaceutical composition The terms “pharmaceutical composition” and "pharmaceutical formulation" are used herein interchangeably and refer to a composition comprising the botanical extract such as FWGE, the probiotic compound, optionally an organic compound such as creatine or Vitamin D and formulated together with one or more pharmaceutically acceptable carriers.

The pharmaceutical formulations described herein may be prepared by any method known or hereafter developed in the art of pharmacology. In general, such preparatory methods include the step of bringing the active ingredient into association with a carrier or one or more other accessory ingredients, and then, if necessary or desirable, shaping or packaging the product into a desired single- or multi-dose unit formulations may comprise additional medicinal agents, pharmaceutical agents, carriers, buffers, adjuvants, dispersing agents, diluents, and the like depending on the intended use and application.

The term "pharmaceutically acceptable carrier" or "pharmaceutically acceptable excipient" as used herein refers to any and all solvents, dispersion media, preservatives, antioxidants, coatings, isotonic and absorption delaying agents, surfactants, fillers, disintegrants, binders, diluents, lubricants, glidants, pH adjusting agents, buffering agents, enhancers, wetting agents, solubilizing agents, surfactants, antioxidants the like, that are compatible with pharmaceutical administration. The use of such media and agents for pharmaceutically active substances is well known in the art. The compositions may contain other active compounds providing supplemental, additional, or enhanced therapeutic functions, solid carriers or excipients such as, for example, lactose, starch or talcum or liquid carriers such as, for example, water, fatty oils or liquid paraffins. Examples of suitable pharmaceutical carriers, excipients and/or diluents are well known in the art and include, but are not limited to, a gum, a starch (e g. com starch, pregeletanized starch), a sugar (e.g., lactose, mannitol, sucrose, dextrose), a cellulosic material (e.g. microcrystalline cellulose), an acrylate (e.g. polymethylacrylate), calcium carbonate, magnesium oxide, talc, or mixtures thereof.

Pharmaceutically acceptable carriers for liquid formulations are aqueous or nonaqueous solutions, suspensions, emulsions or oils, Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, and injectable organic esters such as ethyl oleate. Examples of oils are those of animal, vegetable, or synthetic origin, for example, peanut oil, soybean oil, olive oil, sunflower oil, turmeric oil, fish-liver oil, another marine oil, or a lipid from milk or eggs.

Aqueous carriers include water, alcoholic/aqueous solutions, emulsions or suspensions, including saline and buffered media such as phosphate buffered saline solutions, water, emulsions, such as oil/water emulsions, various types of wetting agents, sterile solutions etc. Formulations comprising such carriers can be formulated by well- known conventional methods. Suitable carriers may comprise any material which, when combined with the biologically active compounds of the disclosure, retains the biological activity. Preparations for parenteral administration may include sterile aqueous or nonaqueous solutions, suspensions, and emulsions. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oils such as olive oil, and injectable organic esters such as ethyl oleate. Aqueous carriers include water, alcoholic/aqueous solutions, emulsions or suspensions, including saline and buffered media. Parenteral vehicles may include sodium chloride solution, Ringer's dextrose, dextrose and sodium chloride, lactated Ringer's, or fixed oils. Intravenous vehicles may include fluid and nutrient replenishes, electrolyte replenishers (such as those based on Ringer's dextrose), and the like. Preservatives and other additives may also be present including, for example, antimicrobials, anti-oxidants, chelating agents, and inert gases and the like, in addition, the pharmaceutical formulations of the present disclosure might comprise proteinaceous carriers, e.g., serum albumin or immunoglobulin, preferably of human origin. The pharmaceutical formulations provided herein may also be administered as controlled- release formulations, i.e. formulations in which the active ingredient is released over a period of time after administration. Controlled- or sustained-release formulations include formulation in lipophilic depots (e.g. fatty acids, waxes, oils). In another embodiment, the formulation is an immediate-release formulation, i.e. a formulation in which all the active ingredient is released immediately after administration.

Further, the pharmaceutical formulations may be administered admixed to food, functional food, beverages, medicinal food.

Although the description of pharmaceutical formulations provided herein are principally directed to pharmaceutical formulations which are suitable for ethical administration to humans, it will be understood by the skilled artisan that such formulations are generally suitable for administration to animals of all sorts, e.g. nonhuman mammals. Modification of pharmaceutical formulations suitable for administration to humans in order to render the formulations suitable for administration to various animals is well understood, and the ordinarily skilled veterinary pharmacologist can design and perform such modification with merely ordinary, if any, experimentation. Subjects to which administration of the pharmaceutical formulations of the disclosure is contemplated include, but are not limited to, humans and other primates, mammals including commercially relevant mammals such as non-human primates, cattle, pigs, horses, sheep, cats, dogs, duck, chicken, and sheep.

Pharmaceutical formulations of the disclosure may be prepared, packaged, or sold in bulk, as a single unit dose, or as a plurality of single unit doses. As used herein, a “unit dose” is discrete amount of the pharmaceutical formulations comprising a predetermined amount of the active ingredient. The amount of the active ingredient is generally equal to the dosage of the active ingredient which would be administered to a subject or a convenient fraction of such a dosage such as, for example, one-half or one-third of such a dosage.

