Non-antibiotic antibacterial composition

Abstract

The present invention relates to non-antibiotic antibacterial compositions, novel uses, and methods of treatment or prevention. In particular, the present invention relates to non-antibiotic antibacterial compositions comprising a zeolite and at least one member of the genus Lactobacillus for the treatment of diseases or conditions associated with or caused by Helicobacter bacteria, such as Helicobacter pylori.

Description

[Technical Field]

[0001] The present invention relates to non-antibiotic antibacterial compositions, novel uses, and methods of treatment or prevention. In particular, the present invention relates to non-antibiotic antibacterial compositions comprising a zeolite and at least one member of the genus Lactobacillus for the treatment of diseases or conditions associated with or caused by Helicobacter bacteria, such as Helicobacter pylori. The present invention further relates to non-antibiotic antibacterial compositions comprising a zeolite and at least one member of the genus Lactobacillus for the treatment of infections caused by urease-active bacteria in the gastrointestinal tract. [Background technology]

[0002] Infectious diseases, including bacterial infections, continue to be a serious health problem worldwide. The Gram-negative (-ve) microorganism Helicobacter pylori was first cultured in 1982, and since then, many closely related species have been identified in humans and other animals. These Helicobacter species (genus Helicobacter) are generally classified according to their ability to invade and colonize gastric or enterohepatic tissues. Colonization of the gastric mucosa appears to be most prevalent in the superficial mucus layer and within gastric glands and parietal cells.

[0003] Gastric Helicobacter species are widespread in mammalian hosts and can often induce inflammatory responses similar to those seen with Helicobacter pylori in humans. Similarly, enterohepatic Helicobacter species are a diverse group of microorganisms present in the intestinal tract and liver of humans, other mammals, and birds. Helicobacter species are often associated with inflammation and malignant transformation in immunocompromised hosts and more severe clinical disease in immunocompromised humans and animals.

[0004] The primary disorder that occurs after Helicobacter pylori colonization is chronic active gastritis. This condition can be observed in all Helicobacter pylori-positive individuals. The distribution and severity of this chronic inflammatory process within the stomach depend on various factors, including the characteristics of the colonizing strain, host genetic factors and immune response, diet, and acid production levels. Helicobacter pylori-induced ulcer disease, gastric cancer, and lymphoma are all complications of this chronic inflammation. Ulcer disease and gastric cancer occur particularly in individuals and at sites where inflammation is most severe (Kusters, Johannes G et al., "Pathogenesis of Helicobacter pylori infection." Clinical microbiology reviews vol.19,3 (2006): 449-90. doi:10.1128 / CMR.00054-05).

[0005] More than 50% of the world's population is infected with this gastric bacterium. The severity of the infection is related to the bacterial load (Celli, Jonathan P et al. "Helicobacter pylori moves through mucus by reducing mucin viscoelasticity." Proceedings of the National Academy of Sciences of the United States of America vol. 106, 34 (2009): 14321-6. doi:10.1073 / pnas.0903438106; Varbanova M, Malfertheiner P. Bacterial load and degree of gastric mucosal inflammation in Helicobacter pylori infection. Dig Dis.2011; 29:592-599. doi: 10.1159 / 000333260).

[0006] Treatment of Helicobacter pylori infection remains challenging due to its side effects. Standard triple therapy (STT), consisting of a proton pump inhibitor and two antibiotics (one of which, most commonly, contains clarithromycin), has been used as first-line treatment worldwide for 20 years, empirically without antimicrobial susceptibility testing (AST). Since 2010, a significant decline in the efficacy of this treatment regimen has been observed worldwide, and it is now recommended to prescribe quadruple therapy with or without bismuth if clarithromycin resistance is greater than 15% (Maastricht V) (Malfertheiner P, Megraud F, O'Morain CA, et al., Management of Helicobacter pylori infection - the Maastricht V / Florence consensus report. Gut 2017; 66: 6-30. doi: 10.1136 / gutjnl-2016-312288 pmid:http: / / www.ncbi.nlm.nih.gov / pubmed / 27707777).

[0007] These regimens have inherent drawbacks, including the empirical use of multiple antibiotics, which may promote further antimicrobial resistance and induce dysbiosis of the gut microbiota (Suzuki Sa · Kusano Cb · Horii Ta · Ichijima Ra · Ikehara Ha et al. The Ideal Helicobacter pylori Treatment for the Present and the Future. Digestion 2022;103:62–68 https: / / doi.org / 10.1159 / 000519413). Antibiotic resistance has been identified by the World Health Organization as one of the top 10 threats to global public health.

[0008] Surveillance of antibiotic resistance in Helicobacter pylori has been carried out at the European level every 10 years since 1998 (Glupczynski Y, Megraud F, Lopez-Brea M, et al., European multicentre survey of in vitro antimicrobial resistance in Helicobacter pylori. Eur J Clin Microbiol Infect Dis 2001; 20: 820-823. doi:10.1007 / s100960100611 pmid:http: / / www.ncbi.nlm.nih.gov / pubmed / 11783701), and again in 2008 (Megraud F, Coenen S, Versporten A, et al., Helicobacter pylori resistance to antibiotics in Europe and its relationship to antibiotic consumption. Gut 2013; 62:34-42. doi:10.1136 / gutjnl-2012-302254) and again in 2018 (Megraud F, Bruyndonckx R, Coenen S The European Helicobacter pylori Antimicrobial Susceptibility Testing Working Group, et al., Helicobacter pylori resistance to antibiotics in Europe in 2018 and its relationship to antibiotic consumption in the community. Gut 2021; 70:1815-1822).

[0009] A recent study in 2018 confirmed a positive correlation between Helicobacter pylori resistance and the corresponding antibiotic consumption in the region. The mean age of patients was 51.2 years (range 17-91 years). The age distribution was consistent with the normal distribution observed in patients undergoing upper gastrointestinal endoscopy. It is believed that many people do not realize they are infected with Helicobacter pylori until later in life. Unless effectively treated, Helicobacter pylori infection usually persists indefinitely and manifests as other, seemingly unrelated disorders or diseases.

[0010] Despite efforts to find effective treatments against Helicobacter species, reports of specific and effective treatments with low resistance and desirable efficacy are rare, if not nonexistent.

[0011] Therefore, there are currently no effective non-antibiotic treatments for Helicobacter spp., and no effective treatments that can be widely adopted and accepted as a long-term, easy-to-use standard of care for Helicobacter spp., without the associated side effects mentioned above.

[0012] Therefore, there is an urgent need not only to develop effective treatments against Helicobacter spp., but also to develop treatments that are widely applicable, non-toxic, and avoid unlimited reliance on antibiotics.

[0013] Helicobacter pylori is a urease-active bacterium. Other urease-positive bacteria have been identified as pathogenic in the gastrointestinal tract. Staphylococcus capitius urealiticum causes urinary tract infections, Klebsiella species, such as Proteus miras (Klebsiella pneumoniae), are associated with gastrointestinal pneumonia, and mycobacteria, such as Mycobacterium tuberculosis, are associated with intestinal tuberculosis. Therefore, there is a need for effective treatments for infections caused by pathogenic urease-positive bacteria that are widely available, nontoxic, and avoid unlimited reliance on antibiotics. Summary of the Invention

[0014] The present invention relates to non-antibiotic antimicrobial compositions, novel uses, and methods of treatment or prevention.

[0015] Specifically, the present invention relates to a non-antibiotic antimicrobial composition comprising a zeolite and at least one member of the genus Lactobacillus for the treatment of a condition or disease associated with or caused by Helicobacter bacteria in a vertebrate.

[0016] Specifically, the present invention further relates to a non-antibiotic antimicrobial composition comprising a zeolite and at least one member of the genus Lactobacillus for the treatment of conditions or diseases associated with or caused by pathogenic urease-active bacteria in vertebrates.

[0017] The pathogenic urease-active bacteria in vertebrates may be one or more selected from the list consisting of Helicobacter, Staphylococcus, Proteus, Klebsiella, and Mycobacterium.

[0018] The pathogenic urease-active bacteria in vertebrates may be one or more selected from the list consisting of Helicobacter pylori, Staphylococcus capitius urealiticum, Proteus mirabilis, Klebsiella pneumoniae, and Mycobacterium tuberculosis.

[0019] In one aspect, there is provided an antibacterial composition comprising a zeolite and at least one member of the genus Lactobacillus for use in treating a condition associated with Helicobacter in a vertebrate.

[0020] In another aspect, there is provided an antibacterial composition comprising a zeolite and at least one member of the genus Lactobacillus for use in treating a condition associated with or caused by pathogenic urease-active bacteria in a vertebrate. The pathogenic urease-active bacteria in a vertebrate may be one or more selected from the list consisting of Helicobacter, Staphylococcus, Proteus, Klebsiella, and Mycobacterium. More specifically, the pathogenic urease-active bacteria in a vertebrate may be one or more selected from the list consisting of Helicobacter pylori, Staphylococcus capitius urealyticum, Proteus mirabilis, Klebsiella pneumoniae, and Mycobacterium tuberculosis.

[0021] In some embodiments, an antimicrobial composition is provided wherein the zeolite is a naturally occurring zeolite or a synthetic zeolite.

[0022] In some embodiments, an antimicrobial composition is provided wherein the zeolite is zeolite clinoptilolite (ZC).

[0023] In some embodiments, an antimicrobial composition is provided wherein the zeolite is activated zeolite clinoptilolite (aZC), preferably doubly activated zeolite clinoptilolite (aZC).

[0024] In some embodiments, at least one member of the genus Lactobacillus is selected from the group consisting of Lactobacillus reuteri, Lactobacillus delbrueckii, Lactobacillus rhamnosus, Lactobacillus casei, Lactobacillus acidophilus, Lacticaseibacillus paracasei, Lactiplantibacillus plantarum, Levilactobacillus brevis, Ligilactobacillus salivarius, Limosilactobacillus fermentum, and the like. fermentum, Lactobacillus bulgaricus, Lactobacillus crispatus, Lactobacillus helveticus, and Lactobacillus johnsonii are provided.

[0025] In some embodiments, an antimicrobial composition is provided, wherein at least one member species of the Lactobacillus genus is Lactobacillus reuteri.

[0026] In some embodiments, an antibacterial composition is provided, wherein the Lactobacillus reuteri is Lactobacillus reuteri DSM17648 or Lactobacillus reuteri UBLRu-87.

[0027] In some embodiments, an antimicrobial composition is provided, wherein a vertebrate is administered a therapeutically effective amount of the antimicrobial composition.

[0028] In some embodiments, an antimicrobial composition is provided, wherein a therapeutically effective amount is achieved by a dosing regimen of zeolite and at least one member species of the Lactobacillus genus.

[0029] In some embodiments, an antibacterial composition is provided, wherein the regimen comprises one or more of zeolite and at least one member species of the genus Lactobacillus.In some embodiments, the administration regimen of the antibacterial composition is suitable for oral administration, rectal administration, topical administration, enteral administration, vaginal administration, buccal administration, orthotopic administration, intratracheal administration, intralesional administration, endoscopic administration, transmucosal administration, sublingual administration, enteral administration, and combinations thereof.In some embodiments, the administration regimen of the antibacterial composition is suitable for oral administration.

[0030] In some embodiments, an antimicrobial agent is provided wherein at least one member species of the Lactobacillus genus is probiotic.

[0031] In some embodiments, antimicrobial compositions are provided that further comprise a bioactive substance or mineral, such as, for example, calcium carbonate, magnesium carbonate, vitamins, such as, for example, vitamin D and vitamin E, pharmaceutically acceptable carriers, additives, and adjuvants, such as antimicrobial chitins, such as chitosan, α-ketoglutarate, citrate, and lactate.

[0032] In some embodiments, an antimicrobial composition is provided, wherein the dosage regimen of zeolite and at least one member species of the genus Lactobacillus is suitable for oral or intragastric administration.

[0033] In some embodiments, an antimicrobial composition is provided that includes activated zeolite clinoptilolite (aZC), Lactobacillus reuteri DSM17648 or Lactobacillus reuteri UBLRu-87, calcium carbonate, and magnesium carbonate.

[0034] In some embodiments, an antibacterial composition is provided wherein the condition is associated with Helicobacter pylori.

[0035] In some embodiments, an antimicrobial agent is provided wherein the vertebrate is a human.

[0036] In some embodiments, antimicrobial compositions are provided that result in a reduction of one or more markers.

[0037] In some embodiments, an antimicrobial composition is provided wherein the marker is a predictive marker selected from the group consisting of urea, gastric ammonia, and urease activity.

[0038] In some embodiments, an antimicrobial composition is provided wherein the predictive marker is gastric ammonia.

