Control of cellular redox levels

Abstract

To provide compositions and methods for regulating redox status and / or reducing oxidative stress in a subject.SOLUTION: The present invention provides methods and compositions comprising TLR agonists comprising bacterial lysates and / or lysate fractions. Also disclosed are compositions and methods comprising bacterial lysates and / or lysate fractions formulated or administered in combination with one or more other therapeutic or pharmaceutical agents.SELECTED DRAWING: None

Description

[Technical field]

[0001] CROSS-REFERENCE TO RELATED APPLICATIONS

[0002] This application claims the benefit of U.S. Provisional Application No. 62 / 253,542, filed November 10, 2015. This application is also a continuation-in-part of U.S. Application No. 14 / 640,075, filed March 6, 2015, which is a continuation-in-part of U.S. Application No. 14 / 034,044, filed September 23, 2013, which claims the benefit of U.S. Provisional Application No. 61 / 704,090, filed September 21, 2012. This application is also a continuation-in-part of U.S. Application No. 13 / 743,194, filed January 16, 2013, which claims the benefit of U.S. Provisional Application No. 61 / 586,975, filed January 16, 2012. All of the above identified applications are incorporated herein by reference in their entireties. [Background technology]

[0003] The innate immune response is one of the pathways that regulate inflammation. Inflammation is stimulated by chemical factors released by damaged cells and serves to establish a physical barrier against the spread of infection and to promote the healing of any damaged tissue after the removal of the pathogen. The acute inflammatory process is initiated by cells already present in all tissues, mainly resident macrophages, dendritic cells, histiocytes, Kupffer cells, and mast cells. These cells present receptors on their surface or contained within their cells (termed pattern recognition receptors (PRRs)) that recognize molecular patterns that are widely shared by pathogens but can be distinguished from those of the host. These molecular patterns are collectively referred to as pathogen-associated molecular patterns (PAMPs). Immune cells undergo activation when one of these PRRs recognizes a PAMP and releases inflammatory mediators in response.

[0004] Thus, PAMPs are structures associated with groups of pathogens that are recognized by cells of the innate immune system. A wide range of different chemical species can act as PAMPs, such as glycans and glycoconjugates. These structures are also referred to as small molecule motifs that are conserved within microbial classes. They are recognized by Toll-like receptors (TLRs) and other PRRs in both plants and animals.

[0005] TLRs are conserved receptors that recognize structures from bacteria, fungi, protozoa, and viruses. Although TLR receptors are located on the surface of the plasma membrane, binding to the receptor is transmitted across the membrane, resulting in a signaling response between cells. TLR signaling ultimately results in the induction or suppression of genes that orchestrate inflammatory responses. For example, activation of certain TLRs initiates a series of intracellular events that result in an immune response characterized by the production of proinflammatory cytokines. TLR signaling originates from the cytoplasmic Toll-interleukin 1 (TIR) ​​domain, which is conserved among all TLRs. The adaptor molecule MyD88, which contains both TIR and death domains, associates with the TIR domains of TLRs and IRAK proteins. Phosphorylation of IRAK leads to its association with TRAF6 and subsequent activation of NF-κB and secretion of proinflammatory cytokines. A52R, an immunomodulatory protein from vaccinia virus, has previously been shown to be an intracellular inhibitor of TIR-dependent signaling. When expressed in HEK293 cells, A52R was shown to inhibit NF-κB activation in response to stimulation with various TLRs, including TLR4, TLR5, and the combination of TLR2 and 6, as well as TLR2 and 1. Furthermore, A52R inhibited NF-κB activation in response to Poly(1:0), a synthetic ligand for TLR3. TLR3 is involved in antiviral innate immune responses.

[0006] One of the main responses of activation is to shift the redox state of the cell. For defensive purposes, reactive oxygen species (ROS) can be generated. The very presence of ROS consumes antioxidants (reducing agents), resulting in a more oxidative redox state. Not only do ROS and oxidative conditions result in cellular damage with the concomitant activation of genes, but the redox state itself controls the expression of genes. For example, as conditions become more oxidative, easily oxidized chemical groups on certain proteins, such as sulfhydryl groups, are oxidized. The oxidative state of these proteins is then recognized, which leads to the activation of certain genes, such as genes that control the redox state and promote or regulate inflammation, or that produce abnormal or disease-promoting proteins.

[0007] Inflammation is part of a complex biological response of body tissues to harmful stimuli, such as pathogens, damaged cells, or irritants. Inflammation can also result from autoimmune diseases, where body tissues are mistakenly perceived as foreign. Inflammation initially acts as a defense response involving immune cells, blood vessels, and molecular mediators. One purpose of inflammation is to eliminate the initial cause of cellular injury, remove necrotic cells and tissues damaged by the original injury and inflammatory process, and initiate tissue repair. It is useful to conceptually distinguish inflammation from infection, because there are many pathological situations in which inflammation is not caused by microbial invasion or infection, such as atherosclerosis, type III hypersensitivity, trauma, and ischemia. There are also pathological situations in which microbial invasion does not result in a classical inflammatory response, such as eosinophilia. Very little inflammation can result in the progressive destruction of tissues by harmful "invaders" (bacteria, viruses, mutated cells) and may compromise the survival of the organism, whereas excessive inflammation (in the case of chronic inflammation) can be a host of diseases such as hay fever, periodontal disease, atherosclerosis, rheumatoid arthritis, and even cancer (e.g., gallbladder cancer).

[0008] The initiation of redox changes and the resulting inflammatory response to pathogens are key components of the innate immune response designed to control infection. Inflammation is usually tightly regulated, as persistent production of inflammatory mediators can lead to chronic inflammation, tissue damage and disease development. Signaling cascades initiated by PAMP / TLR interactions and culminating in gene activation are associated with many pathologies, including sepsis, autoimmune diseases, asthma, heart disease and cancer. For example, sepsis is hypothesized to occur when bacteria and their products activate an uncontrolled network of host-derived mediators, such as proinflammatory cytokines, that can lead to multiple organ failure, cardiovascular collapse and death. Aberrant TLR signaling responses can lead to exaggerated cellular activation responses and contribute to sepsis.

[0009] Inflammation (whether chronic or acute) results from or leads to increased production and release of free radicals and other ROS from damaged and / or inflamed tissues, which in turn contribute to or cause oxidative stress. At the same time, inflammation can result from oxidative stress when ROS damage tissues. For this reason, inflammation and various conditions associated with inflammation can also be considered "oxidative stress-related diseases or conditions". Other stresses, such as psychological stress, can also lead to shifts in redox levels and the resulting oxidative stress, and even inflammation. When redox conditions induce a state of oxidative stress, these positive feedback loops can make the condition self-perpetuating. Oxidative redox conditions and oxidative stress are believed to occur at defined loci for a limited time. When the loci of oxidative redox conditions are inappropriate and / or continue for too long, a pathological or diseased state exists. A wide range of pathological or diseased states are enhanced by inappropriate redox conditions or oxidative stress caused by chronic or acute inflammation, or vice versa.

[0010] Oxidative stress is a pathological form of oxidative redox state that involves the damaging effects of abnormally increased amounts of ROS, such as free radicals. A free radical is a single atom or molecule in which at least one outer electron orbital is "occupied" with one electron ("unpaired") instead of two ("paired"). The presence of the unpaired electron makes free radical compounds specially reactive. They can spontaneously react with and thereby damage a wide variety of important cellular molecules. A certain number of ROS, including free radicals, are naturally produced by the body through cellular metabolism. For example, the synthesis of some hormones involves the production of free radicals, while granulocytes use the production of free radicals as a form of "chemical weapon" to kill bacteria, thereby defending the body against infections. Other free radicals, such as nitric oxide (NO), are fundamental to the body's homeostasis, as they act as chemical messengers that regulate important functions such as vascular tone, platelet aggregation, and cell adhesion.

[0011] Free radicals are potentially dangerous because they tend to spontaneously fill unfilled outer orbitals with a second electron. The presence of two electrons in the same orbit is the condition of maximum stability-minimum energy. Thus, as soon as a free radical collides with a "target molecule" that has one or more "available" electrons, such as a molecule of unsaturated fatty acid (e.g. arachidonic acid), an electron is "extracted" from the target molecule. This effect, "oxidation," causes the original free radical to lose its potential danger and "oxidizes" the newly generated molecule, which then becomes a new free radical, perpetuating the reaction and possibly dampening it in the absence of antioxidants. This reaction can continue with other molecules, such as carbohydrates, lipids, amino acids, peptides, proteins, nucleotides, nucleic acids, etc. ("chain reaction effect"). Such actions by free radicals can result in tissue damage of various degrees and can cause (or conversely result from) an inflammatory response. The initial or primary site of ROS release may be an appropriate response to an invading microorganism, but if the invader is not destroyed or redox homeostasis is not restored after the destruction of the invader, the redox state may spread and a continuing secondary oxidative redox state may result in a chronic injury condition with collateral tissue damage. One example is traumatic brain injury (TBI), which results in local inflammation and an oxidative redox state in the brain. If homeostasis is not re-established, a chronic oxidative redox state may lead to long-term tissue damage and chronic traumatic encephalopathy (CTE).

[0012] Thus, there is a need in the art for compositions and methods for controlling cellular redox levels. Summary of the Invention [Problem to be solved by the invention]

[0013] The methods and compositions disclosed herein are not limited to any particular benefit or function.

[0014] In one aspect, the present disclosure provides a toll-like receptor (TLR) agonist composition for regulating redox status in a subject, the composition comprising: (a) a TLR agonist comprising at least one lysate and / or lysate fraction of a bacterium, the TLR agonist activating at least one or more TLRs or NLRs; (b) an optional promoter for increasing absorption of the composition; and (c) an optional carrier for increasing the volume of the composition, wherein administering an effective amount of the composition to the subject measurably reduces oxidative stress levels in the subject.

[0015] In another aspect, the present disclosure provides a method of modulating redox status in a subject, the method comprising administering a therapeutically effective amount of a lysate composition according to the present disclosure to a subject in need thereof. In some embodiments, the modulation of redox status is assessed by measuring a change in isoprostane concentration in the subject.

[0016] In another aspect, the disclosure provides a method of modulating redox status in a subject, the method comprising: (a) administering to a subject in need thereof repeated doses at intervals in time of a composition comprising: (i) a toll-like receptor (TLR) agonist comprising at least one lysate and / or lysate fraction of a bacterium, wherein the toll-like receptor (TLR) agonist activates at least one or more distinct TLRs or NLRs; (ii) an optional promoter for increasing absorption of the composition; and (iii) an optional carrier for increasing the volume of the composition; and (b) measuring the subject's body fluids to detect changes in oxidative stress levels.

[0017] In another aspect, the disclosure provides a method for reducing the amount of isoprostane in the urine or blood of a subject, the method comprising: (a) determining the level of isoprostane in the urine or blood of the subject; (b) administering to the subject an effective amount of a composition comprising: (i) a toll-like receptor (TLR) agonist comprising at least one bacterial lysate and / or lysate fraction from a bacterium, wherein the TLR agonist activates at least one or more distinct TLRs or NLRs; and (ii) an optional promoter for increasing absorption of the composition; and (c) continuing administration of the composition until the level of isoprostane in the urine or blood of the subject is reduced.

[0018] In another aspect, the disclosure provides a composition comprising: (a) a bacterial lysate and / or lysate fraction capable of activating at least one or more toll-like receptors (TLRs) or Nod-like receptors (NLRs); (b) an optional promoter for increasing absorption of the composition; and (c) an optional carrier for increasing the volume of the composition.

[0019] In another aspect, the present disclosure provides a pharmaceutical formulation comprising a dissolution composition according to the present disclosure, the pharmaceutical formulation being formulated for buccal or sublingual administration. In some embodiments, the pharmaceutical formulation is formulated to dissolve within 1 minute after administration.

[0020] In another aspect, the disclosure provides a method of producing a bacterial lysate comprising: (a) fermenting bacteria in a growth medium to stationary growth phase to produce a fermentation broth, (b) harvesting the bacteria from the fermentation broth, (c) pasteurizing the harvested bacteria, and (d) lysing the pasteurized bacteria with lysozyme to produce a bacterial lysate. In some embodiments, the bacteria are harvested in mid-log, late-log, early stationary, mid-stationary, or late stationary phase.

[0021] In another aspect, the disclosure provides a bacterial lysate produced by a method comprising: (a) fermenting bacteria in a growth medium to stationary growth phase to produce a fermentation broth, (b) harvesting the bacteria from the fermentation broth, (c) pasteurizing the harvested bacteria, and (d) lysing the pasteurized bacteria with lysozyme to produce a bacterial lysate. In some embodiments, the bacteria are harvested in mid-log, late-log, early stationary, mid-stationary, or late stationary phase.

[0022] In another aspect, the disclosure provides a method for reducing one or more oxidative stress-related side effects associated with administration of a pharmaceutical agent, the method comprising administering a therapeutically effective amount of a lysate composition comprising (a) a bacterial lysate and / or a lysate fraction, (b) an optional promoter for increasing absorption of the composition, and (c) an optional carrier for increasing the volume of the composition, in combination with the pharmaceutical agent, wherein the pharmaceutical agent and the lysate composition are administered simultaneously or in any order, and via the same route of administration or different routes of administration.

[0023] In another aspect, the disclosure provides a method for treating an oxidative stress-related disease or condition in a subject, the method comprising administering to the subject a therapeutically effective amount of a composition comprising: (a) a bacterial lysate and / or a lysate fraction capable of activating at least one or more toll-like receptors (TLRs) or Nod-like receptors (NLRs); (b) an optional promoter for increasing absorption of the composition; and (c) an optional carrier for increasing the volume of the composition.

[0024] In another aspect, the disclosure provides a method for reducing oxidative stress in a subject, the method comprising: (a) measuring the level of oxidative stress in the subject by measuring the amount of isoprostanes in the subject's urine or blood; (b) administering to the subject an effective amount of a composition comprising: (i) a toll-like receptor (TLR) agonist comprising at least one lysate and / or lysate fraction from a bacterium, wherein the TLR agonist activates at least one or more TLRs or NLRs; and (ii) an optional promoter for increasing absorption of the composition; and (c) continuing administration of the composition until the level of oxidative stress is reduced, as measured by a decrease in the amount of isoprostanes in the subject's urine.

[0025] In another aspect, the disclosure provides a therapeutic combination comprising: (a) a lysate composition comprising: (i) a bacterial lysate and / or lysate fraction capable of activating at least one or more toll-like receptors (TLRs) or Nod-like receptors (NLRs); (ii) an optional promoter for increasing absorption of the composition; and (iii) an optional carrier for increasing the volume of the composition; and (b) one or more pharmaceutical agents, wherein the lysate composition and the one or more pharmaceutical agents are administered simultaneously or in any order, and the lysate composition and the one or more pharmaceutical agents are administered via the same or different routes of administration.

[0026] In another aspect, the present disclosure discloses a pharmaceutical formulation comprising: (a) a combination of a lysate composition comprising (i) a bacterial lysate and / or a lysate fraction capable of activating at least one or more toll-like receptors (TLRs) or Nod-like receptors (NLRs), (ii) an optional promoter for increasing absorption of the composition, and (iii) an optional carrier for increasing the volume of the composition; and (b) one or more pharmaceutical agents.In some embodiments, the one or more pharmaceutical agents are selected from the group consisting of: antispasmodics, motility stimulants, H2 receptor antagonists, antimuscarinics; chelating agents, prostaglandin analogs, aminosalicylates, corticosteroids, drugs affecting immune response, stimulant laxatives, drugs affecting bile composition and flow, bile acid sequestrants, dopamine antagonists, proton pump inhibitors, opioids, opioid receptor antagonists, analgesics, hypnotics, cardiac glycosides, phosphodiesterase inhibitors, thiazides, diuretics, potassium Reserve diuretics, aldosterone antagonists, osmotic diuretics, drugs for arrhythmias, beta-adrenergic receptor blockers, hypertensive drugs, drugs affecting the renin-angiotensin system, nitrates, calcium blockers, antianginal drugs, peripheral vasodilators, sympathomimetics, anticoagulants, protamines, antiplatelet drugs, fibrinolytic drugs, antifibrinolytic drugs, lipid regulating drugs, omega-3 fatty acid compounds, CNS drugs, anti-infective drugs, or benzothoron, procyclidine, biperiden, amantadine, bromocriptine, pergolide, entacapone, tolcapone , Selegiline, Pramipexole, Budesonide, Formoterol, Quetiapine Fumarate, Olanzapine, Pioglitazone, Montelukast, Zoledronic Acid, Valsartan, Latanoprost, Irbesartan, Clopidogrel, Atomoxetine, Dexamfetamine, Methylphenidate, Modafinil, Bleomycin, Dactinomycin, Daunorubicin, Idarubicin, Mitomycin, Mitoxantrone, Azacitidine, Capecitabine, Cladribine, Clofarabine, Cytara and another agent selected from the group consisting of rifabutin, fludarabine, flourouracil, gemcitabine, mercaptopurine, methotrexate, nelarabine, pemetrexed, raltitrexed, thioguanine, apomorphine, betamethasone, cortisone, deflazacort, dexamethosone, hydrocortisone, methylprednisolone, prednisolone, triamcinolone, cyclosporine, sirolimus, tacrolimus, interferon alpha, and interferon beta.

[0027] In another aspect, the present disclosure provides a formulation comprising: (a) a lysate composition comprising: (i) a bacterial lysate and / or a lysate fraction capable of activating at least one or more toll-like receptors (TLRs) or Nod-like receptors (NLRs); (ii) an optional promoter for increasing the absorption of the composition; and (iii) an optional carrier for increasing the volume of the composition; and (b) an isolated human anti-TNFα antibody or antigen-binding fragment thereof or a TNF inhibitor. In some embodiments, the human anti-TNFα antibody or antigen-binding fragment thereof is adalimumab. In another aspect, the present disclosure provides a use of such a formulation in the manufacture of a medicament for the treatment of chronic rheumatoid arthritis (RA), late-onset RA, or psoriatic arthritis in a subject. In another aspect, the present disclosure provides a method for the treatment of rheumatoid arthritis (RA), late-onset RA, or psoriatic arthritis in a subject, the method comprising administering to the subject a therapeutically effective amount of such a formulation.

[0028] In some embodiments of any of the methods or compositions disclosed herein, the bacteria is a gram-positive or gram-negative bacterium. In some embodiments of any of the methods or compositions disclosed herein, the gram-positive bacterium is selected from the group consisting of bacteria of the Lactobacillaceae family, bacteria of the Streptococcus family, bacteria of the Bifidobacteriaceae family, and bacteria of the Bacillaceae family. In some embodiments, the gram-positive bacterium is selected from the group consisting of Bacillus coagulans, Lactobacillus sporogenes, Streptococcus thermophilus, Bifidobacterium animalis, Bifidobacterium animalis, subspecies animalis, Bifidobacterium infantis, Bifidobacterium longum, Bifidobacterium breve, Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus spp. ... The bacterial strain is selected from the group consisting of Bacillus casei, Lactobacillus delbrueckii, Lactobacillus delbrueckii subsp. bulgaricus, Lactococcus lactis, Lactococcus lactis subsp. lactis, Streptococcus lactis, Streptococcus thermophilus, Bifidobacterium lactis, Bifidobacterium breve, Pediococcus acidilactici and Lactobacillus helveticus.

[0029] In some embodiments of any of the methods or compositions disclosed herein, the Gram-negative bacteria is selected from the group consisting of bacteria of the genera Pseudomonas, Klebsiella, Xanthomonas, Shigella, and Enterobacter, hi some embodiments, the Gram-negative bacteria is selected from the group consisting of Klebsiella oxytoca, Shigella flexneri, Xanthomonas, and Pseudomonas fluorescens.

[0030] In some embodiments of any of the methods or compositions disclosed herein, the TLR agonist, lysate, lysate fraction, or cell wall fraction activates at least one or more of TLR2, TLR3, TLR4, TLR5, TLR7, TLR8, TLR9, NOD1, and NOD2. In some embodiments, the TLR agonist, lysate, lysate fraction, or cell wall fraction activates two or more of TLR2, TLR3, TLR4, TLR5, TLR7, TLR8, TLR9, NOD1, and NOD2. In some embodiments, the TLR agonist, lysate, lysate fraction, or cell wall fraction activates TLR2 and TLR4. In some embodiments, the TLR agonist, lysate, lysate fraction, or cell wall fraction activates three or more of TLR2, TLR3, TLR4, TLR5, TLR7, TLR8, TLR9, NOD1, and NOD2.

[0031] In some embodiments of any of the methods or compositions disclosed herein, the promoter is selected from the group consisting of an amino acid, an amino sugar, and a sugar. In some embodiments, the carrier is selected from the group consisting of a binder, a gum base, and combinations thereof. In some embodiments, the gum base comprises at least one hydrophobic polymer and at least one hydrophilic polymer. In some embodiments, the binder is selected from the group consisting of a sugar, a sugar alcohol, and combinations thereof. In some embodiments, the sugar alcohol is selected from the group consisting of mannitol, sorbitol, xylitol, and combinations thereof.

[0032] In some embodiments, the composition is prepared as a dosage form selected from the group consisting of lozenges, chewing gum, chewable tablets, candies, and dissolving tablets.In some embodiments, the dosage form delivers the TLR agonist to oral mucosa.In some embodiments, the oral mucosa is selected from the group consisting of sublingual mucosa, buccal mucosa, and combinations thereof.

[0033] In some embodiments of any of the methods and compositions disclosed herein, the composition is formulated for oral mucosal delivery.In some embodiments, the composition is formulated for sublingual or buccal delivery.In some embodiments, the composition is formulated to dissolve within 1 minute after administration.

[0034] These and other aspects, advantages and alternatives will become apparent to those of ordinary skill in the art from a reading of the following detailed description, taken together with the appended claims, and where appropriate with reference to the accompanying drawings. [Brief description of the drawings]

[0035] [Figure 1] FIG. 1 is a schematic diagram showing toll-like receptors (TLRs) exhibiting various response pathways. [Diagram 2] 1 shows an exemplary stimulatory effect of compositions of the invention on selected TLR / NLR cell lines. Values ​​on the graph correspond to the average of three screening experiments. [Diagram 3] 1 is a graph showing the results of TLR stimulation of Pediococcus acidilactici lysate compared to Lactobacillus bulgaricus lysate. [Figure 4] FIG. 1 is a graph showing the results of TLR stimulation of Bacillus coagulans lysates at different harvest times. [Diagram 5] Graph showing the results of TLR stimulation of Gram-positive bacterial lysates from Lactobacillus acidophilus, Lactobacillus helveticus, Lactobacillus plantarum, and Streptococcus thermophilus. [Figure 6] Graph showing the results of TLR stimulation of Gram-negative bacterial lysates from Escherichia coli, Klebsiella oxytoca, Shigella flexneri, Xanthomonas campestris, and Pseudomonas fluorescens. [Figure 7] FIG. 1 shows the results of TLR stimulation with Xanthomonas campestris lysate compared to xanthan gum. [Figure 8]1 is a graph showing the reduction in urinary isoprostane levels in the urine of combat veterans diagnosed with PTSD following administration of a composition of the present disclosure. [Figure 9] 1 is a graph showing reduction in sleep disturbance in combat veterans diagnosed with PTSD following administration of a composition of the present disclosure. [Figure 10] 1 is a graph showing the reduction in neurological symptoms in combat veterans diagnosed with PTSD following administration of a composition of the present disclosure. [Figure 11] 1 is a graph showing improvement in overall mood in combat veterans diagnosed with PTSD following administration of a composition of the present disclosure. [Figure 12] 1 is a graph showing an increase in overall energy levels in combat veterans diagnosed with PTSD following administration of a composition of the present disclosure. [Figure 13] 1 is a graph showing increased overall satisfaction with joint health in combat veterans diagnosed with PTSD following administration of a composition of the present disclosure. [Figure 14] 1 is a graph showing increased satisfaction with digestive health in combat veterans diagnosed with PTSD following administration of a composition of the present disclosure. [Figure 15] 1 is a graph showing reduced irritability levels in combat veterans diagnosed with PTSD following administration of a composition of the present disclosure. [Figure 16] 1 is a graph showing an increase in overall levels of self-reported satisfaction with sexual functioning in combat veterans diagnosed with PTSD following administration of a composition of the present disclosure. [Figure 17] 1 is a graph showing reduction in daytime sleepiness in combat veterans diagnosed with PTSD following administration of a composition of the present disclosure. [Figure 18] 1 is a graph showing an increase in overall levels of self-reported satisfaction with life in combat veterans diagnosed with PTSD following administration of a composition of the present disclosure. [Figure 19] 1 is a graph showing reduced depression levels in combat veterans diagnosed with PTSD following administration of a composition of the present disclosure. [Figure 20] 1 is a graph showing the reduction in isoprostane levels in individuals with a history of concussion following administration of a lysate of the present disclosure. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0036] All publications, patents, and patent applications cited herein are hereby expressly incorporated by reference for all purposes.