The relative amounts of an active ingredient, a pharmaceutically acceptable carrier, and any additional ingredients in a pharmaceutical formulation of the disclosure will vary, depending upon the identity, size, and condition of the subject treated and further depending upon the route by which the formulation is to be administered. By way of example, the formulation may comprise between 0.1% and 100% (w/w) active ingredient. In addition to the active ingredients, a pharmaceutical formulation of the disclosure may further comprise one or more additional pharmaceutically active agents.

Controlled- or sustained-release pharmaceutical formulations may be made using conventional technology.

Formulations of the present disclosure may also further comprise additional prebiotic component. The term “prebiotic” includes substances or compounds that are fermented by the intestinal flora of the pet and hence promote the growth or development of lactic acid bacteria in the gastro-intestinal tract of the pet at the expense of pathogenic bacteria. The result of this fermentation can be a release of fatty acids, in particular shortchain fatty acids in the colon. This release can have the effect of reducing the pH value in the colon. Non-limiting examples of suitable prebiotics include oligosaccharides, such as inulin and its hydrolysis products commonly known as fructo-oligosaccharides, galacto-oligosaccharides, xylo-oligosaccharides, or oligo derivatives of starch (such as pectin, beta-glucan, and resistant starch). The prebiotics may be provided in any suitable form.

For example, the prebiotic may be provided in the form of plant material that contains the fiber. Suitable plant materials include asparagus, artichokes, onions, wheat or chicory, or residues of these plant materials. Alternatively, the prebiotic fiber may be provided as an inulin extract, for example extracts from chicory are suitable. Suitable inulin extracts may be obtained from Orafti SA of Tirlemont 3300, Belgium under the trade mark RAFTILINE®. For example, the inulin may be provided in the form of Raftiline (g) ST which is a fine white powder, which contains about 90 to about 94% by weight of inulin, up to about 4% by weight of glucose and fructose, and about 4 to 9% by weight of sucrose. Alternatively, the fiber may be in the form of a fructo-oligosaccharide such as obtained from Orafti SA of Tirlemont 3300, Belgium under the trade mark RAFTILOSE®. For example, the inulin may be provided in the form of Raftilose (g) P95. Otherwise, the fructo-oligosaccharides may be obtained by hydrolyzing inulin, by enzymatic methods, or by using micro-organisms.

The term "pharmaceutical formulations" also include nutritional formulations, such as oral nutritional formulations for oral consumption and optionally for enteral adsorption, wherein the nutritional formulation includes the compounds of the present disclosure.

If the nutritional formulations are formulated to be administered orally, the formulations may be a liquid oral nutritional formulation. In some embodiments, the oral nutritional formulation in an incomplete nutritional formulation. In some embodiments, the oral nutritional formulation is a complete nutritional formulation. In this manner, the nutritional formulations may be administered in any known form including, for example, tablets, capsules, liquids, chewables, soft gels, sachets, powders, syrups, liquid suspensions, emulsions and solutions in convenient dosage forms.

A nutritional formula encompasses any nutritionally complete or nutritionally incomplete (for e.g., a supplementary formulation, a nutritional supplement). As used herein, “nutritionally complete” are preferably nutritional products that contain sufficient types and levels of macronutrients (protein, fats and carbohydrates) and micronutrients to be sufficient to be a sole source of nutrition for the subject to which it is being administered to. Patients can receive 100% of their nutritional requirements from such complete nutritional formulations. According to some embodiments, the nutritional formula is a supplementary formulation providing supplementary nutrition. A “supplementary formula” may not be nutritionally complete, but preferably contains specific nutrients that are supportive, for example in combination with physical exercise, which further the beneficial effects of the disclosure, and/or which address specific or additional needs of the subject.

The nutritional formula may be a generally applicable nutritional formula, for example adapted to subjects of a specific age, for example a formula for children, but it may also be a formula for elderly patients, for intensive care patients, or a specially adapted formula for patients suffering from a specific disease, for example. Any nutritional formula may be reconstitutable, that is, present in a substantially dried, for example powdered form, or ready -to-drink, in the form of liquid formulas, for example. According to any one of the above embodiments, the compositions of the present invention is an enteric coated composition.

The term "enteric coating" comprises any pharmaceutically or edible acceptable coating preventing the release of the active agent in the stomach and sufficiently disintegrating in the intestine tract (by contact with approximately neutral or alkaline intestine juices) to allow the resorption of the active agent through the walls of the intestinal tract. Various in vitro tests for determining whether or not a coating is classified as an enteric coating have been published in the pharmacopoeia of various countries.

The composition of the present invention may be adjusted according to applications. In particular, the pharmaceutical composition may be formulated using a method known in the art so as to provide rapid, continuous or delayed release of the active ingredient after administration to mammals. For example, the formulation may be any one selected from among plasters, granules, lotions, liniments, lemonades, aromatic waters, powders, syrups, ophthalmic ointments, liquids and solutions, aerosols, extracts, elixirs, ointments, fluidextracts, emulsions, suspensions, decoctions, infusions, ophthalmic solutions, tablets, suppositories, injections, spirits, capsules, creams, troches, tinctures, pastes, pills, and soft or hard gelatin capsules.

According to some embodiments, the composition of the present invention is a tablet. According to other embodiments, the composition is a capsule. According to some embodiments, the composition of the present invention is a powder, granules or pellets. According to other embodiments, the composition is a sachet. The compositions of the present invention may be administered in any known conventional way.