[0039] In some embodiments, an antibacterial composition is provided that results in a reduction of intragastric ammonia levels in a vertebrate infected with Helicobacter pylori.

[0040] In some embodiments, antimicrobial compositions are provided that reduce gastric ammonia by at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% or more compared to placebo.

[0041] In some embodiments, antimicrobial compositions are provided that reduce gastric ammonia by at least 0.1, 0.25, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, or 10 times or more compared to placebo.

[0042] In some embodiments, antimicrobial compositions are provided wherein the reduction in predictive markers is statistically significant compared to placebo.

[0043] In some embodiments, antimicrobial compositions are provided that result in a statistically significant reduction in gastric ammonia marker levels in humans infected with pathogenic urease-active bacteria.

[0044] In some embodiments, an antibacterial composition is provided that produces a statistically significant reduction in gastric ammonia marker levels in humans infected with Helicobacter pylori.

[0045] In some embodiments, antimicrobial compositions are provided that result in statistically significant clinical improvement in humans infected with pathogenic urease-active bacteria.

[0046] In some embodiments, antibacterial compositions are provided that result in a statistically significant clinical improvement in humans infected with Helicobacter pylori.

[0047] In one aspect of the invention, there is provided use of an antibacterial composition for the preparation or manufacture of a pharmaceutical formulation, for separate, combined or simultaneous administration, designed to treat, alleviate or prevent a GIT disease or disorder, gastritis, gastric ulcer, duodenal ulcer, gastric cancer or duodenal cancer in a vertebrate, including mammals and birds, in need of such treatment, alleviation or prevention.

[0048] In one aspect of the invention, there is provided use of an antibacterial composition for the preparation or manufacture of a pharmaceutical formulation, for separate, combined or simultaneous administration, designed for the treatment, reduction or prevention of urinary tract infections (particularly those associated with Staphylococcus or Proteus infections), intestinal pneumonia, or intestinal tuberculosis in vertebrates, including mammals and birds, in need of such treatment, reduction or prevention.

[0049] In one aspect of the present invention, there is provided a use of an antimicrobial composition for the preparation or manufacture of a pharmaceutical formulation for separate, combined, or simultaneous administration designed to treat, reduce, or prevent liver disease, kidney disease, or other diseases in vertebrates, including mammals and birds, in need thereof, both of which are associated with high levels of ammonium in the gastrointestinal tract.

[0050] In some embodiments, the present invention is directed to the use of antimicrobial compositions wherein the pharmaceutical formulation is in a form suitable for administering a daily dose of the antimicrobial composition, either in a single dose or as divided doses or subdoses administered at appropriate intervals.

[0051] In some embodiments, the present invention is directed to the use of antibacterial compositions wherein the pharmaceutical formulation is administered as one, two, three, four or more doses or subdoses per day or administration.

[0052] In some embodiments, the present invention provides a pharmaceutical formulation comprising 10 7 ~10 9 The antibacterial composition comprises CFU / g of Lactobacillus reuteri DSM 17648 or Lactobacillus reuteri UBLRu-87, and a zeolite, wherein the zeolite is present in an amount of at least 50 mg / gram of the formulation.

[0053] In some embodiments, the invention is directed to the use of an antimicrobial composition, wherein the pharmaceutical formulation further comprises calcium carbonate and magnesium carbonate, wherein the calcium carbonate and magnesium carbonate are each present in an amount of at least 0.5 mg per gram of formulation.

[0054] In some embodiments, the present invention is directed to the use of antimicrobial compositions wherein the dose of the antimicrobial composition is formulated as a capsule, lyophilized formulation, liquid, pill, powder, or gel.

[0055] In some embodiments, the present invention is directed to the use of antibacterial compositions where treatment results in complete remission of a disease or disorder caused by or associated with infection with Helicobacter.

[0056] In some embodiments, the present invention is directed to an antimicrobial composition, as described in the paragraph above, for use in reducing ammonia in the gastrointestinal tract, preferably in reducing ammonia concentrations in gastric fluids.

[0057] The inventors have surprisingly and unexpectedly determined that the non-antibiotic antimicrobial compositions of the present invention are effective in inactivating and treating Helicobacter pylori infections where established antibiotic-based therapies have failed. In particular, the inventors have surprisingly and unexpectedly determined that the non-antibiotic antimicrobial compositions of the present invention are effective in inactivating and treating Helicobacter pylori infections where established antibiotic-based therapies have failed, by reducing predictive markers such as urea, gastric ammonia, acid reflux, bloating, and urease activity. [Brief explanation of the drawings]

[0058] [Figure 1] FIG. 1. In vitro urea and ammonia reduction assay using the antimicrobial composition of the present invention. [Figure 2] FIG. 2 is a graph showing in vivo clinical trial data. [Figure 3]Figure 3 shows the blank-corrected absorbance of H. pylori strains 83 and ATCC 700824 after 1:10 or 1:100 dilutions in brain heart infusion broth + 10 mM urea and 0.1% YE and incubation for 120 hours under microaerobic conditions. [Figure 4] Figure 4 is a graph showing the total viable counts of Helicobacter pylori strains 83 and ATCC 700824 after 1:10 or 1:100 dilutions in brain heart infusion broth + 10 mM urea and 0.1% YE and incubation for 144 hours under microaerophilic conditions. [Figure 5A] Figure 5 is a graph showing the pH and ammonium concentration in brain heart infusion broth + 10% [v / v] FBS, 0.1% [w / v] yeast extract, and 0 mM, 10 mM, or 50 mM urea, initially adjusted to pH 2 (Figure 5A), adjusted to pH 5 (Figure 5B), or left unadjusted (pH 7) (Figure 5C), after 8 hours of incubation under microaerobic conditions. [Figure 5B] Figure 5 is a graph showing the pH and ammonium concentration in brain heart infusion broth + 10% [v / v] FBS, 0.1% [w / v] yeast extract, and 0 mM, 10 mM, or 50 mM urea, initially adjusted to pH 2 (Figure 5A), adjusted to pH 5 (Figure 5B), or left unadjusted (pH 7) (Figure 5C), after 8 hours of incubation under microaerobic conditions. [Figure 5C] Figure 5 is a graph showing the pH and ammonium concentration in brain heart infusion broth + 10% [v / v] FBS, 0.1% [w / v] yeast extract, and 0 mM, 10 mM, or 50 mM urea, initially adjusted to pH 2 (Figure 5A), adjusted to pH 5 (Figure 5B), or left unadjusted (pH 7) (Figure 5C), after 8 hours of incubation under microaerobic conditions. [Figure 6A]Figure 6 is a graph showing the pH and total viable count of Helicobacter pylori ATCC 700824 in brain heart infusion broth + 10% [v / v] FBS, 0.1% [w / v] yeast extract, and 0 mM, 10 mM, or 50 mM urea, initially adjusted to pH 2 (Figure 6A), pH 5 (Figure 6B), or unadjusted (pH 7) (Figure 6C) after 8 hours of incubation under microaerobic conditions. [Figure 6B] Figure 6 is a graph showing the pH and total viable count of Helicobacter pylori ATCC 700824 in brain heart infusion broth + 10% [v / v] FBS, 0.1% [w / v] yeast extract, and 0 mM, 10 mM, or 50 mM urea, initially adjusted to pH 2 (Figure 6A), pH 5 (Figure 6B), or unadjusted (pH 7) (Figure 6C) after 8 hours of incubation under microaerobic conditions. [Figure 6C] Figure 6 is a graph showing the pH and total viable count of Helicobacter pylori ATCC 700824 in brain heart infusion broth + 10% [v / v] FBS, 0.1% [w / v] yeast extract, and 0 mM, 10 mM, or 50 mM urea, initially adjusted to pH 2 (Figure 6A), pH 5 (Figure 6B), or unadjusted (pH 7) (Figure 6C) after 8 hours of incubation under microaerobic conditions. DETAILED DESCRIPTION OF THE INVENTION

[0059] Throughout this disclosure, various scientific publications, patents and published patent specifications are referenced by an identifying citation. The disclosures of these publications, patents and published patent specifications are incorporated by reference into this disclosure in order to more fully describe the state of the art to which this disclosure pertains.

[0060] Certain terms used herein may have the following defined meanings.

[0061] As used in this specification and the appended claims, the singular forms "a," "an," and "the" include both the singular and the plural unless the context clearly dictates otherwise. For example, the term "zeolite" includes silicate-containing materials that have an affinity for ammonia (NH4+). In the context of the present invention, a zeolite material can be a microporous or mesoporous silicate, also known as a molecular sieve, in granular or powder form for separate, combined, or simultaneous administration, either once daily or for several days, weeks, months, or years, or twice or more times daily for several days to years, alone or in combination with at least one member of the genus Lactobacillus, for use in the treatment, alleviation, or prevention of GIT diseases or disorders, gastritis, gastric ulcers, duodenal ulcers, gastric cancer, duodenal cancer in vertebrates, including mammals and birds, in need thereof.

[0062] As used herein, the term "substance" refers to zeolites, pharmaceutical agents, therapeutically active ingredients, and / or mineral salts, such as calcium carbonate and / or magnesium carbonate, and combinations thereof.

[0063] As used herein, the term "antibacterial" refers to destroying, killing, or inhibiting the growth of microorganisms, particularly pathogenic microorganisms, such as pathogenic bacteria such as Helicobacter.

[0064] As used herein, the abbreviation "(w / w)" means weight percent calculated based on the weight of the component and the total weight of the composition or formulation. Generally, the terminology used in this application is well known to those skilled in the art.

[0065] As used herein, all numerical values ​​or figures indicating, for example, amounts, proportions, physical properties, and / or uses of materials or substances shall be understood to be modified or altered by the term "about," unless expressly indicated otherwise.

[0066] As used herein, the term "about" includes the stated number or numerical value and ±10% from the stated number or numerical value. As a non-limiting example, the term "about ten (10)" includes nine (9) to eleven (11) or 9 to 11.

[0067] As used herein, the term "subject" refers to any animal, e.g., a vertebrate, preferably a mammal such as a human, to which a substance, compound, or composition according to an embodiment of the invention is or has been administered. Preferably, the subject is in need of treatment or prevention of Helicobacter infection, or has been the subject of observation or experimentation for the treatment or prevention of such infection. Preferably, the subject is in need of treatment or prevention of the following Helicobacter infections, or has been the subject of observation or experimentation for the treatment or prevention of such infections: Helicobacter canis, Helicobacter felis, Helicobacter heilmannii, Helicobacter mustelae, Helicobacter bizzozeronii, Helicobacter 10 cinonyx, Helicobacter fenneliae, Helicobacter rappini, Helicobacter hepaticus, Helicobacter pleurorum. pullorum, Helicobacter bilis, Helicobacter rodentium, Helicobacter trogontum, Helicobacter cinaedi, Helicobacter muridarum, Helicobacter pametensis, Helicobacter cholecystus, Helicobacter pylori infections, etc.Preferably, the subject is in need of treatment or prevention of Helicobacter pylori infection, or has been the subject of observation or experimentation for the treatment or prevention thereof.

[0068] As used herein, the term "symptoms" refers to, for example, vomiting, gastritis, peptic ulcers, gastric neoplasms, lymphoma, fever, GI disorders, and hormonal imbalances associated with or caused by Helicobacter bacteria.