[0037] The redox (oxidation-reduction) state of cells is subject to great changes. Oxidation and reduction reactions are important for cellular biological energetics. Usually, when the oxidation of food molecules results in the transport of electrons and the eventual capture of energy as energy-rich molecules such as NADP (nicotinamide adenine dinucleotide phosphate) and ATP (adenosine triphosphate), TLRs are activated in such a manner that downstream oxidation / reduction reactions are in equilibrium. As used herein, the term "equilibrium" refers to a homeostatic equilibrium, i.e., a situation in which the amount of oxidation is equal to the amount of reduction in a given system, but rather oxidation and reduction are immunologically and therefore metabolically homeostatic for the host. However, there are multiple cellular states in which the redox state changes. Normally, cells are armed with antioxidant molecules, but when such molecules are depleted, the redox state of cells changes. One possible cause of such changes is the deliberate generation of reactive oxygen species (ROS), such as O2 (superoxide radical), OH (hydroxyl radical), and H2O2 (hydrogen peroxide), for defensive or similar purposes. When the redox balance is shifted, oxidative stress can ensue. Oxidative stress is a pathological condition caused by the damaging effects of abnormally increased ROS levels on the body's cells and tissues. Oxidative stress is a direct result of immunologically uncontrolled increased production of ROS and / or decreased bioactivity of antioxidant defenses against excess ROS. Inflammation (whether chronic or acute) as well as other stresses and infections can lead to increased production and release of ROS from damaged and / or inflamed tissues, which shifts the cellular redox balance and thus contributes to oxidative stress. At the same time, inflammation can result from oxidative stress when ROS damage tissues. A wide range of diseases and disease states are associated with changes in redox state and oxidative stress brought about by chronic or acute inflammation or vice versa. Current therapies for treating chronic or acute inflammation are not available without adverse side effects. Described herein are compositions and methods for altering redox levels in the treatment of oxidative stress and related conditions.

[0038] Before describing the disclosed methods and compositions in detail, some terms are defined. As used herein, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. For example, "nucleic acid" means one or more nucleic acids.

[0039] For purposes of describing and defining the present invention, it should be noted that the term "substantially" is used herein to express the inherent degree of uncertainty that can be attributed to any quantitative comparison, value, measurement, or other representation. The term "substantially" is also utilized herein to express the extent to which a quantitative representation may vary from the stated standard without resulting in a change in the basic functionality of the subject matter in question.

[0040] As mentioned above, pathogen-associated molecular patterns (PAMPs) can activate the innate immune response by stimulating TLRs, which are generally activated by conserved non-self biochemical structures and thus protect the host from infection. Bacterial lipopolysaccharide (LPS) is found on the bacterial cell membrane of some bacteria and is considered to be the prototypic PAMP. LPS is specifically recognized by TLR4, a recognition receptor of the innate immune system. Other PAMPs include bacterial flagellin (recognized by TLR5), lipoteichoic acid, peptidoglycan, and nucleic acid variants that are typically associated with viruses, such as double-stranded RNA (dsRNA) recognized by TLR3, or unmethylated CpG motifs recognized by TLR9.

[0041] However, in some cases, PAMPs reduce inflammation. EPS (exopolysaccharides), a material that typically stimulates immune responses, has been shown to result in reduced inflammatory responses by stimulating negative regulators of TLRs. More specifically, EPS has been shown to stimulate the expression of immunoglobin IL-1-related receptor, toll-interacting protein, B-cell lymphoma 3-encoded protein, A20, mitogen-activated protein kinase phosphate-1, and interleukin-related kinase M, resulting in negative regulation of TLRs and inflammation.

[0042] These seemingly contradictory effects of PAMPs are explained, at least in part, by the interplay between the innate immune system and the microbiome. The immune system does not exist in a vacuum. Even when the organism is not mounting an active inflammatory response, immune cells are responding to an onslaught of PAMPs, especially from the microbiome environment. Given that the various TLR receptors that are activated create downstream redox conditions that are in immunological and metabolic homeostasis, this system, by issuing an “all clear” signal, avoids inadvertent responses that could damage essential microbiome components. Thus, presenting the cells of the innate immune system with the right combination of PAMPs can control the entire redox system, down- or up-regulating it to achieve or, in some cases, restore immunological and metabolic homeostasis.

[0043] Figure 1 shows a schematic diagram of a transmembrane TLR. While the ligand-receiving (stimulating) portion of the molecule is located on the surface of the plasma membrane, the transmembrane domain of the protein can transmit a signal to the cytoplasmic surface of the membrane via a conformational change that occurs when a ligand is bound. At the cytoplasmic surface, this signal (arrow) is coupled to several different signaling pathways. The proliferation / differentiation and stress response pathways are shown. Note that one TLR type does not control both pathways simultaneously, as in this generalized diagram. Rather, some TLRs control one pathway or set or pathways, while other TLRs control a different pathway or set of pathways. Furthermore, one type of TLR may control different pathways depending on the cell type in which it is placed. For this reason, delivering the correct balance of TLR agonists is important to maintain homeostasis.

[0044] Downstream single transduction mechanisms may be shared to a greater or lesser extent. In Figure 1, the growth response primarily uses the ERK pathway, whereas the stress response uses the MEKK and TAK pathways. In each pathway, the signal molecule is phosphorylated and a phosphorylation cascade may exist to amplify the signal. Finally, the phosphorylated protein enters the nucleus (through nuclear pores) where the phosphorylated intermediates alter both transcription and translation. In this way, TLRs can control a whole set of genes. In total, thousands of genes are activated by TLR signaling, and collectively TLRs constitute one of the most multifaceted and tightly regulated gateways of genetic polymorphism.

[0045] One of the primary responses of TLR activation is to shift the downstream redox state of the cell, if warranted. Initiation of redox changes and the resulting inflammatory response to pathogens are key components of the innate immune response designed to control infection. Inflammation (whether chronic or acute) results from or leads to increased production and release of free radicals and other ROS from damaged and / or inflamed tissues, which in turn contribute to or cause oxidative stress. At the same time, inflammation can result from oxidative stress when ROS damage tissues. For this reason, inflammation and various conditions associated with inflammation can also be considered "oxidative stress-associated diseases or conditions."

[0046] Oxidative stress, a biochemical condition, does not generally manifest itself with any specific clinical symptoms or signs, except for the specific pathological condition it induces, and may remain generally unknown, with attendant damage to the patient, until a clinician suspects its existence and decides to assay for oxidative stress.

[0047] Various common diseases and / or conditions are often associated with oxidative stress. One example is Alzheimer's disease. Studies have shown that chronic oxidative stress increases tau phosphorylation levels, a known biomarker of Alzheimer's disease. Studies have also shown that oxidative stress leads to tau-induced neurodegeneration in Alzheimer's disease models.

[0048] Other known "oxidative stress-related diseases or conditions" include, but are not limited to, acerobacteremia, acute and chronic alcoholic liver disease, acute autoimmune myocarditis, sickle cell disease acute chest syndrome, acute pancreatitis, acute respiratory distress syndrome, alcoholic liver disease, amyotrophic lateral sclerosis, arterial / systemic hypertension, asbestosis, asthma, ataxia telangiectasia, atherosclerosis, atopic dermatitis, cerebral ischemia, bronchopulmonary dysplasia, burns, some cancers, cardiopulmonary bypass, cardiovascular disease, cataracts, cellulitis, chemotherapy side effects, chronic fatigue syndrome, chronic hepatitis C, chronic kidney disease, chronic obstructive pulmonary disease, chronic renal failure, colitis, coronary artery disease, Creutzfeldt-Jakob disease, and the like. Torjakop disease, Crohn's disease, cutaneous leishmaniasis, cystic fibrosis, type 1 diabetes, type 2 diabetes, dyslipidemia, Down's syndrome, eclampsia, end-stage renal disease, erectile dysfunction, Friedreich's ataxia, headache, heart failure, Helicobacter pylori infection / inflammation, side effects of hemodialysis, liver cirrhosis, human immunodeficiency virus infection, Huntington's disease, hyperbaric oxygen therapy-related diseases, hypercholesterolemia, hyperhomocysteinemia, hyperlipidemia, idiopathic pulmonary fibrosis, interstitial lung disease, ischemia / reperfusion injury, juvenile chronic arthritis, kidney transplant failure, leukemia, lung cancer, pulmonary disorders, macular degeneration, male infertility, Meniere's disease, meningitis, mild cognitive impairment, multiple sclerosis, myelodysplastic syndrome syndromes), myocardial infarction, myocarditis, neonatal bronchopulmonary dysplasia, obesity, osteoarthritis, osteoporosis, pancreatitis, Parkinson's syndrome, periodontal disease, side effects of peritoneal dialysis, photoageing, post-traumatic stress disorder, preeclampsia, primary biliary cirrhosis, broncopulmonary disease, progeria, psoriasis, psoriatic arthritis, pulmonary hypertension, side effects of radiation therapy, reactive arthritis, renal cell carcinoma, respiratory distress syndrome, retinopathy of prematurity, retrolenticular fibroplasia fibroplasy, rheumatoid disease, rheumatoid arthritis, sarcoidosis, sepsis, sickle cell disease, sleep apnea, spherocytosis, spinal cord injury, stroke, synucleinopathy, systemic amyloidosis, systemic lupus erythematosus, systemic sclerosis, thrombophilia, tauopathy, traumatic stress tuberculosis, variable angina, uremia, venous insufficiency, Werner's syndrome, and Zellweger's syndrome.

[0049] Oxidative stress mediates the pathological symptoms and control of redox levels of a great many disorders, and controlling redox levels, and therefore oxidative stress, prevents many tissue damages, and allows easier control of the underlying disease when reducing oxidative stress is insufficient to achieve disease control. For example, myocardial infarction is the death or damage of myocardium caused by vascular occlusion. Except for the role that oxidative stress may play in causing vascular occlusion, once occlusion occurs, the affected muscle cells become anoxic and eventually die. However, if the occlusion is quickly reversed (e.g., by thrombolytic drugs), circulation is restored and, theoretically, the affected muscle cells are rescued. However, in many cases, despite the rapid restoration of circulation, the initial injury causes an inflammatory response, which results in oxidative stress and muscle cell damage. By controlling this oxidative stress, damage to myocardial cells can be virtually eliminated. In all of these diseases and disorders, the regulation of oxidative stress can alleviate or treat the symptoms and / or causes of the disease or disorder.

[0050] Changes in redox state can result in the oxidation of sulfhydryl groups on key proteins, which themselves are often difficult to measure. However, chronic or acute increases in oxidative stress or redox changes can alter several other cellular components, either by oxidation of sulfhydryl groups as described above, or by other chemical oxidation mechanisms such as peroxidation, resulting in the transfer of electrons.

[0051] Isoprostanes are prostaglandin-like compounds formed in vivo from free radical-catalyzed peroxidation of essential fatty acids (mainly arachidonic acid) without the direct action of cyclooxygenase (COX) enzymes normally involved in the formation of prostaglandins, which is the normal mechanism of prostaglandin formation. These compounds have potent biological activity as inflammatory mediators that increase pain perception. These molecules are regulated by at least two different pathways. One pathway is mediated by COX enzymes, which convert lipids to isoprostanes in response to gene activation and signaling molecules. In another pathway, lipids are directly oxidized to isoprostanes in response to high oxidative redox conditions. As isoprostanes are mediators of inflammation, they form part of a positive feedback loop that may maintain damaging oxidative redox conditions.

[0052] Therefore, isoprostanes such as F2-isoprostane are accurate markers of oxidative redox state in both animal and human oxidative stress models, and measurement of isoprostanes has emerged as one of the most reliable approaches to assess oxidative stress in vivo due to their genetic stability and ease of measurement in body fluids (such as urine and blood). This ease and stability has made measurement of isoprostanes an important and reliable tool in exploring the role of oxidative stress in the pathogenesis of human diseases. Isoprostane levels directly correlate with oxidative redox state and the resulting oxidative stress. Even if the site of oxidative redox state is limited in range, isoprostanes generated in the range can be measured in body fluids distant from the site of oxidative stress, such as in urine, and in the resulting inflammation.

[0053] Other biological compounds that can be oxidized include, but are not limited to, proteins, metalloproteins, enzymes, lipids, fatty acids, carbohydrates, neurotransmitters, DNA, vitamins, polyphenols, antioxidants, and coenzymes. Thus, components such as, but not limited to, superoxide dismutase (SOD), peroxidase, glutathione, and forms of cellular components that are sufficiently oxidized by exposure to reactive species such as, but not limited to, advanced oxidative proteins, malondiadehyde, and 8-hydroxydeoyguanosine, are other potential biomarkers of oxidative stress.

[0054] Given the importance of inflammation in immune responses and the deleterious consequences it can cause through oxidative stress, oxidative stress-related diseases or conditions, and / or other processes, it is important that organisms maintain tight control over their redox state. Also, given the intertwined relationship between oxidative stress and inflammation, biomarkers that measure inflammation have been proposed as indirect biomarkers of oxidative stress. These biomarkers, such as but not limited to C-reactive protein, serum amyloid A, and cytokines, are limited in their ability to pick up low-grade changes in inflammation and, as a result, low-grade changes in redox. Alternative methods focus on biomarkers such as isopropylamine to measure redox state and mechanisms for altering redox state.

[0055] Redox can be altered by either increasing reduced species (antioxidants) or decreasing oxidants. There has been little success in controlling harmful redox levels via added antioxidants or antioxidant therapy (see Ho et al., Biological markers of oxidative stress: Applications to cardiovascular research and practice. 2013 Redox Biology). There is evidence for the long-term use of dietary antioxidants, possibly as preventative agents. However, if the immune system cannot achieve or maintain homeostatic redox levels, added antioxidants are generally ineffective in correcting the equilibrium.

[0056] Another route to affect redox status is to target downstream targets. Such strategies include the use of aspirin and glucocorticoids to block activation of NF-κB, and targeting specific inflammatory mediators such as TNF-α. Given their role in mediating innate immune responses and inflammation, TLRs present another target for controlling innate and inflammatory responses.

[0057] As mentioned above, TLRs help mediate the innate immune response by inducing or suppressing genes that regulate the inflammatory response. TLRs recognize and respond to a variety of signals. These signals, upon binding to specific TLRs, promote or block signaling pathways that can induce or suppress genes that mediate inflammation. Targeting specific TLRs with specific agents can mediate the occupation of immune responses and inflammation.

[0058] Inhibiting multiple TLR-dependent responses or activating specific responses by targeting common signaling components may prove to be an effective approach in controlling inflammatory responses. Thus, the compositions disclosed herein can be used to treat oxidative stress disorders associated with TLR signaling pathways (e.g., TLR-induced inflammation), the method comprising administering a therapeutically effective amount of the compositions described herein, wherein the affected TLR is one or more of TLR2, TLR4, TLR5, TLR7 and TLR9.

[0059] As used herein, the term "therapeutically effective amount" refers to the amount administered to a subject sufficient to elicit a desired effect in the subject.

[0060] As used herein, the terms "pharmaceutical formulation" and "pharmaceutical composition" are preparations that are in a form that effectively activates the biological activity of the active ingredient and that do not contain any additional ingredients that are unacceptably toxic to the subject to which the formulation is administered. The terms "product," "supplement," and "composition" are used interchangeably herein with respect to pharmaceutical compositions that include the bacterial lysates, lysate fractions, and / or cell wall fractions of the present disclosure.

[0061] The compositions described herein can be modified to target individual TLR or groups of TLRs.By targeting individual TLR or groups of TLRs, the compositions described herein can be used to mediate acute or chronic inflammation cases.Furthermore, the compositions described herein can be used to suppress abnormal inflammation cases and restore healthy levels of inflammation.

[0062] Thus, compositions and methods for regulating oxidative stress are described herein. The compositions can activate various TLRs, as assessed by measuring the expression of NF-κB in various cell lines. The compositions described herein can also reduce or downregulate the activity of TLRs, thereby reducing or regulating inflammation levels. When the appropriate balance of TLR activation is achieved, the redox state of cells is optimized, and oxidative stress is reduced. This can be achieved by providing a composition that contains PAMPs that simultaneously bind to two or more different types of TLRs.

[0063] In some cases, the lysate of a single type of microorganism may act as an agonist for more than one TLR. As disclosed herein, the precise method of processing the microbial lysate can affect which TLR is stimulated by the lysate. In some embodiments, the activation of a panel of TLRs can be achieved by combining different lysates or fractions from a single type of microorganism, or lysates or fractions from two or more microorganisms.

[0064] There are many different but similar PAMPs that bind to a given TLR type, but with different affinities. There are also PAMP molecules that stimulate completely different TLR types. How the lysate or fraction is processed can affect the population of PAMPs present in the final product. Thus, different types of microorganisms present different palettes of PAMPs to act on. As disclosed herein, providing a composition that simultaneously stimulates (acts as an agonist) specific TLR types results in a dramatic downregulation of oxidative stress.

[0065] Unexpectedly, stimulating TLRs does not necessarily lead to increased oxidative stress. Rather, constant stimulation of TLRs by the microbiome is a normal situation, without resulting in inappropriate redox states and oxidative stress. Part of the explanation may be related to the location of a given TLR receptor as well as the type of TLR receptor. Activation of a TLR receptor located in a mucosa leads to different results than activation of the same TLR receptor located in the internal tissues of the body. Without being bound to a particular explanation or principle, the present disclosure provides a method to apply or enhance normal microbiome signals, reassessing homeostasis and reconstructing the overall system by disrupting positive feedback loops that lead to abnormal redox states and oxidative stress.

[0066] The therapeutically active compositions of the present disclosure comprise a non-synthetic biologically active agent, preferably one or more cell wall fractions of one or more microorganisms, including gram-positive bacteria, gram-negative bacteria, or a combination thereof, in a form such as a lysate, together with a promoter and, optionally, one or more other additives, including controlled release components, that allow the composition to be absorbed into or interact with the mucosal wall of a subject in need of treatment.

[0067] As used herein, the terms "dose" and "administration" include bolus or loading doses, and also include chronic or maintenance doses. According to the present invention, the active therapeutic agent is a lysate, lysate fraction, or cell wall fraction of bacteria, such as gram-positive or gram-negative bacteria, and the amount ranges from about 0.01 mg of therapeutic agent per kilogram of body weight to about 100 mg of therapeutic agent per kg of body weight as required to act as an agonist for two or more TLRs depending on the particular therapeutic application. In some embodiments, the active therapeutic agent of the present disclosure is administered at a dosage of about 0.01 mg of therapeutic agent per kg of body weight to about 10 gm of therapeutic agent per kg of body weight. In some embodiments, the active therapeutic agent of the present disclosure is administered at a dosage of about 0.01 mg of therapeutic agent per kg of body weight to about 1 gm of therapeutic agent per kg of body weight. In some embodiments, the active therapeutic agent of the present disclosure is administered at a dosage of about 0.01 mg of therapeutic agent per kg of body weight to about 50 mg of therapeutic agent per kg of body weight. In some embodiments, the active therapeutic agent of the present disclosure is administered at a dosage of about 0.05 mg of therapeutic agent per kg of body weight to about 30 mg of therapeutic agent per kg of body weight. In some embodiments, the active therapeutic agent of the present disclosure is administered at a dosage of about 0.05 mg of therapeutic agent per kg of body weight to about 5 mg of therapeutic agent per kg of body weight.In some embodiments, an active therapeutic agent disclosed herein is administered at a dose of about 0.1 mg of therapeutic agent per kg body weight, or about 0.2 mg of therapeutic agent per kg body weight, or about 0.3 mg of therapeutic agent per kg body weight, or about 0.4 mg of therapeutic agent per kg body weight, or about 0.5 mg of therapeutic agent per kg body weight, or about 0.6 mg of therapeutic agent per kg body weight, or about 0.7 mg of therapeutic agent per kg body weight, or about 0.8 mg of therapeutic agent per kg body weight, or about 0.9 mg of therapeutic agent per kg body weight, or about 1 mg of therapeutic agent per kg body weight, or about 2 mg of therapeutic agent per kg body weight, or about 3 mg of therapeutic agent per kg body weight, or about 4 mg of therapeutic agent per kg body weight, or about 5 mg of therapeutic agent per kg body weight, or about 6 mg of therapeutic agent per kg body weight, or about 7 mg of therapeutic agent per kg body weight, or about 8 mg of therapeutic agent per kg body weight, or about 9 mg of therapeutic agent per kg body weight, or about 10 mg of therapeutic agent per kg body weight. In some embodiments, the dosage amounts above describe the total amount of active ingredient administered to a subject daily, which may be divided into two or more administrations per day, or may be administered in a single daily dose.

[0068] As used herein, the terms "therapeutic agent" and "active ingredient" refer to the lysate, lysate fraction, and / or cell wall fraction of the present disclosure, or a combination thereof, as opposed to non-active ingredients contained in a composition or formulation.

[0069] In some embodiments, the pharmaceutical compositions disclosed herein are administered in formulations, such as oral formulations, such as tablet formulations, which contain about 0.01 mg to about 10 gm of active ingredient per dose, or about 0.5 mg to about 50 mg of active ingredient per dose, or about 3 mg to about 30 mg of active ingredient per dose, or about 10 to about 30 mg of active ingredient per dose. In some embodiments, the pharmaceutical formulations are formulated and / or administered containing about 0.5 mg, or about 1 mg, or about 5 mg, or about 10 mg, or about 15 mg, or about 20 mg, or about 25 mg, or about 30 mg of active ingredient per dose. In some embodiments, the pharmaceutical compositions disclosed herein are formulated as tablets, each tablet containing about 0.5 mg to about 30 mg of active ingredient per tablet. In some embodiments, each tablet contains about 0.5 mg of active ingredient. In some embodiments, each tablet contains about 1 mg of active ingredient. In some embodiments, each tablet contains about 5 mg of active ingredient. In some embodiments, each tablet contains about 10 mg of active ingredient. In some embodiments, each tablet contains about 15 mg of active ingredient. In some embodiments, each tablet contains about 25 mg of active ingredient. In some embodiments, each tablet contains about 40 mg of active ingredient. In some embodiments, each tablet contains about 50 mg of active ingredient. In some embodiments, each tablet contains about 1 gm of active ingredient. In some embodiments, each tablet contains about 10 gm of active ingredient.

[0070] In some embodiments, the pharmaceutical formulation of the present disclosure is administered once a day to three times a day. In some embodiments, the pharmaceutical formulation of the present disclosure is administered once a day. In some embodiments, the pharmaceutical formulation of the present disclosure is administered twice a day. In some embodiments, the pharmaceutical formulation of the present disclosure is administered three times a day.

[0071] In some embodiments, the pharmaceutical formulation is administered at a dose level and frequency such that the subject receives a total amount of about 0.01 mg to about 10 gm of active therapeutic agent per day, or a total amount of about 1 mg to about 1 gm of active therapeutic agent per day, or a total amount of about 5 mg to about 1 gm of active therapeutic agent per day, or a total amount of about 5 mg to about 500 mg of active therapeutic agent per day, or a total amount of about 12 mg to about 375 mg of active therapeutic agent per day. In some embodiments, the pharmaceutical formulations of the present disclosure are administered at a dose level and frequency such that a subject receives a total amount of about 1 mg, or about 2 mg, or about 3 mg, or about 4 mg, or about 5 mg, or about 6 mg, or about 7 mg, or about 8 mg, or about 9 mg, or about 10 mg, or about 11 mg, or about 12 mg, or about 13 mg, or about 14 mg, or about 15 mg, or more of active ingredient (i.e., lysate, lysate fraction, and / or cell wall fraction, or a combination thereof) per day, as needed. In some embodiments, the pharmaceutical formulations of the present disclosure are administered at a dose level and frequency such that a subject receives a total amount of about 12 mg, or about 24 mg, or about 36 mg, or about 48 mg, or about 60 mg, or about 72 mg, or about 84 mg, or about 96 mg, or about 108 mg, or about 120 mg, or about 132 mg, or about 144 mg, or about 156 mg, or about 168 mg, or about 180 mg, or more of active ingredient (i.e., lysate, lysate fraction, and / or cell wall fraction, or a combination thereof) per day, as needed. In some embodiments, the pharmaceutical formulations of the present disclosure are administered at a dose level and frequency such that a subject receives a total amount of about 15 mg, or about 30 mg, or about 45 mg, or about 60 mg, or about 75 mg, or about 90 mg, or about 105 mg, or about 120 mg, or about 135 mg, or about 150 mg, or about 165 mg, or about 180 mg, or about 195 mg, or about 210 mg, or about 225 mg, or more, of active ingredient (i.e., lysate, lysate fraction, and / or cell wall fraction, or a combination thereof) per day, as needed.In some embodiments, the pharmaceutical formulations of the present disclosure are administered at a dosage level and frequency such that a subject receives a total amount of about 25 mg, or about 50 mg, or about 75 mg, or about 100 mg, or about 125 mg, or about 150 mg, or about 175 mg, or about 200 mg, or about 225 mg, or about 250 mg, or about 275 mg, or about 300 mg, or about 325 mg, or about 350 mg, or about 375 mg, or about 400 mg, or more, of active ingredient (i.e., lysate, lysate fraction, and / or cell wall fraction, or a combination thereof), as needed, per day.

[0072] Any and all of the above doses and dosage amounts are applicable to both the single administration of the active ingredients of the present disclosure, as well as to embodiments in which the active ingredients of the present disclosure are administered in combination with one or more other therapeutic or pharmaceutical agents. active ingredient

[0073] The active ingredients used in the compositions and methods of the present disclosure include bacterial lysates, lysate fractions, or cell wall fractions. The active ingredients may be produced, generated, or derived from any gram-positive or gram-negative bacterial organism.

[0074] As used herein, the term "lysate" refers to a composition prepared from lysed cells, such as bacterial cells. A lysate contains the entire cell contents and, in some embodiments, associated surface components, such as exopolysaccharides, depending on the precise process conditions used to generate the lysate.

[0075] A non-limiting example of a method for producing an active ingredient according to the present disclosure is described in Example 1 and generally includes the steps of: (1) fermenting bacteria in a growth medium; (2) harvesting the bacteria therefrom by centrifuging the bacterial suspension; (3) washing and sterilizing the harvested bacteria; (4) disrupting the bacterial cell walls to lyse the bacteria; and (4) lyophilizing the resulting mixture to obtain the active ingredient, such as a bacterial lysate, a lysate fraction, or a cell wall fraction.