The present disclosure relates to the unexpected discovery that certain compositions comprising the botanical extract and probiotic bacteria are beneficial to a subject’s immune system. In some embodiments, compositions comprising the botanical extract and probiotic bacteria improve a subject’s immune system function. In some embodiments, compositions comprising the botanical extract and probiotic bacteria enhance a subject’s immune system function. In some embodiments, compositions comprising the botanical extract and probiotic bacteria restore a subject’s dysfunctional immune system. In some embodiments, compositions comprising the botanical extract and probiotic bacteria control a subject’s overactive immune system. In some embodiments, compositions comprising the botanical extract and probiotic bacteria reduce, inhibit or prevent the proliferation of cancer cells and/or tumors. According to some embodiments, the combination or the composition of the present invention may modulate mitochondria energy metabolism. Without being bound to any particular theory, the modulation of the mitochondria energy metabolism may be conferred by FWGE which is combined with the effect of the probiotic bacteria.

According to another aspect, the present invention provides the combination or the composition of the present invention for use in treating a disease or condition involving the immune system.

In some aspects, the combination or the composition of the present invention is for use in enhancement of an immune system function. In some embodiments, the combination or the composition of the present invention is for use in enhancing the immune response to vaccination. In some embodiments, the combination or the composition of the present invention is for use treating a disease or disorder that is associated with dysfunctional immune response. In some embodiments, the combination or the composition of the present invention is for use in enhancing the immune response in cancer treatment. In some embodiments, the combination or the composition of the present invention is for use in enhancing the immune response to instances where the immune system is challenged or compromised, such as in therapy, surgery and the likes. According to yet another embodiment, the combination or the composition is for use in modulating mitochondria energy metabolism.

According to some embodiments, modulating mitochondria energy metabolism comprises supporting production of SCFA in the colon. In one embodiment, the combination or the composition of the presented in the invention is for use in enhancing and supporting immune system of a subject during radiation and/or chemotherapy.

In some embodiments, the use comprises administering the combination of the composition of the present invention to a subject. In some embodiments of the method, the subject is a mammal. In some embodiments of the method, the subject is a human. In some embodiments, the botanical extract is a fermented wheat germ extract. In some embodiments, the botanical extract comprises at least one polypeptide found in fermented wheat germ extract. In some embodiments of the combination, the probiotic component is selected from the group consisting of Lactobacillus rhamnosus, Lactobacillus rhamnosus GG, Lactobacillus reuteri, Bifidobacterium animalis subsp. lactis (BB12)), Bifidobacterium bifidum, and Bifidobacterium longum.

According to another aspect, the present invention provides a method for treating a disease or condition involving an immune system in a subject in need thereof comprising administering to said subject a therapeutically effective amount of botanical extract, a probiotic component and optionally an organic compound to said subject. According to one embodiment, the botanical extract is FWGE. According to some embodiments, the method comprises administering the composition of the present invention.

In some embodiments, the present invention provides a methods a for enhancing the immune system comprising administering to said subject a therapeutically effective amount of botanical extract, a probiotic component and optionally an organic compound to said subject. According to some embodiments, the method comprises administering the composition of the present invention to the subject. In some embodiments, the methods relate to methods of enhancing the immune response to vaccination. In some embodiments, the method relates to enhancing the immune response to treat a disease or disorder that is associated with dysfunctional immune response. In some embodiments, the methods relate to methods of enhancing the immune response to treat cancer. In some embodiments, the methods relate to methods of enhancing the immune response to instances where the immune system is challenged or compromised, such as in therapy, surgery and the likes.

According to any one of the aspects and embodiments, the disease or condition involving immune system is selected from cancer, dysfunctional or compromised immune system.

In some embodiments, the disclosure relates to generically treating diseases or disorders associated with dysfunctional immune response whereby having one or more beneficial effect on the immune system is a desired therapeutic outcome. Non-limiting examples of beneficial effects on a subject’s immune system include increase in red blood cell count, increase in hemoglobin content, increase in hematocrit, improved antigen presentation and improved immunoglobulin secretion. Other non-limiting examples of beneficial effects on a subject’s immune system macrophage antigen presentation activity and inhibition of tumor-derived immune suppressive factors, enhance macrophage proliferation and activation, promote differentiation of Thl cells; increase macrophage production of IL-12, increase NK activity; promote apoptosis of cancer cells. AHCC in cancer patients has been reported to increase TNF-a, y-interferon, interleukin- 12 and decrease immunosuppressive acidic protein (IAP) and tumor growth factor (TGF)-alpha.