[0069] As used herein, the term "condition" refers to, for example, hyperammonemia associated with or caused by Helicobacter bacteria. The term also relates to symptoms associated with infection with urease-active bacteria in the gastrointestinal tract. As used herein, the term "treatment" or "treating" refers to the improvement, prevention, or reversal of a disease or disorder, or at least one discernible symptom or condition thereof. It is also contemplated that treatment based on the data described herein will result in one or more of the following in a patient: clinical improvement, reduction in the severity of symptoms or disease, and reduction of one or more markers of one or more markers. In some embodiments, "treatment" or "treating" refers to the improvement, prevention, or reversal of at least one measurable physical parameter associated with the disease or disorder being treated, although not necessarily discernible in or by the mammal. In some embodiments, "treatment" or "treating" refers to inhibiting or slowing the progression of a disease, disorder, or condition, either physically, e.g., by stabilizing a discernible symptom, physiologically, e.g., by stabilizing a physical parameter, or both. In some embodiments, "treatment" results in partial or complete remission of a disease or disorder. In some embodiments, "treatment" results in clinical improvement in a subject infected or afflicted with Helicobacter, such as Helicobacter pylori. In some embodiments, "treatment" results in a reduction in disease severity in a subject infected or afflicted with Helicobacter, such as Helicobacter pylori. In some embodiments, "treatment" results in a reduction in viral load in a subject infected or afflicted with Helicobacter, such as Helicobacter pylori. Without wishing to be bound by theory, elevated blood levels of different markers, such as inflammatory markers, CBC markers, and predictive markers, have been associated with disease states or conditions associated with Helicobacter, such as Helicobacter pylori. Thus, in some embodiments of the present invention, the predictive marker is selected from the group consisting of: The marker is a predictive marker selected from the group consisting of urea, gastric ammonia, acid reflux, bloating, and urease activity.In some embodiments, the predictive marker is gastric ammonia. In some embodiments of the invention, treatment results in a reduction of one or more predictive markers selected from the group consisting of urea, gastric ammonia, acid reflux, bloating, and urease activity. In some embodiments of the invention, treatment results in a decrease in the predictive marker gastric ammonia. In some embodiments of the invention, treatment results in a reduction or alleviation of hyperammonemia. Helicobacter pylori is strongly urease positive. The ability of Helicobacter pylori to cleave urea within 30 seconds distinguishes Helicobacter pylori from other Helicobacter species. Bacteriology - Identification | ID 26 | Issue no: 3 | Issue date: 03.07.15 | Page: 18 of 27. UK Standards for Microbiology Investigations | Issued by the Standards Unit, Public Health England. In some embodiments, "treatment" results in clinical improvement in a subject infected with or suffering from a urease-active bacterium, which may be selected from Helicobacter, Staphylococcus, Proteus, Klebsiella, and Mycobacterium, more specifically Helicobacter pylori, Staphylococcus capitius urealiticum, Proteus mirabilis, Klebsiella pneumoniae, and / or Mycobacterium tuberculosis.

[0070] The term "urease-active bacteria" refers to one or more bacteria that can convert urea into ammonia and carbon dioxide, resulting in an increase in urinary pH. These organisms include Proteus, Nocardia, Ureaplasma, Helicobacter pylori, Klebsiella, Cryptococcus, Staphylococcus epidermidis, and Staphylococcus saprophyticus. These organisms are sometimes referred to as urease-positive bacteria.

[0071] The present invention relates to novel compositions and the use of such compositions in methods of treatment that alleviate, eliminate, or otherwise reduce or cure any one or more symptoms caused by or associated with infection with Helicobacter, such as Helicobacter pylori. Specifically, the present invention relates to novel compositions and the use of such compositions in treating subjects infected with Helicobacter, such as Helicobacter pylori. In some embodiments, the treatment results in a substantial cure of a disease or disorder caused by or associated with infection with Helicobacter, such as Helicobacter pylori. Specifically, the present invention relates to novel compositions and the use of such compositions as palliatives for subjects infected with Helicobacter, such as Helicobacter pylori.

[0072] While this disclosure focuses on subjects, one of skill in the art will appreciate that the present invention also applies to non-human subjects (i.e., vertebrates), such as, for example, livestock (e.g., cattle, horses, pigs, and sheep), exotic animals (e.g., big cats such as pandas, tigers, lions, and pumas, elephants, bats, mice, rats, and similar animals), companion animals (e.g., dogs and cats), and particularly when the disease or condition is caused by Helicobacter canis, Helicobacter felis, Helicobacter heilmannii, Helicobacter mustelae, Helicobacter bizzozeronii, Helicobacter 12 synonychus, Helicobacter 12 ... cinonyx, Helicobacter fenneliae, Helicobacter rappini, Helicobacter hepaticus, Helicobacter pullorum, Helicobacter bilis, Helicobacter rodentium, Helicobacter trogontum, Helicobacter cinaedi, Helicobacter muridarum, Helicobacter pametensis, Helicobacter cholecyctus, and Helicobacter pylori It will be readily appreciated that the present invention is equally applicable and effective when the disease or condition is associated with or caused by a Helicobacter infection, such as Helicobacter canis. In some embodiments of the present invention, the disease or condition is associated with or caused by Helicobacter canis.

[0073] The inventors have surprisingly and unexpectedly found that the non-antibiotic antimicrobial compositions of the present invention are effective in inactivating and treating Helicobacter pylori infections where established antibiotic-based therapies have failed. In particular, the inventors have surprisingly and unexpectedly determined that the non-antibiotic antimicrobial compositions of the present invention are effective in inactivating and treating Helicobacter pylori infections where antibiotic-based therapies have failed, for example, due to antibiotic resistance, by reducing predictive markers such as urea, gastric ammonia, acid reflux, bloating, and urease activity.

[0074] As used herein, the term "non-antibiotic" refers to not using, relying on, or containing antibiotics. In some embodiments, the compositions of the present invention do not use, rely on, or contain antibiotics.

[0075] As used herein, the term "antibiotic" refers to an antibacterial substance (e.g., penicillin, cephalosporin, metronidazole, clarithromycin, furazolidone, amoxicillin, tetracycline, ciprofloxacin, levofloxacin, etc.) isolated from cultures of specific microorganisms (e.g., fungi) or of semisynthetic or synthetic origin, administered orally, topically, or by injection, used to treat or prevent infection by killing or inhibiting the growth of bacteria in or on a subject. Currently, standard triple therapy consisting of two antibiotics and a proton pump inhibitor is reported as the first-line regimen. Alternatively, quadruple therapy including bismuth, sequential therapy, or simultaneous quadruple therapy without bismuth is also used in current treatments. Levofloxacin-containing triple therapy is recommended as rescue therapy for Helicobacter pylori infection after first-line therapy has failed. However, the rapid development of antibiotic resistance dramatically reduces the effectiveness of any antibiotic regimen, inevitably resulting in acute and long-term suffering and associated, often unpredictable, side effects for affected subjects. Helicobacter pylori is known to be increasingly resistant to metronidazole and clarithromycin. Bacteriology - Identification | ID 26 | Issue no: 3 | Issue date: 03.07.15 | Page: 10 of 27. UK Standards for Microbiology Investigations | Issued by the Standards Unit, Public Health England.

[0076] As contemplated in the context of the present invention, antibiotics can be used to treat or prevent Helicobacter infections, such as Helicobacter pylori infections. In some embodiments, the bacterium selected from the group consisting of Helicobacter canis, Helicobacter felis, Helicobacter heilmannii, Helicobacter mustelae, Helicobacter bizzozeronii, Helicobacter 14 cinonyx, Helicobacter fenneliae, Helicobacter rappini, Helicobacter hepaticus, Helicobacter pullorum, Helicobacter bilis, and Helicobacter rodentium. Helicobacter bacteria, such as Helicobacter rodentium, Helicobacter trogontum, Helicobacter cinaedi, Helicobacter muridarum, Helicobacter pametensis, Helicobacter cholecyctus, and Helicobacter pylori, are resistant to antibiotic treatment. In some embodiments, the Helicobacter pylori is resistant to antibiotic treatment.

[0077] As used herein, the term "administration" is understood to include, for example, oral, intravenous, parenteral, inhalation, pulmonary, rectal, nasal, topical, intravesical, intrathecal, enteral, intralymphatic, intracavity, vaginal, urethral, ​​intradermal, aural, intramammary, buccal, orthotopic, intratracheal, intralesional, transdermal, endoscopic, transmucosal, sublingual, enteral administration, and combinations thereof. In some embodiments, the administration regimen of the antimicrobial composition is suitable for oral, rectal, topical, enteral, vaginal, buccal, orthotopic, intratracheal, intralesional, endoscopic, transmucosal, sublingual, enteral administration, and combinations thereof. In some embodiments, the administration regimen or formulation of the antimicrobial composition is oral.

[0078] In some embodiments, the zeolite is a silicate-containing material that has an affinity for ammonia (NH4+). In some embodiments, the zeolite is a synthetic zeolite that has an affinity for ammonia (NH4+). As used herein, "synthetic zeolite" refers to a zeolite that is manufactured or synthesized by one or more chemical reactions involving the breaking and / or creation of chemical bonds. The physicochemical properties and characteristics of zeolites include, for example, different pore sizes, channels, and ion exchange capacities. The channels are filled with water and exchangeable cations, balancing the negative charge in the zeolite's structural framework. As a result of their highly porous structure and substitution, zeolites such as clinoptilolite have large cation exchange capacities (CECs), on the order of 100-300 cmol charge / kg. However, as with most zeolites, exchange selectivity is limited by channel size. Larger ions may be more selectively rejected. This "ion sieving" property of zeolites is an important property that is exploited in many industrial applications: in the presence of ammonium-rich liquids, NH4+-N is exchanged for cations present in the framework.

[0079] In some embodiments, antimicrobial compositions are provided wherein the zeolite is zeolite clinoptilolite (ZC). In some embodiments, antimicrobial compositions are provided wherein the zeolite clinoptilolite includes clinoptilolite-K, clinoptilolite-Na, and clinoptilolite-Ca. In some embodiments, antimicrobial compositions are provided wherein the zeolite is activated zeolite clinoptilolite (aZC). In some embodiments, antimicrobial compositions are provided wherein the activated zeolite clinoptilolite (aZC) is doubly activated zeolite clinoptilolite (aZC). The clinoptilolite series includes three known species, clinoptilolite-K, clinoptilolite-Na, and clinoptilolite-Ca, and their predominant chemical elemental component, i.e., K, is a stoichiometrically unsaturated fatty acid. + , Na + , and Ca ++ These chemical elements preferentially exchange heavy metals, toxins, mycotoxins, and ammonia present in the environment, e.g., GIT, during cation exchange, and exhibit a high chemical affinity for zeolites such as clinoptilolite-K, clinoptilolite-Na, and clinoptilolite-Ca.

[0080] In some embodiments, the activated zeolite clinoptilolite is a tribomechanically activated zeolite clinoptilolite.

[0081] In some embodiments, the tribomechanically activated zeolite clinoptilolite is (TMAZ®), whose chemical formula will be known to those skilled in the art: SiO2, 65.0-71.3%; Al2O3, 11.5-13.1%; CaO, 2.7-5.2%; K2O, 2.2-3.4%; Fe2O3, 0.7-1.9%; MgO, 0.6-1.2%; Na2O, 0.2-1.3%; TiO2, 0.1-0.3%; Si / Al ratio, 4.8-5.4.

[0082] The empirical formula of zeolite is (Ca, K2, Na2, Mg)4A18 Si 40 O 96 It will be understood by those skilled in the art that the specific molecular mass may be 2.2-2.5 g / cm; the porosity may be 32-40%; and the effective pore size may be 0.4 nm or greater, for example, 0.5 nm, 0.75 nm, 1 nm, 1.5 nm, 2 nm, 10 nm, 25 nm, 50 nm, 100 nm, 150 nm, 250 nm, 0.5 μm or greater in diameter.

[0083] In some embodiments, the zeolite has an ion exchange capacity of 1.2 to 1.5 mol / kg; 2+ , 0.64~0.98mol / kg;Mg 2+ , 0.06~0.19mol / kg;K + , 0.22~0.45mol / kg;Na + , 0.01~0.19mol / kg, showing ion exchange selectivity Cs>NH4+>Pb2+>K+>Na+>Mg2+>Ba2+>Cu2+>Zn2+.

[0084] In some embodiments, the chemicals or toxins adsorbed to the zeolite are NH3, C1-C4 hydrocarbons, CO2, H2S, SO2, NO X , containing aldehydes.

[0085] Preferably, as contemplated within the context of the compositions or formulations of the present invention, the zeolite is non-toxic, hi some embodiments, the zeolite is generally recognized as safe (GRAS) under the U.S. Code of Federal Regulations (21 CFR 182, Subpart C).

[0086] It is proposed that antibacterial compositions and their use in treatment methods can alleviate, eliminate, or otherwise reduce or cure any one or more symptoms caused by or associated with Helicobacter infection, such as Helicobacter pylori, by restoring the natural acidic pH value in a subject's GIT.

[0087] The mineral salts in the compositions of the present invention, such as calcium carbonate and magnesium carbonate, are further proposed to enhance or contribute to the ion exchange capacity of the zeolite, thereby enhancing the absorption and / or neutralization of ammonium, such as ammonium, in the GIT.