[0076] The structures present in the lysates, lysate fractions, and / or cell wall fractions of the present disclosure that are responsible for TLR activation and reduced oxidative stress are structures that are common and conserved in all bacterial microorganisms. Thus, in principle, any bacterial organism can be used to produce the active ingredients (i.e., lysates, lysate fractions, and / or cell wall fractions) of the present disclosure.

[0077] In some embodiments, the bacterial organism used to produce the active ingredients of the present disclosure is a gram-positive bacterium. In some embodiments, the gram-positive bacterium is Lactobacillus acidophilus, Lactobacillus buhinerii, Lactobacillus casei, Lactobacillus catenaforme, Lactobacillus cellobiosus, Lactobacillus crispatus, Lactobacillus calvertus, Lactobacillus delbrueckii, Lactobacillus delbrueckii subsp. bulgaricus, Lactobacillus delbrueckii subsp. lactis, Lactobacillus helveticus, Lactobacillus jensen, Lactobacillus reichmannii, Lactobacillus spp. Lactobacillus minutus, Lactobacillus paracasei, Lactobacillus plantarum, Lactobacillus rhamnosus, Lactobacillus logosae, Lactobacillus salivarius, Lactobacillus sporogenes (also known as Bacillus coagulans), Lactobacillus brevis, Lactobacillus gasseri, Lactobacillus fermentum, Bifidobacterium adolescentis, Bifidobacterium animalis (especially B. animalis subsp. animalis), Bifidobacterium angulatum, Bifidobacterium bifidum, Bifidobacterium breve, Bifidobacterium catenulatum, Bifidobacterium dentium, Bifidobacterium erixonyi, Bifidobacterium infantis, Bifidobacterium l lactis (Bifidobacterium animalis subsp. lactis), Bifidobacterium longum, Bifidobacterium plantarum, Bifidobacterium pseudocatenulatum, Bifidobacterium spp. lactis), Streptococcus raffinolactis, Acidaminococcus fermentus, Cytophaga fermentus, Rhodoferax fermentus, Cellulomonas fermentus, Zymomonas mobilus, Pediococcus acidilactici or Streptococcus thermophilus, and functionally equivalent variants thereof.

[0078] In some embodiments, the bacterial organisms used to produce the active ingredients of the present disclosure are gram-negative bacteria. In some embodiments, the gram-negative bacteria is selected from the group consisting of Acinetobacter baumannii, Actinobacillus, Acetobacter xylinus, Bacteroides thetaiotaomicron, Bacteroides fragilis, Bordetella pertussis, Brucella abortus, Campylobacter jejuni, Citrobacter freundii, Enterobacter cloacae, Enterobacter sakazakii, Cyanobacteria, Erwinia amylovora, Escherichia coli, Francisella tularensis, Helicobacter pylori, Haemophilus influenzae, Legionella pneumophila, Moraxella catarrhalis, Neisseria gonorrhoeae, Proteus mirabilis, Pseudomonas aeruginosa, Pseudomonas fluorescens, Salmonella enteritidis, Salmonella typhi, Serratia marcescens, Shigella flexneri, Vibrio cholerae, Vibrio algenily, Vibrio algenily, Mycobacterium tuberculosis, Mycobacterium kansasii, Klebsiella oxytoca, and Klebsiella pneumoniae, and Xanthomonas campestris. In some embodiments, the gram-negative bacterium is selected from the genera Pseudomonas, Klebsiella, Xanthomonas, Shigella, and Enterobacter. In some embodiments, the gram-negative bacterium is one or more of Escherichia coli, Klebsiella oxytoca, Shigella flexneri, Pseudomonas fluorescens, and Xanthomonas campestris, and functionally equivalent variants thereof.

[0079] All of the above bacteria are believed to produce immune stimulatory cellular components, including, but not limited to, EPS, which are distinct from the LPS (lipopolysaccharide) found in many bacteria. Other species of bacteria can also be used in the compositions and methods of the present disclosure, such as those disclosed in the state of the art and generally available in microorganism collections such as the European Collection of Cell Cultures (ECACC), the American Society for Testing and Materials (ASTM), the American Type Culture Collection (ATCC), and the German Collection of Microorganisms and Cell Cultures (DSM).

[0080] With respect to the fermentation step to produce a lysate, lysate fraction, or cell wall fraction according to the present disclosure, acceptable growth media used for fermentation will depend on the particular bacteria being grown. Typical growth media include those that include a nitrogen source (1-4%) which may include one or more of the following: yeast extract, milk protein and casein hydrolysates, soy and soy hydrolysates, meat extract, peptones or ammonia salts, simple sugars or ingredients including, but not limited to, simple sugars or hydrolyzed carbohydrates that produce simple sugars such as glucose or lactose (0.5-3%), and minerals (0.05-0.3%), which may include salts of sodium, manganese, magnesium, calcium, and potassium. Surfactants, cysteine ​​HCL, and ribonucleotides (0.001-0.75%) may be added to support cell growth. The growth medium is adjusted to a pH of 6-8. In some embodiments, the pH of the growth medium is in the range of about 6.0-6.5, depending on the particular bacteria being grown. In some embodiments, the pH of the growth medium is in the range of about 6.5-7.0. In some embodiments, the pH of the growth medium ranges from about 7.0 to 7.5.

[0081] In some embodiments, the seed medium is incubated at 30-50°C during fermentation. In some embodiments, the seed medium is incubated at 30-40°C during fermentation. In some embodiments, the seed medium is incubated at about 30°C, about 31°C, about 32°C, about 33°C, about 34°C, about 35°C, about 36°C, about 37°C, about 38°C, about 39°C, about 40°C, about 45°C. In some embodiments, the seed medium is incubated at about 30°C. In some embodiments, the seed medium is incubated at about 33°C. In some embodiments, the seed medium is incubated at about 35°C. In some embodiments, the seed medium is incubated at about 37°C. In some embodiments, the seed medium is incubated at about 40°C. In some embodiments, the seed medium is incubated at about 45°C.

[0082] In some embodiments, fermentation is continued for about 6 hours to about 120 hours before harvesting the bacteria. In some embodiments, fermentation is continued for about 12 hours to about 48 hours before harvesting the bacteria. In some embodiments, fermentation is continued for about 12 hours to about 24 hours before harvesting the bacteria. In some embodiments, fermentation is continued for about 14 hours, or about 15 hours, or about 16 hours, or about 17 hours, or about 18 hours, or about 19 hours, or about 20 hours, or about 22 hours, or about 24 hours, or about 48 hours. In some embodiments, fermentation is continued until bacterial growth reaches mid-log phase, late log phase, early stationary phase, mid-stationary phase, or late stationary phase. In some embodiments, fermentation is continued until the bacteria reaches stationary growth phase. In some embodiments, fermentation can be held for 14 days or more prior to downstream processing as warranted using techniques known to those skilled in the art, such as, but not limited to, cooling and pH control.

[0083] After fermentation, the broth is typically cooled and the bacteria are harvested by centrifugation. In some embodiments, the broth is cooled to about 1° C. to about 25° C. after fermentation. In some embodiments, the broth is cooled to about 1° C., or about 2° C., or about 3° C., or about 4° C., or about 5° C., or about 6° C., or about 7° C., or about 8° C., or about 10° C. In some embodiments, the broth is cooled to about 4° C. to about 7° C.

[0084] After cooling, centrifugation is performed to separate the cells from the surrounding growth medium. The cells are then washed in fresh medium, deionized water, water, or other solution via repeated centrifugation and resuspension, and then resuspended in fresh medium or other solution in preparation for the pasteurization step.

[0085] In some embodiments, the washed bacteria are pasteurized. In some embodiments, the bacteria are pasteurized at about 75° C. to about 85° C. for 30 minutes to 60 minutes. In some embodiments, the harvested bacteria are pasteurized at about 80° C. In some embodiments, pasteurization proceeds for about 30 minutes, or about 45 minutes, or about 60 minutes.

[0086] After pasteurization, the cell concentrate is treated to disrupt the cell walls, thereby exposing the TLR agonist. Disruption of the cell walls can be accomplished by using chelating agents, detergents, surfactants, and hydrolases. Examples of hydrolases that can be used include, but are not limited to, lysozymes, such as hen (egg) egg white lysozyme (e.g., INOVPUR®, LYSOLAC®, DELVOZYME®, LYSOVIN®, LYSOBAC®), lysins, endolysins, and hydrolases. In some embodiments, lysozyme is added to the sterilized bacterial cell suspension to a final concentration of 0.01-4% by volume. In some embodiments, lysozyme is added to the sterilized bacterial cell suspension to a final concentration of about 0.5% by volume, or about 1% by volume, or about 2% by volume, or about 3% by volume, or about 4% by volume. In some embodiments, treatment of the pasteurized bacterial suspension with lytic enzymes continues for about 1 to about 10 hours, or about 6 to about 8 hours, or about 5 hours, or about 6 hours, or about 7 hours, or about 8 hours, or about 9 hours, or about 10 hours. In some embodiments, treatment of the pasteurized bacterial suspension with lytic enzymes is carried out at a temperature of about 25° C. to about 50° C., or about 30° C. to about 45° C., or about 35° C., or about 37° C., about 40° C., about 42° C., about 45° C. After lysis, the lysate is typically frozen and lyophilized.

[0087] In some embodiments, the lyophilized material is then blended with a promoter, such as N-Acetyl D-Glucosamine HCl (NAG), as described in more detail below. If necessary, other formulation excipients can be added to appropriately produce a solid form pill or powder.

[0088] In some embodiments, the specific TLR(s) activated or stimulated by a particular lysate can be altered by changing the raw materials, process materials, or process conditions. The term "TLR specificity" as used herein refers to the specific TLR(s) activated by a given composition of the present disclosure, including bacterial lysate, lysate fraction, and / or cell wall fraction. In some embodiments, TLR specificity varies depending on the density at which the bacterial cells grow during fermentation. In some embodiments, the specific bacterial species or subspecies in which the active ingredient is produced also results in differences in TLR specificity. In some embodiments, the specific enzymes used to lyse the bacterial cells after pasteurization result in differences in TLR specificity.

[0089] As used herein, when used in reference to a receptor or other biomolecular target, the terms "activation," "stimulation," "targeting," and "agonism" are synonymous and refer to binding to and activating a receptor, such as a toll-like receptor, such that the signaling cascade downstream of the receptor is altered, regulated, or otherwise affected.

[0090] It will be appreciated that cells can be fractionated prior to lysis, or the lysate produced above can be fractionated by well-known biochemical procedures, such as, for example, differential centrifugation or column chromatography (e.g., gel permeation chromatography, ion exchange chromatography, chromatography on hydrophobic media, or precipitation, etc.). In some embodiments, lysate fractionation produces fractions with different TLR specificities. For example, fractions can be produced that target fewer TLRs than complete lysate.

[0091] In some embodiments, components including exopolysaccharides may be combined with the lysates, lysate fractions, and / or cell wall fractions of the present disclosure. Exopolysaccharides can activate TLRs. Thus, for example, xanthan gum (from Xanthomonas campestris) may be used in combination with the lysates, lysate fractions, and / or cell wall fractions of the present disclosure to increase TLR activation and / or modify the TLR specificity of the composition. In some embodiments, for example, in embodiments where the xanthan gum is derived from a lysate produced from fermentation of Xanthomonas campestris by fractionation of the Xanthomonas campestris lysate, the xanthan gum is itself a lysate fraction. Other ingredients and delivery forms

[0092] The therapeutic composition of the present disclosure may further and optionally include one or more promoters to aid in therapeutic delivery of active agents across biological membranes.The promoters useful according to the present disclosure may be amino acids, N-alkylated peptides, sugars, amino sugars or amino sugar chelates.The amino sugar chelates include one or more amino sugar ligands, one or more saturated hydroxylated carboxylic acid ligands, and nutritionally acceptable metals, wherein at least one of the one or more amino sugar ligands is glucosamine, the metal is selected from the group consisting of manganese, magnesium, sodium, potassium and zinc, the one or more saturated hydroxylated carboxylic acid ligands are gluconic acid, the ratio of glucosamine ligand to nutritionally acceptable metal is 2:1, and the nutritional metal is non-ferrous.

[0093] According to one embodiment of the present disclosure, the therapeutic formulation may comprise one or more acetylated or deacetylated amino sugars selected from the group consisting of NAG, galactosamine, N-acetylgalactosamine, mannosamine, and N-acetylmannosamine in their monomeric, oligomeric, and / or polymeric forms, such as chitin, and human glycosaminoglycans, and derivatives thereof. As used herein with respect to amino sugars, the term "derivatives thereof" refers to derivatives of amino sugars that have the same or essentially the same ability to form cytotoxic degradation products during sterilization. According to selected further aspects of the disclosure, the promoter is a member selected from the group consisting of poly-L-lysine, glucosamine, poly-L-arginine, galactosamine, N-acetylmannosamine (NAM; N-Ac-Man), N-15 acetylglucosamine (NAG; N-Ac-Glc), N,N'-diacetylglucosamine (NAG-NAG; N,N'-diacetylchitobiose), N,N',N",N'"-tetraacetylglucosamine (NAG-NAG-NAG-NAG; N,N',N",N'"-tetraacetylchitotetraose), and mixtures thereof.

[0094] Optionally, similarly acceptable promoters may be acylated glycosyloxysaccharides of 2-12, a-1,2 and / or a-1,6 linked sugars or optionally acylated oligoglycosyloxysaccharide moieties, where the sugar(s) are selected from the group consisting of D-mannose, D-galactose, D-glucose, D-glucosamine, N-acetylglucosamine and 6-deoxy-L-mannose, and the oligoglycosyloxysaccharide moieties may contain the same or different sugars.

[0095] In another embodiment, the composition of the present invention is in a dosage form selected from the group consisting of lozenges, chewing gums, chewable tablets, and dissolving tablets, e.g., slow dissolving tablets, fast dissolving tablets, or controlled release tablets, or other suitable sustained release formulations.

[0096] In one embodiment, the active agent of the present disclosure is delivered to the oral mucosa of a subject, the oral mucosa being selected from the group consisting of the sublingual mucosa, the buccal mucosa, and combinations thereof, and the composition is administered sublingually, thereby allowing the active ingredient to be delivered across the sublingual mucosa.

[0097] In another embodiment, the carrier is typically a solid, semi-solid or liquid, such as a binder, a gum base, or a combination thereof. Binders suitable for use in the compositions of the present invention include, but are not limited to, sugar alcohols such as mannitol, sorbitol, and xylitol; sugars such as lactose, dextrose, sucrose, glucose, powdered sugar, and other materials such as inositol, molasses, maltodextrin, starch, cellulose, microcrystalline cellulose, polyvinylpyrrolidone, acacia gum, guar gum, tragacanth gum, alginates, iris moss extract, bread gum, ghatti gum, mucilage of isagol (psyllium) husk, VEEGUM®, larch arabogalactan, gelatin, methylcellulose, ethylcellulose, carboxymethylcellulose, hydroxypropylmethylcellulose, polyacrylic acid (e.g., carbopol), calcium silicate, calcium phosphate, dicalcium phosphate, calcium sulfate, kaolin, sodium chloride, polyethylene glycol, and combinations thereof. Suitable gum bases for use in the compositions of the present invention include, for example, materials selected from many water-insoluble and saliva-insoluble gum base materials known in the art. In some examples, the gum base includes at least one hydrophobic polymer and at least one hydrophilic polymer. Non-limiting examples of hydrophobic and hydrophilic polymers suitable for gum bases include both natural and synthetic polymers, such as elastomers, rubbers, and combinations thereof. Examples of suitable natural polymers include, but are not limited to, materials of plant origin, such as chicle, jelutong, gutta-percha, crown gum, and combinations thereof. Examples of suitable synthetic polymers include butadiene-styrene copolymers, isobutylene and isoprene copolymers (e.g., butyl rubber), polyethylene, polyisobutylene, polyvinyl esters (e.g., polyvinyl acetate and polyvinyl acetate phthalate), and combinations thereof.In another example, the gum base includes a mixture of butyl rubber (ie, an isobutylene and isoprene copolymer), polyisobutylene, and optionally polyvinyl acetate (eg, having a molecular weight of about 12,000).

[0098] In yet another embodiment, the compositions of the present invention may further comprise a sweetener, a flavoring agent, a protectant, a plasticizer, a wax, an elastomer solvent, a filler, a preservative, or a combination thereof. In yet another embodiment, the compositions of the present invention may further comprise a lubricant, a wetting agent, an emulsifier, a solubilizer, a suspending agent, a colorant, a disintegrant, or a combination thereof. In one embodiment, the average particle size of the drug in the compositions described herein is about 20 micrometers, as compared to a typical average drug particle size of about 75 to about 100 micrometers. In another embodiment, the average particle size of the drug in the compositions described herein is equal to or less than the average particle size of the carrier components (e.g., gum base, binder, etc.).

[0099] In one embodiment of the present disclosure, the therapeutic composition can optionally include a buffer system to raise the pH of saliva to about 8.0 to about 11, regardless of the starting pH of the saliva in the oral cavity of the subject to be treated.The therapeutic agent suitable for use in the present invention is described above.The carbonate salt and bicarbonate salt suitable for use in the buffer system of the present invention are also described above.In some cases, the composition further includes a non-biological therapeutic agent, such as an NSAID.

[0100] Suitable citrate, phosphate, and borate salts include, but are not limited to, any salt of citric acid, phosphoric acid, or boric acid known in the art. For example, in some embodiments, the citrate salt is selected from the group consisting of sodium citrate, potassium citrate, calcium citrate, magnesium citrate, and ammonium citrate.

[0101] In other embodiments, the phosphate salt is selected from the group consisting of sodium monohydrogen phosphate, sodium dihydrogen phosphate, potassium monohydrogen phosphate, potassium dihydrogen phosphate, calcium monohydrogen phosphate, calcium dihydrogen phosphate, magnesium monohydrogen phosphate, magnesium dihydrogen phosphate, ammonium monohydrogen phosphate, and ammonium dihydrogen phosphate. In yet other embodiments, the borate salt is selected from the group consisting of sodium borate, potassium borate, calcium borate, magnesium borate, and ammonium borate. In some cases, the buffer system includes a carbonate salt, a bicarbonate salt, and / or a citrate salt. In certain other examples, the buffer system includes a carbonate salt, a bicarbonate salt, and / or a phosphate salt. In further examples, the buffer system includes a carbonate salt, a bicarbonate salt, and / or a borate salt.

[0102] In addition to buffer systems that include carbonate salts, bicarbonate salts, and / or metal oxides, other buffer systems are suitable for use in the compositions of the present invention. For example, in another embodiment, a ternary buffer system includes a carbonate salt, a bicarbonate salt, a citrate salt, a phosphate salt, or a borate salt. In another alternative embodiment, the buffer system includes a carbonate salt or a bicarbonate salt, and two or more buffering agents selected from the group consisting of a metal oxide, a citrate salt, a phosphate salt, and a borate salt. In yet another alternative embodiment, the buffer system is a binary buffer system that includes a carbonate salt or a bicarbonate salt and a metal oxide. In yet another alternative embodiment, the buffer system is a binary buffer system that includes a carbonate salt or a bicarbonate salt and a citrate salt, a phosphate salt, or a borate salt. In a further alternative embodiment, the buffer system is a binary buffer system that includes a metal oxide and a citrate salt, a phosphate salt, or a borate salt. In yet another alternative embodiment, the buffer system is a binary buffer system comprising a carbonate salt and a bicarbonate salt. Mode of delivery

[0103] Therapeutic compositions of the present invention may take the form of solid, semi-solid, lyophilized powder or liquid dosage forms, such as tablets (e.g., chewable, slow dissolving, fast dissolving), pills, capsules, lozenges, candies, gums, powders, solutions, suspensions, emulsions, aerosols, etc. The dosage form can be a chewing gum, a fast dissolving tablet, a candy, or a lozenge.

[0104] As used herein, the term "dosage form" refers to a physically discrete unit suitable as a unitary dose for human subjects, mammals, and other non-mammalian animals, each unit containing a predetermined amount of a therapeutic agent calculated to produce a desired onset, tolerability, and therapeutic effect, in combination with one or more suitable pharmaceutical excipients, such as carriers. Methods for preparing such dosage forms are known or will be apparent to those skilled in the art. For example, in some embodiments, the chewing gum dosage forms of the present invention can be prepared according to industry standard procedures. In other embodiments, the tablet, lozenge, or candy dosage forms (e.g., suckers or roll pops) of the present invention may be prepared according to procedures found, for example, in Remington "The Science and Practice of Pharmacy" 20th Edition Ed., [Lippincott, Williams & Wilkins (2003), and "Pharmaceutical Dosage Forms" Vol. 1: Tablets, 2nd Edition, Marcel Dekker, Inc., New York, NY (1989)]. In either case, the dosage form administered will contain a quantity of the active therapeutic agent that, when administered in accordance with the teachings of the present invention, is a therapeutically effective amount for the relief of the condition being treated.

[0105] As used herein, the term "carrier" refers to a typical inert material used as a diluent or vehicle for drugs such as therapeutic agents. This term also includes a typical inert material that imparts cohesiveness to the composition. Suitable carriers for use in the compositions of the present invention include solids, semi-solids, or liquids, such as, but not limited to, binders or gum bases. Examples of binders are known to those skilled in the art. The binder can be pretreated to improve flowability and taste by methods known in the art, such as lyophilization (see, e.g., "Fundamentals of Freeze-Drying," Pharm. Biotechnol., 14th ed., pp. 281-360 (2002); "Lyophilization of Unit Dose Pharmaceutical Dosage Forms," ​​Drug. Dev. Ind. Pharm., Vol. 29, pp. 595-602 (2003)), solid solution preparation, and lubricant dusting, and wet granulation preparation with a suitable lubricant (see, e.g., "Remington: The Science and Practice of Pharmacy," supra). When a binder is included in the formulation, the composition of the present invention can include about 15% to about 90% by weight of the binder, and about 35% to about 80% by weight of the binder. However, one skilled in the art will understand that the composition of the present invention can be prepared without any binder, for example, to produce a very friable dosage form. tablet

[0106] When the dosage form is a tablet, such as a dissolving tablet (i.e., a disintegrating tablet) or a chewable tablet, the composition of the present invention comprises one or more of the therapeutic agents described herein derived from bacteria, or pharma- ceutically acceptable salts thereof, a carrier such as a promoter, a binder, and a buffer system, such as a binary or ternary buffer system. The tablet composition may further comprise a lubricant, a wetting agent, an emulsifier, a suspending agent, a preservative, a sweetener, a flavoring agent, a colorant, and a disintegrant. Typically, the tablet composition of the present invention comprises about 0.001% to about 10.0% by weight, more typically about 1.0% to about 5.0% by weight of the active therapeutic agent (in any selected form measured by its free base form). One skilled in the art will appreciate that the aforementioned percentages will vary depending on the particular source of active therapeutic agent utilized, the amount of active therapeutic agent desired in the final formulation, and the particular release rate of the active therapeutic agent desired. The buffer system of the tablet composition provides a final saliva pH of at least about 8.0 or greater, at least about 9.5, and / or in the range of about pH 9.9 to about pH 11.

[0107] In certain embodiments, the tablet is a dissolving tablet, such as a slow-dissolving or fast-dissolving tablet, that is dissolved by the subject's saliva without the need for chewing. For example, a dissolving tablet placed on the subject's tongue can be used for buccal delivery of a therapeutic agent. Alternatively, a dissolving tablet placed under the subject's tongue can be used for sublingual delivery of a therapeutic agent. This type of dosage form can be particularly desirable for pediatric and geriatric patients, as children and the elderly often have difficulty chewing certain items. Typically, the dissolving tablet is formulated to dissolve within about 1 to about 15 minutes, within about 2 to about 10 minutes, for example, within about 2, 3, 4, 5, 6, 7, 8, 9, or 10 minutes after administration. Those skilled in the art will appreciate that a fast-dissolving tablet dissolves faster than a slow-dissolving tablet, and is typically dissolved gradually, rather than rapidly, by the subject's saliva. In one embodiment, the slow-dissolving or fast-dissolving tablet delivers a therapeutic agent to the sublingual mucosa for a period of more than about 1 minute.

[0108] In certain other embodiments, the tablet is a chewable tablet formulated to be chewed by the subject and dissolve quickly or slowly. For example, a chewable placed on the subject's tongue can be used for buccal delivery of a therapeutic agent. During chewing, the chewable tablet can be moved around in the mouth and can even be placed between the gum and cheek or under the tongue. As a result, at least a portion of the therapeutic agent contained within the chewable tablet can be delivered sublingually (i.e., across the sublingual mucosa). Typically, the chewable tablet is formulated to dissolve within about 1 to about 15 minutes, within about 2 to about 10 minutes, and within 1 minute or more, such as within about 2, 3, 4, 5, 6, 7, 8, 9, or 10 minutes, after administration.

[0109] As noted above, the dissolving and chewable tablets of the present invention are typically formulated to dissolve within about 1-15 minutes after administration and within about 1 minute after administration. However, while these time frames are subject to the maximum exposure of the therapeutic agent to the oral mucosa (e.g., exposure to the sublingual and / or buccal mucosa), they are not always subject to the user's compliance (e.g., the user may swallow too frequently, preventing maximum transmucosal absorption). Thus, in some cases, it may be desirable to strike a balance between patient compliance and the maximum exposure time of the therapeutic agent to the oral mucosa. This can be accomplished, for example, by reducing the tablet size (e.g., from about 700-800 mg to about 200-300 mg) without reducing the concentration or amount per unit dose of the buffer system or therapeutic agent. Additionally, minor changes to the tablet formulation may be used to reduce saliva production, for example, substituting one flavoring for another (e.g., chocolate for spearmint) or substituting one binder or sweetener for another (e.g., lactose for mannitol or sorbitol). Combination with other medications

[0110] Many pharmacologic active agents cause adverse side effects, which are driven by ROS generated as the active agent progresses through biochemical and other metabolic reactions. Thus, the compositions described herein are attractive candidates for the treatment of oxidative stress and oxidative stress-related diseases and conditions, and can be combined with one or more pharmaceutical agents known to those skilled in the art, such as those listed in the Physicians' Desk Reference (available at http: / / www.pdr.net).