In some embodiments, the methods or use relates to the administration of a therapeutic amount of the combination or compositions of the present invention. For example, in certain instances, a subject may be administered from 0.5 mg to 100 g of the compositions of the disclosure. Preferably, subject may be administered from 1 mg to 12 g of the compositions of the disclosure. According to some embodiments, the administration of botanical extract, the probiotic and optionally the organic compound is as described in any one of the above aspects and embodiments. In some embodiments, the FWGE is administered in amount of from about 0.5 mg/day to about 15 g/day. In some embodiments, the amount of FWGP is administered in amount of from about 0.5 mg to about 500 mg/day. In other embodiments, the amount of FWGP is administered in amount of from about 1 mg to about 450 mg/day, from 10 to 400 mg/day, from 20 to 300 mg/day, from 30 to 200 mg/day, from 35 to 150 mg/day, from 40 to 120 mg/day, from 50 to 100 mg/day, from 60 to 80 mg/day, from 20 to 80 mg/day or from 30 to 60/day mg of FWGP. In other embodiments, the amount of FWGP is administered in amount of from 60 to 320 mg/day or from 100 to 250 mg/day. In other embodiments, the probiotic is administered in the amount of from about 0.5 million CFU/day to about 300 billion CFU/day. In other embodiments, the probiotic is administered in the amount of from about 20 billion CFU/day to about 250 billion CFU/day, from 40 billion to 200 billion CFU/day or from 60 billion to 150 billion to CFU/day. In some embodiments, creatine is administered in the amount of from about 0.5 mg to about 5 g. In some embodiments of the combination, the probiotic component is selected from the group consisting of Lactobacillus rhamnosus, Lactobacillus rhamnosus GG, Lactobacillus reuteri, Bifidobacterium animalis subsp. lactis (BB12)), Bifidobacterium bifidum, and Bifidobacterium longum. In one embodiment, the probiotic component is Lactobacillus rhamnosus, Lactobacillus reuteri, or a combination thereof.

According to some embodiments, the methods or use comprises administering from 1 to 8 dosage forms per day of the composition of the present invention. According to some embodiments, the composition is in form of capsules. Thus, according to some embodiments, the use comprises administering from 1 to 8, from 2 to 6 or from 2 to 4 capsules comprising the composition of the present inventions. According to some embodiments, the capsule comprises from 3 to 60 mg or FWGP and from 30 billion to 60 billion CFU/day of the probiotic compound. In some embodiments of the combination, the probiotic component is selected from the group consisting of Lactobacillus rhamnosus, Lactobacillus reuteri, Bifidobacterium animalis subsp. lactis (BB12)), Bifidobacterium bifidum, and Bifidobacterium longum. In one embodiment, the probiotic component is Lactobacillus rhamnosus, Lactobacillus reuteri, or a combination thereof. In certain embodiments, the capsule further comprises creatine.

In some embodiments, the combination of botanical extract, e.g. FWGE and probiotic produces an additive beneficial effect on the immune system, such that the prebiotic and probiotic ingredients in the combination behave as they are observed to behave when given by themselves - where neither ingredient modifies the effect of the other. In some embodiments, the combination of FWGE and probiotic produces an unexpected and surprising synergistic effect. Such synergistic effects are greater than additive effects. An additive effect is observed when the effect on the immune system is equal to the sum of the individual effects of the prebiotic component and the probiotic component. A synergistic effect is observed when the potentiation is greater than the sum of the individual effects of the prebiotic component and the probiotic component. Synergistic effect, additive effect or both can occur in human patients, non-human patients, non-patient human volunteers, in vivo models, ex vivo models, in vitro models, etc. Potentiation can range from about <1 to about 100-fold. In some embodiments, the synergistic effect is about 2 to about 75-fold. In some embodiments, the potentiation ranges from <1, 1, >1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 20, 30, 40, 50, 60, 70, 80, 90, or 100-fold, or within a range defined by any two of the aforementioned values. A synergistic effect allows one or all of the components of a composition to be used in amounts and/or doses that are lower that the amounts and/or doses of doses of the components used individually. Thus, in some embodiments, the FWGE can potentiate the effect of the probiotic on the immune system. In some embodiments, the probiotic can potentiate the effect of the FWGE on the immune system. In some embodiments, the prebiotic can potentiate the effect of the probiotic and the FWGE can potentiate the effect of the prebiotic on the immune system. The potentiation can occur in several ways. Non-limiting examples include enhancing the effectiveness of an already effective FWGE and/or probiotic, making an ineffective prebiotic and/or probiotic effective (the prebiotic and/or FWGE could also have been previously effective but become ineffective following long term and/or short term use in a patient), increasing the length of time for which a FWGE and/or probiotic is effective, decreasing the effective dose of administration of a FWGE and/or probiotic, decreasing the duration of time for which a FWGE and/or probiotic is administered, decreasing the frequency of administration of a prebiotic and/or probiotic, and/or enabling the administration of a prebiotic and/or probiotic via one or more amenable route.

Immune Activation / Methods of Enhancing Immune System

The synergistic effects of the composition or the combination of the present invention provide synergistic activation of multiple types of immune cells, synergistic production of immune-activating, anti-inflammatory, and regenerative biomarkers, and synergistic reduction in breast cancer cell viability.