[0088] As used herein, "palliative" refers to any form of medical care or treatment that does not result in a cure but is dedicated to reducing the severity of symptoms of disease or slowing its progression. Palliatives aim to improve quality of life, particularly reducing or eliminating pain. This definition specifically focuses on the lack of a cure in general and emphasizes the proactive, total care of patients with illnesses that do not respond to curative treatments. Without wishing to be bound by theory, Helicobacter canis, Helicobacter felis, Helicobacter heilmannii, Helicobacter mustelae, Helicobacter bizzozeronii, Helicobacter 16 cinonyx, Helicobacter fenneliae, Helicobacter rappini, Helicobacter hepaticus, Helicobacter pullorum, Helicobacter bilis, Helicobacter Helicobacter bilis, Helicobacter rodentium, Helicobacter trogontum, Helicobacter cinaedi, Helicobacter muridarum, Helicobacter pametensis, Helicobacter cholecyctus, and Helicobacter pylori infection. Diseases or symptoms associated with or caused by Helicobacter infection are characterized by increased or higher than normal levels of urea, gastric ammonia, acid reflux, bloating, and urease activity. Gastric Helicobacter infection has been reported to be associated with hyperammonemia.Helicobacter pylori, H. bizzozeroni, H. cynogastricus, H. salomonis, H. suis, H. bilis, and H. heilmannii are urease positive. Bacteriology - Identification | ID 26 | Issue no: 3 | Issue date: 03.07.15 | Page: 21 of 27. UK Standards for Microbiology Investigations | Issued by the Standards Unit, Public Health England.

[0089] As used herein, the term "hyperammonemia," or other terms used to denote higher-than-normal ammonia levels, means a level higher than normal, where normal is the normal range for a particular laboratory, as determined in blood from healthy, disease-free subjects using an analytical methodology selected by the laboratory performing the assay (which may be venous blood, arterial blood, or a portion of whole blood, including plasma or serum), and processed according to laboratory procedures that specify whether the blood should be placed immediately on ice, the time frame for delivery to the laboratory, and the use of designated preservatives, if any.

[0090] Lactic acid bacteria, particularly lactobacilli, are the most commonly used microorganisms as "Generally Recognized As Safe" (GRAS) probiotics. Acidity, ammonia, bile salts, and the presence of pancreatic enzymes are some of the major stresses that orally ingested probiotics experience in the GIT. Apart from being viable, probiotic strains of the present invention must also be able to adhere to and subsequently (at least temporarily) colonize the intestinal tract. Because the GIT is a dynamic environment, bacteria that do not adhere to the intestinal mucosa may be washed away by the flow of gastrointestinal contents. Therefore, probiotic strains with adherence capabilities have a greater chance of colonizing the GIT and providing beneficial effects. In some embodiments, lactobacilli can tolerate and preferably remain active or biochemically effective in the elevated gastric ammonia levels observed in Helicobacter infections, such as Helicobacter pylori infection.

[0091] Various methods for producing probiotics have been reported and are well documented. Some of these methods describe the production of mixed microbial cultures in liquid growth media. The production of such mixed cultures is fraught with difficulties due to competition between microorganisms within the diverse population in the medium. One method for producing mixed microbial cultures is to grow different strains separately and then combine the grown strains in the desired ratio.

[0092] Other methods for growing probiotics focus on the quorum-sensing ability of microorganisms, particularly bacteria such as lactic acid bacteria. Lactic acid bacteria increase the release of various antibacterial metabolites when they experience metabolic stress. In particular, the combination of metabolic stress and quorum-sensing ability of microorganisms maximizes the release of various antibacterial metabolites (WO 2020 / 177858).

[0093] In the context of the present invention, at least one member of the genus Lactobacillus is produced by a method known in the art.In some embodiments, at least one member of the genus Lactobacillus is produced by a method known in the art.

[0094] In some embodiments, at least one member species of the genus Lactobacillus is grown or produced using a combination of metabolic stress and quorum sensing capabilities of at least one member species of the genus Lactobacillus.

[0095] In some embodiments, at least one member of the genus Lactobacillus is selected from the group consisting of Lactobacillus reuteri, Lactobacillus delbrueckii, Lactobacillus rhamnosus, Lactobacillus casei, Lactobacillus acidophilus, Lacticaseibacillus paracasei, Lactiplantibacillus plantarum, Levilactobacillus brevis, Ligilactobacillus salivarius, Limosilactobacillus fermentum, and the like. fermentum, Lactobacillus bulgaricus, Lactobacillus crispatus, Lactobacillus helveticus, and Lactobacillus johnsonii. In some embodiments, at least one member of the genus Lactobacillus is selected from the group consisting of Lactobacillus reuteri, Lactobacillus delbrueckii, Lactobacillus rhamnosus, Lactobacillus casei, Lactobacillus acidophilus, Lacticaseibacillus paracasei, Lactiplantibacillus plantarum, Levilactobacillus brevis, Lactobacillus casei, Lactobacillus casei, Lactobacillus acidophilus, Lacticaseibacillus paracasei, Lactiplantibacillus plantarum, Lactobacillus casei, Lactobacillus casei, Lactobacillus casei, Lactobacillus casei, Lactobacillus casei, Lactobacillus casei, Lactiplantib ...Lactobacillus brevis, Ligilactobacillus salivarius, Limosilactobacillus fermentum, Lactobacillus bulgaricus, Lactobacillus crispatus, Lactobacillus helveticus, and Lactobacillus johnsonii; Lactobacillus reuteri, Lactobacillus delbrueckii, Lactobacillus rhamnosus, Lactobacillus casei, Lactobacillus acidophilus The bacterial strains are grown or produced using a combination of metabolic stress and quorum sensing ability of at least one constituent species of Lactobacillus acidophilus, Lacticaseibacillus paracasei, Lactiplantibacillus plantarum, Levilactobacillus brevis, Ligilactobacillus salivarius, Limosilactobacillus fermentum, Lactobacillus bulgaricus, Lactobacillus crispatus, Lactobacillus helveticus, and Lactobacillus johnsonii. In some embodiments, at least one member of the genus Lactobacillus is selected from the group consisting of Lactobacillus reuteri, Lactobacillus delbrueckii, Lactobacillusdelbrueckii, Lactobacillus rhamnosus, Lactobacillus casei, Lactobacillus acidophilus, Lacticaseibacillus paracasei, Lactiplantibacillus plantarum, Levilactobacillus brevis, Ligilactobacillus salivarius, Limosilactobacillus fermentum, Lactobacillus bulgaricus, Lactobacillus crispatus The strain is selected from the group consisting of Lactobacillus crispatus, Lactobacillus helveticus, and Lactobacillus johnsonii.

[0096] In some embodiments, at least one member species of the Lactobacillus genus is Lactobacillus reuteri. In some embodiments, at least one member species of Lactobacillus reuteri is Lactobacillus reuteri DSM17648. In some embodiments, Lactobacillus reuteri DSM17648 is produced or grown using a combination of metabolic stress and quorum sensing capabilities of Lactobacillus reuteri DSM17648.

[0097] Additionally, WO 2007 / 073709 identifies several other suitable Lactobacillus strains, which are listed at the German Collection of Microorganisms and Cell Cultures GmbH (DSMZ), Mascheroder Weg 1b, D-38124, Braunschweig, Germany, namely: DSM 17646, DSM 17647, DSM 17649, DSM 17650, DSM 17651, DSM 17652, and DSM 17653. In some embodiments, at least one member of the Lactobacillus genus is Lactobacillus reuteri UBLRu-87 (MTCC 5403).

[0098] Antimicrobial metabolites can be toxic to pathogenic microorganisms, such as bacterial pathogens. In some embodiments, a composition or formulation of the invention comprises an antimicrobial metabolite that is toxic to Helicobacter, such as Helicobacter pylori.

[0099] In some embodiments, at least one species of Lactobacillus in the compositions of the present invention is lyophilized. In some embodiments, at least one species of Lactobacillus in the compositions of the present invention is freeze-dried. In some embodiments, at least one species of Lactobacillus in the compositions of the present invention is live.

[0100] As used in the present invention, a therapeutically effective amount of a composition comprising the compound of the present invention can be delivered to a patient as part of a regimen.

[0101] In some embodiments, an antibacterial composition is provided, wherein the regimen comprises one or more administration methods of zeolite and at least one member species of the genus Lactobacillus.In some embodiments, the administration regimen of the antibacterial composition is suitable for oral administration, rectal administration, topical administration, enteral administration, vaginal administration, buccal administration, orthotopic administration, intratracheal administration, intralesional administration, endoscopic administration, transmucosal administration, sublingual administration, enteral administration, and combinations thereof.In some embodiments, the antibacterial composition is suitable for oral administration.

[0102] As used herein, the term "regimen" refers to a schedule or set of rules for different possible routes or modes of administration, preferably to achieve a therapeutically effective amount of the antibacterial composition of the present invention. Those skilled in the art will recognize that different regimens may be employed in the context of the present invention.

[0103] In some embodiments, regimens of administering ivermectin, avermectin, doramectin, selamectin, moxidectin, emamectin, eprinomectin, milbemectin, abamectin, milbemycin oxime, nemadection, and their macrolide derivatives lacking a sugar residue attached to the 13 carbon, in free form or in the form of a physiologically acceptable derivative and / or salt thereof, are carried out using methods known in the art.

[0104] As used herein, the term "administration" should be understood to include, for example, oral administration, rectal administration, topical administration, enteral administration, vaginal administration, buccal administration, orthotopic administration, intratracheal administration, intralesional administration, endoscopic administration, transmucosal administration, sublingual administration, enteral administration, and combinations thereof.

[0105] Oral administration: The compositions of the present invention can be formulated to take the form of tablets or capsules prepared by conventional means using one or more pharmaceutically acceptable carriers (e.g., excipients such as binders, fillers, lubricants, and disintegrants). The inventors have found that the compositions can be conveniently administered to subjects by oral route, particularly in the form of tablets or capsules (e.g., tablets). In some embodiments, the specific dosage regime contemplated by the present invention is particularly suitable for oral administration in the form of tablets or capsules formulated to release the compounds used in the present invention, such as zeolite, and at least one member species of the Lactobacillus genus, calcium carbonate, and magnesium carbonate. The term "modified" or "modified release" used herein in connection with the compositions of the present invention or in any other context means a release that is not immediate, and is considered to include controlled release, sustained release, prolonged release, timed release, retarded release, extended release, and delayed release.

[0106] Controlled-release administration: Controlled-release (or sustained-release) formulations can be formulated to extend the activity of a substance and reduce the frequency of administration. Controlled-release formulations can also be used to affect other characteristics, such as the time of onset of action or blood levels of the substance, and thereby affect the occurrence of side effects.

[0107] Controlled-release formulations can be designed to initially release an amount of substance that produces a desired therapeutic effect, followed by a gradual, continuous release of other amounts of substance to maintain that level of therapeutic effect over an extended period of time. To maintain a near-constant level of substance in the body, the substance can be released from the dosage form at a rate that replenishes the amount of the composition of the present invention that is metabolized and / or excreted from the body. Controlled-release can be stimulated by various inducers, such as changes in pH, temperature, enzymes, water, and / or other physiological conditions, or molecules such as ammonia.

[0108] Controlled-release systems can include, for example, infusion pumps, which can be used to administer the composition in a manner similar to that typically used to deliver insulin or chemotherapy to the body or to specific organs or tumors. Typically, using such systems, the composition is administered in combination with a biodegradable, biocompatible polymeric implant that releases the composition at a selected site over a controlled period of time. Exemplary polymeric materials include polyanhydrides, polyorthoesters, polyglycolic acid, polylactic acid, polyethylene vinyl acetate, and copolymers and combinations thereof. Furthermore, controlled-release systems can be placed near the therapeutic target, such as a GIT ulcer, thereby requiring only a fraction of the oral dose.

[0109] The compositions of the present invention can be administered by other controlled-release means or delivery devices known to those skilled in the art. These include, for example, hydropropylmethyl cellulose, other polymer matrices, gels, permeable membranes, osmotic systems, multilayer coatings, microparticles, liposomes, microspheres, or the like, or combinations of any of the above to provide the desired release profile in varying proportions. Other methods of controlled-release delivery of the compositions or formulations of the present invention are known to those skilled in the art and are within the scope of the present invention. Those skilled in the art will understand that the amounts of the various components of the compositions of the present invention required for the uses and methods of the present invention will vary depending on the nature or severity of the condition being treated, the age, weight, existing treatments, and overall condition of the subject.

[0110] In some embodiments, antimicrobial compositions are provided that further comprise a bioactive substance or mineral, such as, for example, calcium carbonate, magnesium carbonate, vitamins, such as, for example, vitamin D and vitamin E, pharmaceutically acceptable carriers, additives, and adjuvants, such as antimicrobial chitins, such as chitosan, α-ketoglutarate, citrate, and lactate.

[0111] In some embodiments, an antimicrobial composition is provided, wherein the dosage regimen of zeolite and at least one member species of the Lactobacillus genus is suitable for oral or intragastric administration.

[0112] In some embodiments, an antimicrobial composition is provided that includes activated zeolite clinoptilolite (aZC), Lactobacillus reuteri DSM17648 or Lactobacillus reuteri UBLRu-87, calcium carbonate, and magnesium carbonate.