[0111] In some embodiments, the active ingredient of the present disclosure may be combined with known pharmaceutical agents that target oxidative stress, inflammation, immune response and / or oxidative stress-related diseases or conditions, or may be combined with pharmaceutical agents that produce oxidative stress as a side effect.If a pharmaceutical agent targets some other disease state but produces oxidative stress as a side effect, combining with the composition of the present invention reduces the damage caused by oxidative stress and improves the efficacy of the pharmaceutical agent by allowing high doses without too much damage caused by oxidative stress.It is understood that optimal results with a particular combination may require adjustment of the lysate to optimally target the correct TLR in the presence of the added pharmaceutical agent.

[0112] Thus, in one aspect, the present disclosure provides a method for reducing or alleviating one or more oxidative stress-related side effects associated with administration of a pharmaceutical agent, the method comprising administering a lysate, lysate fraction, and / or cell wall fraction of the present disclosure in combination with a pharmaceutical agent. In some embodiments, the pharmaceutical agent and the lysate composition are administered simultaneously or in any order, through the same or different routes of administration. In some embodiments, the pharmaceutical agent is any pharmaceutical agent having one or more oxidative stress-related side effects, for example, any pharmaceutical agent having one or more oxidative stress-related side effects listed available in the Physicians' Desk Reference (http: / / www.pdr.net). In some embodiments, the pharmaceutical agent is selected from the group consisting of antirheumatic drugs, anti-inflammatory drugs, chemotherapeutic drugs, radiotherapeutic drugs, immunosuppressants, interferons, interferon-based chemotherapeutic drugs, and cytotoxic drugs. In some embodiments, the oxidative stress related side effect is selected from the group consisting of aceruloplasminemia, arterial / systemic hypertension, arthritis, asthma, atherosclerosis, atopic dermatitis, cancer, bladder cancer, leukemia, uterine cancer, cervical cancer, dizziness, nausea, vomiting, constipation, diarrhea, insomnia, drowsiness, lightheadedness, decreased libido, memory loss, tremors, jaundice, arrhythmia, increased heart rate, decreased heart rate, hives, depression, clinical depression, cerebral ischemia, bronchopulmonary dysplasia, cardiovascular disease, cataracts, cellulitis, chemotherapy side effects, chronic fatigue syndrome, colitis, coronary artery disease, dyslipidemia, eclampsia, erectile dysfunction, ataxia, headache, heart failure, hemodialysis side effects, liver The condition is selected from cirrhosis, hypercholesterolemia, hyperhomocysteinemia, hyperlipidemia, interstitial lung disease, lung injury, macular degeneration, male infertility, mild cognitive impairment, myocardial infarction, myocarditis, myocardial degeneration, neuropathy, obesity, osteoarthritis, osteoporosis, pancreatitis, periodontal disease, side effects of peritoneal dialysis, post traumatic stress disorder, pre-eclampsia, psoriasis, psoriatic arthritis, pulmonary hypertension, side effects of radiation therapy, reactive arthritis, respiratory distress syndrome, rhabdomyolysis, rheumatic disease, sepsis, sleep apnea, stroke, suicidal thoughts, amyloidosis, thrombophily, tauopathy, unstable angina, uremia, or venous insufficiency.

[0113] In some embodiments, the pharmaceutical agent having one or more oxidative stress-related side effects is selected from the group consisting of: antispasmodics selected from the group consisting of atropine sulfate, dicycloverine hydrochloride, hyoscine butyl bromide, propantheline bromide, alverine citrate, and mebeverine hydrochloride; motility stimulants selected from the group consisting of metoclopramide and domperidone; H2-receptor antagonists selected from the group consisting of cimetidine, famotidine nizatidine, and ranitidine; antimuscarinics; tripotassium bismuth dicitrate and sucralfate. chelates; prostaglandin analogues; aminosalicylates selected from the group consisting of valsazid sodium, mesalazine, olsalazine, and sulfasalazine; corticosteroids selected from the group consisting of beclomethasone dipropionate, budenoside, hydrocortisone, and prednisolone; those affecting the immune system selected from the group consisting of cyclosporine, mercaptopurine, methotrexate, adalimumab, and infliximab; stimulant laxatives selected from the group consisting of bisacodyl, danthron, docusate, and sodium picosulfate; drugs affecting bile composition and flow;Cholestyramine, oxyphencyclimine, camylofine, mebeverine, trimebutine, rosiverine, dicycloverine, dihexyverine, difemerine, piperidolate, benzilon, mepenzolate, pipenzolate, glycopyrronium, oxyphenonium, pentienate, methantheline, propantheline, otilonium bromide, tridihexetyl, isopropamide, hexocyclium, polzine, bevonium, diphemanil, thiemonium iodide, prifinium bromide, thimenium bromide Pygium, fenpiverinium, papaverine, drotaverine, moxaverine, 5-HT3 antagonist, 5-HT4 agonist, fenpiprane, diisopromine, chlorbenzoxamine, pinaverium, fenoberine, idanpramine, proxazole, alverine, trepibutone, isometheptene, caroverine, phloroglucinol, silicone, trimethyldiphenylpropylamine, atropine, hyoscyamine, scopolamine, butylscopolamine, methylscopolamine, methyl Bile acid sequestrants selected from the group consisting of atropine, phentonium, cimetropium bromide, and primarily dopamine blockers; proton pump inhibitors selected from the group consisting of omeprazole, lansoprazole, pantoprazole, esomeprazole, and rabeprazole sodium; opioids and opioid receptor antagonists; acetaminophen, diclofenac, diflunisal, etodolac, fenoprofen, flurbiprofen, ibuprofen, indomethacin, analgesic selected from the group consisting of ketoprofen, ketorolac, meclofenamate, mefenamic acid, meloxicam, nabumetone, naproxen, oxaprozin, phenylbutazone, piroxicam, sulindac, tolmetin, celecoxib, buprenorphine, butorphanol, codeine, hydrocodone, hydromorphone, levorphanol, meperidine, methadone, morphine, nalbuphine, oxycodone, oxymorphone, pentazocine, propoxyphene, and tramadol;Sleeping pills selected from the group consisting of nitrazepam, flurazepam, loprazolam, lormetazepam, temazepam, zaleplon, zolpidem, zopiclone, chloral hydrate, triclofos, clomethiazole, quazepam, triazolam, estazolam, clonazepam, alprazolam, eszopiclone, rozerem, trazodone, amitriptyline, doxepin, benzodiazepines, melatonin, diphenhydramine, and herbal remedies; cardiac glycosides selected from the group consisting of digoxin and digitoxin, phosphodiesterase inhibitors selected from the group consisting of enoximone and milrinone, bendroflumethiazide, chlorthalidone, cyclopenthiazide, inapamide, methadone ... thiazides and thiazide-related diuretics selected from the group consisting of thalasone, thalasone, and oxamide; diuretics selected from the group consisting of furosemide, bumetanide, and trasemide; potassium-sparing diuretics and aldosterone antagonists selected from the group consisting of amiloride hydrochloride, triamterene, weplerenone, and spironolactone; osmotic diuretics; drugs for arrhythmia selected from the group consisting of adenosine, amiodarone hydrochloride, disopyramide, flecainide acetate, propafenone hydrochloride, and lidocaine hydrochloride, propanalol, atenolol, acebutolol, bisprolol fumarate fumarate), carvedilol, celiprolol, esmolol, levatolol, metoprolol tartrate, nadolol, nebivolol, oxyprenolol, pindolol, solatol, and timolol; a hypertensive drug selected from the group consisting of ambrisentan, bosentan, diazoxide, hydralazine, iloprost, minoxidil, sildenafil, sitaxsentan, sodium nitroprusside, clonidine, methyldopa, moxonidine, guanethidine monosulfate, doxazosin, indoramin, prazosin, terazosin, phenoxybenzamine, and phentolamine mesylate;Drugs affecting the renin-angiotensin system selected from the group consisting of captopril, cilazapril, enalapril maleate, fosinopril, imidapril, lisinopril, moexipril, perindopril erbumine, quinapril, ramipril, trandolapril, candesartan cilexetil, eprosartan, irbesartan, losartan, olmesartan medoxomil, telmisartan, valsartan, and aliskiren; glycerin trinitrate, isosorbide dinitrate, isosorbide mononitrate, amlodipine, diltiazem, felodipine, isradipine, nitrates, calcium channel blockers, and antianginal agents selected from the group consisting of valproate, lacidipine, lercanidipine, nicardipine, nifedipine, nimodipine, verapamil, ivaprazine, nicorandil, and ranolazine; peripheral vasodilators, and related agents selected from the group consisting of cilostazol, inositol nicotinate, moxisylyte, naftidrofuryl oxalate, and pentoxifylline; dopamine, dopexamine, ephedrine, metaraminol, noradrenaline tartrate, norephedrine bitartrate sympathomimetics selected from the group consisting of heparin, bemiparin, dalteparin, enoxaparin, tinzaparin, danaparoid, bivalirudin, lepirudin, epoprostenol, fondaprinux, warfarin, acenocoumarol, phenindione, dabigatran etexilate, rivaroxaban, and protamine sulfate; antiplatelet agents selected from the group consisting of abciximab, aspirin, clopidogrel, dipyridamole, eptifibatide, prasugrel, and tirofiban; fibrinolytics and antifibrinolytics selected from the group consisting of alteplase, reteplase, streptokinase, tenecteplase, urokinase, ethamsylate, and tranexamic acid;a lipid-modulating agent selected from the group consisting of atorvastatin, fluvastatin, pravastatin, rosuvastatin, simvastatin, colesevelam, cholestyramine, colestipol, ezetimibe, bezafibrate, ciprofibrate, fenofibrate, gemfibrozil, acipimox, nicotinic acid, omega-3 fatty acids, ethanolamine oleate, and sodium tetradecyl sulfate;Benperidol, Chlorpromazine, Flupenthixol, Haloperidol, Levomepromazine, Pericyazine, Perphenazine, Pimozide, Prochlorperazine, Promazine, Sulpiride, Trifluoperazine, Zuclopenthixol, Amisulpiride, Aripiprazole, Clozapine, Olanzapine, Paliperidone, Quetiapine, Riperidone, Sertindole, Zotepine, Flupenthixol, Fluphenazine, Olanzapine Emboate (Olanzapine Pamoate), Pipotiazine Palmitate, Risperidone, Zuclopenthixol Decanoate valproate, carbamazepine, valproate, valproic acid, lithium carbonate, lithium citrate, amitriptyline, clomipramine, dosulepin, imipramine, lofepramine, nortriptyline, trimipramine, mianserin, trazodone, phenelzine, isocarboxazid, tranylcypromine, moclobemide, citalopram, escitalopram, fluoxetine, fluvoxamine, paroxetine, sertraline, agomelatine, duloxetine, flupentixol, mirtazapine, reboxetine, trytophan, benflaxine Venflaxine, Atomoxetine, Dexamethamine, Methylphenidate, Modafinil, Eslicarbazepine, Ocarbazepene, Ethosuximide, Gabapentin, Pregabalin, Lacosamide, Lamotrigine, Levetiracetam, Phenobarbital, Primidone, Phenytoin, Rufinamide, Tiagabine, Topiramate, Vigabatrin, Zonisamide, Ropinirole, Rotigotine, Co-Beneldopa, Levodopa, CNS drugs selected from the group consisting of Co-Careldopa, Rasagiline, Selegiline, Entacapone, Tolcapone, Amantidine, Orphenadrine, Procyclidine, Trihexyphenidyl, Haloperidol, Piracetam, Riluzole, Tetrabenazine, Acamprosate, Disulfiram, Bupropion, Varenicillin, Buprenorphine, Lofexidine, Donepezil, Galantamine, Memantine, and Rivastidimine;Benzylpenicillin, Phenoxymethylpenicillin, Flucloxacillin, Temocillin, Amoxicillin, Ampicillin, Co-amoxiclav, Co-fluampicil, Piperacillin, Ticarcillin, Pivmecillinam, Cephalosporins, Cefaclor, Cefadroxil, Cephalexin, Cefixime, Cefotaxime, Cephradine, Ceftazidime, Cefuroxime, Ertapenem, Imipenem, Meropenem, Aztreonam, Tetracycline, Demeclocycline, Doxocycline, Lymecycline, Minocycline, Oxytetracycline, Tigecycline, Gentamicin, Amikacin, Neomycin, Tobramycin, Erythromycin, Azithromycin, Clarithromycin, Telithromycin, Clindamycin , Chloramphenicol, Fusidic acid, Vancomycin, Teicoplanin, Daptomycin, Linezolid, Quinupristin, Colistin, Co-trimoxazole, Sulpadiazine, Trimethoprim, Capreomycin, Cycloserine, Ethambutol, Isoniazid, Pyrazinamide, Rifabutin, Rifampicin, Streptomycin, Dapsone, Clofazimine, Metronidazole, Tinidazole, Ciprofloxacin, Levofloxacin, Moxifloxacin, Nalidixic acid, Norflaxine, Orflaxacin, Nitrofurantoin, Methenamine hippurate, Amphotericin, Anidulafungin, Caspofungin, Fluconazole, Flucytosine, Griseofulvin, Itraconazole; Itraconzole, Ketoconazole, Micafungin, Nystatin, Posaconazole, Terbinafine, Voriconazole, Abacavir, Didanosine, Emtricitabine, Lamivudine, Stavudine, Tenofovir Disoproxil, Zidovudine, Atazanavir, Darunavir, Fosamprenavir, Indinavir, Lopinair, Nelfinavir, Ritonavir, Saquinavir, Tipranavir, Efavirenz, Etravirine, Nevarapine, Enfuvirtide, Maraviroc, Raltegravir, Acyclovir, Famciclovir, Inosine Pranobex, Valacyclovir, Cidofovir, Gangciclovir an anti-infective selected from the group consisting of Gangciclovir, Foscarnet, Valgangciclovir, Adefovir Dipivoxil, Entecavir, Telbivudine, Amantadine, Oseltamivir, Zanamivir, Palivizumab, Ribavirin, Artemether, Chloroquine, Mefloquine, Primaquine, Proguanil, Pyrimethamine, Quinine, Doxycycline, Diloxanide Furoate, Metronidaziole, Tinidazole, Mepacrine, Sodium Stibogluconate, Atovaquone, Pentamidine Isethionate, Mebendazole, and Piperazine;and benzothorone, procyclidine, biperiden, amantadine, bromocriptine, pergolide, entacapone, tolcapone, selegiline, pramipexole, budesonide, formoterol, quetiapine fumarate, olanzapine, pioglitazone, montelukast, zoledronic acid, valsartan, latanoprost, irbesartan, clopidogrel, atomoxetine, dexamfetamine, methylphenidate, modafinil, bleomycin, dactinomycin, daunorubicin, idarubicin, mitomycin, mitoxantrone, azacitidine, kathrylamide, ketamine ... Other drugs selected from the group consisting of Pecitabine, Cladribine, Clofarabine, Cytarabine, Fludarabine, Flourouracil, Gemcitabine, Mercaptopurine, Methotrexate, Nelarabine, Pemetrexed, Raltitrexed, Thioguanine, Apomorphine, Betamethasone, Cortisone, Deflazacort, Dexamethosone, Hydrocortisone, Methylprednisolone, Prednisolone, Triamcinolone, Cyclosporine, Sirolimus, Tacrolimus, Interferon alpha, and Interferon beta;

[0114] Thus, the therapeutic formulation of the present disclosure may further include one or more additional therapeutic agents, such as any of the therapeutic agents described below. In some embodiments, the composition is supplied as part of a sterile pharmaceutical composition that includes a pharma- ceutically acceptable carrier. Such compositions that include additional therapeutic agents may be in any suitable form (depending on the desired method of administering the therapeutic agent to the patient). In some embodiments, the active ingredient of the present disclosure is co-administered with one or more additional therapeutic agents, but not necessarily combined with one or more additional therapeutic agents in a single formulation. In some embodiments, the active ingredient(s) of the present disclosure are administered through one route of administration, while the co-administered additional therapeutic agent(s) are administered via a second route of administration. For example, the active ingredient of the present disclosure may be administered orally, mucosally, sublingually, bucally, etc., while the co-administered additional therapeutic agent(s) are administered parenterally, intravenously, etc.

[0115] Examples of compounds that can be combined with the compositions of the present invention include anti-rheumatic drugs, anti-inflammatory drugs, chemotherapeutic drugs, radiotherapeutic drugs, immunosuppressants, interferons, interferon-based chemotherapeutic drugs, or cytotoxic drugs.

[0116] Antirheumatic drugs include, but are not limited to, auranofin, azathioprine, chloroquine, D-penicillamine, gold sodium thiomalate hydroxychloroquine, myocrisin, and sulfasalazine methotrexate.

[0117] Anti-inflammatory agents include, but are not limited to, dexamethasone, pentasa, mesalazine, asacol, codeine phosphate, benorylate, fenbufen, naprosyn, diclofenac, etodolac and indomethacin, aspirin and ibuprofen, and other nonsteroidal anti-inflammatory drugs (NSAIDS).

[0118] Chemotherapeutic agents include, but are not limited to, radioactive molecules, toxins also referred to as cytotoxins or cytotoxic agents. This includes any agent that is detrimental to the viability of cells, agents, and liposomes or other vesicles containing the chemotherapeutic compound. Examples of suitable chemotherapeutic agents include, but are not limited to, 1-dehydrotestosterone, 5-fluorouracil dacarbazine, 6-mercaptopurine, 6-thioguanine, actinomycin D, adriamycin, aldesleukin, alkylating agents, allopurinol sodium, altretamine, amifostine, anastrozole, anthramycin (AMC), mitotic inhibitors, cis-dichlorodiamineplatinum(II) (DDP) cisplatin, diaminodichloroplatinum, anthracyclines, antibiotics, antimetabolites, asparaginase, live BCG (intravesical), betamethasone sodium phosphate, and betamethasone acetate, bicalutamide, bleomycin sulfate, busulfan, leucovorin ... Calcium phosphate, calicheamicin, capecitabine, carboplatin, lomustine (CCNU), carmustine (BSNU), chlorambucil, cisplatin, cladribine, colchicine, conjugated estrogen, cyclophosphamide, cytarabine, cytochalasin B, cytoxan, dacarbazine, dactinomycin, dactinomycin (formerly actinomycin), daunirubicin HCL, daunorubicin citrate, denileukin difitox, dexrazoxane, dibromomannitol, dihydroxy anthracin dione, docetaxel, dolasetron mesylate, doxorubicin HCL, dronabinol, Escherichia coli L-asparaginase, emetine, epoetin alfa, ErwiniaL-asparaginase, esterified estrogens, estradiol, estramustine sodium phosphate, ethidium bromide, ethinyl estradiol, etidronate, etoposide citrovorum factor, etoposide phosphate, filgrastim, floxuridine, fluconazole, fludarabine phosphate, fluorouracil, flutamide, folinic acid, gemcitabine HCL, glucocorticoids, goserelin acetate, gramicidin D, granisetron HCL, hydroxyurea, idarubicin HCL, ifosfamide, interferon alpha-2b, irinotecan HCL, letrozole, calcium leucovorin, leuprolide acetate, levamisole HCL, lidocaine, lomustine, maytansinoids, mechlorethamine HCL, medroxyprogesterone acetate, megestrol acetate, melphalan HCL, mercaptopurine, mesna, methotrexate, Methyltestosterone, mithramycin, mitomycin C, mitotane, mitoxantrone, nilutamide, octreotide acetate, ondansetron HCL, paclitaxel, pamidronate disodium, pentostatin, pilocarpine HCL, primycin, carmustine implant, porfimer sodium, procaine, procarbazine HCL, propranolol, rituximab, sargramostim, streptozotocin, tamoxifen, taxol, tenoposide, testolactone, tetracaine, thioepa, chlorambucil, thioguanine, thiotepa, topotecan HCL, toremifene citrate, trastuzumab, tretinoin, valrubicin, vinblastine sulfate, vincristine sulfate, and vinorelbine tartrate.

[0119] In yet another embodiment of the present disclosure, the active ingredient of the present disclosure is administered in combination with a TNF-α antagonist or an anti-TNF-α antibody. Examples of such TNF-α antagonists include, but are not limited to, soluble TNF-α receptors; etanercept (ENBREL®; Immunex) or a fragment, derivative or analog thereof; infliximab (REMICADE®; Centacor) or a derivative, analog or antigen-binding fragment thereof; adalimumab (Humira and Exemptia, known to block IL-10, TNF-α production via interferon-γ-activated macrophages, TNFR-IgG; mouse product TBP-1; vaccine CytoTAb (Protherics); antisense molecule 104838 (ISIS); peptide RDP-58 (SangStat); thalidomide (Celgene); CDC-801 (Celgene); DPC-333 (Dupont); VX-745 (Vertex); AGIX-4207 (AtheroGenics); ITF-2357 (Italfarmaco); NPI-13021-31 (Nereus); SCIO-469 (Scios); TACE targeter (Immunix / AHP); CLX-120500 (Calyx); Thiazolopyrim (Dynavax); Auranofin (Ridaura) (SmithKline Beecham Pharmaceuticals); quinacrine (mepacrine dichlorohydrate); tenidap (Enablex); melanin (Large Scale Biological); and anti-p38 MAPK agents (Uriach).

[0120] In a further embodiment of the present disclosure, the active ingredient of the present disclosure is administered in combination with rapamycin or similar macrocyclic antibiotic.As used herein, rapamycin includes rapamycin and all its analogs, derivatives and congeners, and other immunophilins that have the same pharmacological properties as rapamycin include the inhibition of TOR or mTOR (mammalian target of rapamycin) (e.g., act as TOR kinase inhibitors).Other immunosuppressants that can be used as one or more pharmaceutical or therapeutic agents include, but are not limited to, cyclosporine, tacrolimus (FK-506), azatropine and mycophenolate mofetil.

[0121] Additional therapeutic agents that may be combined with the active ingredients of the present disclosure include angiogenic agents such as vascular endothelial growth factor (VEGF) and fibroblast growth factor (FGF); angiotensin receptor blockers; nitric oxide donors; antisense oligonucleotides and combinations thereof; cell cycle inhibitors, mTOR inhibitors and growth factor receptor signaling kinase inhibitors; retinoids; cyclin 1 CDK inhibitors; HMG coenzyme reductase inhibitors (e.g., statins); and protease inhibitors. TNF inhibitors

[0122] Tumor necrosis factor (TNF) inhibitors, particularly TNFα inhibitors, used in the methods and compositions of the invention include any agent that interferes with TNF-α activity. In a preferred embodiment, the TNFα inhibitor is capable of neutralizing TNFα activity, particularly the deleterious TNFα activity associated with oxidative stress diseases and disorders (see definitions) and associated complications and conditions.

[0123] As used herein, the term "human TNF-α" (abbreviated herein as hTNFα or simply hTNF) is intended to refer to a human cytokine that exists as a 17 kD secreted form and a 26 kD membrane-associated form, the biologically active form of which is composed of a trimer of non-covalently linked 17 kD molecules. For example, the structure of hTNFα is further described in Pennica, D., et al. (1984) Nature 312:724-729, Davis, JM, et al. (1987) Biochemistry 26:1322-1326, and Jones, EY, et al. (1989) Nature 338:225-228. The term human TNFα is intended to include recombinant human TNFα (rhTNFα), which can be prepared by standard recombinant expression methods or purchased commercially (e.g., from R&D Systems, Minneapolis, Minn.). TNF-α is also referred to equivalently as TNF herein.

[0124] The term "TNFα inhibitor" refers to an agent that interferes with TNFα activity. This term also includes each of the anti-TNFα human antibodies and antibody portions described herein, as well as those described in U.S. Patent Nos. 6,090,382, 6,258,562, 6,509,015, and 7,223,394. In one embodiment, TNFα inhibitors for use in the present invention include anti-TNFα antibodies or fragments thereof, such as infliximab (Remicade®, Johnson and Johnson; U.S. Pat. No. 5,656,272, incorporated herein by reference); protease inhibitors (U.S. Pat. No. 5,656,272, incorporated herein by reference), CDP571 (humanized monoclonal anti-TNF-α IgG4 antibody), CDP870 (humanized monoclonal anti-TNF-α antibody fragment), anti-TNF dAb (Peptech), CNTO148 (golimumab (Medarex and Centocor), see WO 02 / 12502), and adalimumab (Humira®, described as "D2E7" in U.S. Pat. No. 6,090,382, Abbott Laboratories, human anti-TNF mAb). Additional TNF antibodies that can be used in the present invention are described in U.S. Patent Nos. 6,593,458, 6,498,237, 6,451,983, and 6,448,380, each of which is incorporated herein by reference.In other embodiments, the TNFα inhibitor is a TNF fusion protein, such as etanercept (Enbrel®), Amgen; described in WO91 / 03553 and WO09 / 406,476, which are incorporated herein by reference).In another embodiment, the TNF-α inhibitor is recombinant TNF binding protein (r-TBP-1) (Serono).