In some embodiments, the composition or the combination of the present invention enhances the production of one or more cytokines by the one or more immune cells. In some embodiments, the composition or the combination of the present invention activates one or more immune cells and induces and/or enhances the production of one or more cytokines by the one or more immune cells. In some embodiments, the composition or the combination of the present invention activates one or more immune cells and induces and/or enhances the production of one or more cytokines by the one or more immune cells. In some embodiments, the one or more immune cells activated by the composition or the combination of the present invention are the same. In some embodiments, the one or more immune cells activated by the composition or the combination of the present invention are different. In some embodiments, the one or more immune cells activated by the composition or the combination of the present invention are overlapping

In some embodiments, an immune activation by a combination of FWGE and probiotic is additive. In some embodiments, an immune activation by a combination of FWGE and probiotic is synergistic. In some embodiments, the probiotic is whole cells (e.g., whole bacteria), such as Lactobacillus rhamnosus GG. In some embodiments, the probiotic is one or more cellular fractions of the probiotic. Non-limiting examples, of one or more cellular fractions include cell wall preparation/cell wall fraction, cellular metabolites fraction, cytoplasmic fraction, or combinations thereof. In some embodiments, one or more commercially available probiotic bacteria are cultured to obtain a secreted probiotic metabolites fraction and/or cell wall fraction for testing. In some embodiments, metabolites and cell walls are isolated by one or more techniques and/or methods known in the art. In some embodiments, after the metabolites were harvested, the probiotic cell wall fraction was isolated.

In some embodiments, an immune modulation by a combination of FWGE and probiotic cell wall fraction is additive. In some embodiments, an immune modulation by a combination of FWGE and probiotic cell wall fraction is synergistic. In some embodiments, an immune modulation by a combination of FWGE and probiotic cellular metabolites fraction is additive. In some embodiments, an immune modulation by a combination of FWGE and probiotic cellular metabolites fraction is synergistic. In some embodiments, an immune modulation by a combination of FWGE and probiotic cellular metabolites fraction is additive. In some embodiments, an immune modulation by a combination FWGE and probiotic cellular metabolites fraction is synergistic. In some embodiments, the immune modulation includes immune activation, inhibition and/or regulation of an immune response. In some embodiments, the immune modulation results in a beneficial effect, for example, suppression of breast cancer cell viability, killing of cancer cells, etc.

In some embodiments, immune cell activation includes one or more of cytokine induction, upregulating of expression of activation markers on immune cells, and induction of anti-inflammatory cytokines. In some embodiments, the immune cell activation is associated with reduced cellular viability of cancer cells.

In some embodiments, provided herein are methods for activating immune cells and/or inducing cytokines in a subject. In some embodiments, the subject is a human. In some embodiments, the subject is a non-human. In some embodiments, the subject is a non-human mammal. In some embodiments, the method comprising administering to the subject an effective amount of a FWGE. In some embodiments, the method comprising administering to the subject an effective amount of a probiotic. In some embodiments, the method comprising administering to the subject a combination of an effective amount of a FWGE and an effective amount of one or more probiotics.

The term “patient”, “subject”, or “individual” are used interchangeably and refer to either a human or a non-human animal. These terms include mammals, such as humans, primates, livestock animals (including bovines, porcines, etc.), companion animals (e.g., canines, felines, etc.) and rodents (e.g., mice and rats). According to some embodiments the non-human mammal is selected from the group consisting of livestock animals, domestic pets, rodents, wild animals and primate.

According to some embodiments, the subject is a healthy subject. In some embodiments, the present invention provides a composition or combination for use in enhancing immune system. For example, in such embodiments, the subject may be a healthy subject. According to some embodiments, the condition involving immune system is a normal state of the subject.

In some embodiments, the probiotic is an extract, for example, an isolated cell wall fraction, an isolated metabolite produced by a probiotic, or a combination thereof.

In some embodiments, the present disclosure includes methods of enhancing the immune system by administering the compositions of the disclosure to a subject. The methods include methods of potentiating an immune response in normal subjects, in subjects who are immunocompromised, or in subjects who are at risk of infection due to disease, hospitalization, age or other predisposing medical factors. A method for activating immune cells and/or inducing cytokines in a subject in need thereof, the method comprising administering to the composition or the combination of the present invention.

The methods and use of the present disclosure are effective in generally boosting the immune system in normal subjects. Normal subjects have a normally functioning immune system but may wish to enhance their immune system. For instance, normal subjects may use the methods of the present disclosure to maintain their health or as prophylaxis against possible immune system challenges.

In some embodiments, a subject’s immune system is boosted by (i.e., a beneficial effect on a subject’s immune system is by) an increase in red blood cell count, increase in hemoglobin content, increase in hematocrit, improved antigen presentation and improved immunoglobulin secretion.

In some embodiments, a beneficial effect on subject’s immune system is observed in about 30 days to about 60 days. In some embodiments, a beneficial effect on subject’s immune system is observed in about 10 days to about 30 days.

In some embodiments, a beneficial effect on subject’s immune system lasts for about 60 days to about 360 days. In some embodiments, a beneficial effect on subject’s immune system lasts for about 180 days to about 720 days. In some embodiments, a beneficial effect on subject’s immune system lasts for more than about 720 days.

In some embodiments, the compositions disclosed herein cause an increase in monocytes by about 30%. In some embodiments, the increase in monocytes ranges from about 5% to about 95%.

The methods and the use of the present disclosure are effective in boosting the immune response, for example, of subjects who are injured, immunocompromised or protein malnourished. Immunocompromised subjects generally exhibit an attenuated or reduced ability to mount a normal cellular or humoral defense to challenge by infectious agents, e.g., viruses, bacteria, fungi and protozoa. Protein malnourished subjects generally have a serum albumin level of less than about 3.2 grams per deciliter (g/dl) and/or unintentional weight loss of greater than 10% of usual body weight.