[0113] In some embodiments, antimicrobial compositions are provided that result in a reduction of one or more markers.

[0114] In some embodiments, an antimicrobial composition is provided wherein the marker is a predictive marker selected from the group consisting of urea, gastric ammonia, acid reflux, bloating, and urease activity.

[0115] In some embodiments, an antimicrobial composition is provided wherein the predictive marker is gastric ammonia.

[0116] In some embodiments, an antibacterial composition is provided that results in a reduction of intragastric ammonia levels in a vertebrate infected with Helicobacter pylori.

[0117] In some embodiments, antimicrobial compositions are provided that result in a reduction of gastric ammonia levels in vertebrates infected with Staphylococcus, e.g., Staphylococcus capitius urealiticum infection; Proteus, e.g., Proteus mirabilis infection; Klebsiella, e.g., Klebsiella pneumoniae infection, and Mycobacterium, e.g., Mycobacterium tuberculosis infection.

[0118] In some embodiments, antimicrobial compositions are provided that reduce gastric ammonia by at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% or more compared to placebo.

[0119] In some embodiments, antimicrobial compositions are provided that reduce gastric ammonia by at least 0.1, 0.25, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, or 10 times or more compared to placebo.

[0120] In some embodiments, antimicrobial compositions are provided wherein the reduction in predictive markers is statistically significant compared to placebo.

[0121] In some embodiments, antibacterial compositions are provided that result in a statistically significant reduction in gastric ammonia marker levels in humans infected with Helicobacter pylori. In some embodiments, antibacterial compositions are provided that result in a statistically significant clinical improvement in humans infected with Helicobacter pylori.

[0122] The term "regression" can be used interchangeably with the terms "reduction," "decrease," or "reducing" a disease or disorder, viral load, or at least one discernible symptom or marker caused by or associated with a Helicobacter infection, such as Helicobacter pylori. In some embodiments, the reduction is at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% or more compared to a control or placebo. In some embodiments, the reduction is at least 0.1, 0.25, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, or 10-fold or more compared to a control or placebo. In some embodiments, the regression or reduction is statistically significant. In some embodiments, the efficacy or treatment of the present invention is assessed by a quantification step. In some embodiments, the regression is assessed by a quantification step. In another embodiment, the quantification step is performed on a sample. In another embodiment, the quantification step is performed by immunoassay. In some embodiments, the sample is one of a plasma sample, a blood sample, a sputum sample, a lavage fluid, a synovial fluid, or a combination thereof.

[0123] In some embodiments, regression refers to a disease or disorder, or at least one identifiable symptom, caused by or associated with a Helicobacter infection, such as Helicobacter pylori. In some embodiments, regression refers to a disease or disorder, or at least one identifiable symptom, caused by or associated with a urease-active bacterial infection, including a Staphylococcus infection, e.g., Staphylococcus capitius urealiticum infection; a Proteus infection, e.g., Proteus mirabilis infection; a Klebsiella infection, e.g., Klebsiella pneumoniae infection; and a Mycobacterium infection, e.g., Mycobacterium tuberculosis infection. In some embodiments, regression is used, for example, to provide an indication of the degree to which a disease, disorder, or symptom changes, e.g., in terms of frequency of occurrence, intensity, and severity. In some embodiments, regression means, for example, a decrease, drop, lowering, or alleviation of fever in a treated subject. In some embodiments, reduction means a decrease, drop, lowering, or alleviation of viral load in a treated patient. In some embodiments, the treatment accelerates or speeds up the reduction in viral load in a treated patient compared to placebo. In some embodiments, the reduction in viral load is statistically significant.

[0124] As used herein, the term "significant" refers to, for example, a reduction, enhancement, amelioration, improvement, prevention, or reversal that is statistically significant, is not due to mere chance, and has a p-value of 0.05 or less. In particular, the term "significant" refers to, for example, a reduction, enhancement, amelioration, improvement, prevention, or reversal of a disease, disorder, or symptom caused by or associated with infection with a Helicobacter species, such as Helicobacter pylori, when compared to the level or frequency of implantation or pregnancy in one or more untreated patients, or compared to the level or frequency of implantation or pregnancy in the same patients observed at an earlier time point (e.g., compared to a "baseline" level or placebo), and may have a p-value of less than 0.05, 0.04, 0.03, 0.01, 0.005, 0.001, etc. Those skilled in the art will be familiar with various statistical computation approaches. Examples include t-tests, z-tests, sample tests, and the O'Brien-Fleming method for normally distributed data.

[0125] As used herein, the term "significant" refers to, for example, a reduction, enhancement, remission, improvement, prevention, or reversal that is statistically significant and not due to chance alone, with a p-value of 0.05 or less. In particular, the term "significant" refers to, for example, a Staphylococcus spp., e.g., Staphylococcus capitius urealiticum infection; a Proteus spp., e.g., Proteus mirabilis infection; a Klebsiella spp., e.g., Klebsiella pneumoniae infection; and a Mycobacterium spp., e.g., Mycobacterium tuberculosis infection. When referring to a reduction, enhancement, amelioration, improvement, prevention, or reversal of a disease, disorder, or symptom caused by or associated with a HIV-infected (tuberculosis) infection, for example, when compared to the level or frequency of implantation or pregnancy in one or more untreated patients, or compared to the level or frequency of implantation or pregnancy in the same patients observed at an earlier time point (e.g., compared to a "baseline" level or placebo), it may have a p-value of less than 0.05, less than 0.04, less than 0.03, less than 0.01, less than 0.005, less than 0.001, etc. Those skilled in the art will be familiar with various statistical calculation approaches. Examples include t-tests, z-tests, sample tests, the O'Brien-Fleming method for normally distributed data, etc.

[0126] In some embodiments, significant reduction, enhancement, amelioration, improvement, prevention, or reversal refers to a statistically significant reduction, enhancement, amelioration, improvement, prevention, or reversal of a disease, disorder, or symptom caused by or associated with infection with a Helicobacter bacterium, such as Helicobacter pylori.

[0127] In one aspect of the invention, there is provided use of an antibacterial composition for the preparation or manufacture of a pharmaceutical formulation, for separate, combined or simultaneous administration, designed to treat, alleviate or prevent a GIT disease or disorder, gastritis, gastric ulcer, duodenal ulcer, gastric cancer or duodenal cancer in a vertebrate, including mammals and birds, in need of such treatment, alleviation or prevention.

[0128] Without wishing to be bound by theory, the inventors hypothesize that the body naturally produces excess stomach acid in an inflammatory response to combat Helicobacter pylori. The body also recognizes the need for buffering when this stomach acid becomes too much and uses calcium and magnesium stores to achieve this. Over time, excess acidity and depletion of magnesium and calcium stores can lead to numerous other illnesses in addition to the gastric disorders caused by autoimmune responses and Helicobacter pylori colonization. The use of the non-antibiotic antimicrobial composition of the present invention has been experimentally demonstrated to effectively reduce ammonia and urea levels in affected subjects, such as humans, as well as completely eliminate the pathogen Helicobacter pylori, thereby alleviating and / or preventing symptoms, diseases, and conditions associated with Helicobacter pylori.

[0129] Furthermore, it has been hypothesized that the same situation applies to other urease-positive bacterial infections in the gastrointestinal tract.

[0130] In some embodiments, the present invention is directed to the use of antimicrobial compositions wherein the pharmaceutical formulation is in a form suitable for administering a daily dose of the antimicrobial composition, either in a single dose or as divided doses or subdoses administered at appropriate intervals.

[0131] In some embodiments, the present invention is directed to the use of antibacterial compositions wherein the pharmaceutical formulation is administered as one, two, three, four or more doses or subdoses per day or administration.

[0132] In some embodiments, the present invention provides a pharmaceutical formulation comprising 10 7 ~10 9 The antibacterial composition comprises CFU / g of Lactobacillus reuteri DSM 17648 or Lactobacillus reuteri UBLRu-87, and a zeolite, wherein the zeolite is present in an amount of at least 50 mg / gram of the formulation.

[0133] In some embodiments, the zeolite is typically administered in the range of about 0.1-2000 mg / kg body weight, about 0.15-1750 mg / kg body weight, about 0.2-1700 mg / kg body weight, about 0.3-1500 mg / kg body weight, about 0.5-1250 mg / kg body weight, about 1-1000 mg / kg body weight, about 2-900 mg / kg body weight, about 3-800 mg / kg body weight, about 4-700 mg / kg body weight, about 5-600 mg / kg body weight, or about 10-500 mg / kg body weight, administered as one, two, three, four, or more doses or subdoses per day or administration. In some embodiments, the dose is 400 mg / kg body weight, administered as one, two, three, four, or more doses or subdoses per day or administration.

[0134] In some embodiments, the invention is directed to the use of an antimicrobial composition, wherein the pharmaceutical formulation further comprises calcium carbonate and magnesium carbonate, wherein the calcium carbonate and magnesium carbonate are each present in an amount of at least 0.5 mg per gram of composition or formulation.

[0135] In some embodiments, calcium carbonate and magnesium carbonate are typically within the range of about 0.1-200 mg / kg body weight, about 0.15-150 mg / kg body weight, about 0.2-120 mg / kg body weight, about 0.3-100 mg / kg body weight, about 0.5-75 mg / kg body weight, about 1-50 mg / kg body weight, about 2-40 mg / kg body weight, about 3-30 mg / kg body weight, about 4-20 mg / kg body weight, or about 5-10 mg / kg body weight, and the daily dose or administration may be administered as one, two, three, four, or more doses or subdoses.

[0136] In some embodiments, the present invention is directed to the use of antimicrobial compositions wherein the dose of the antimicrobial composition is formulated as a capsule, lyophilized formulation, liquid, pill, powder, or gel.

[0137] In some embodiments, the present invention is directed to the use of antibacterial compositions where treatment results in complete remission of a disease or disorder caused by or associated with infection with Helicobacter.

[0138] In some embodiments, the composition comprises at least one member species of the genus Lactobacillus in an amount of about 10 10 In some embodiments, the inoculated aqueous medium contains at least one member species of the genus Lactobacillus at about 10 cells / ml. 9 In some embodiments, the inoculated aqueous medium contains at least one member species of the genus Lactobacillus at about ____ cells / ml. In some embodiments, the inoculated aqueous medium contains at least one member species of the genus Lactobacillus at about ____ cells / ml. In some embodiments, the inoculated aqueous medium contains at least one member species of the genus Lactobacillus at about ____ cells / ml. 7 In some embodiments, the inoculated aqueous medium contains at least one member species of the genus Lactobacillus at about ____ cells / ml. In some embodiments, the inoculated aqueous medium contains at least one member species of the genus Lactobacillus at about ____ cells / ml. In some embodiments, the inoculated aqueous medium contains at least one member species of the genus Lactobacillus at about ____ cells / ml. 5 Contains cells / ml.

[0139] In some embodiments, an antimicrobial agent is provided wherein at least one member species of the Lactobacillus genus is probiotic.

[0140] Those skilled in the art will know that microbial cell viability can be assessed or estimated using colony forming units per milliliter (CFU / mL) if a liquid is being tested, or colony forming units per gram (CFU / g) if a solid is being tested.