[0125] The term "antibody" as used herein is intended to refer to an immunoglobulin molecule consisting of four polypeptide chains, including two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. Each heavy chain consists of a heavy chain variable region (abbreviated herein as HCVR or VH) and a heavy chain constant region. The heavy chain constant region consists of three domains, CH1, CH2 and CH3. Each light chain consists of a light chain variable region (abbreviated herein as LCVR or VL) and a light chain constant region. The light chain constant region consists of one domain, CL. The VH and VL regions can be further subdivided into hypervariable regions, termed complementarity determining regions (CDRs), interspersed with regions that are more conserved and termed framework regions (FRs). Each VH and VL is composed of three CDRs and four FRs arranged from amino-terminus to carboxy-terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The antibodies of the present invention are described in US Pat. Nos. 6,090,382, 6,258,562, and 6,509,015, each of which is incorporated herein by reference in its entirety.

[0126] The term "human antibody" as used herein is intended to include antibodies having variable and constant regions derived from human germline immunoglobulin sequences. The human antibody of the present invention may contain amino acid residues (e.g., mutations introduced randomly or by site-specific mutagenesis in vitro or by somatic mutation in vivo), e.g., in the CDRs and especially in CDR3, that are not encoded by human germline immunoglobulin sequences. However, the term "human antibody" as used herein is not intended to include antibodies in which CDR sequences derived from the germline of another mammalian species, such as a mouse, are grafted onto human framework sequences.

[0127] The term "recombinant human antibody" as used herein is intended to include all human antibodies prepared, expressed, produced or isolated by recombinant means, such as antibodies expressed using a recombinant expression vector transfected into a host cell (described in further detail below), antibodies isolated from a recombinant combinatorial human antibody library (described in further detail below), antibodies isolated from an animal (e.g., a mouse) that is transgenic for human immunoglobulin genes (see, e.g., Taylor et al. (1992) Nucl. Acids Res. 20:6287), or antibodies prepared, expressed, produced or isolated by any other means including splicing of human immunoglobulin gene sequences into other DNA sequences. Such recombinant human antibodies have variable and constant regions derived from human germline immunoglobulin sequences. However, in certain embodiments, such recombinant human antibodies are subjected to in vitro mutagenesis (or, when animals transgenic for human Ig sequences are used, in vivo somatic mutagenesis) and thus the amino acid sequences of the VH and VL regions of the recombinant antibodies are derived from and related to human germline VH and VL sequences, but may not naturally occur within the human antibody germline repertoire in vivo.

[0128] In one embodiment, the term "TNFα inhibitor" does not include "infliximab." In one embodiment, the term "TNFα inhibitor" does not include adalimumab. In another embodiment, the term "TNFα inhibitor" does not include adalimumab and infliximab.

[0129] In one embodiment, the term "TNFα inhibitor" does not include etanercept, and optionally adalimumab, infliximab, and adalimumab and infliximab.

[0130] In one embodiment, the term "TNFα antibody" does not include infliximab. In one embodiment, the term "TNFα antibody" does not include adalimumab. In another embodiment, the term "TNFα antibody" does not include adalimumab and infliximab.

[0131] In one embodiment, the invention features uses and compositions for treating, treating, or determining the efficacy of TNFα inhibitors for the treatment of Crohn's disease. The TNFα antibody is an isolated human antibody or antigen-binding portion thereof that binds to human TNFα with high affinity and low dissociation rate and has high neutralizing capacity. Preferably, the human antibody used in the invention is a recombinant neutralizing human anti-hTNFα antibody. The most preferred recombinant neutralizing antibody of the invention is referred to herein as "D2E7", also referred to as HUMIRA® or adalimumab (the amino acid sequence of the D2E7VL region is shown in SEQ ID NO:1, and the amino acid sequence of the D2E7VH region is shown in SEQ ID NO:2). The properties of D2E7 (adalimumab / HUMIRA®) have been described, each of which is incorporated herein by reference. The methods of the invention can also be performed using chimeric and humanized murine anti-hTNFα antibodies that have undergone clinical trials for the treatment of rheumatoid arthritis (see, e.g., Elliott, MJ, et al. (1994) Lancet 344:1125-1127; Elliot, MJ, et al. (1994) Lancet 344:1105-1110; Rankin, EC, et al. (1995) Br. J. Rheumatol. 34:334-342).

[0132] Thus, in one aspect, the present disclosure provides a toll-like receptor (TLR) agonist composition for regulating redox status in a subject. The composition comprises (a) a TLR agonist comprising at least one or more lysates and / or lysate fractions of bacteria, the TLR agonist activating at least one or more TLRs or NLRs, (b) an optional promoter for increasing the absorption of the composition, and (c) an optional carrier for increasing the volume of the composition, and administering an effective amount of the composition to the subject measurably reduces the oxidative stress level in the subject. In some embodiments, the agonist activates at least two different TLRs and / or NLRs. In some embodiments, the bacteria is a gram-positive or gram-negative bacterium.

[0133] In some embodiments, the composition is administered in combination with a pharmaceutical agent to enhance the activity of the pharmaceutical agent. In some embodiments, the composition is administered in combination with a pharmaceutical agent to reduce a side effect of the pharmaceutical agent. In some embodiments, the side effect of the pharmaceutical agent is oxidative stress.

[0134] In another aspect, the present disclosure provides a method of modulating redox status in a subject, comprising administering a therapeutically effective amount of any of the lysate or lysate fraction compositions disclosed herein to a subject in need thereof. In some embodiments, the modulation of redox status is ideally assessed by measuring changes in isoprostanes concentration or other methods, such as, but not limited to, gene expression in the subject. In some embodiments, the subject is a mammal. In some embodiments, the mammal is a human. In some embodiments, the subject is a non-mammalian animal, such as, but not limited to, fish, poultry, crustaceans, and insects such as fruit flies. In some embodiments, the redox status is oxidative, resulting in oxidative stress. In some embodiments, oxidative stress in humans is associated with post-traumatic stress disorder.

[0135] In another aspect, the present disclosure provides a method of regulating redox status in a subject, the method comprising: (a) administering to a subject in need thereof, at intervals in time, a dose of a composition comprising (i) at least one lysate and / or lysate fraction of a toll-like receptor (TLR) agonist, the toll-like receptor (TLR) agonist activating at least one or more different TLRs; (ii) an optional promoter for increasing absorption of the composition; and (iii) an optional carrier for increasing the volume of the composition; and (b) measuring the subject's body fluid to detect changes in oxidative stress levels. In some embodiments, the TLR agonist activates at least three different TLRs. In some embodiments, the TLR agonist comprises lysate(s) and / or lysate fraction(s) and / or cell wall fraction(s) from a single species of bacteria. In some embodiments, the TLR agonist comprises lysate(s) and / or lysate fraction(s) from at least two species or strains of bacteria.

[0136] In some embodiments, the change in isoprostane concentration in the subject is assessed by measuring body fluids. In some embodiments, the change in isoprostane concentration is measured indirectly by methods such as, but not limited to, gene expression in the subject. In some embodiments, the subject is a mammal. In some embodiments, the mammal is a human. In some embodiments, the subject is a non-mammalian animal, such as, but not limited to, fish, poultry, crustaceans, and insects such as fruit flies. In some embodiments, oxidative stress in humans is associated with post-traumatic stress disorder.

[0137] In another aspect, the disclosure provides a method for reducing the amount of isoprostane in the urine or blood of a subject, the method comprising: (a) determining the level of isoprostane in the urine or blood of the subject; (b) administering to the subject an effective amount of a composition comprising: (i) a toll-like receptor (TLR) agonist comprising at least one bacterial lysate and / or lysate fraction from a bacterium, wherein the TLR agonist activates at least one or more TLRs or NLRs; and (ii) an optional promoter for increasing absorption of the composition; and (c) continuing administration of the composition until the level of isoprostane in the urine or blood of the subject is reduced.

[0138] In another aspect, the disclosure includes providing a composition comprising: (a) a bacterial lysate and / or lysate fraction capable of activating at least one or more toll-like receptors (TLRs) or Nod-like receptors (NLRs); (b) an optional promoter for increasing absorption of the composition; and (c) an optional carrier for increasing the volume of the composition.

[0139] In another aspect, the present disclosure provides a pharmaceutical formulation comprising any of the compositions disclosed herein.In some embodiments, the pharmaceutical formulation is formulated for buccal or sublingual administration.In some embodiments, the pharmaceutical formulation is formulated to dissolve within 1 minute after administration.

[0140] In another aspect, the disclosure provides a method for producing a bacterial lysate comprising: (a) fermenting Gram-positive or Gram-negative bacteria in a growth medium to stationary growth phase to produce a fermentation broth; (b) harvesting the bacteria from the fermentation broth; (c) pasteurizing the harvested bacteria; and (d) lysing the pasteurized bacteria with lysozyme to produce a bacterial lysate. In some embodiments, the lysate is harvested at different times in the growth cycle after the start of any particular fermentation. In some embodiments, the bacteria are fermented in a characteristically defined medium.

[0141] In another aspect, the disclosure provides a bacterial lysate produced by a process comprising: (a) fermenting Gram-positive or Gram-negative bacteria in a growth medium to stationary growth phase to produce a fermentation broth; (b) harvesting the bacteria from the fermentation broth; (c) pasteurizing the harvested bacteria; and (d) lysing the pasteurized bacteria with lysozyme to produce a bacterial lysate. In some embodiments, the lysate is harvested at different times in the growth cycle after the start of any particular fermentation. In some embodiments, the bacteria are fermented in a characteristically defined medium.

[0142] In another aspect, the present disclosure provides a method for reducing one or more oxidative stress-related side effects associated with administration of a pharmaceutical agent, the method comprising administering a therapeutically effective amount of a composition comprising (a) a bacterial lysate and / or lysate fraction, (b) an optional promoter for increasing the absorption of the composition, and (c) an optional carrier for increasing the volume of the composition, in combination with the pharmaceutical agent, wherein the pharmaceutical agent and the lysate composition are administered simultaneously or in any order, and via the same or different routes of administration. In some embodiments, the lysate and / or lysate fraction activates at least one or more TLRs or NLRs. In some embodiments, the lysate and / or lysate fraction activates at least two TLRs and / or NLRs. In some embodiments, the lysate and / or lysate fraction activates at least three TLRs and / or NLRs.

[0143] In some embodiments, the one or more oxidative stress related side effects are selected from the group consisting of aceruloplasminemia, arterial / systemic hypertension, arthritis, asthma, atherosclerosis, atopic dermatitis, cancer, bladder cancer, leukemia, uterine cancer, cervical cancer, dizziness, nausea, vomiting, constipation, diarrhea, insomnia, drowsiness, lightheadedness, decreased libido, memory loss, tremors, jaundice, arrhythmia, increased heart rate, decreased heart rate, hives, depression, clinical depression, cerebral ischemia, bronchopulmonary dysplasia, cardiovascular disease, cataracts, cellulitis, chemotherapy side effects, chronic fatigue syndrome, colitis, coronary artery disease, dyslipidemia, eclampsia, erectile dysfunction, ataxia, headache, heart failure, hemodialysis side effects, liver The condition is selected from the group consisting of cirrhosis, hypercholesterolemia, hyperhomocysteinemia, hyperlipidemia, interstitial lung disease, lung injury, macular degeneration, male infertility, mild cognitive impairment, myocardial infarction, myocarditis, myocardial degeneration, neuropathy, obesity, osteoarthritis, osteoporosis, pancreatitis, periodontal disease, side effects of peritoneal dialysis, post traumatic stress disorder, pre-eclampsia, psoriasis, psoriatic arthritis, pulmonary hypertension, side effects of radiation therapy, reactive arthritis, respiratory distress syndrome, rhabdomyolysis, rheumatic disease, sepsis, sleep apnea, stroke, suicidal thoughts, amyloidosis, thrombophily, tauopathy, unstable angina, uremia, and venous insufficiency.

[0144] In another aspect, the disclosure provides a method for treating an oxidative stress-related disease or condition in a subject, the method comprising administering to the subject a therapeutically effective amount of a composition comprising: (a) a bacterial lysate and / or a lysate fraction capable of activating at least one or more toll-like receptors (TLRs) or Nod-like receptors (NLRs); (b) an optional promoter for increasing absorption of the composition; and (c) an optional carrier for increasing the volume of the composition.

[0145] In some embodiments, the oxidative stress associated condition is aceruloplasminemia, acute and chronic alcoholic liver disease, acute autoimmune myocarditis, sickle cell disease acute chest syndrome, acute pancreatitis, acute respiratory distress syndrome, alcoholic liver disease, amyotrophic lateral sclerosis, arterial / systemic hypertension, asbestosis, asthma, ataxia telangiectasia, atherosclerosis, atopic dermatitis, cerebral ischemia, bronchopulmonary dysplasia, burns, some cancers, cardiopulmonary bypass, cardiovascular disease, cataracts, cellulitis, chemotherapy side effects, chronic fatigue syndrome, chronic hepatitis C, chronic kidney disease, chronic obstructive pulmonary disease, chronic renal failure, colitis, coronary artery disease, Creutzfeldt-Jakob disease, Chronic Kidney Disease, Chronic Hepatitis C, Chronic Kidney Disease ... Rohn's disease, cutaneous leishmaniasis, pancreatic cystic fibrosis, type 1 diabetes, type 2 diabetes, dyslipidemia, Down's syndrome, eclampsia, end-stage renal disease, erectile dysfunction, Friedreich's ataxia, headache, heart failure, Helicobacter pylori infection / inflammation, side effects of hemodialysis, liver cirrhosis, human immunodeficiency virus infection, Huntington's disease, hyperbaric oxygen therapy-related diseases, hypercholesterolemia, hyperhomocysteinemia, hyperlipidemia, idiopathic pulmonary fibrosis, interstitial lung disease, ischemia / reperfusion injury, juvenile chronic arthritis, kidney transplant failure, leukemia, lung cancer, pulmonary disorders, macular degeneration, male infertility, Meniere's disease, meningitis, mild cognitive impairment, multiple sclerosis, myelodysplastic syndrome syndromes), myocardial infarction, myocarditis, neonatal bronchopulmonary dysplasia, obesity, osteoarthritis, osteoporosis, pancreatitis, Parkinson's syndrome, periodontal disease, side effects of peritoneal dialysis, photoageing, post-traumatic stress disorder, preeclampsia, primary biliary cirrhosis, broncopulmonary disease, progeria, psoriasis, psoriatic arthritis, pulmonary hypertension, side effects of radiation therapy, reactive arthritis, renal cell carcinoma, respiratory distress syndrome, retinopathy of prematurity, retrolental fibroplasia (RETF), olenticolarfibroplasy, rheumatoid disease, rheumatoid arthritis, sarcoidosis, sepsis, sickle cell disease, sleep apnea, spherocytosis, spinal cord injury, stroke, synucleinopathy, systemic amyloidosis, systemic lupus erythematosus, systemic sclerosis, thrombophilia, tauopathy, traumatic stress tuberculosis, variable angina, uremia, venous insufficiency, Werner's syndrome, or Zellweger's syndrome.

[0146] In another aspect, the disclosure provides a method for reducing oxidative stress in a subject, the method comprising: (a) measuring the level of oxidative stress in the subject by measuring the amount of isoprostanes in the urine or blood of the subject; (b) administering to the subject an effective amount of a composition comprising: (i) a toll-like receptor (TLR) agonist comprising at least one lysate and / or lysate fraction from a gram-negative or gram-positive bacterium, wherein the TLR agonist activates at least one or more different TLRs or NLRs; and (ii) an optional promoter for increasing absorption of the composition; and (c) continuing administration of the composition until the level of oxidative stress is reduced, as measured by a decrease in the amount of isoprostanes in the subject's urine.

[0147] In some embodiments of any of the above aspects, administration of the bacterial lysate continues until the amount of isoprostanes in the subject's urine is less than about 3 ng / mg creatinine, less than about 2 ng / mg creatinine, less than about 1 ng / mg creatinine, or less than about 0.5 ng / mg creatinine.

[0148] In another aspect, the disclosure provides a pharmaceutical formulation comprising: (a) a lysate composition comprising: (i) a bacterial lysate and / or a lysate fraction capable of activating at least one or more toll-like receptors (TLRs) or Nod-like receptors (NLRs); (ii) an optional promoter for increasing absorption of the composition; and (iii) an optional carrier for increasing the volume of the composition; and (b) one or more pharmaceutical agents.In some embodiments, the one or more pharmaceutical agents are selected from the group consisting of: antispasmodics, motility stimulants, H2 receptor antagonists, antimuscarinics; chelating agents, prostaglandin analogs, aminosalicylates, corticosteroids, drugs affecting immune response, stimulant laxatives, drugs affecting bile composition and flow, bile acid sequestrants, dopamine antagonists, proton pump inhibitors, opioids, opioid receptor antagonists, analgesics, hypnotics, cardiac glycosides, phosphodiesterase inhibitors, thiazides, diuretics, potassium Reserve diuretics, aldosterone antagonists, osmotic diuretics, drugs for arrhythmias, beta-adrenergic receptor blockers, hypertensive drugs, drugs affecting the renin-angiotensin system, nitrates, calcium blockers, antianginal drugs, peripheral vasodilators, sympathomimetics, anticoagulants, protamines, antiplatelet drugs, fibrinolytic drugs, antifibrinolytic drugs, lipid regulating drugs, omega-3 fatty acid compounds, CNS drugs, anti-infective drugs, or benzothoron, procyclidine, biperiden, amantadine, bromocriptine, pergolide, entacapone, tolcapone , Selegiline, Pramipexole, Budesonide, Formoterol, Quetiapine Fumarate, Olanzapine, Pioglitazone, Montelukast, Zoledronic Acid, Valsartan, Latanoprost, Irbesartan, Clopidogrel, Atomoxetine, Dexamfetamine, Methylphenidate, Modafinil, Bleomycin, Dactinomycin, Daunorubicin, Idarubicin, Mitomycin, Mitoxantrone, Azacitidine, Capecitabine, Cladribine, Clofarabine, Cytara and another agent selected from the group consisting of rifabutin, fludarabine, flourouracil, gemcitabine, mercaptopurine, methotrexate, nelarabine, pemetrexed, raltitrexed, thioguanine, apomorphine, betamethasone, cortisone, deflazacort, dexamethosone, hydrocortisone, methylprednisolone, prednisolone, triamcinolone, cyclosporine, sirolimus, tacrolimus, interferon alpha, and interferon beta.

[0149] In another aspect, the disclosure provides a formulation, such as a medical formulation, comprising: (a) a lysate composition comprising: (i) a bacterial lysate and / or a lysate fraction capable of activating at least one or more toll-like receptors (TLRs) or Nod-like receptors (NLRs); (ii) an optional promoter for increasing the absorption of the composition; and (iii) an optional carrier for increasing the volume of the composition; and (b) an isolated human anti-TNFα antibody or antigen-binding fragment thereof or a TNF inhibitor. In some embodiments, the human anti-TNFα antibody or antigen-binding fragment thereof is adalimumab. In some embodiments, the disclosure provides a use of a combination formulation of this aspect in the manufacture of a medicament for the treatment of chronic rheumatoid arthritis (RA) or late-onset RA in a subject. In some embodiments, the disclosure provides a method for the treatment of rheumatoid arthritis (RA) or late-onset RA in a subject, the method comprising administering to the subject a therapeutically effective amount of a combination formulation of this aspect, as disclosed herein. In some embodiments, the subject is 50 years of age or older. In some embodiments, the human anti-TNFα antibody or antigen-binding fragment thereof is administered to the subject in a biweekly dosing regimen. In some embodiments, the human anti-TNFα antibody or antigen-binding fragment thereof is administered to the subject at a dose of 30 mg or more. In some embodiments, the TNFα inhibitor is a TNFα fusion protein. In some embodiments, the TNFα fusion protein is etanercept. In some embodiments, the anti-TNFα antibody or antigen-binding fragment thereof is infliximab or golimumab. In some embodiments, the anti-TNFα antibody or antigen-binding fragment thereof is adalimumab.

[0150] In another aspect, the disclosure provides a method for inhibiting the progression of an oxidative stress related disease or disorder in a human subject having an oxidative stress related disease or disorder associated with the disorder in which TNFα activity is detrimental, the method comprising administering to a subject having an oxidative disease or disorder a lysate, lysate fraction and / or cell wall fraction of the disclosure, and an isolated human anti-TNFα antibody, or antigen-binding portion thereof, such that progression of the oxidative stress disease is inhibited in the subject, and the human anti-TNFα antibody, or antigen-binding portion thereof, is at least 1×10 in a standard in vitro L929 assay. -7 M or lower IC 50 In some embodiments, the lysate of the present disclosure and the human anti-TNF-α or antigen-binding portion thereof are administered at different times under different dose regimens. In some embodiments, the human anti-TNFα antibody is golimumab or an antigen-binding portion thereof. In some embodiments, the human anti-TNFα antibody or an antigen-binding portion thereof is adalimumab or an antigen-binding portion thereof. In some embodiments, the human anti-TNFα antibody or an antigen-binding portion thereof is administered to the subject in a biweekly dosing regimen. In some embodiments, the method further comprises administering an additional therapeutic agent to the subject. In some embodiments, the lysate of the present disclosure is administered for a period of at least 24 weeks. In some embodiments, the anti-TNFα antibody or an antigen-binding portion thereof of the present disclosure is administered for a period of at least 24 weeks. In some embodiments, the lysate of the present disclosure, the anti-TNFα antibody, or an antigen-binding portion thereof, or both, repair or prevent oxidative damage to the subject by a combination of one or more different mechanisms.

[0151] In some embodiments of any of the methods or compositions disclosed herein, the bacteria is a gram-positive or gram-negative bacterium. In some embodiments of any of the methods or compositions disclosed herein, the gram-positive bacterium is selected from the group consisting of bacteria of the Lactobacillaceae family, bacteria of the Streptococcus family, bacteria of the Bifidobacteriaceae family, and bacteria of the Bacillaceae family. In some embodiments, the gram-positive bacterium is selected from the group consisting of Bacillus coagulans, Lactobacillus sporogenes, Streptococcus thermophilus, Bifidobacterium animalis, Bifidobacterium animalis, subspecies animalis, Bifidobacterium infantis, Bifidobacterium longum, Bifidobacterium breve, Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus spp. ... The bacterial strain is selected from the group consisting of Bacillus casei, Lactobacillus delbrueckii, Lactobacillus delbrueckii subsp. bulgaricus, Lactococcus lactis, Lactococcus lactis subsp. lactis, Streptococcus lactis, Streptococcus thermophilus, Bifidobacterium lactis, Bifidobacterium breve, Pediococcus acidilactici and Lactobacillus helveticus.

[0152] In some embodiments of any of the methods or compositions (including formulations) disclosed herein, the Gram-negative bacteria is selected from the group consisting of bacteria of the genera Pseudomonas, Klebsiella, Xanthomonas, Shigella, and Enterobacter. In some embodiments, the Gram-negative bacteria is selected from the group consisting of Klebsiella oxytoca, Shigella flexneri, Xanthomonas, and Pseudomonas fluorescens.

[0153] In some embodiments of any of the methods or compositions (including formulations) disclosed herein, the TLR agonist, lysate, lysate fraction, or cell wall fraction activates at least one or more of TLR2, TLR3, TLR4, TLR5, TLR7, TLR8, TLR9, NOD1, and NOD2. In some embodiments, the TLR agonist, lysate, lysate fraction, or cell wall fraction activates two or more of TLR2, TLR3, TLR4, TLR5, TLR7, TLR8, TLR9, NOD1, and NOD2. In some embodiments, the TLR agonist, lysate, lysate fraction, or cell wall fraction activates TLR2 and TLR4. In some embodiments, the TLR agonist, lysate, lysate fraction, or cell wall fraction activates three or more of TLR2, TLR3, TLR4, TLR5, TLR7, TLR8, TLR9, NOD1, and NOD2.

[0154] In some embodiments of any of the methods or compositions (including formulations) disclosed herein, the promoter is selected from the group consisting of an amino acid, an amino sugar, and a sugar. In some embodiments, the carrier is selected from the group consisting of a binder, a gum base, and combinations thereof. In some embodiments, the gum base comprises at least one hydrophobic polymer and at least one hydrophilic polymer. In some embodiments, the binder is selected from the group consisting of a sugar, a sugar alcohol, and combinations thereof. In some embodiments, the sugar alcohol is selected from the group consisting of mannitol, sorbitol, xylitol, and combinations thereof.

[0155] In some embodiments, the composition is prepared as a dosage form selected from the group consisting of lozenges, chewing gum, chewable tablets, candies, and dissolving tablets.In some embodiments, the dosage form delivers the TLR agonist to oral mucosa.In some embodiments, the oral mucosa is selected from the group consisting of sublingual mucosa, buccal mucosa, and combinations thereof.

[0156] In some embodiments of any of the methods or compositions (including formulations) disclosed herein, the composition is formulated for oral mucosal delivery. In some embodiments, the composition is formulated for sublingual or buccal delivery. In some embodiments, the composition is formulated to dissolve within 1 minute after administration. Working Example

[0157] The following examples are illustrative of certain embodiments of the invention and various uses thereof, they are provided for illustrative purposes only and should not be construed as limiting the invention. Example 1: Preparation of active ingredients for TLR agonist compositions

[0158] An example of a method for formulating the composition includes the following process.

[0159] Active Ingredients. The active ingredients were derived from the bacterial fermentation and cell isolation process described below. Lactobacillus delbrueckii subsp. bulgaricus (referred to herein as L. bulgaricus) was the organism used in this example, however the bacterial organism may be any gram-positive or gram-negative bacterium, or both.

[0160] Lactobacillus delbrueckii subsp. bulgaricus was fermented in a growth medium containing 1.5% casein hydrolysate, 1% yeast extract, 3% lactose, 0.2% sodium acetate, 0.02% sodium formate, 0.01% 5'-disodium inosinate, 0.01% manganese sulfate, 0.05% magnesium sulfate, and 0.05% polysorbate 80 at pH 6.4. The seed medium was incubated at 37 °C until the fermentation reached stationary growth as determined by cessation of metabolism. The fermentation broth was cooled to 4 °C and harvested by centrifugation at approximately 4000–4500 × g.