The methods of the use of the present disclosure can be used to therapeutically or prophylactically treat subjects who are at a heightened risk of infection due to imminent surgery, injury, illness, radiation or chemotherapy, or other condition which deleteriously affects the immune system. The composition of the present invention are useful in any condition that is stressful for immune system, such as viral infections. The method is useful to treat subjects who have a disease or disorder which causes the normal metabolic immune response to be reduced or depressed, such as HIV infection (AIDS). For example, the method can be used to pre-initiate the metabolic immune response in subjects who are undergoing chemotherapy or radiation therapy, or who are at a heightened risk for developing secondary infections or post-operative complications because of a disease, disorder or treatment resulting in a reduced ability to mobilize the body's normal metabolic responses to infection. Treatment with the compositions of the disclosure has been shown to be particularly effective in mobilizing the host's normal immune defenses, thereby engendering a measure of protection from infection in the treated host. In a further embodiment, the components of the compositions may be entrapped in a liposome. Liposomes are vesicular structures characterized by a phospholipid bilayer membrane and an inner aqueous medium. Multilamellar liposomes have multiple lipid layers separated by aqueous medium. They form spontaneously when phospholipids are suspended in an excess of aqueous solution. The lipid components undergo selfrearrangement before the formation of closed structures and entrap water and dissolved solutes between the lipid bilayers.

In another embodiment, the components of the compositions may be immobilized on the surface of a substrate. The substrate surface may be any surface capable of having an agent/ligand bound thereto or integrated into and that is biocompatible. The biocompatible surface may be biodegradable or non-biodegradable. The surface may be natural or synthetic, and a synthetic surface may be a polymer. The surface may comprise collagen, purified proteins, purified peptides, polysaccharides, glycosaminoglycans, or extracellular matrix compositions. A polysaccharide may include for example, cellulose, agarose, dextran, chitosan, hyaluronic acid, or alginate. Other polymers may include polyesters, polyethers, polyanhydrides, polyalkylcyanoacryllates, polyacrylamides, polyorthoesters, polyphosphazenes, polyvinyl acetates, block copolymers, polypropylene, polytetrafluorethylene (PTFE), or polyurethanes. The polymer may be lactic acid or a copolymer. A copolymer may comprise lactic acid and glycolic acid (PLGA). Non- biodegradable surfaces may include polymers, such as poly(dimethylsiloxane) and poly(ethylene-vinyl acetate). Biocompatible surfaces include for example, glass (e.g., bioglass), collagen, metal, hydroxyapatite, aluminate, bioceramic materials, hyaluronic acid polymers, alginate, acrylic ester polymers, lactic acid polymer, glycolic acid polymer, lactic acid/glycolic acid polymer, purified proteins, purified peptides, or extracellular matrix compositions. Other polymers comprising a surface may include glass, silica, silicon, hydroxyapatite, hydrogels, collagen, acrolein, polyacrylamide, polypropylene, polystyrene, nylon, or any number of plastics or synthetic organic polymers, or the like.

In some embodiments, the compositions may be formulated in a form suitable for delivery to a subject having a condition in need of treatment or a condition at risk of development in need of prevention. In some embodiments, the subject is a human. In some embodiments, the subject is a non-human. In some embodiments, the disclosed methods of producing the compositions further comprise formulating the compositions into a form suitable for delivery to a human and a non-human. In some embodiments, the compositions are formulated in a form suitable for delivery as a dietary supplement, a nutraceutical, a medical food, and/or an animal feedstuff. According to some embodiments, the compositions or the combinations of the present invention are orally administered.

In some embodiments, the compositions are formulated in the form of a dietary supplement tablet or capsule. In some embodiments, the method further comprises formulating the compositions with other components to be used as a nutraceutical or medical food for human consumption or formulated with animal feed material as a substitute for antibiotics. Examples including formulating the compositions with dietary supplements, food additives, nutrients, micronutrients, vitamins, minerals, additional active agents, as well as conventional excipients used in oral delivery formulations.

The components of the combination may be administered orally, and thus be formulated in a form suitable for oral administration, i.e. as a solid or a liquid preparation. Suitable solid oral formulations include tablets, capsules, pills, granules, pellets, sachet and the like. Suitable liquid oral formulations include solutions, suspensions, dispersions, emulsions, oils and the like. If formulated in form of a capsule, the compositions of the present disclosure comprise, in addition to the active compound and the inert carrier or diluent, a hard gelatin capsule. In some embodiments, the dried product is formulated for oral administration in any dosage form that is suitable for oral ingestion. Non-limiting examples include liquid compositions such as elixir, suspension, syrup, emulsion, ampoule, etc., solid compositions such as gel, gum, drop, powder, granule, pill, sugar- coated tablet, film-coated tablet, capsule, package agent, sustained-release compositions such as gel-coated compositions, multi-coated compositions, localized release compositions, and the like.

The compositions may also, for example, be formulated as suppositories, containing conventional suppository bases for use in human or veterinary medicine or as pessaries, for example, containing conventional pessary bases.

In various embodiments, the immunostimulatory agent(s) may be co-administered with various other compounds (cytokines, chemotherapeutic and/or antiviral drugs, among many others). Alternatively, the immunostimulatory agent(s) may be administered an hour, a day, a week, a month, or even more, in advance of an immunogenic composition, or any permutation thereof. Further, the immunostimulatory agent may be administered an hour, a day, a week, or even more, after administration of an immunogenic composition, or any permutation thereof. The frequency and administration regimen will be readily apparent to the skilled artisan and will depend upon any number of factors such as, but not limited to, the type and severity of the disease being treated, the age and health status of the animal, the identity of the compound or compounds being administered, the route of administration of the various immunostimulatory agent and the immunogenic composition, and the like.