[0141] In some embodiments, when the composition of at least one Lactobacillus species is liquid, the concentration of viable cells can be estimated via colony forming units per milliliter (CFU / mL or growth medium). In some embodiments, the concentration of viable cells in the composition is 0.5×10 6 ~1×10 9 CFU / mL, 0.5 × 10 6 ~500×10 6 CFU / mL, 0.5 × 10 6 ~400×10 6 CFU / mL, 0.5 × 10 6 ~300×10 6 CFU / mL, 0.5 × 10 6 ~200×10 6 CFU / mL, 0.5 × 10 6 ~150×10 6 CFU / mL, 0.5 × 10 6 ~125×10 6 CFU / mL, 0.5 × 10 6 ~100×10 6 CFU / mL, 0.5 × 10 6 ~75×10 6 CFU / mL, 0.5 × 10 6 ~50×10 6 CFU / mL, 0.5 × 10 6 ~10×10 6 CFU / mL, 0.5 × 10 6 ~5×10 6 CFU / mL, 0.5 × 10 6 ~1×10 6 CFU / mL, 1 × 10 6 ~1×10 9CFU / mL、1×10 6 ~500×10 6 CFU / mL、1×10 6 ~400×10 6 CFU / mL、1×10 6 ~300×10 6 CFU / mL、1×10 6 ~200×10 6 CFU / mL、1×10 6 ~150×10 6 CFU / mL、1×10 6 ~125×10 6 CFU / mL、1×10 6 ~100×10 6 CFU / mL、1×10 6 ~75×10 6 CFU / mL、1×10 6 ~50×10 6 CFU / mL、1×10 6 ~10×10 6 CFU / mL、1×10 6 ~5×10 6 CFU / mL、5×10 6 ~1×10 9 CFU / mL、5×10 6 ~500×10 6 CFU / mL、5×10 6 ~400×10 6 CFU / mL、5×10 6 ~300×10 6 CFU / mL、5×10 6 ~200×10 6 CFU / mL、5×10 6 ~150×10 6 CFU / mL、5×10 6 ~125×10 6 CFU / mL、5×10 6 ~100×10 6 CFU / mL、5×10 6 ~75×10 6 CFU / mL、5×10 6 ~50×10 6 CFU / mL、5×10 6 ~10×10 6 CFU / mL、10×10 6 ~1×10 9CFU / mL、10×10 6 ~500×10 6 CFU / mL、10×10 6 ~400×10 6 CFU / mL、10×10 6 ~300×10 6 CFU / mL、10×10 6 ~200×10 6 CFU / mL、10×10 6 ~150×10 6 CFU / mL、10×10 6 ~125×10 6 CFU / mL、10×10 6 ~100×10 6 CFU / mL、10×10 6 ~75×10 6 CFU / mL、10×10 6 ~50×10 6 CFU / mL、50×10 6 ~1×10 9 CFU / mL、50×10 6 ~500×10 6 CFU / mL、50×10 6 ~400×10 6 CFU / mL、50×10 6 ~300×10 6 CFU / mL、50×10 6 ~200×10 6 CFU / mL、50×10 6 ~150×10 6 CFU / mL、50×10 6 ~125×10 6 CFU / mL、50×10 6 ~100×10 6 CFU / mL、50×10 6 ~75×10 6 CFU / mL、100×10 6 ~1×10 9 CFU / mL、100×10 6 ~500×10 6 CFU / mL、100×10 6 ~400×10 6 CFU / mL、100×10 6 ~300×10 6 CFU / mL、100×106 ~200×10 6 CFU / mL、100×10 6 ~150×10 6 CFU / mL、100×10 6 ~125×10 6 CFU / mL、125×10 6 ~1×10 9 CFU / mL、125×10 6 ~500×10 6 CFU / mL、125×10 6 ~400×10 6 CFU / mL、125×10 6 ~300×10 6 CFU / mL、125×10 6 ~200×10 6 CFU / mL、125×10 6 ~150×10 6 CFU / mL、150×10 6 ~1×10 9 CFU / mL、150×10 6 ~500×10 6 CFU / mL、150×10 6 ~400×10 6 CFU / mL、150×10 6 ~300×10 6 CFU / mL、150×10 6 ~200×10 6 CFU / mL、200×10 6 ~1×10 9 CFU / mL、200×10 6 ~500×10 6 CFU / mL、200×10 6 ~400×10 6 CFU / mL、200×10 6 ~300×10 6 CFU / mL、300×10 6 ~1×10 9 CFU / mL、300×10 6 ~500×10 6 CFU / mL、300×10 6 ~400×10 6 CFU / mL、400×10 6 ~1×10 9 CFU / mL、400×106 ~500×10 6 CFU / mL, or 500 x 10 6 ~1×10 9 CFU / mL or more (10 x 10 9 CFU / ml, etc.

[0142] In some embodiments, when the composition is solid, the concentration of viable cells of at least one member species of the genus Lactobacillus can be estimated via colony forming units per gram (CFU / g). In some embodiments, the concentration of at least one member species of the genus Lactobacillus is 0.5×10 6 ~1×10 9 CFU / g, 0.5 × 10 6 ~500×10 6 CFU / g, 0.5 × 10 6 ~400×10 6 CFU / g, 0.5 × 10 6 ~300×10 6 CFU / g, 0.5 × 10 6 ~200×10 6 CFU / g, 0.5 × 10 6 ~150×10 6 CFU / g, 0.5 × 10 6 ~125×10 6 CFU / g, 0.5 × 10 6 ~100×10 6 CFU / g, 0.5 × 10 6 ~75×10 6 CFU / g, 0.5 × 10 6 ~50×10 6 CFU / g, 0.5 × 10 6 ~10×10 6 CFU / g, 0.5 × 10 6 ~5×10 6 CFU / g, 0.5 × 10 6 ~1×10 6 CFU / g, 1 × 10 6 ~1×10 9 CFU / g, 1 × 10 6 ~500×10 6 CFU / g, 1 × 10 6 ~400×10 6 CFU / g, 1 × 106 ~300×10 6 CFU / g、1×10 6 ~200×10 6 CFU / g、1×10 6 ~150×10 6 CFU / g、1×10 6 ~125×10 6 CFU / g、1×10 6 ~100×10 6 CFU / g、1×10 6 ~75×10 6 CFU / g、1×10 6 ~50×10 6 CFU / g、1×10 6 ~10×10 6 CFU / g、1×10 6 ~5×10 6 CFU / g、5×10 6 ~1×10 9 CFU / g、5×10 6 ~500×10 6 CFU / g、5×10 6 ~400×10 6 CFU / g、5×10 6 ~300×10 6 CFU / g、5×10 6 ~200×10 6 CFU / g、5×10 6 ~150×10 6 CFU / g、5×10 6 ~125×10 6 CFU / g、5×10 6 ~100×10 6 CFU / g、5×10 6 ~75×10 6 CFU / g、5×10 6 ~50×10 6 CFU / g、5×10 6 ~10×10 6 CFU / g、10×10 6 ~1×10 9 CFU / g、10×10 6 ~500×10 6 CFU / g、10×10 6 ~400×10 6 CFU / g、10×10 6 ~300×106 CFU / g、10×10 6 ~200×10 6 CFU / g、10×10 6 ~150×10 6 CFU / g、10×10 6 ~125×10 6 CFU / g、10×10 6 ~100×10 6 CFU / g、10×10 6 ~75×10 6 CFU / g、10×10 6 ~50×10 6 CFU / g、50×10 6 ~1×10 9 CFU / g、50×10 6 ~500×10 6 CFU / g、50×10 6 ~400×10 6 CFU / g、50×10 6 ~300×10 6 CFU / g、50×10 6 ~200×10 6 CFU / g、50×10 6 ~150×10 6 CFU / g、50×10 6 ~125×10 6 CFU / g、50×10 6 ~100×10 6 CFU / g、50×10 6 ~75×10 6 CFU / g、100×10 6 ~1×10 9 CFU / g、100×10 6 ~500×10 6 CFU / g、100×10 6 ~400×10 6 CFU / g、100×10 6 ~300×10 6 CFU / g、100×10 6 ~200×10 6 CFU / g、100×10 6 ~150×10 6 CFU / g、100×10 6 ~125×10 6 CFU / g、125×106 ~1×10 9 CFU / g, 125×10 6 ~500×10 6 CFU / g, 125×10 6 ~400×10 6 CFU / g, 125×10 6 ~300×10 6 CFU / g, 125×10 6 ~200×10 6 CFU / g, 125×10 6 ~150×10 6 CFU / g, 150×10 6 ~1×10 9 CFU / g, 150×10 6 ~500×10 6 CFU / g, 150×10 6 ~400×10 6 CFU / g, 150×10 6 ~300×10 6 CFU / g, 150×10 6 ~200×10 6 CFU / g, 200×10 6 ~1×10 9 CFU / g, 200×10 6 ~500×10 6 CFU / g, 200×10 6 ~400×10 6 CFU / g, 200×10 6 ~300×10 6 CFU / g, 300×10 6 ~1×10 9 CFU / g, 300×10 6 ~500×10 6 CFU / g, 300×10 6 ~400×10 6 CFU / g, 400×10 6 ~1×10​​​​​​​​​​​​​​ In some embodiments, the present invention provides a pharmaceutical formulation comprising 10 7 ~10 9 The antibacterial composition comprises CFU / g of Lactobacillus reuteri DSM 17648 or Lactobacillus reuteri UBLRu-87, and a zeolite, wherein the zeolite is present in an amount of at least 50 mg / gram of the formulation.

[0144] In some embodiments, the invention is directed to the use of an antimicrobial composition, wherein the pharmaceutical formulation further comprises calcium carbonate and magnesium carbonate, wherein the calcium carbonate and magnesium carbonate are each present in an amount of at least 0.5 mg per gram of formulation.

[0145] In some embodiments, the present invention is directed to the use of antimicrobial compositions wherein the dose of the antimicrobial composition is formulated as a capsule, lyophilized formulation, liquid, pill, powder, or gel.

[0146] In some embodiments, the present invention is directed to the use of antibacterial compositions where treatment results in complete remission of a disease or disorder caused by or associated with infection with Helicobacter.

[0147] In some embodiments, the present invention is directed to the use of antibacterial compositions wherein the treatment results in complete remission of a disease or disorder caused by or associated with Staphylococcus, e.g., Staphylococcus capitius urealiticum infection; Proteus, e.g., Proteus mirabilis infection; Klebsiella, e.g., Klebsiella pneumoniae infection, and Mycobacterium, e.g., Mycobacterium tuberculosis infection.

[0148] Quorum Sensing Without wishing to be bound by theory, quorum sensing is generally thought to represent a response to fluctuations in cell population density. Quorum-sensing microorganisms, such as bacteria, produce and release chemical signal molecules called autoinducers, whose concentrations increase as a function of cell density. When a minimum threshold concentration of autoinducer is detected, gene expression changes occur. Microorganisms, including Gram-positive and Gram-negative bacteria, use quorum-sensing signaling pathways to regulate a wide variety of physiological activities. These processes include symbiosis, virulence, competence, conjugation, antibiotic production, motility, sporulation, and biofilm formation. Generally, Gram-negative bacteria use acylated homoserine lactones as autoinducers, while Gram-positive bacteria process and use oligopeptides for signaling. Recent advances in this field have demonstrated that autoinducer-mediated intercellular communication occurs both within and between bacterial species.

[0149] The desired level of antimicrobial metabolites may depend on the desired subsequent use of the antimicrobial composition.

[0150] In some embodiments, the desired level of antimicrobial metabolite is at the peak of the exponential phase.

[0151] In some embodiments, growth or proliferation of the antimicrobial culture is substantially reduced or stopped once the peak log phase is reached.

[0152] In some embodiments, an antimicrobial metabolite can be toxic to a pathogenic microorganism, such as a bacterial pathogen. As used herein, the term "toxic" refers to a toxin or other microbial substance or metabolite that induces an immune response, particularly the production of antibodies.

[0153] In various embodiments, the grown antimicrobial cultures are useful for treating mammalian, particularly human, diseases caused by microorganisms such as bacteria through inhibition of the bacterial quorum-sensing cascade, rendering the pathogen avirulent. Such diseases include endocarditis, respiratory and pulmonary infections (preferably in immunocompromised patients), bacteremia, skin conditions, vaginal infections, colon infections, central nervous system infections, ear infections including otitis externa, eye infections, bone and joint infections, urinary tract infections, gastrointestinal infections, and skin and soft tissue infections including wound infections, pyoderma, and atopic dermatitis, all of which can be induced by Helicobacter species, such as Helicobacter pylori.

[0154] In some embodiments, the grown antimicrobial cultures are freeze-dried. As used herein, the term "lyophilization" refers to preserving grown antimicrobial cultures by very quickly freezing them and then subjecting them to vacuum or sublimation to remove the ice. In some embodiments, the freeze-dried antimicrobial cultures are stored long-term. In some embodiments, the freeze-dried antimicrobial cultures contain viable microbial cells. In some embodiments, freeze-drying can be used to prepare dosage forms that are reconstituted for injection. In some embodiments, the grown antimicrobial cultures are dehydrated or otherwise dried.

[0155] In some embodiments, the freeze-dried antimicrobial culture is a freeze-dried antimicrobial composition. In some embodiments, the grown antimicrobial culture is in the form of a powder.

[0156] In some embodiments, the freeze-dried antimicrobial culture is a freeze-dried antimicrobial composition. In some embodiments, the freeze-dried antimicrobial composition comprises zeolite, and at least one member species of the Lactobacillus genus is Lactobacillus reuteri. In some embodiments, the freeze-dried antimicrobial composition comprises zeolite and Lactobacillus reuteri DSM 17648. In some embodiments, the freeze-dried antimicrobial composition comprises zeolite, Lactobacillus reuteri DSM 17648, and mineral salts. In some embodiments, the freeze-dried antimicrobial composition comprises zeolite, Lactobacillus reuteri DSM 17648, calcium carbonate, and magnesium carbonate. In these embodiments, the Lactobacillus reuteri strain may be UBLRu-87.