[0161] The concentrated cells were washed twice successively with cold water and the cells were centrifuged again. The washed cell concentrate was pasteurized at 80° C. for 45 min.

[0162] After pasteurization, the cell concentrate was treated to disrupt the cell walls, thereby lysing the bacteria and exposing the TLR agonists. Disruption of the bacterial cell walls was achieved with hen egg white lysozyme at a concentration of 3% by weight for 7 hours at 40° C. The resulting lysate was frozen and lyophilized.

[0163] The freeze-dried material was blended with a promoter such as N-acetylglucosamine HCl (NAG) to form a mixture of dissolved Lactobacillus delbrueckii subsp. bulgaricus and NAG. Optionally, other formulation excipients were added appropriately to produce a solid form pill or powder. This product was then used in the following screening test. Example 2: TLR screening assay

[0164] Toll-like receptor (TLR) stimulation was tested by assessing activation of NF-κB in HEK293 cells expressing a given TLR or Nod-like receptor (NLR). Activity of samples was tested on seven different human TLRs: TLR2, 3, 4, 5, 7, 8 and 9 (Invivogen, San Diego, CA), and two different human NLRs, nucleotide-binding oligomerization domain-containing proteins 1 and 2 (NOD1 and NOD2). Each ligand was tested on TLR or NLR cells at a final concentration of 1 / 100 of the stock solution and compared to the control ligands described below. This step was performed in triplicate.

[0165] The control ligands, control cell lines and sample products used in the examples are as shown in Table 1. [Table 1]

[0166] General procedure. TLR stimulation in the screen is tested by assessing the activation of NF-κB in HEK293 cells expressing a given TLR. A secreted alkaline phosphatase reporter is under the control of a promoter inducible by the transcription factor NF-κB. TLR stimulation in the screen was tested by assessing the activation of NF-κB in HEK293 cells expressing a given TLR or NLR. This reporter gene allows the monitoring of TLR / NLR-mediated signaling based on the activation of NF-κB. In a 96-well plate (total volume 200 μL) containing the appropriate cells (50,000-75,000 cells / well), 20 μL of sample (lysate) or positive control ligand is placed in the wells. The medium added to the wells is designed to detect the expression of NFκB-induced SEAP (secreted alkaline phosphatase). After 16-20 hours of incubation, the OD (optical density) at 650 nm was read and recorded with a Molecular Devices Spectra Max 340PC absorbance detector.

[0167] Screening results from these experiments are shown graphically in Figure 2. These results indicate that the exemplary lysates generated as described above are potent agonists for at least TLR2 (and to a lesser extent TLR4) and NOD2. Example 3: Effect of changing process variables on TLR signaling

[0168] It should be noted that TLR signaling patterns can be altered by observing the effects of altering specific processes on TLR signaling.

[0169] Figure 3 shows the differences in cell morphology versus TLR signaling for Gram-positive bacteria. Lysates from the coccus organism Pediococcus acidilactici produced more TLR4 and less NOD2 signal compared to the bacillus L. bulgaricus, and the TLR2 signal produced by P. acidilactici was only slightly more than that produced by L. bulgaricus. NOD2 activation was significantly lower in P. acidilactici reflecting the fewer muramyl peptides found in coccus.

[0170] As shown in Figure 4, the TLR signal pattern was also affected by the culture harvest time of Bacillus coagulans lysates. Bacillus coagulans was grown using standard yeast extract / glucose medium with pH 6.5 in shake flasks incubated at 45 °C and 250 rpm. Culture harvesting followed the lysate generation process of Example 1, but by varying the bacterial harvest time after fermentation according to the growth phase: mid-logarithmic, late-logarithmic, early stationary, mid-stationary, and late stationary, as determined by OD, substrate exhaustion, metabolite production, and EPS. In general, these data indicate that the specificity of TLR activation can be altered by varying the bacterial harvest time after fermentation. For example, lysates generated from cultures harvested at mid-logarithmic phase activated all targets tested except TLR4, with the highest signal for TLR2, 3, 5, 8, 9, and NOD1. TLR signal intensity decreased as the cultures left the logarithmic phase and entered stationary phase, with TLR signaling ceasing completely at late stationary phase. Cell cultures were plated at all harvest stages and cell numbers were 5 × 10 at all time points. 8 Note that the numbers are greater than CFU / ml. Visual inspection of the cultures did not reveal endospores. The TLR effect seen in the lysates was from vegetative cells. Large amounts of EPS were seen in the side walls of late stationary cultures.

[0171] Figure 5 shows the TLR signal patterns obtained from lysates of selected Gram-positive bacteria. Streptococcus thermophilus, Lactobacillus helveticus, Lactobacillus acidophilus, Lactobacillus plantarum, Bifidobacterium animalis subsp. lactis and Bacillus subtilis were grown on standard yeast extract / lactose / casein hydrolysate medium at pH 6.5 and incubated at 37°C and 110 rpm in shake flasks. Lactobacillus plantarum, Bifidobacterium animalis subsp. lactis (B. lactis), and Bacillus subtilis were grown on yeast extract medium / glucose medium at pH 6.5 in shake flasks and incubated at 37°C and 110 rpm, while B. subtilis was incubated at 37°C and 250 rpm. For B. lactis, 0.05% cysteine ​​HCL was added to the medium. Harvesting of the cultures followed the lysate production process of Example 1. S. thermophilus (coccus organism) produced a similar TLR signal pattern (high TLR2 and TLR4, and low NOD2) to another coccus culture, P. acidilactici, as described in Example 4. However, B. lactis produced a similar TLR pattern, although morphologically different, to L. bulgaricus. There was little similarity in the TLR patterns of the Gram-positive bacillus cultures of L. helveticus, L. acidophilus, L. plantarum, and B. subtilis. L. helveticus produced a TLR pattern similar to that of L. bulgaricus. L. acidophilus produced results similar to the pattern produced by coccus, although with lower signal intensity. When the cultures were harvested and concentrated, L. acidophilus did not produce any significant EPS, whereas L. bulgaricus, L. helveticus and B. lactis all produced significant EPS and all produced similar TLR signal patterns. L. plantarum lysates had a very different TLR signal pattern than other Gram-positive bacteria, all of which produced weaker TLR signals. This muting of the signal pattern was not evident from the fermentation characteristics, but L. plantarum is known to produce a number of metabolites that may explain the reduced TLR signal.B. subtilis had a similar TLR pattern to log-phase harvested cultures of B. coagulans except that B. subtilis produced higher TLR4 activation. Overall, the TLR signaling patterns across Gram-positive bacteria varied with organism morphology, EPS production, and bacterial metabolic products.

[0172] Figure 6 shows the TLR signal patterns obtained from lysates of selected Gram-negative bacteria. E. coli, Klebsiella oxytoca, Shigella flexneri, Pseudomonas fluorescens, and Xanthomonas campestris were grown in shake flasks with tryptic soy broth. Klebsiella oxytoca, E. coli, and S. flexneri were incubated at 37°C and 250 rpm, and P. fluorescens and Xanthomonas campestris were incubated at 30°C and 250 rpm. Xanthomonas campestris was grown in yeast extract / glucose medium supplemented with 1% calcium carbonate. Before harvesting, CaCO3 was removed by centrifugation at 500 x g for 3 min. Cultures were harvested from the fermentation broth and then subjected to the lysate generation process of Example 1. Surprisingly and unexpectedly, the TLR signal patterns of lysates from Gram-negative bacteria such as E. coli were much lower than those of Gram-positive bacteria when tested at similar concentrations, except for TLR5 and 9. In general, the membrane / cell wall of Gram-negative bacteria is more easily damaged than that of Gram-positive bacteria, which may explain the higher TLR9 signal. K. oxytoca and X. campestris were observed to produce EPS during fermentation, which also resulted in relatively high activation of TLR3, 4 and 9. K. oxytoca resulted in the highest activation of TLR2 and NOD1 among the strains tested. EPS appears to alter the TLR signal pattern compared to a non-EPS producing strain (Shigella flexneri). Interestingly, all Gram-negative bacteria had significant activation of NF-κB in null cells in the TLR assay. This NF-κB activity was subtracted from the TLR signal, explaining the slightly negative activity for TLR7 and NOD2 of P. fluorescens.

[0173] Finally, Figure 7 shows the TLR signaling of a lysate composition containing xanthan gum. These data show that the lysate produced from the Xanthomonas campestris strain as well as the EPS produced by the Xanthomonas campestris strain, xanthan gum itself, both result in TLR activation. There is a strong stimulation by the strain and the EPS xanthan gum at TLR5 and moderate activity at TLR4 and 9. These data suggest that the lysate from a given bacterial strain as well as the corresponding EPS from that strain (xanthan gum in this example) can both be used or blended into a formulation to target TLR activity.

[0174] Surprisingly and unexpectedly, these data indicate that the TLR signal pattern resulting from a given lysate, as well as the strength of the TLR signal, is dependent on the manufacturing process and choice of raw materials, including but not limited to, media, microorganisms, etc. These data indicate that the specificity of TLR activation can be altered and tuned by varying process, bacterial organisms, raw materials, and reagent parameters. Example 4: Indicators of oxidative stress, cognition, and quality of life in combat veterans diagnosed with post-traumatic stress disorder (PTSD)

[0175] PTSD is an oxidative stress-related disease or condition. The objectives of this study were fourfold: (1) to evaluate the effectiveness of the composition of the present invention in quantitatively reducing the level of isoprostanes, a marker of oxidative stress, in combat veterans diagnosed with PTSD; (2) to evaluate the effectiveness of the composition of the present invention in quantitatively reducing the level of nitric oxide in combat veterans diagnosed with PTSD; (3) to evaluate the effectiveness of the composition of the present invention in improving word finding and word recall in combat veterans diagnosed with PTSD; and (4) to evaluate the effectiveness of the composition of the present invention in improving quality of life parameters in combat veterans diagnosed with PTSD.

[0176] Emerging evidence-based literature supports a link between oxidative stress and post-traumatic stress disorder (PTSD). Oxidative stress occurs in the body and at the cellular level as a result of an imbalance between circulating oxidants and antioxidants. Many studies have shown that this cellular oxidative stress leads to free radical effects on the neurophysiology of the body. Nitric oxide has been shown to play a key role in protecting against oxidative stress.

[0177] Neurocognition is often affected in individuals who have suffered traumatic brain injury and in combat veterans. Closed head injury / traumatic brain injury (CHI / TBI) and PTSD are often co-occurring disorders.

[0178] Inclusion Criteria

[0179] The expected enrollment in this study was a minimum of 10 and a maximum of 15 patients.

[0180] The expected gender distribution was as close as possible to 50% male and 50% female. Due to the large number of male veterans diagnosed with PTSD, it is possible that there was a slightly higher proportion of males in the participant pool.

[0181] Pregnant women were not allowed to participate in this study.

[0182] The only age restriction was due to the minimum recruitment age for the military, 18 years, so the expected age range for the study was from 18 to approximately 80 years, with the upper limit determined by health level.

[0183] There were no restrictions based on race or ethnicity. The intended proportions based on race reflected those of the larger Houston area from which participants were recruited. These proportions were mostly approximately 44% Hispanic, 26% White, 23% African American, 6% Asian, and 1% other races.

[0184] Study inclusion and exclusion criteria are outlined in Table 2 below. [Table 2] Methods and Procedures

[0185] Participants, with informed consent, were evaluated to assess parameters such as levels of cellular oxidative stress, nitric oxide levels, language / word-finding ability, levels of anxiety and depression, and current quality of life.

[0186] Participants were provided with the composition for 15 days, which is hypothesized to reduce oxidative stress, as indicated by, among other things, reduced urinary concentrations of isoprostanes. Two 12 mg product tablets were taken sublingually twice daily for a total of 48 mg / day.

[0187] To minimize measurement variability, no placebo was used and subjects served as their own measurement controls. All subjects were assigned to the same study groups and underwent the same procedures.

[0188] Oxidative stress was measured by urine testing for isoprostanes using a commercially available assay (Oxford Biomedical Research). Collected samples were standardized for creatine to control for differences in urinary concentration levels using a creatinine assay kit (Oxford Biomedical Research).

[0189] Each participant had their urine tested for isoprostane levels at least on days 1 and 15. A consistent group of patients was asked to provide urine samples for each of the 15 study days to establish data points as a means to examine possible changes in isoprostane over time. A third party (blind to the study objectives) was retained to perform the isoprostane assays.

[0190] All participants completed three language tests in a private room on days 1 and 15. The language tests used were the ROWPVT-4 (Receptive One-Word Picture Vocabulary Test, 4th edition), the EOWPVT-4 (Expressive One-Word Picture Vocabulary Test, 4th edition), and the TONI-4 (Test of Non-Verbal Intelligence, 4th edition).

[0191] In addition, each participant was asked to complete the Zung Depression Self-Rating Inventory, the Satisfaction with Life Scale, and the Eppworth Sleepiness Scale on days 1 and 15. These forms were completed in a private room and coded using standard methods that concealed participant identities.

[0192] Each participant completed a daily Quality of Life (QOL) questionnaire that was emailed each morning at 4 a.m. To familiarize each participant with the QOL survey format and establish a more robust database, each participant began recording their scores on the QOL questionnaire for a minimum of four days before beginning product use. Data Analysis and Monitoring

[0193] After the above parameters were collected over a 15-day period, the study data were analyzed for trends in identification of oxidative stress and nitric oxide levels, word-finding ability, presence of anxiety / depression, and overall quality of life scores. The statistical analysis method used was simple, with only one quantitative parameter, isoprostanes, assessed. Data were analyzed using appropriate multivariate statistical tests. Risk / benefit assessment

[0194] Risk Category: The risk category for this endeavor was "minimal." Therefore, consent regarding injury language on the consent form was not required and thus no Clinical Trial Agreement (CTA) was required. The minimal risk category was due to the fact that the product being tested was recognized as a food-grade GRAS compound found in yogurt. Additionally, no invasive procedures were performed as urine and saliva were the only bodily fluids collected.

[0195] At the time of data collection, there were approximately 35 individuals of both genders who had taken the product. At the time of the study, the individual with the longest history of use had begun using the product 17 months prior and had not reported any negative side effects from using the product. There were no reports of negative side effects among the approximately 35 participants who were administered the product. Methods of subject identification and recruitment

[0196] This study used a convenience sample and the recruitment method of participants was based primarily on the researcher's personal and professional contacts. Target Ability

[0197] If an individual was deemed to lack sufficient capacity to consent, they could not take part in the study. The study did not include minors and no one was required to provide consent or have a legal representative. Understanding of the subject

[0198] To assess participants' proper understanding, they were asked to explain the study to the person conducting the consent process before enrolling in the study. Subjects confirmed that they had answered all questions appropriately. result

[0199] In the study, the effects of the supplements were assessed by measuring / analysing: 1) isoprostane levels, 2) quality of life survey results, 3) Zung / psychological survey results, and 4) speech-language outcomes.

[0200] As shown in Figure 8, treatment with the supplements caused a decrease in urinary isoprostane levels. Isoprostanes are prostaglandin-like compounds formed in vivo from free radical-catalyzed peroxidation of essential fatty acids (mainly arachidonic acid) without the direct action of the cyclooxygenase (COX) enzyme normally involved in the production of prostaglandins. These compounds are accurate markers of lipid peroxidation in both animal and human models of oxidative stress. At day 15, most subjects showed a decrease in isoprostane levels. By day 70, all subjects showed lower levels of isoprostane than on day 1.

[0201] As seen in Tables 3A and 3B, treatment with the composition significantly reduced the amount of isoprostane after only 15 days of treatment. Table 2A shows the raw data of the study, and Table 3B shows the results after calculating the percentage reduction and improvement of oxidative stress markers. In Tables 3A and 3B, "PTSD+" are participants diagnosed with PTSD. "PTSD-" are participants not diagnosed with PTSD. "TX+" are participants who used the composition before the study. "TX-" are participants who did not use the composition.

[0202] As can be seen in Table 3B, subjects demonstrated a 60% reduction in isoprostane levels after 15 days of treatment, followed by an additional 3.18% reduction in isoprostane levels after 70 days of treatment. Additionally, subjects demonstrated a 469% improvement in oxidative stress biomarker levels after 15 days of treatment, followed by an additional 53.19% improvement in oxidative stress biomarker (isoprostane) levels after 70 days of treatment. [Table 3] [Table 4]

[0203] Subjects were asked to self-assess and rate their sleep. The rating was on a scale of 1 to 10, with a value of 1 corresponding to 1 sleep deprivation and a value of 10 corresponding to no sleep deprivation. As seen in FIG. 9, by day 70, all subjects showed a reduction in sleep deprivation compared to their self-assessed sleep quality rating on day 1.

[0204] Subjects were asked to self-assess and rate their neuropathic symptoms. The rating was on a scale of 1 to 10, with a value of 1 corresponding to major discomfort and pain from neuropathic symptoms and a value of 10 corresponding to minimal discomfort and pain from neuropathic symptoms. As seen in FIG. 10, at day 15, all subjects except two (P03 and P04) showed minimal discomfort and pain from neuropathic symptoms compared to day 1. At day 70, all patients except one (P03) showed minimal discomfort and pain from neuropathic symptoms or, in the case of P04, no difference in discomfort compared to day 1 (note that P04 showed minimal discomfort and pain from neuropathic symptoms at the start of the study).

[0205] Subjects were asked to self-assess and rate their overall mood. Ratings were on a scale of 1 to 10, with a value of 1 corresponding to a generally unhappy or anxious mood and a value of 10 corresponding to a generally happy or relaxed mood. As can be seen in FIG. 11, at day 15, all subjects except two showed improvement in overall mood. At day 70, all patients except one (P04) showed improvement in overall mood compared to day 1.

[0206] Subjects were asked to self-assess and rate their overall energy levels on a scale of 1 to 10, with a value of 1 corresponding to a low level of energy and a value of 10 corresponding to a high level of energy. As can be seen in FIG. 12, on days 15 and 70, all subjects (except P03) showed an increase in energy levels compared to day 1.

[0207] Subjects were asked to self-assess and rate their overall satisfaction with their joint health (as determined by assessing several factors, such as flexibility, stiffness, and pain). The ratings were on a scale of 1 to 10, with a value of 1 corresponding to a low level of satisfaction and a value of 10 corresponding to a high level of satisfaction. As can be seen in FIG. 13, at day 15, all subjects (except P03 and P04) showed improved levels of joint satisfaction compared to day 1. This trend continued at day 70.

[0208] Subjects were asked to self-assess and rate their overall satisfaction with their digestive health. The rating was on a scale of 1 to 10, with a value of 1 corresponding to a low level of satisfaction and a value of 10 corresponding to a high level of satisfaction. As seen in FIG. 14, on day 15, all subjects (except P01 and P04) showed improved levels of satisfaction with their digestive health compared to day 1. This trend continued on day 70.

[0209] Subjects were asked to self-assess and rate their overall level of irritability on a scale of 1 to 10, with a value of 1 corresponding to a high level of irritability and a value of 10 corresponding to the lowest level of irritability. As can be seen in Figure 15, at day 70, all subjects showed a decrease in their level of irritability compared to day 1.

[0210] Subjects were asked to self-assess and rate their overall satisfaction level with sexual function. The rating was on a scale of 1 to 10, with a value of 1 corresponding to a low level of satisfaction and a value of 10 corresponding to a high level of satisfaction. As can be seen in FIG. 16, at day 70, all subjects (except P04 and P05) showed an increase in the level of satisfaction with sexual performance compared to day 1.

[0211] Subjects were asked to self-assess and rate their overall level of daytime sleepiness according to the guidelines of the Epworth Sleepiness Scale (ESS). The ESS is a self-administered questionnaire that includes eight questions. This questionnaire provides a measure of a person's general level of daytime sleepiness, or average sleep tendency in daily life. It is the global standard method for making this assessment. The ESS asks subjects to rate, on a four-point scale (0-3), their usual opportunities to doze off or sleep in eight different situations or activities that most people engage in as part of their daily lives, but which do not necessarily occur daily. It is not asking how often they doze off in each situation. This will depend largely on how frequently these situations occurred. Rather, it is asking whether the opportunity was always an opportunity to doze off when they were in each situation. This requires mental judgment, which most people are likely to be able to produce in a meaningful way. The total ESS score is the sum of the scores of the eight items and can range from 0 to 24. The higher the score, the higher the level of daytime sleepiness. The total ESS score provides an estimate of each participant's general characteristic, i.e., their average level of sleepiness in everyday life.

[0212] As can be seen in Figure 17, all subjects experienced a decrease in daytime sleepiness by Day 15. At Day 70, all subjects who completed the study (except P03) demonstrated a decrease in daytime sleepiness compared to Day 1.

[0213] Subjects were asked to self-assess and rate their overall level of life satisfaction. Ratings were on a scale of 1 to 40, with a value of 1 corresponding to a low level of satisfaction and a value of 40 corresponding to a high level of satisfaction. As can be seen in FIG. 18, at day 15, all subjects showed increased levels of life satisfaction compared to day 1. This trend continued through day 70.

[0214] Subjects were asked to self-assess and rate their overall anxiety levels according to the guidelines of the Zung Self-Rating Depression Scale, designed by Duke University psychiatrist Dr. William W. K. Zung (1929-1992) to assess levels of depression in patients diagnosed with depressive disorders.

[0215] The Zung Self-Rating Depression Scale is a short, self-administered survey to quantify a patient's state of depression. There are 20 items on the scale that rate the emotional, psychological, and physical symptoms associated with depression. There are 10 positive-word questions and 10 negative-word questions. Each question is scored on a scale of 1 to 4 (based on the following answers: "almost never," "sometimes," "most of the time," and "most of the time"). The range of scores on the test is 20 to 80. Scores are classified into four ranges: 20 to 44 is associated with the normal range, 45 to 59 is associated with a mildly depressed state, 60 to 69 is associated with a moderately depressed state, and 70 or above is associated with a severely depressed state. As can be seen in Figure 19, on day 15, all subjects except P01 and P05 showed a decrease in depression levels compared to day 1. On day 70, subjects P02, P03, P04, P06, and P07 all showed a decrease in anxiety levels compared to day 1. Example 5: Reduction of oxidative stress-related side effects

[0216] Relief from headaches.A 17-year-old male who had suffered three concussions in the 1.5 years prior to treatment was administered the lysate of the present disclosure at a dose of 45 mg twice daily. On day 14 of treatment, the dose was increased to 75 mg of active ingredient twice daily. Prior to treatment, the patient reported frequent headaches. Pretreatment F2-isoprostanes (F2IsoP) were measured in urine at 2.4 ng / mg creatinine. Figure 20 shows that isoprostanes are decreasing at day 85 as measured in the patient's urine, with an average of 0.82 ng isoprostanes / mg creatinine from days 72 to 85. Letters A-H in FIG. 20 indicate the following events: (A) baseline F2IsoP levels on days 1-3, (B) severe headache on day 3, peak F2IsoP of 9.99 ng / mg, (C) days 1-13, using lysate 45 mg bid, with daily headaches, (D) day 13, patient begins using lysate 75 mg bid to address headaches. (E) days 18-89, headache resolved as of now. Continued for resolution. (F) day 30, new impact event from soccer. 18 hour headache with visual and neurological impairment. (G) day 81, situational stress event. Elevated F2IsoP levels, but no headache resulted. (H) days 1-89, trend line for isoprostanes showing a 98% reduction over day 89. Overall, these data suggest a concomitant reduction in oxidative stress in response to lysate administration. Additionally, subjects reported a significant reduction in headaches following lysate administration.

[0217] Reduction of other oxidative stress-related side effects. A patient experiencing one or more side effects associated with administration of a pharmaceutical agent is co-administered with the bacterial lysate of the present disclosure at a dose of 45-100 mg bid. Prior to initiating co-administration of the lysate, F2-isoprostane levels are measured in blood or urine. Co-administration of the lysate results in a repeat decrease in F2IsoP levels over time, with an overall decrease of about 50% in F2IsoP levels over about 45 days, and an increase of about 95% in F2IsoP levels over about 90 days, with a concomitant decrease in drug-associated oxidative stress-related side effects. Example 6: Lysate / adalimumab combination is effective in treating skin disease in psoriatic arthritis.

[0218] This study is conducted to evaluate the efficacy of a therapeutic composition comprising adalimumab in combination with the bacterial lysate composition of the present disclosure, administered together or separately, against skin disease, oxidative stress disorders in patients with psoriatic arthritis (PsA).

[0219] A randomized phase III study of adalimumab will be conducted in patients with active PsA (swollen joints ≥ 3, tender joints ≥ 3) who have failed NSAID treatment. Randomization will be stratified by methotrexate (MTX) use and extent of psoriasis at baseline (< 3% or ≥ 3% of body surface area [BSA]). Patients who complete 24 weeks will be eligible to continue in an open-label extension study.

[0220] Patients were eligible if they had a history of psoriasis, were 18 years of age or older, had 3 or more swollen joints and 3 or more tender joints, and had an inadequate response to NSAID therapy. Exclusion criteria included previous anti-TNF therapy, alefacept within 12 weeks prior to study enrollment, other biologics within 6 weeks prior to study enrollment, DMARDs (except MTX) within 4 weeks prior to study enrollment, systemic therapy for psoriasis within 4 weeks prior to study enrollment, and phototherapy and topical therapy within 2 weeks prior to study enrollment.