Any order of administration can be used for the combination of FWGE and probiotic and optionally the organic compound in a combination. For example, the one or more prebiotics and the one or more probiotics can be administered simultaneously or sequentially. For example, all components of the combination are administered simultaneously, or only some of the components of the combination are administered simultaneously and the rest are administered sequentially. In some embodiments, none of the components are administered simultaneously, i.e., all the components are administered sequentially. When administering sequentially, any order of administration can be used.

The compositions of the present invention comprising FWGE, probiotic compound and optionally the organic compound may be co-administered with other compositions comprising FWGE.

In some embodiments, a frequency of administration of the compositions of the present invention or of the FWGE and/or probiotic and/or organic compound can be varied depending various parameters such as level of potentiation, prognosis following administration of a combination provided herein, patient compliance, side effects, etc., for example, daily, weekly, biweekly, monthly, bimonthly. FWGE can be administered along with probiotics daily, weekly, biweekly, monthly, bimonthly. In some embodiments, the FWGE is administered less frequently compared to the probiotic, or more frequently compared to the probiotic.

In some embodiments, administration can be daily, or 1, 2, 3, 4, 5, 6 or more times weekly, or more or less frequently as required. In some embodiments, administration can be provided as a single dose or as divided doses, such that a daily dose may be given in 2, 3, 4, or more portions in a single day.

In some embodiments, co-administration of the components of a combination may comprise administering the components simultaneously, or within about 1, 5, 15, 30, 45 or 60 minute of one another, within about 1 hour to within about 6 hours of one another, or as desired by one of ordinary skill in the art.

Kits The disclosure also includes one or more kits comprising any of the compositions or pharmaceutical formulations useful within the methods of the disclosure and an instructional material that describes, for instance, the method of administering FWGE, and probiotic compound as described elsewhere herein, or the method of administering FWGE, probiotics and an organic compound selected from creatine and Vitamin D as described elsewhere herein.

According to some embodiments, the kit comprises a botanical extract, probiotic and instructions to administer said compounds. According to some embodiments, the kit comprises a botanical extract, probiotic, and an organic compound and instructions to administer said compounds. According to some embodiments, the botanical extract is FWGE. According to another embodiment, the probiotic component is selected from comprises Lactobacillus rhamnosus, Lactobacillus rhamnosus GG, Lactobacillus reuteri, Bifidobacterium animalis subsp. lactis (BB12)), Bifidobacterium bifidum, and Bifidobacterium longum. According to one embodiment, the probiotic component is Lactobacillus rhamnosus. According to one embodiment, the organic compound is selected from Vitamin D and creatine.

In some embodiments, pharmaceutical compositions comprising FWGE and probiotics of the present disclosure in combination with one or more other therapeutic agents such as an anti-tumor agent, a chemotherapeutic agent, an anti-cell proliferation agent, an anti-tumor vaccine and the like are also contemplated. For example, in some embodiments, pharmaceutical compositions comprising FWGE and Lactobacillus rhamnosus in combination with one or more other therapeutic agents such as an antitumor agent, a chemotherapeutic agent, an anti-cell proliferation agent, an anti-tumor vaccine and the like are also contemplated. Likewise, in some embodiments, pharmaceutical compositions comprising FWGE, Lactobacillus rhamnosus and at least one organic compound selected from Vitamin D and creatine in combination with one or more other therapeutic agents such as an anti-tumor agent, a chemotherapeutic agent, an anti-cell proliferation agent, an anti-tumor vaccine and the like are also contemplated.

Having now generally described the invention, the same will be more readily understood through reference to the following examples, which are provided by way of illustration and are not intended to be limiting of the present invention. Examples

Example 1 - Effects of a nutraceutical blend on selected immune parameters

A randomized, double-blinded, placebo controlled study is conducted, in which 50 healthy subjects ingested either a placebo or a composition comprising FWGP (about 105 mg e.g. METRATOL®) with a probiotic product Lactobacillus rhamnosus GG (about 40 billion CFU), 2-5 times a day for 8 weeks. At the 4 week time point, the subjects are administered a flu vaccine. The ingredients of the composition are blended, then encapsulated. A color-matched placebo was made using inert coloring substances and the same excipients as used in the active product. Blood is drawn at the 0, 4, and 8 week time points and analyzed for several parameters including red blood cell numbers, hemoglobin content, hematocrit, and numbers of various subsets of circulating immune cells at the various time points.

Example 2

A group of 50 patients with cancer are selected. Prior to administration of a composition comprising FWGE/FWGP and probiotic according to the present disclosure laboratory tests are performed to obtain baseline cytokine production, K cell activity, macrophage numbers, dendritic cell numbers, and T cell numbers. The group is split into subgroups. One subgroup is administered a composition comprising FWGE and probiotic (test composition 1), another subgroup is administered a composition comprising FWGE (test composition 2), a third subgroup is administered a composition comprising a probiotic (test composition 3), and a fourth subgroup is administered placebo.