[0157] In some embodiments, a grown antimicrobial culture is provided in combination with one or more pharmaceutically acceptable ingredients, hi some embodiments, a grown antimicrobial culture is provided in combination with one or more pharmaceutically acceptable excipients or additives.

[0158] In the context of this specification, a "pharmaceutically acceptable excipient" refers to an inert substance added to a pharmaceutical composition to further facilitate administration of a grown antimicrobial culture, e.g., a lyophilized culture. Examples of excipients include, but are not limited to, calcium carbonate, calcium phosphate, various sugars and types of starch, cellulose derivatives, gelatin, vegetable oils, and polyethylene glycols.

[0159] In some embodiments, the pharmaceutical composition comprises an antibacterial agent. In some embodiments, the pharmaceutical composition comprises an antibacterial agent.

[0160] Thus, the present invention also relates to compositions, including pharmaceutical compositions, comprising a therapeutically effective amount of a grown antimicrobial culture, e.g., a lyophilized culture, as described herein. As used herein, a grown antimicrobial culture, e.g., a lyophilized culture, exhibits a therapeutic effect if it is capable of affecting a target microbial concentration.

[0161] Preferably, a grown antimicrobial culture, e.g., a lyophilized culture or formulation, exhibits a therapeutic effect if it is capable of affecting a target microbial concentration that is capable of treating or preventing a microbial-associated disease, such as a bacterial-associated disease or disorder, in a subject after the composition, e.g., a lyophilized composition, is administered to the subject. In some embodiments, the disease or disorder is associated with a pathogen.

[0162] In some embodiments, the pathogen is a Helicobacter. In some embodiments, the pathogen is Helicobacter canis, Helicobacter felis, Helicobacter heilmannii, Helicobacter mustelae, Helicobacter bizzozeronii, Helicobacter acinonyx, Helicobacter fenneliae, Helicobacter rappini, Helicobacter hepaticus, Helicobacter pullorum, Helicobacter bilis, Helicobacter rodentium, Helicobacter rodentium, Helicobacter trogontum, Helicobacter cinaedi, Helicobacter muridarum, Helicobacter pametensis, Helicobacter cholecyctus, and Helicobacter pylori.

[0163] In some embodiments of the invention, the disease or condition is associated with or caused by Helicobacter canis.

[0164] In some embodiments, the pathogen is Helicobacter pylori.

[0165] In some embodiments, the pathogen is a Staphylococcus spp., e.g., Staphylococcus capitius urealiticum infection; a Proteus spp., e.g., Proteus mirabilis infection; a Klebsiella spp., e.g., Klebsiella pneumoniae infection, and a Mycobacterium spp., e.g., Mycobacterium tuberculosis infection.

[0166] In another embodiment, the grown antimicrobial cultures, e.g., lyophilized cultures, of the present invention can be administered directly to animals, preferably mammals, especially humans, as a non-antibiotic antibiotic per se, in admixture with one another, or in the form of a pharmaceutical formulation which allows for enteral or parenteral use and which contains an effective amount of the grown antimicrobial culture, e.g., lyophilized culture, as an active ingredient in addition to conventional pharmaceutical excipients and additives.

[0167] As mentioned above, in addition to the grown antibacterial culture, the culture may further contain conventional, usually inert, carrier substances, additives, or excipients. That is, the culture may contain additives or adjuvants commonly used in galenic preparations, such as fillers, extenders, disintegrants, binders, glidants, wetting agents, stabilizers, emulsifiers, preservatives, sweeteners, colorants, flavorings or aromatizers, buffer substances, solvents or solubilizers, or agents for achieving a depot effect, as well as salts for modifying osmotic pressure, coating agents, or antioxidants. The culture may also contain two or more cultures and may also contain other therapeutically active substances, such as antivirals, antifungals, or antibiotics.

[0168] Thus, the cultures of the invention can be used alone, in combination with other compounds of the invention, or in combination with other active compounds, for example active ingredients already known for the treatment of the aforementioned diseases, in the latter case advantageous additive effects being observed.

[0169] In another aspect, the present invention provides a process for preparing a product selected from a food, a beverage, a nutritional product, a dietary supplement, and an animal feed, the process comprising combining one or more ingredients with a grown antimicrobial culture.

[0170] In some embodiments, the grown antimicrobial culture product can incorporate an inert excipient, an inorganic excipient, or an organic excipient.

[0171] In some embodiments, for preparing pills, powders, tablets, coated tablets, and hard gelatin capsules, for example, lactose, corn starch or its derivatives, talc, stearic acid or its salts, etc., can be used. Excipients for soft gelatin capsules and suppositories include fats, waxes, semisolid and liquid polyols, natural or hardened oils, etc. Excipients suitable for producing solutions and syrups include water, alcohol, sucrose, invert sugar, glucose, polyols, etc.

[0172] Materials and Methods 1. Growth and maintenance of Helicobacter pylori in the laboratory

[0173] Various microbiological techniques and methods for maintaining and growing Helicobacter pylori will be known to those skilled in the art.

[0174] An example of such an experimental technique employed in the present invention can be found at the following link: https: / / www.ncbi.nlm.nih.gov / pmc / articles / PMC3357201 / #:~:text=CULTURE%20OF%20H ELICOBACTER%20ORGANISMS%20ON,as%20horse%2C%20ox%2C%20or%20sheep

[0175] 2. Urease output test

[0176] Urease activity assay kit (Sigma-Aldrich)

[0177] The urease test identifies organisms capable of hydrolyzing urea to produce ammonia and carbon dioxide. The urease test is primarily used to distinguish urease-positive Proteus species from other Enterobacteriaceae.

[0178] Method 1

[0179] The ureolytic activity assay using M9U medium allows for cell-based and cell-free screening of urease activity. This method continuously monitors and screens urease activity from both bacterial cells and pure urease in a urea-containing growth medium and in a plate reader configuration in real time. Chemically defined M9-based urea (M9U) medium was found to be more sensitive and suitable for a plate reader configuration than both Christensen's urea broth (CUB) and Stewart's urea broth (SUB), the well-established and well-known complex urea media that formed the principle foundation of M9U. Furthermore, urease activity measurement using M9U medium in a plate reader-based method allows for a reliable, high-throughput screening system for urease inhibitors.

[0180] Development of an M9-based urea medium (M9U) for sensitive real-time monitoring of bacterial and cell-free urease ureolytic activity Jens Jakob Sigurdarson, Simon Svane, Henrik Karring: https: / / doi.org / 10.1002 / mbo3.976

[0181] Method 2

[0182] The developed assay provides a robust cellular model for testing urease inhibitors directly in a cellular environment. The effectiveness of co-expressed peptides was observed to affect the interaction between UreF and UreD, two accessory proteins required for urease activation. This phenomenon involves a folding-mediated process upon binding, suggesting the importance of intrinsically disordered hotspots at protein interfaces.

[0183] Targeting Helicobacter pylori urease activity and maturation: In-cell high-throughput approach for drug discovery CinziaTarsia, AlbertoDanielli, FrancescaFlorini, PaoloCinelli, StefanoCiurli, Barbara Zambelli.

[0184] Two media are commonly used to detect urease activity.

[0185] Christensen's urea agar is used to detect urease activity in various microorganisms. Stuart's urea broth is primarily used for the differentiation of Proteus species. With Christensen's urea agar, urease production appears as a bright pink (fuchsia) color on the slant, which may spread to the bottom after 1 to 6 hours of incubation.

[0186] If the organism is urease negative, the medium will remain yellowish in color.

[0187] In Stuart's Urea Broth, urease production is indicated by the broth turning a bright pink (fuchsia) color throughout.

[0188] Ammonia testing - exemplary predictive markers

[0189] Ammonium test kit Quantofix (Sigma-Aldrich) Ammonia Assay Kit (Sigma-Aldrich).

[0190] Four solutions with the following concentrations of N-NH4+: 50 mg / L, 100 mg / L, 300 mg / L, and 500 mg / L were prepared from a standard ammonium chloride solution with a concentration of 1 g / L. These solutions were subjected to pH analysis performed by potentiometry.

[0191] Clinoptilolite samples of 1 g, 2 g, and 3 g were weighed and added to 100 ml of N-NH solution. The prepared samples were shaken for 30, 60, and 180 minutes, after which the pH was measured. The samples were then filtered to measure the concentration of ammoniacal nitrogen.

[0192] Each solution was prepared in triplicate.

[0193] The ammonium concentration in the aqueous phase was measured by standard distillation. Prior to measuring ammoniacal nitrogen, the filtered samples were distilled (BUCHI 323) and the N-NH4+ concentration was measured using a spectrophotometer (Hach).

[0194] Testing Antimicrobial Compositions A, B, C, and D for Reduction of Ammonia, Ammonium, and Urease (Exemplary Predictive Markers)

[0195] FIG. 1 provides data from in vitro urea and ammonia reduction assays using antimicrobial compositions of the present invention.

[0196] Fixed-bed ion exchange column test To test the feasibility of natural zeolite (stilbite) as an adsorbent for removing nitrate from groundwater, a continuous application mode in a glass column (60 cm × 0.8 cm) was tested.

[0197] Natural zeolite (stilbite) was suspended in distilled water for approximately 10 minutes before use in the column test. Glass wool was placed at the bottom of the column to prevent loss of the adsorbent due to the flow of the nitric acid solution. The adsorbent was then transferred onto the glass wool inside the column. The nitric acid solution was pumped into the column at a flow rate of 10 ml / min using a peristaltic pump. Journal of Water Resource and Hydraulic Engineering Dec. 2014, Vol.b, Iss. 4, PP. 74-80 - 77 - To measure the exhaustive capacity, 50 ml of effluent was taken from the bottom of the column.

[0198] This process was continued until the amount of nitrate in the effluent was equal to that in the feed. The pH was measured using a pH meter (Erico Model LI-107). The effect of pH was investigated in the range of 2 to 7 by adjusting the pH of the nitric acid solution using dilute HCl and NaOH solutions.

[0199] The effect of flow rate was investigated by varying the flow rate between 1 and 5 mL / min using a peristaltic pump. The effect of different initial concentrations was investigated using nitric acid solutions with different concentrations ranging from 40 to 240 mg / L.

[0200] After each exchange service cycle, a regeneration cycle was performed by passing a 30 g / L sodium nitrate solution through the zeolite fixed bed from the top of the column. The regeneration process was stopped when the nitrate ion concentration in the effluent reached .

[0201] Testing the ability of antibacterial compositions A, B, C, and D to effectively reduce Helicobacter pylori load

[0202] In a model gut test using high ammonia / ammonium, what effect do antimicrobial compositions A, B, C, and D each exhibit?

[0203] Ammonia Exchange Capacity (AEC), Optimized

[0204] The inhibitory effect of Ca2+ on NH4+ exchange efficiency was fitted to a competitive inhibition Mono model with a half-saturation rate constant of 134.7 mg / L. The addition of Ca2+ decreased the NH4+ removal rate and prolonged the exchange equilibrium time of the zeolite.

[0205] Periodic precipitation of Ca2+ from the used regenerant in the form of calcium carbonate reduced the inhibitory effect of Ca2+, excellently maintained the NH4+ removal efficiency, and extended the service life of the zeolite.

[0206] Compared with the Bohart-Adams and Thomas models, the Dose-Response model could predict the breakthrough curve well, and the fitted parameters further confirmed that NaClO-NaCl regeneration and periodic Ca2+ removal were effective methods to maintain efficient NH4+ removal from wastewater by zeolite. [Example]

[0207] Helicobacter pylori infection is known to be associated with elevated ammonia and / or urea levels, which allows some of this pathogenic bacteria to survive and grow in the high pH environment sometimes found in the GIT.

[0208] However, removing or reducing the levels of ammonia and / or urea may adversely affect the survival of Helicobacter pylori when exposed to physiologically normal gastric acid conditions. The ability of the antibacterial compositions of the present invention to remove or reduce the levels of ammonia and / or urea produced by Helicobacter spp., such as Helicobacter pylori, was tested.

[0209] The Gram-positive (+ve) bacterium Lactobacillus L. reuteri clumps against exposed Helicobacter pylori and, together with active mineral salts such as calcium carbonate and / or magnesium carbonate, helps to remove the pathogen from the subject.

[0210] Carbonates help soothe stomach discomfort and help balance the stomach's natural functions.

[0211] Lactobacillius L. reuteri can survive the normal acidic conditions of the stomach and contribute to a more favorable microflora.