[0221] Patients will be stratified by methotrexate use (yes / no) and severity of psoriasis (including <3% and ≥3% BSA) and will receive oral 20-50 mg of lysate daily, 40 mg every other week, or placebo in combination with adalimumab for 24 weeks.

[0222] Efficacy measures used will include: ACR response criteria (co-primary endpoint, ACR20 response at week 12), Psoriasis Area and Severity Index (PASI) in patients with significant psoriasis at study enrollment, and Physician's Global Assessment of Psoriasis (PGA). Patients will be examined according to the severity of psoriasis at baseline: PASI<10 vs. PASI≥10.

[0223] Thus, at baseline, efficacy measures in psoriasis patients with BSA ≥ 3% include PASI, PGA for psoriasis, and DLQI. ACR response criteria are also used as efficacy measures. Post-hoc analyses will be performed for patients with baseline PASI < 10 vs. patients with PASI ≥ 10. PASI analysis will be by NRI, and PGA and DLQI scores will be calculated as LOCF.

[0224] The treatment group receiving the co-administered lysate experienced improved injection site reactions (such as pain, redness or irritation) and reduced overall oxidative stress-related side effects, allowing for improved compliance with treatment, which in turn provides a greater opportunity for improving the primary disease. Example 7: Lysate / statin / ACE inhibitor combination is effective in treating hypertension.

[0225] A human subject being treated for hypertension and receiving a combination of 20 mg pravastatin (QD) and 20 mg lisonopril (QD) experienced oxidative stress-related side effects of skin rash and insomnia.

[0226] When the patient was given an additional 45 mg solution (bid) encompassed by the disclosure herein, the side effects of skin rash and insomnia resolved within 60 days. Example 8: Lysate / SSRI combination is effective in treating depression.

[0227] Human subjects being treated for depression and receiving 60 mg duloxetine (CYMBALTA®) QD experienced oxidative stress-related side effects of anorgasmia, dizziness, numbness, and irritability.

[0228] In patients supplemented with 45 mg (bid) of a solution encompassed by the disclosure herein, anorgasmia resolved within 10 days, dizziness resolved within 7 days, numbness resolved within 10 days, and irritability resolved within 5 days. Example 9: Lysate / hormone combination is effective in treating hypothyroidism.

[0229] Human subjects being treated for hypothyroidism and receiving 150 mcg levothyroxine (SYNTHROID®) QD experienced an oxidative stress-related side effect: hair loss.

[0230] When the patient was given an additional 75 mg solution (bid) encompassed by the disclosure herein, the hair loss side effect resolved within 180 days. Example 10: Lysate / reverse transcriptase inhibitor combination is effective in treating HIV infection.

[0231] Human subjects undergoing treatment for HIV (non-AIDS) receiving efavirenz / emtricitabine / tenofovir disoproxil fumarate (ATRIPLA®) one tablet (QD) were experiencing oxidative stress-related side effects, including erectile dysfunction, neuropathy, insomnia, and brain fog.

[0232] When the patient was given an additional dose of 45 mg (bid) of a dissolve encompassed by the disclosure herein, erectile dysfunction resolved within 10 days, neuropathy resolved within 10 days, insomnia resolved within 3 days, and brain fog resolved within 3 days. Example 11: Lysate / direct acting antiviral combination is effective in treating Hepatitis C.

[0233] Human subjects being treated for Hepatitis C and receiving ledipastatvir / sofosviravir (HARVONI®) one tablet (QD) experienced oxidative stress-related side effects such as brain fog, headaches, and arthritis.

[0234] In patients receiving the additional dose of 75 mg (bid) of a solution contained within the present disclosure, brain fog resolved within 3 days, headache resolved within 10 days, and arthritis resolved within 14 days. Example 12: Lysate / anticancer drug combination is effective in treating cancer.

[0235] Human subjects undergoing treatment for cancer and receiving topoisomerase I inhibitors / vinca alkaloids / alkylating antineoplastic agents (IRINOTECAN®, VINCRISTIN®, TEMODAR®) have experienced oxidative stress-related side effects such as brain fog, fatigue, insomnia, and rashes.

[0236] In patients supplemented with 75 mg (bid) of a solution encompassed by the disclosure herein, brain fog resolved within 3 days, fatigue resolved within 5 days, insomnia resolved within 5 days, and rash resolved within 14 days. Example 13: Lysate / insulin analog combination is effective in treating type 2 diabetes.

[0237] Human subjects undergoing treatment for type 2 diabetes and receiving daily doses of the insulin analog insulin glargine (LANTUS®) have experienced oxidative stress-related side effects of rash (injection site), irritation, and headache.

[0238] In patients receiving a supplemental dose of 45 mg (bid) of a solution contained within the present disclosure, irritation resolved within 5 days, headache resolved within 7 days, and rash (injection site) resolved within 10 days. Example 14: Lysate / antibiotic combination is effective in treating rheumatoid arthritis.

[0239] A human subject diagnosed with rheumatoid arthritis and undergoing treatment with a TNF inhibitor had also been treated with antibiotics causing grade 3 diarrhea as determined by National Cancer Institute (NCI) criteria for the past 10 months. Concurrently, the subject was receiving additional antibiotics to combat a Clostridium difficile infection caused by severe diarrhea. Without changing the current pharmacological therapy, the subject was given an additional dose of 45 mg of the solution twice daily. The diarrhea began to resolve within 2 weeks and improved to NCI grade 2 criteria in 1 month. Example 15: Lysate / radiation combination is effective in treating cancer.

[0240] The human subject was being treated for prostate cancer and received radiation treatments of increasing intensity and / or duration over a 40-day regimen as follows: 8 sessions of 180cGy (1cGy=1 rad) in 2 bursts, one for 8 seconds and one for 5 seconds, during weekdays, in 9 points; 8 sessions of 180cGy, bursts for 12 and 9 seconds; 9 sessions of 180cGy, bursts for 14 and 10 seconds on the right 2 segments, followed by bursts for 12 and 9 seconds on the left segment; 16 sessions of 200cGy, 2 consecutive full circumferential passes for 80 seconds, and 2 consecutive full circumferential passes in the opposite direction in every other session. The radiation dose change was always on Mondays, and every Tuesday thereafter, the patient saw a doctor for an assessment of side effects. Each increase caused nausea for 3 hours, about 2 hours after the treatment, and complete recovery within 6 hours after every treatment session. In combination with radiation treatment, the patient was administered the lysate of the present disclosure at a dose of 75 mg, three times a day. The patient was not receiving any other treatment therapy such as nausea, vomiting, or bleeding. On a quality of life scale of 1-10 (with "1" being the worst and "10" being the best), the patient reported an 8 throughout the beginning of the final escalation of exposure. The patient reported that nausea was never worse than 7 at maximum exposure, and continued to function daily at work during treatment.

[0241] Considering the diversity of the mechanisms of action of the above drugs co-administered with the lysate composition of the present disclosure, and the mechanism of action of the lysate itself, the lysate composition disclosed herein may, with reasonable extrapolation, result in beneficial reduction of side effects with most drugs. For example, HUMIRA®, an injectable a-TNF, when co-administered with the lysate, results in improved injection site irritation. As per the package insert, this drug has been identified as causing arthritis, as seen with the direct acting antiviral drug HARVONI®. When patients use the lysate of the present disclosure together with a combined HCV drug such as HARVONI®, side effects are reduced during treatment and do not persist after treatment.

[0242] It should be understood that the configurations and functions described herein are presented for illustrative purposes only, and that many variations are possible. For example, elements may be added, omitted, combined, distributed, rearranged, or otherwise modified. Therefore, the following claims should be understood to include what is specifically illustrated and described above, what is conceptually equivalent, what can be obviously substituted, and what essentially incorporates the essential idea of ​​the present invention. Those skilled in the art will understand that various adaptations and modifications of the above-described embodiments can be made without departing from the scope of the present invention. The illustrated embodiments are set forth for illustrative purposes only and should not be construed as limiting the present invention. Therefore, it should be understood that within the scope of the appended claims, the present invention may be practiced otherwise than as specifically described herein.

Claims

1. 1. A toll-like receptor (TLR) agonist composition for regulating redox status in a subject, comprising: (a) a TLR agonist comprising at least one lysate and / or lysate fraction of a bacterium, the TLR agonist activating at least one or more TLRs or NLRs; (b) an optional promoter for increasing absorption of the composition; (c) a carrier for increasing the volume of the composition; A composition, wherein administration of an effective amount of the composition to the subject measurably reduces oxidative stress levels in the subject.

2. The composition of claim 1 , wherein the agonist activates at least two different TLRs and / or NLRs.

3. The composition of claim 1 , wherein the agonist activates at least three different TLRs and / or NLRs.

4. The composition of claim 1 , wherein the bacteria is a gram-positive or gram-negative bacterium.

5. 5. The composition of claim 4, wherein the at least one Gram-positive bacterium is selected from the group consisting of bacteria of the Lactobacillaceae family, Streptococcus family, Bifidobacterium family, and Bacillaceae family, and the at least one Gram-negative bacterium is selected from the group consisting of bacteria of the genera Pseudomonas, Klebsiella, Xanthomonas, Shigella, and Enterobacter.

6. The at least one Gram-positive bacterium is selected from the group consisting of Bacillus coagulans, Lactobacillus sporogenes, Streptococcus thermophilus, Bifidobacterium animalis, Bifidobacterium animalis subsp. animalis, Bifidobacterium infantis, Bifidobacterium longum, Bifidobacterium breve, Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus purpuratus ... Lactobacillus casei, Lactobacillus delbrueckii, Lactobacillus delbrueckii subsp. bulgaricus, Lactococcus lactis, Lactococcus lactis subsp. lactis, Streptococcus lactis, Streptococcus thermophilus, Bifidobacterium lactis, Bifidobacterium breve, Pediococcus acidilactici and Lactobacillus helveticus; 5. The composition of claim 4, wherein the at least one gram-negative bacterium is selected from the group consisting of Klebsiella oxytoca, Shigella flexneri, Xanthomonas campestris, and Pseudomonas fluorescens.

7. 2. The composition of claim 1, wherein the TLR agonist activates at least one or more of TLR2, TLR3, TLR4, TLR5, TLR7, TLR8, TLR9, NOD1 and NOD2.

8. The composition of claim 2 , wherein the TLR agonist activates TLR2 and TLR4.

9. 2. The composition of claim 1, wherein the TLR agonist activates two or more of TLR2, TLR3, TLR4, TLR5, TLR7, TLR8, TLR9, NOD1 and NOD2.

10. 2. The composition of claim 1, wherein the TLR agonist activates three or more of TLR2, TLR3, TLR4, TLR5, TLR7, TLR8, TLR9, NOD1 and NOD2.

11. The composition of claim 1 , wherein the promoter is selected from the group consisting of an amino acid, an amino sugar, and a sugar.

12. The composition of claim 1 , wherein the carrier is selected from the group consisting of a binder, a gum base, and combinations thereof.

13. The composition of claim 12, wherein the gum base comprises at least one hydrophobic polymer and at least one hydrophilic polymer.

14. The composition of claim 12 , wherein the binder is selected from the group consisting of sugars, sugar alcohols, and combinations thereof.

15. 15. The composition of claim 14, wherein the sugar alcohol is selected from the group consisting of mannitol, sorbitol, xylitol, and combinations thereof.

16. 10. The composition of claim 1, wherein the composition is manufactured in a dosage form selected from the group consisting of a lozenge, a chewing gum, a chewable tablet, a candy, and a dissolving tablet.

17. The composition of claim 16 , wherein the dosage form delivers the agonist to the oral mucosa.

18. 18. The composition of claim 17, wherein the oral mucosa is selected from the group consisting of sublingual mucosa, buccal mucosa, and combinations thereof.

19. The composition of claim 1 , wherein the composition is administered in combination with a pharmaceutical agent to enhance the activity of the pharmaceutical agent.

20. The composition of claim 1 , wherein the composition is administered in combination with a pharmaceutical agent to reduce the side effects of the pharmaceutical agent.

21. 21. The composition of claim 20, wherein the side effect of the pharmaceutical agent is oxidative stress.

22. 13. A method for regulating redox status in a subject, comprising administering a therapeutically effective amount of a composition according to claim 1 to a subject in need thereof.

23. 23. The method of claim 22, wherein modulation of redox status is assessed by measuring changes in isoprostane concentration in the subject.

24. 24. The method of claim 23, wherein the subject is a mammal.

25. 25. The method of claim 24, wherein the mammal is a human.

26. 24. The method of claim 23, wherein the subject is a non-mammal.

27. 27. The method of claim 26, wherein the subject is a fish, poultry, crustacean or insect.

28. 28. The method of claim 27, wherein the insect is a Drosophila.

29. 26. The method of claim 25, wherein the redox condition is oxidative, resulting in oxidative stress.

30. 30. The method of claim 29, wherein the oxidative stress in the human is associated with post-traumatic stress disorder.

31. 1. A method of regulating redox status in a subject, comprising: (a) to subjects in need thereof, at intervals in time; (i) a toll-like receptor (TLR) agonist comprising at least one lysate and / or lysate fraction of a bacterium, the TLR agonist activating at least one or more different TLRs or NLRs; (ii) an optional promoter to increase absorption of the composition; (iii) administering repeatedly in doses comprised of a composition comprising a carrier to increase the volume of said composition; (b) measuring the subject's bodily fluids to detect changes in oxidative stress levels.

32. 32. The method of regulating the redox status in a subject according to claim 31, wherein the agonist activates at least two different TLRs and / or NLRs.

33. 33. The method of regulating redox status in a subject according to claim 32, wherein the agonist activates at least three different TLRs and / or NLRs.

34. 32. The method of regulating the redox status in a subject according to claim 31, wherein the bacteria is a gram-positive or gram-negative bacterium.

35. the at least one gram-positive bacterium is selected from the group consisting of bacteria from the family Lactobacillaceae, Streptococcusceae, Bifidobacteriumceae, and Bacillaceae; 35. The method of regulating the redox status in a subject according to claim 34, wherein the at least one gram-negative bacterium is selected from the group consisting of bacteria of the genera Pseudomonas, Klebsiella, Xanthomonas, Shigella and Enterobacter.

36. The at least one Gram-positive bacterium is selected from the group consisting of Bacillus coagulans, Lactobacillus sporogenes, Streptococcus thermophilus, Bifidobacterium animalis, Bifidobacterium animalis subsp. animalis, Bifidobacterium infantis, Bifidobacterium longum, Bifidobacterium breve, Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus purpuratus ... Lactobacillus casei, Lactobacillus delbrueckii, Lactobacillus delbrueckii subsp. bulgaricus, Lactococcus lactis, Lactococcus lactis subsp. lactis, Streptococcus lactis, Streptococcus thermophilus, Bifidobacterium lactis, Bifidobacterium breve, Pediococcus acidilactici and Lactobacillus helveticus; 35. The method of regulating the redox status in a subject according to claim 34, wherein the at least one gram-negative bacterium is selected from the group consisting of Klebsiella oxytoca, Shigella flexneri, Xanthomonas spp., and Pseudomonas fluorescens spp.

37. 32. The method of regulating redox status in a subject of claim 31, wherein the TLR agonist activates at least one or more of TLR2, TLR3, TLR4, TLR5, TLR7, TLR8, TLR9, NOD1 and NOD2.

38. 32. The method of regulating redox status in a subject according to claim 31, wherein the TLR agonist activates two or more of TLR2, TLR3, TLR4, TLR5, TLR7, TLR8, TLR9, NOD1 and NOD2.

39. 32. The method of regulating redox status in a subject according to claim 31, wherein the TLR agonist activates three or more of TLR2, TLR3, TLR4, TLR5, TLR7, TLR8, TLR9, NOD1 and NOD2.

40. 40. The method of regulating redox status in a subject of claim 38, wherein the TLR agonist activates TLR2 and TLR4.

41. 32. The method of regulating the redox state in a subject according to claim 31, wherein the promoter is selected from the group consisting of amino acids, amino sugars and sugars.

42. 32. The method of regulating a redox status in a subject of claim 31, wherein the carrier is selected from the group consisting of a binder, a gum base, and combinations thereof.

43. 43. The method of regulating the redox status in a subject according to claim 42, wherein the gum base comprises at least one hydrophobic polymer and at least one hydrophilic polymer.

44. 43. The method of regulating a redox state in a subject of claim 42, wherein the binding agent is selected from the group consisting of a sugar, a sugar alcohol, and combinations thereof.

45. 45. The method of regulating redox status in a subject of claim 44, wherein the sugar alcohol is selected from the group consisting of mannitol, sorbitol, xylitol, and combinations thereof.

46. 32. The method of regulating a redox status in a subject of claim 31, wherein the composition is manufactured as a dosage form selected from the group consisting of a lozenge, a chewing gum, a chewable tablet, a candy, and a dissolving tablet.

47. 47. The method of regulating the redox state in a subject according to claim 46, wherein the dosage form delivers the agonist to the oral mucosa.

48. 32. The method of regulating redox status in a subject according to claim 31, wherein the oral mucosa is selected from the group consisting of sublingual mucosa, buccal mucosa, and combinations thereof.

49. 32. The method of regulating redox status in a subject of claim 31, wherein performing measurements of bodily fluids comprises assessing changes in isoprostane concentration in the subject.

50. 32. The method of regulating the redox state in a subject according to claim 31, wherein the subject is a mammal.

51. 51. The method of regulating redox status in a subject according to claim 50, wherein the mammal is a human.

52. 32. The method of claim 31 , wherein the subject is a non-mammal.

53. 53. The method of claim 52, wherein the subject is a fish, poultry, crustacean or insect.

54. 54. The method of claim 53, wherein the insect is a Drosophila.

55. 55. The method of regulating redox status in a subject according to claim 54, wherein the oxidative stress in the human is associated with post-traumatic stress disorder.

56. 1. A method for decreasing the amount of isoprostane in the urine or blood of a subject, comprising: (a) determining the level of isoprostanes in the urine or blood of the subject; (b) (i) a toll-like receptor (TLR) agonist comprising at least one bacterial lysate and / or lysate fraction from a bacterium, the TLR agonist activating at least one or more different TLRs or NLRs; (ii) administering to the subject an effective amount of a composition comprising an optional promoter for increasing absorption of the composition; (c) continuing administration of the composition until the level of isoprostane in the urine or blood of the subject is reduced.

57. 57. The method of claim 56, wherein the bacterium is a gram-positive or gram-negative bacterium.

58. The bacterium (a) Bacillus coagulans, Lactobacillus sporogenes, Streptococcus thermophilus, Bifidobacterium animalis, Bifidobacterium animalis, subspecies animalis, Bifidobacterium infantis, Bifidobacterium longum, Bifidobacterium breve, Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus casei, Lactobacillus bulgaricus, Lactococcus lactis, Lactococcus lactis subsp. lactis, Streptococcus lactis, Streptococcus thermophilus, Bifidobacterium lactis, Bifidobacterium breve, Pediococcus acidilactici and Lactobacillus helveticus, or 58. The method of claim 57, wherein (b) the bacterium is a gram-negative bacterium selected from the group consisting of Klebsiella oxytoca, Shigella flexneri, Xanthomonas campestris, and Pseudomonas fluorescens.

59. 1. A composition comprising: (a) a bacterial lysate and / or lysate fraction capable of activating at least one or more toll-like receptors (TLRs) or Nod-like receptors (NLRs); (b) an optional promoter for increasing absorption of the composition; (c) an optional carrier for increasing the volume of the composition.

60. 60. The pharmaceutical formulation comprising the composition of claim 59, wherein the pharmaceutical formulation is formulated for buccal or sublingual administration.

61. 61. The pharmaceutical formulation of claim 60, wherein the pharmaceutical formulation is formulated to dissolve in 1 minute or more after administration.

62. 60. The composition of claim 59, wherein the bacterial lysate and / or lysate fraction is a gram-negative or gram-positive bacterial lysate and / or lysate fraction.

63. The bacterial lysate and / or lysate fraction comprises: a Gram-positive bacterial lysate and / or lysate fraction selected from the group consisting of a Lactobacillus family bacterial lysate and / or lysate fraction, a Streptococcus bacterial lysate and / or lysate fraction, a Bifidobacterium bacterial lysate and / or lysate fraction, or a Bacillus bacterial lysate and / or lysate fraction, or 63. The composition of claim 62, which is a gram-negative bacterial lysate or lysate fraction selected from the group consisting of Pseudomonas bacterial lysate or lysate fraction, Klebsiella bacterial lysate or lysate fraction, Xanthomonas bacterial lysate or lysate fraction, Shigella bacterial lysate or lysate fraction, or Enterobacter bacterial lysate or lysate fraction.

64. The bacterial lysate and / or lysate fraction may be (a) Bacillus coagulans, Lactobacillus sporogenes, Streptococcus thermophilus, Bifidobacterium animalis, Bifidobacterium animalis subspecies animalis, Bifidobacterium infantis, Bifidobacterium longum, Bifidobacterium breve, Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus casei, Lactobacillus delbrueckii, Lactobacillus bulgaricus, Lactococcus lactis, Lactococcus lactis subsp. lactis, Streptococcus lactis, Streptococcus thermophilus, Bifidobacterium lactis, Bifidobacterium breve, Pediococcus acidilactici or Lactobacillus helveticus lysates and / or lysate fractions, or 63. The composition of claim 62, wherein (b) is a gram-negative bacterial lysate and / or lysate fraction selected from the group consisting of Klebsiella oxytoca, Shigella flexneri, Xanthomonas campestris, or Pseudomonas fluorescens, lysates and / or lysate fractions.

65. 1. A method for producing a bacterial lysate, comprising: (a) fermenting bacteria in a growth medium to stationary growth phase to produce a fermentation broth; (b) harvesting the bacteria from the fermentation broth; (c) pasteurizing the harvested bacteria; (d) lysing the pasteurized bacteria with lysozyme to produce a bacterial lysate.

66. 66. The method of claim 65, wherein the bacteria are harvested in mid-log phase, late-log phase, early stationary phase, mid-stationary phase, or late stationary phase.

67. 66. The method of claim 65, wherein the bacterium is a gram-positive or gram-negative bacterium.

68. the gram-positive bacterium is selected from the group consisting of bacteria of the Lactobacillaceae family, bacteria of the Streptococcus family, bacteria of the Bifidobacteriaceae family, and bacteria of the Bacillaceae family; 68. The method of claim 67, wherein the gram-negative bacteria is selected from the group consisting of bacteria of the genera Pseudomonas, Klebsiella, Xanthomonas, Shigella and Enterobacter.

69. 68. The method of claim 67, comprising: The gram-positive bacteria is selected from the group consisting of Bacillus coagulans, Lactobacillus sporogenes, Streptococcus thermophilus, Bifidobacterium animalis, Bifidobacterium animalis subsp. animalis, Bifidobacterium infantis, Bifidobacterium longum, Bifidobacterium breve, Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus spp. casei, Lactobacillus delbrueckii, Lactobacillus delbrueckii subsp. bulgaricus, Lactococcus lactis, Lactococcus lactis subsp. lactis, Streptococcus lactis, Streptococcus thermophilus, Bifidobacterium lactis, Bifidobacterium breve, Pediococcus acidilactici and Lactobacillus helveticus; The method, wherein the gram-negative bacterium is selected from the group consisting of Klebsiella oxytoca, Shigella flexneri, Xanthomonas campestris, and Pseudomonas fluorescens.

70. 70. A bacterial lysate produced according to the method of any one of claims 65 to 69.

71. 1. A method for reducing one or more oxidative stress related side effects associated with administration of a pharmaceutical agent, the method comprising, in combination with the pharmaceutical agent, (a) a bacterial lysate and / or a lysate fraction; (b) an optional promoter for increasing absorption of the composition; (c) administering a therapeutically effective amount of a lysate composition comprising an optional carrier to increase the volume of said composition; The method, wherein the pharmaceutical agent and the lysate composition are administered simultaneously or in any order, through the same route of administration or through different routes of administration.

72. 72. The method of claim 71, wherein the lysate and / or lysate fraction activates at least one or more TLRs or NLRs.

73. 72. The method of claim 71, wherein the lysate and / or lysate fraction activates at least two TLRs and / or NLRs.

74. 72. The method of claim 71, wherein the lysate and / or lysate fraction activates at least three TLRs or NLRs.

75. 72. The method of claim 71, wherein the lysate and / or lysate fraction activates at least one or more of TLR2, TLR3, TLR4, TLR5, TLR7, TLR8, TLR9, NOD1 and NOD2.

76. 72. The method of claim 71, wherein the lysate and / or lysate fraction activates TLR2 and TLR4.

77. 72. The method of claim 71, wherein the lysate and / or lysate fraction activates two or more of TLR2, TLR3, TLR4, TLR5, TLR7, TLR8, TLR9, NOD1 and NOD2.

78. 72. The method of claim 71, wherein the lysate and / or lysate fraction activates three or more of TLR2, TLR3, TLR4, TLR5, TLR7, TLR8, TLR9, NOD1 and NOD2.

79. 72. The method of claim 71, wherein the bacterium is a gram-positive or gram-negative bacterium.

80. the gram-positive bacterium is selected from the group consisting of bacteria of the Lactobacillaceae family, bacteria of the Streptococcus family, bacteria of the Bifidobacteriaceae family, and bacteria of the Bacillaceae family; 80. The method of claim 79, wherein the gram-negative bacteria is selected from the group consisting of bacteria of the genera Pseudomonas, Klebsiella, Xanthomonas, Shigella and Enterobacter.