After about 30 days of administration of test compositions, a second round of laboratory tests are performed to compare cytokine production, NK cell activity, macrophage numbers, dendritic cell numbers, and T cell numbers in the test and placebo subgroups. The probiotic used is Lactobacillus rhamnosus GG, Lactobacillus reuteri and combination thereof.

Similarly, after about 60 days of administration of either the test compositions, a second round of laboratory tests are performed to compare cytokine production, NK cell activity, macrophage numbers, dendritic cell numbers, and T cell numbers in the test and placebo subgroups.

Laboratory tests on days 30 and 60 show significant increases in cytokine production, NK cell activity, macrophage numbers, dendritic cell numbers, and T cell numbers in the test subgroups as compared to placebo subgroup suggesting an increase in innate and adaptive immune responses in test subgroups. Additionally, significant reduction in cancer is observed in the test subgroups as compared to the placebo subgroup.

Furthermore, the increases in cytokine production, NK cell activity, macrophage numbers, dendritic cell numbers, and T cell numbers, and reduction in cancer in the test subgroup that is administered test composition 1 is greater than the sum of the increases in cytokine production, NK cell activity, macrophage numbers, dendritic cell numbers, and T cell numbers, and reduction in cancer observed in test subgroups administered either test composition 2 alone or test composition 3 alone. Thus, a synergistic effect is observed when a combination of FWGE and probiotic used.

Although the present invention has been described herein above by way of preferred embodiments thereof, it can be modified, without departing from the spirit and nature of the subject invention as defined in the appended claims.

Claims

1. A combination comprising a probiotic component and a fermented wheat germ extract (FWGE), wherein the combination enhances immune system function.

2. The combination according to claim 1, wherein the probiotic component comprises bacteria selected from Lactobacillus rhamnosus, Lactobacillus rhamnosus GG, Lactobacillus reuteri, Bifidobacterium animalis subsp. lactis (BB12), Bifidobacterium bifidum, Bifidobacterium longum and any combination thereof.

3. The combination according to claim 2, wherein the probiotic component comprises Lactobacillus rhamnosus GG.

4. The combination according to any one of claims 1 to 3, wherein the combination further comprises an organic compound.

5. The combination according to claim 4, wherein the organic compound is selected from Vitamin D and creatine.

6. The combination according to claim 1, comprising FWGE, Lactobacillus rhamnosus GG and creatine.

7. The combination according to any one of claims 1 to 6, wherein the combination provides a synergistic enhancement of immune system function.

8. The combination according to any one of claims 1 to 7, wherein the combination is formulated as a single formulation, wherein the probiotic component and the FWGE are separated by a barrier.

9. A composition comprising a probiotic component and a fermented wheat germ extract (FWGE), wherein the probiotic component and the FWGE are separated by a barrier that is a water impervious or water resistant coating.

10. The composition according to claim 9, wherein the probiotic component comprises bacteria selected from Lactobacillus rhamnosus, Lactobacillus rhamnosus GG, Lactobacillus reuteri, Bifidobacterium animalis subsp. lactis (BB12)), Bifidobacterium bifidum, Bifidobacterium longum and any combination thereof.

11. The composition according to claim 10, wherein the probiotic component comprises Lactobacillus rhamnosus GG.

12. The composition according to any one of claims 9 to 11, wherein the composition further comprises an organic compound.

13. The composition according to claim 12, wherein the organic compound is selected from Vitamin D and creatine.

14. The composition according to claim 9, comprising FWGE, Lactobacillus rhamnosus GG and at least one compound selected from Vitamin D creatine.

15. The composition according to any one of claims 9 to 14, wherein the barrier is configured to prevent contact of FWGE and probiotic compound.

16. The composition according to claim 15, wherein the barrier is a water impermeable barrier.

17. The composition according to claim 15, wherein the barrier has a cut-off size of equal or below 3 kD.

18. The composition according to claim 15, wherein the barrier is selected from a vegetable oil coating and an enteric coating.

19. The composition according to any one of claims 9 to 18, wherein the composition is in a form of capsules, tablets, powder, granules, pellets or sachet.

20. The composition according to any one of claims 9 to 19, wherein the composition is an enteric coated composition.

21. The composition according to any one of claims 9 to 20, wherein the composition enhances immune system function.

22. The composition according to claim 21, wherein the composition provides a synergistic enhancement of immune system function.

23. The combination according to any one of claims 1 to 8, or the composition according to any one of claims 9 to 22, for use in treating a disease or condition involving an immune system.

24. The combination or the composition for use according to claim 23, therein the disease is cancer.

25. The combination or the composition for use according to claim 23, comprising enhancing immune system function of a subject.

26. A method for in treating a disease or condition involving an immune system in a subject in need thereof comprising administering to said subject a therapeutically effective amount of FWGE, a probiotic component and optionally an organic compound to said subject.

27. A method for enhancing immune system function comprising administering to a subject a therapeutically effective amount of FWGE, a probiotic component and optionally an organic compound.

28. The method according to claim 25 or 26, wherein the organic compound is selected from Vitamin D creatine.

29. The method according to any one of claims 26 to 28, comprising administering the composition according to any one of claims 9 to 22.

30. A kit comprising FWGE and a probiotic component, and instructions for use.

31. The kit according to claim 30, further comprising at least one organic compound selected from Vitamin D and creatine.