81. The gram-positive bacteria is selected from the group consisting of Bacillus coagulans, Lactobacillus sporogenes, Streptococcus thermophilus, Bifidobacterium animalis, Bifidobacterium animalis subsp. animalis, Bifidobacterium infantis, Bifidobacterium longum, Bifidobacterium breve, Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus spp. casei, Lactobacillus delbrueckii, Lactobacillus delbrueckii subsp. bulgaricus, Lactococcus lactis, Lactococcus lactis subsp. lactis, Streptococcus lactis, Streptococcus thermophilus, Bifidobacterium lactis, Bifidobacterium breve, Pediococcus acidilactici and Lactobacillus helveticus; 80. The method of claim 79, wherein the gram-negative bacteria is selected from the group consisting of Klebsiella oxytoca, Shigella flexneri, Xanthomonas campestris, and Pseudomonas fluorescens.

82. The one or more oxidative stress related side effects are aceruloplasminemia, arterial / systemic hypertension, arthritis, asthma, atherosclerosis, atopic dermatitis, cancer, bladder cancer, leukemia, uterine cancer, cervical cancer, dizziness, nausea, vomiting, constipation, diarrhea, insomnia, drowsiness, lightheadedness, decreased libido, memory loss, tremors, jaundice, arrhythmia, increased heart rate, decreased heart rate, urticaria, depression, clinical depression, cerebral ischemia, bronchopulmonary dysplasia, cardiovascular disease, cataracts, cellulitis, chemotherapy side effects, chronic fatigue syndrome, colitis, coronary artery disease, dyslipidemia, eclampsia, erectile dysfunction, ataxia, headache, heart failure, hemodialysis side effects, liver cirrhosis, high cholesterol 73. The method of claim 72, wherein the condition is selected from the group consisting of hypertension, hyperhomocysteinemia, hyperlipidemia, interstitial lung disease, lung injury, macular degeneration, male infertility, mild cognitive impairment, myocardial infarction, myocarditis, myocardial degeneration, neuropathy, obesity, osteoarthritis, osteoporosis, pancreatitis, periodontal disease, side effects of peritoneal dialysis, post traumatic stress disorder, preeclampsia, psoriasis, psoriatic arthritis, pulmonary hypertension, side effects of radiation therapy, reactive arthritis, respiratory distress syndrome, rhabdomyolysis, rheumatic disease, sepsis, sleep apnea, stroke, suicidal thoughts, amyloidosis, thrombophilia, tauopathy, unstable angina, uremia, and venous insufficiency.

83. 1. A method for treating an oxidative stress-related disease or condition in a subject, comprising: (a) a bacterial lysate and / or lysate fraction capable of activating at least one or more toll-like receptors (TLRs) or Nod-like receptors (NLRs); (b) an optional promoter for increasing absorption of the composition; (c) administering to the subject a therapeutically effective amount of the composition, and optionally a carrier to increase the volume of the composition.

84. The oxidative stress-related conditions are selected from the group consisting of aceruloplasminemia, acute and chronic alcoholic liver disease, acute autoimmune myocarditis, acute chest syndrome of sickle cell disease, acute pancreatitis, acute respiratory distress syndrome, alcoholic liver disease, amyotrophic lateral sclerosis, arterial / systemic hypertension, asbestosis, asthma, ataxia telangiectasia, atherosclerosis, atopic dermatitis, cerebral ischemia, bronchopulmonary dysplasia, burns, some cancers, cardiopulmonary bypass, cardiovascular disease, cataracts, cellulitis, chemotherapy side effects, chronic fatigue syndrome, chronic hepatitis C, chronic kidney disease, chronic obstructive pulmonary disease, chronic renal failure, colitis, coronary artery disease, Creutzfeldt-Jakob disease, Crohn's disease, Cutaneous leishmaniasis, cystic fibrosis, type 1 diabetes, type 2 diabetes, dyslipidemia, Down's syndrome, eclampsia, end-stage renal disease, erectile dysfunction, Friedreich's ataxia, headache, heart failure, Helicobacter pylori infection / inflammation, side effects of hemodialysis, liver cirrhosis, human immunodeficiency virus infection, Huntington's disease, hyperbaric oxygen therapy-related diseases, hypercholesterolemia, hyperhomocysteinemia, hyperlipidemia, idiopathic pulmonary fibrosis, interstitial lung disease, ischemia / reperfusion injury, juvenile chronic arthritis, kidney transplant failure, leukemia, lung cancer, pulmonary disorders, macular degeneration, male infertility, Meniere's disease, meningitis, mild cognitive impairment, multiple sclerosis, myelodysplastic syndrome syndromes), myocardial infarction, myocarditis, neonatal bronchopulmonary dysplasia, obesity, osteoarthritis, osteoporosis, pancreatitis, Parkinson's syndrome, periodontal disease, side effects of peritoneal dialysis, photoaging, post-traumatic stress disorder, preeclampsia, primary biliary cirrhosis, broncopulmonary disease, progeria, psoriasis, psoriatic arthritis, pulmonary hypertension, side effects of radiation therapy, reactive arthritis, renal cell carcinoma, respiratory distress syndrome, retinopathy of prematurity, retrolental fibroplasia 84. The method of claim 83, wherein the condition is selected from the group consisting of rheumatoid arthritis, sarcoidosis, sepsis, sickle cell disease, sleep apnea, spherocytosis, spinal cord injury, stroke, synucleinopathy, systemic amyloidosis, systemic lupus erythematosus, systemic sclerosis, thrombophilia, tauopathy, traumatic stress tuberculosis, variable angina, uremia, venous insufficiency, Werner's syndrome, and Zellweger's syndrome.

85. The bacterium (a) Bacillus coagulans, Lactobacillus sporogenes, Streptococcus thermophilus, Bifidobacterium animalis, Bifidobacterium animalis, subspecies animalis, Bifidobacterium infantis, Bifidobacterium longum, Bifidobacterium breve, Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus casei, Lactobacillus bulgaricus, Lactococcus lactis, Lactococcus lactis subsp. lactis, Streptococcus lactis, Streptococcus thermophilus, Bifidobacterium lactis, Bifidobacterium breve, Pediococcus acidilactici and Lactobacillus helveticus, or The method of claim 83, wherein (b) the bacterium is a gram-negative bacterium selected from the group consisting of Klebsiella oxytoca, Shigella flexneri, Xanthomonas campestris, and Pseudomonas fluorescens.

86. 1. A method for reducing oxidative stress in a subject, comprising: (a) determining the level of oxidative stress in the subject by measuring the amount of isoprostanes in the urine or blood of the subject; (b) (i) a toll-like receptor (TLR) agonist comprising at least one lysate and / or lysate fraction from a bacterium, the TLR agonist activating at least one or more TLRs or NLRs; (ii) administering to the subject an effective amount of a composition comprising an optional promoter for increasing absorption of the composition; (c) continuing administration of the composition until the level of oxidative stress is reduced as measured by a decrease in the amount of isoprostanes in the subject's urine.

87. 87. The method of claim 86, wherein administration of the bacterial lysate continues until the amount of isoprostanes in the subject's urine is less than about 3 ng / mg creatinine, less than about 2 ng / mg creatinine, less than about 1 ng / mg creatinine, or less than about 0.5 ng / mg creatinine.

88. 87. The method of claim 86, wherein the bacterium is a gram-negative bacterium or a gram-positive bacterium.

89. The bacterium (a) Bacillus coagulans, Lactobacillus sporogenes, Streptococcus thermophilus, Bifidobacterium animalis, Bifidobacterium animalis, subspecies animalis, Bifidobacterium infantis, Bifidobacterium longum, Bifidobacterium breve, Lactobacillus acidophilus, Lactobacillus plantarum, Lactobacillus casei, Lactobacillus bulgaricus, Lactococcus lactis, Lactococcus lactis subsp. lactis, Streptococcus lactis, Streptococcus thermophilus, Bifidobacterium lactis, Bifidobacterium breve, Pediococcus acidilactici and Lactobacillus helveticus, or The method of claim 86, wherein (b) the bacterium is a gram-negative bacterium selected from the group consisting of Klebsiella oxytoca, Shigella flexneri, Xanthomonas campestris, and Pseudomonas fluorescens.

90. 87. The method of claim 86, wherein the lysate and / or lysate fraction activates at least one or more TLRs or NLRs.

91. 87. The method of claim 86, wherein the lysate and / or lysate fraction activates at least two TLRs and / or NLRs.

92. 87. The method of claim 86, wherein the lysate and / or lysate fraction activates at least three TLRs or NLRs.

93. 87. The method of claim 86, wherein the lysate and / or lysate fraction activates at least one or more of TLR2, TLR3, TLR4, TLR5, TLR7, TLR8, TLR9, NOD1 and NOD2.

94. 87. The method of claim 86, wherein the lysate and / or lysate fraction activates TLR2 and TLR4.

95. 87. The method of claim 86, wherein the lysate and / or lysate fraction activates two or more of TLR2, TLR3, TLR4, TLR5, TLR7, TLR8, TLR9, NOD1 and NOD2.

96. 87. The method of claim 86, wherein the lysate and / or lysate fraction activates three or more of TLR2, TLR3, TLR4, TLR5, TLR7, TLR8, TLR9, NOD1 and NOD2.

97. 1. A therapeutic combination comprising: (a) (i) a bacterial lysate and / or lysate fraction capable of activating at least one or more toll-like receptors (TLRs) or Nod-like receptors (NLRs); (ii) an optional promoter to increase absorption of the composition; (iii) an optional carrier for increasing the volume of the composition; (b) one or more pharmaceutical agents; A therapeutic combination, wherein the lysate composition and the one or more pharmaceutical agents are administered simultaneously or in any order, and wherein the lysate composition and the one or more pharmaceutical agents are administered by the same or different routes of administration.

98. 98. The therapeutic combination of claim 97, wherein the lysate composition is administered sublingually or bucally.

99. 1. A pharmaceutical formulation comprising: (a) (i) a bacterial lysate and / or lysate fraction capable of activating at least one or more toll-like receptors (TLRs) or Nod-like receptors (NLRs); (ii) an optional promoter to increase absorption of the composition; (iii) an optional carrier for increasing the volume of the composition; (b) A pharmaceutical formulation comprising a combination of one or more pharmaceutical agents.

100. wherein the one or more pharmaceutical agents are Antispasmodics, motility stimulants, H2 receptor antagonists, antimuscarinics; chelating agents, prostaglandin analogues, aminosalicylates, corticosteroids, drugs affecting immune response, stimulant laxatives, drugs affecting bile composition and flow, bile acid sequestrants, dopamine antagonists, proton pump inhibitors, opioids, opioid receptor antagonists, analgesics, hypnotics, cardiac glycosides, phosphodiesterase inhibitors, thiazides, diuretics, potassium sparing diuretics, aldosterone antagonists, osmotic diuretics, drugs for arrhythmias , beta-adrenergic receptor blockers, hypertensive drugs, drugs affecting the renin-angiotensin system, nitrates, calcium blockers, antianginal drugs, peripheral vasodilators, sympathomimetics, anticoagulants, protamines, antiplatelet drugs, fibrinolytic drugs, antifibrinolytic drugs, lipid regulating drugs, omega-3 fatty acid compounds, CNS drugs, anti-infective drugs, or benzothrone, procyclidine, biperiden, amantadine, bromocriptine, pergolide, entacapone, tolcapone, selegiline, pramipexole, budesonide, formoterol, quetiapine fumarate , Olanzapine, Pioglitazone, Montelukast, Zoledronic Acid, Valsartan, Latanoprost, Irbesartan, Clopidogrel, Atomoxetine, Dexamfetamine, Methylphenidate, Modafinil, Bleomycin, Dactinomycin, Daunorubicin, Idarubicin, Mitomycin, Mitoxantrone, Azacitidine, Capecitabine, Cladribine, Clofarabine, Cytarabine, Fludarabine, Flourouracil, Gemcitabine, Mercapto The therapeutic combination of claim 97 or the pharmaceutical formulation of claim 99, wherein the additional agent is selected from the group consisting of topramine, methotrexate, nelarabine, pemetrexed, raltitrexed, thioguanine, apomorphine, betamethasone, cortisone, deflazacort, dexamethosone, hydrocortisone, methylprednisolone, prednisolone, triamcinolone, cyclosporine, sirolimus, tacrolimus, interferon alpha, and interferon beta.

101. wherein the one or more pharmaceutical agents are Antispasmodics selected from the group consisting of atropine sulfate, dicycloverine hydrochloride, hyoscine butyl bromide, propantheline bromide, alverine citrate, and mebeverine hydrochloride; a motility stimulant selected from the group consisting of metoclopramide and domperidone; H2-receptor antagonists selected from the group consisting of cimetidine, famotidine, nizatidine, and ranitidine; Antimuscarinic; A chelate selected from the group consisting of tripotassium bismuth dicitrate and sucralfate; Prostaglandin analogues; an aminosalicylates selected from the group consisting of valsazide sodium, mesalazine, olsalazine, and sulfasalazine; Corticosteroids selected from the group consisting of beclomethasone dipropionate, budenoside, hydrocortisone and prednisolone; drugs that affect the immune system selected from the group consisting of cyclosporine, mercaptopurine, methotrexate, adalimumab, and infliximab; a stimulant laxative selected from the group consisting of bisacodyl, danthron, docusate, and sodium picosulfate; Drugs that affect bile composition and flow; Cholestyramine, oxyphencyclimine, camylofine, mebeverine, trimebutine, rosiverine, dicycloverine, dihexyverine, difemerine, piperidolate, benzilon, mepenzolate, pipenzolate, glycopyrronium, oxyphenonium, pentienate, methantheline, propantheline, otilonium bromide, tridihexetyl, isopropamide, hexocyclium, polzine, bevonium, diphemanil, thiemonium iodide, prifinium bromide, timepidium bromide, fenpiverinium, papaverine, drotaverine, moxa bile acid sequestrants selected from the group consisting of verine, 5-HT3 antagonists, 5-HT4 agonists, fenpiprane, diisopromine, chlorbenzoxamine, pinaverium, fenoberin, idanpramine, proxazole, alverine, trepibutone, isometheptene, caroverine, phloroglucinol, silicones, trimethyldiphenylpropylamine, atropine, hyoscyamine, scopolamine, butylscopolamine, methylscopolamine, methylatropine, phenthonium, cimetropium bromide, and dopamine blockers; A proton pump inhibitor selected from the group consisting of omeprazole, lansoprazole, pantoprazole, esomeprazole, and rabeprazole sodium; Opioids and opioid receptor antagonists; an analgesic selected from the group consisting of acetaminophen, diclofenac, diflunisal, etodolac, fenoprofen, flurbiprofen, ibuprofen, indomethacin, ketoprofen, ketorolac, meclofenamate, mefenamic acid, meloxicam, nabumetone, naproxen, oxaprozin, phenylbutazone, piroxicam, sulindac, tolmetin, celecoxib, buprenorphine, butorphanol, codeine, hydrocodone, hydromorphone, levorphanol, meperidine, methadone, morphine, nalbuphine, oxycodone, oxymorphone, pentazocine, propoxyphene, and tramadol; a sleep medication selected from the group consisting of nitrazepam, flurazepam, loprazolam, lormetazepam, temazepam, zaleplon, zolpidem, zopiclone, chloral hydrate, triclofos, clomethiazole, quazepam, triazolam, estazolam, clonazepam, alprazolam, eszopiclone, rozerem, trazodone, amitriptyline, doxepin, benzodiazepines, melatonin, diphenhydramine, and herbal remedies; a cardiac glycoside selected from the group consisting of digoxin and digitoxin; a phosphodiesterase inhibitor selected from the group consisting of enoximone and milrinone; Thiazides and thiazide-related diuretics selected from the group consisting of bendroflumethiazide, chlorthalidone, cyclopenthiazide, inapamide, metolazone, and oxamide; a diuretic selected from the group consisting of furosemide, bumetanide, and trasemide; a potassium-sparing diuretic and aldosterone antagonist selected from the group consisting of amiloride hydrochloride, triamterene, weplerenone, and spironolactone; Osmotic diuretics; a drug for arrhythmia selected from the group consisting of adenosine, amiodarone hydrochloride, disopyramide, flecainide acetate, propafenone hydrochloride, and lidocaine hydrochloride; beta-adrenergic receptor blockers selected from the group consisting of propanalol, atenolol, acebutolol, bisprolol fumarate, carvedilol, celiprolol, esmolol, revatolol, metoprolol tartrate, nadolol, nebivolol, oxyprenolol, pindolol, solatol, and timolol; hypertension drugs selected from the group consisting of ambrisentan, bosentan, diazoxide, hydralazine, iloprost, minoxidil, sildenafil, sitaxsentan, sodium nitroprusside, clonidine, methyldopa, moxonidine, guanethidine monosulfate, doxazosin, indoramin, prazosin, terazosin, phenoxybenzamine, and phentolamine mesylate; Drugs affecting the renin-angiotensin system selected from the group consisting of captopril, cilazapril, enalapril maleate, fosinopril, imidapril, lisinopril, moexipril, perindopril erbumine, quinapril, ramipril, trandolapril, candesartan cilexetil, eprosartan, irbesartan, losartan, olmesartan medoxomil, telmisartan, valsartan, and aliskiren; nitrates, calcium channel blockers, and antianginal agents selected from the group consisting of nitroglycerin, isosorbide dinitrate, isosorbide mononitrate, amlodipine, diltiazem, felodipine, isradipine, lacidipine, lercanidipine, nicardipine, nifedipine, nimodipine, verapamil, ivaprazine, nicorandil, and ranolazine; Peripheral vasodilators selected from the group consisting of cilostazol, inositol nicotinate, moxisylyte, naftidrofuryl oxalate, and pentoxifylline, and related drugs; Sympathomimetics selected from the group consisting of Dopamine, Dopexamine, Ephedrine, Metaraminol, Noradrenaline Bitartrate, Norephedrine Bitartrate, and Phenylephrine; An anticoagulant and protamine selected from the group consisting of heparin, bemiparin, dalteparin, enoxaparin, tinzaparin, danaparoid, bivalirudin, lepirudin, epoprostenol, fondaprinux, warfarin, acenocoumarol, phenindione, dabigatran etexilate, rivaroxaban, and protamine sulfate; an antiplatelet agent selected from the group consisting of abciximab, aspirin, clopidogrel, dipyridamole, eptifibatide, prasugrel, and tirofiban; Fibrinolytic and antifibrinolytic agents selected from the group consisting of alteplase, reteplase, streptokinase, tenecteplase, urokinase, ethamsylate, and tranexamic acid; lipid regulating agents selected from the group consisting of atorvastatin, fluvastatin, pravastatin, rosuvastatin, simvastatin, colesevelam, cholestyramine, colestipol, ezetimibe, bezafibrate, ciprofibrate, fenofibrate, gemfibrozil, acipimox, nicotinic acid, omega-3 fatty acids, ethanolamine oleate, and sodium tetradecyl sulfate; Benperidol, Chlorpromazine, Flupenthixol, Haloperidol, Levomepromazine, Pericyazine, Perphenazine, Pimozide, Prochlorperazine, Promazine, Sulpiride, Trifluoperazine, Zuclopenthixol, Amisulpiride, Aripiprazole, Clozapine, Olanzapine, Paliperidone, Quetiapine, Riperidone, Sertindole, Zotepine, Flupenthixol, Fluphenazine, Olanzapine Emboate (Olanzapine Pamoate), Pipotiazine Palmitate, Risperidone, Zuclopenthixol Decanoate valproate, carbamazepine, valproate, valproic acid, lithium carbonate, lithium citrate, amitriptyline, clomipramine, dosulepin, imipramine, lofepramine, nortriptyline, trimipramine, mianserin, trazodone, phenelzine, isocarboxazid, tranylcypromine, moclobemide, citalopram, escitalopram, fluoxetine, fluvoxamine, paroxetine, sertraline, agomelatine, duloxetine, flupentixol, mirtazapine, reboxetine, trytophan, benflaxine Venflaxine), atomoxetine, dexamethamine, methylphenidate, modafinil, eslicarbazepine, ocarbazepine, ethosuximide, gabapentin, pregabalin, lacosamide, lamotrigine, levetiracetam, phenobarbital, primidone, phenytoin, rufinamide, tiagabine, topiramate, vigabatrin, zonisamide, ropinirole, rotigotine, co-beneldopa, levodopa, CNS drugs selected from the group consisting of Co-Careldopa, Rasagiline, Selegiline, Entacapone, Tolcapone, Amantidine, Orphenadrine, Procyclidine, Trihexyphenidyl, Haloperidol, Piracetam, Riluzole, Tetrabenazine, Acamprosate, Disulfiram, Bupropion, Varenicilline, Buprenorphine, Lofexidine, Donepezil, Galantamine, Memantine, and Rivastidine; Benzylpenicillin, Phenoxymethylpenicillin, Flucloxacillin, Temocillin, Amoxicillin, Ampicillin, Co-amoxiclav, Co-fluampicil, Piperacillin, Ticarcillin, Pivmecillinam, Cephalosporins, Cefaclor, Cefadroxil, Cephalexin, Cefixime, Cefotaxime, Cephradine, Ceftazidime, Cefuroxime, Ertapenem, Imipenem, Meropenem, Aztreonam, Tetracycline, Demeclocycline, Doxocycline, Rimonam, Cycline, Minocycline, Oxytetracycline, Tigecycline, Gentamicin, Amikacin, Neomycin, Tobramycin, Erythromycin, Azithromycin, Clarithromycin, Telithromycin, Clindamycin, Chloramphenicol, Fusidic Acid, Vancomycin, Teicoplanin, Daptomycin, Linezolid, Quinupristin, Colistin, Co-trimoxazole, Sulpadiazine, Trimethoprim, Capreomycin, Cycloserine, Ethambutol, Isoniazid, Pyrazinamide, Rifabutin, Rifampicin, Streptomycin, Dapsone, Clofazimine, Metronidazole, Tinidazole, Ciprofloxacin, Levofloxacin, Moxifloxacin, Nalidixic Acid, Norflaxine, Orflaxacin, Nitrofurantoin, Methenamine Hippurate, Amphotericin, Anidulafungin, Caspofungin, Fluconazole, Flucytosine, Griseofulvin, Itraconzole le), Ketoconazole, Micafungin, Nystatin, Posaconazole, Terbinafine, Voriconazole, Abacavir, Didanosine, Emtricitabine, Lamivudine, Stavudine, Tenofovir Disoproxil, Zidovudine, Atazanavir, Darunavir, Fosamprenavir, Indinavir, Lopinair, Nelfinavir, Ritonavir, Saquinavir, Tipranavir, Efavirenz, Etravirine, Nevarapine, Enfuvirtide, Maraviroc, Raltegravir, Acyclovir, Famciclovir,an anti-infective selected from the group consisting of Inosine Pranobex, Valacyclovir, Cidofovir, Gangciclovir, Foscarnet, Valgangciclovir, Adefovir Dipivoxil, Entecavir, Telbivudine, Amantadine, Oseltamivir, Zanamivir, Palivizumab, Ribavirin, Artemether, Chloroquine, Mefloquine, Primaquine, Proguanil, Pyrimethamine, Quinine, Doxycycline, Diloxanide Furoate, Metronidaziole, Tinidazole, Mepacrine, Sodium Stibogluconate, Atovaquone, Pentamidine Isethionate, Mebendazole, and Piperazine; and Benzotone, procyclidine, biperiden, amantadine, bromocriptine, pergolide, entacapone, tolcapone, selegiline, pramipexole, budesonide, formoterol, quetiapine fumarate, olanzapine, pioglitazone, montelukast, zoledronic acid, valsartan, latanoprost, irbesartan, clopidogrel, atomoxetine, dexamfetamine, methylphenidate, modafinil, bleomycin, dactinomycin, daunorubicin, idarubicin, mitomycin, mitoxantrone, azacitidine, capecitabine, cladribine, clofarabine , Cytarabine, Fludarabine, Flourouracil, Gemcitabine, Mercaptopurine, Methotrexate, Nelarabine, Pemetrexed, Raltitrexed, Thioguanine, Apomorphine, Betamethasone, Cortisone, Deflazacort, Dexamethosone, Hydrocortisone, Methylprednisolone, Prednisolone, Triamcinolone, Cyclosporine, Sirolimus, Tacrolimus, Interferon alpha, and Interferon beta.

102. A formulation comprising: (a) (i) a bacterial lysate and / or lysate fraction capable of activating at least one or more toll-like receptors (TLRs) or Nod-like receptors (NLRs); (ii) an optional promoter for increasing absorption of the composition; (iii) an optional carrier for increasing the volume of the composition; and (b) an isolated human anti-TNFα antibody or antigen-binding fragment thereof, or a TNF inhibitor.

103. The formulation of claim 102, wherein the human anti-TNFα antibody or antigen-binding fragment thereof is adalimumab.

104. 104. Use of the formulation of claim 103 in the manufacture of a medicament for the treatment of rheumatoid arthritis (RA), late-onset RA, or psoriatic arthritis in a subject.

105. 103. A method for the treatment of rheumatoid arthritis (RA), late-onset RA, or psoriatic arthritis in a subject, comprising administering to the subject a therapeutically effective amount of the formulation of claim 102.

106. The method of claim 105, wherein the human anti-TNFα antibody or antigen-binding fragment thereof is administered to the subject on a biweekly dosing regimen.

107. The method of claim 105, wherein the human anti-TNFα antibody or antigen-binding fragment thereof is administered to the subject at a dose of 30 mg or more.

108. 106. The method of claim 105, wherein the TNFα inhibitor is a TNF alpha fusion protein.

109. 109. The method of claim 108, wherein the TNFα fusion protein is etanercept.

110. The method of claim 105, wherein the anti-TNFα antibody or antigen-binding fragment thereof is infliximab or golimumab.

111. The method of claim 105, wherein the anti-TNFα antibody or antigen-binding fragment thereof is adalimumab.

112. The method of claim 105, wherein the anti-TNFα antibody or antigen-binding fragment thereof is adalimumab.

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