[0212] If the subject has any of the following symptoms of Helicobacter pylori: feeling of fullness, abdominal pain that worsens when the stomach is empty, nausea, shortness of breath, acid reflux, high ammonia / ammonium levels, ulcers, gastritis, stomach cancer, or black tarry stools.

[0213] Example - Antibacterial composition A used to treat Helicobacter pylori pathogen infection: -3.5g of dual activated zeolite clinoptilolite -10×10 9 Probiotic Lactobacillus reuteri DSM17648 culture.

[0214] Example - Antibacterial composition B used in the study to treat Helicobacter pylori pathogen infection: -2g calcium carbonate -2g magnesium carbonate -3.5g of dual activated zeolite clinoptilolite -10×10 9 Probiotic Lactobacillus reuteri DSM17648 culture.

[0215] Example - Antibacterial composition C for treating acid reflux and bloating associated with Helicobacter pylori infection. -1.5g of dual activated zeolite clinoptilolite -5×10 9 Probiotic Lactobacillus reuteri DSM17648 culture.

[0216] Example - Antibacterial Composition D for Treating Acid Reflux and Bloating Associated with Helicobacter pylori Infection: -1g calcium carbonate -1g magnesium carbonate -1.5g of dual activated zeolite clinoptilolite -5×10 9 Probiotic Lactobacillus reuteri DSM17648 culture.

[0217] Antimicrobial Compositions A, B, C, or D can be used in a regimen for treating a Helicobacter pylori pathogen infection in a subject, such as a human. -Sachets of powder, such as freeze-dried cultures of selected antimicrobial compositions - to be dissolved in water; -Suspension in lactulose -Tablet format

[0218] Antibacterial composition A, B, C, or D for use in a regimen for treating a Helicobacter pylori pathogen infection in a subject, such as an animal. -Sachets of powder, such as freeze-dried cultures of selected antibacterial compositions - to be dissolved in water; -Suspension in lactulose

[0219] Composition and physicochemical properties of the tribomechanically activated zeolite clinoptilolite (TMAZ®)

[0220] Chemical composition SiO2, 65.0~71.3%; Al2O3, 11.5~13.1%; CaO 2.7~5.2%; K2O, 2.2~3.4%; Fe2O3, 0.7~1.9%; MgO, 0.6~1.2%; Na2O, 0.2~1.3%; TiO2, 0.1~0.3%; Si / Al ratio, 4.8~5.4.

[0221] Empirical formula: (Ca, K2, Na2, Mg)4Al8Si40O96×24H2O -Physicochemical properties specific mass, 2.2~2.5g / cm 3 ;Porosity, 32~40%;Effective pore diameter, 0.4nm -Ion exchange capacity Total exchange capacity: 1.2-1.5 mol / kg; Ca 2+ , 0.64~0.98mol / kg;Mg 2+ , 0.06~0.19mol / kg;K + , 0.22~0.45mol / kg;Na + , 0.01~0.19mol / kg Ion exchange selectivity Cs>NH 4+ >Pb 2+ >K + >Na + >Mg 2+ >Ba 2+ >Cu 2+ >Zn 2+

[0222] Absorbed chemicals NH3, hydrocarbons C1-C4, CO2, H2S, SO2, NO X ,aldehyde -Toxicity Non-toxic; Generally Recognized as Safe (GRAS) under the US Code of Federal Regulations (21 CFR 182, Subpart C).

[0223] Patients Effective dosing regimen against Helicobacter pylori - regimen duration 5 days - Figure 2 - Pre- and post-treatment cfu shown. Days 1 to 5 Do not eat before your 8:00 AM treatment 8:00 AM: Antibacterial composition of choice (antibacterial composition B was used here) in 500 ml of water, taken on an empty stomach 9 AM: Plain toast, lots of water 10:00 AM Antibacterial Composition B in 500 ml of water 11:00 AM: Plain toast, lots of water 12:00 PM Antibacterial Composition B in 500 ml of water After 1pm, you can eat and drink as normal.

[0224] Effective in vivo testing on subjects such as human patients in a clinical setting.

[0225] Prior to May 2020, the patient was a healthy, active 45-year-old woman and mother of three children.

[0226] Her stomach had never been abnormal before, and after giving birth to all three children, it returned to normal quickly and she had a consistently flat belly.

[0227] In May 2020, the patient experienced severe bloating and visited her primary care physician, who advised her that her symptoms were simply age- and hormone-related, and that no tests were necessary, simply a manual examination of her abdomen would suffice.

[0228] However, the patient's vision began to deteriorate, his joints became stiff and painful, and the bloating in his stomach continued and worsened, putting pressure on his lungs and making it difficult to breathe.

[0229] On July 1, 2021, an x-ray confirmed mild to moderate degeneration of the patient's right metatarsophalangeal joint.

[0230] A blood test was performed on June 29, 2021, and the serum vitamin B12 level (XE2pf) was reported to be abnormally low at 107 ng / L (150-1000), indicating that the patient had mild hypothyroidism.

[0231] A series of six or more B12 injections was administered as a loading dose within a two-week period from July 12th to 26th.

[0232] After a stool test on 26 July 2021 confirmed Helicobacter pylori infection, the patient was prescribed NHS protocol triple therapy (amoxicillin 500mg, clarithromycin 500mg, lansoprazole 30mg) for seven days.

[0233] The patient completed the standard antibiotic course prescribed for the treatment of Helicobacter pylori infection and had a repeat stool test in September 2021. However, the test results remained positive and there was no discernible improvement in symptoms.

[0234] The patient was treated for 5 days with a combination of calcium carbonate, magnesium carbonate, the zeolite clinoptilolite, and Lactobacillus reuteri (antimicrobial composition B detailed in section Patients Administration and effective regimen or course for treatment against Helicobacter pylori - duration of regimen 5 days).

[0235] The patient completed the course and had a retest of the stool on October 8, 2021, which came back negative.

[0236] November 30th: Serum thyroid-stimulating hormone (TSH) level 5.6 mlU / L (0.3-4.5) Hypothyroidism Abnormal result - before treatment.

[0237] January 31st: Serum TSH level 4.06 mlU / L (0.3-4.5) Results: Normal - after treatment with the antibacterial composition of the present invention.

[0238] It is hypothesized that the body of a subject, such as a human, naturally produces excess acid to combat Helicobacter bacteria, such as Helicobacter pylori, during an inflammatory response. The body also recognizes the need for buffering when acid levels become too high and achieves this with the aid of calcium and magnesium stores. However, prolonged infection with Helicobacter bacteria, such as Helicobacter pylori, over a period of time (e.g., weeks, months, or even years), can lead to excess acid and depletion of magnesium and calcium stores, potentially resulting in gastric disorders caused by autoimmune responses or colonization by the pathogen, as well as other illnesses and associated symptoms. The use of the non-antibiotic antimicrobial composition of the present invention not only effectively reduced ammonia and urea levels, but also eliminated Helicobacter pylori in the patient's body, preventing symptoms, diseases, or conditions associated with Helicobacter infection, such as Helicobacter pylori. Similar results are expected with Lactobacillus reuteri UBLRu-87.

[0239] Further experimental studies

[0240] Additional experimental studies were performed to examine the survival of Helicobacter pylori at various pH levels.

[0241] Studies were conducted to culture Helicobacter pylori strains.

[0242] All test organisms were cultured on brain heart infusion agar (BHIA) + 5% blood (blood should be approximately 2 weeks old) and incubated at 37°C for up to 7 days in a DG250 microaerobic cabinet supplied with 10% CO2, 5% O2, and 85% N2.

[0243] For each test strain, 2 McFarland (1–10 × 10 7An inoculum equivalent to 1000 CFU / mL was prepared in 5 mL of sterile phosphate-buffered saline (PBS). Each inoculum suspension was diluted 1:10 (2 mL inoculum + 18 mL medium) or 1:100 (200 μL + 19.8 mL medium) into the respective growth medium plus 10% [v / v] fetal bovine serum (FBS). A medium-only (sterile) control was also included.

[0244] Immediately after inoculation, a 0.5 mL sample was taken and placed in a sterile Eppendorf tube. Total viable counts were determined from 1 mL samples on brain heart infusion (BHI) agar containing 5% blood using the Miles and Misra methodology. Plates were incubated under microaerophilic conditions for approximately 5 days until adequate growth could be quantified. Ammonium levels were also detected by adding two drops of test reagent to a 1 mL sample and quantifying the levels present in the sample using urine test strips (Quantofix lot 315250).

[0245] From the remaining samples, 200 μL was transferred to a sterile 96-well flat-bottom microdilution plate (Corning Incorporated; #3370), and the absorbance at 550 nm was recorded using a plate reader. Sampling was repeated 2, 4, 8, 24, 48, and 124 hours after inoculation. The results are shown in Figure 3 (blank-corrected absorbance for H. pylori strain 83 and ATCC 700824) and Figure 4 (total viable counts for H. pylori strain 83 and ATCC 700824).

[0246] The effects of pH on total viable cell count (TVC) and ammonium concentration were investigated.

[0247] Helicobacter pylori ATCC700824 was cultured in brain heart infusion (BHI) plus 5% blood (blood should be approximately 2 weeks old) and incubated at 37°C for up to 7 days in a DG250 microaerobic cabinet supplied with 10% CO2, 5% O2, and 85% N2.

[0248] 2 McFarland (1-10 x 10 7An inoculum equivalent to 100 CFU / mL was prepared in sterile phosphate-buffered saline (PBS) and diluted 1:10 in BHI plus 10% [v / v] fetal bovine serum (FBS) and 0.1% [w / v] yeast extract (20 mL inoculum + 180 mL medium) and incubated under microaerophilic conditions for 96 h.

[0249] The culture medium was divided into 920 mL aliquots, centrifuged at 3500 rpm for 10 minutes, and the supernatant was removed. The cell pellet was resuspended in 20 mL of buffered BHI plus 10% [v / v] fetal bovine serum (FBS) and 0.1% [w / v] yeast extract, adjusted to pH 2, pH 5, or pH 7 with 5 M HCl, and supplemented with 0 mM, 10 mM, or 50 mM urea. A total of nine samples (plus a sterility control for each urea concentration) were tested.

[0250] Samples were removed at 0, 0.5, 1, 2, 4, and 8 hours after inoculation and total viable counts were determined on BHI agar containing 5% blood using the Miles and Misra methodology. The results can be seen in Figure 5. Plates were incubated under microaerobic conditions for approximately 5 days until adequate growth could be quantified. Ammonium concentration (Quantofix, lot no. 315250) and pH (Fisherbrand, lot no. 18F0131) were measured using semiquantitative colorimetric strips. The results can be seen in Figure 6.

[0251] This study confirmed that Helicobacter pylori cannot survive in the acidic gastrointestinal tract without some kind of survival mechanism. Without wishing to be bound by theory, it is hypothesized that Helicobacter pylori survives in the periplasm of the gastrointestinal tract, particularly in the stomach (Bury-Mone, S., Skouloubris, S., Labigne, A. and De Reuse, H. (2001), The Helicobacter pylori UreI protein: role in adaptation to acidity and identification of residues essential for its activity and for acid activation. Molecular Microbiology, 42: 1021-1034 and Wen Y, Scott DR, Vagin O, Tokhtaeva E, Marcus EA, Sachs G. Measurement of Internal pH in Helicobacter pylori by Using Green Fluorescent Protein Fluorimetry. J Bacteriol.2018 Jun 25;200(14):e00178-18. doi: 10.1128 / JB.00178-18. PMID: 29735759; PMCID: PMC6018362. The periplasm allows the bacteria to adapt to the pH of their environment and survive through the hydrolysis of urea. The production of ammonium from this urea hydrolysis regulates the pH, creating optimal conditions for Helicobacter pylori survival. Other studies have also demonstrated this survival mechanism, but it differs from the extensive literature suggesting that Helicobacter pylori survives hidden beneath the mucus layer in the gastrointestinal tract. The present inventors hypothesize that both theories are correct. Therefore, they propose a dual-action treatment to eradicate this bacterium from the gastrointestinal tract. Lactobacillus reuteri (or similar lactic acid bacteria) binds to the bacterium and helps remove it from the mucus layer of the gastrointestinal tract, but this does not necessarily efficiently remove it from the body.Zeolites, such as clinoptilolite or Na-clinoptilolite, absorb Helicobacter pylori and Lactobacillus reuteri (or other lactic acid bacteria). The cation absorption capacity of zeolites, such as clinoptilolite, is hypothesized to exchange sodium, calcium, and / or magnesium ions for ammonium. The inventive composition of zeolite and at least one member species of Lactobacillus is insoluble and therefore efficiently excreted from the body, along with Helicobacter pylori. The inventive composition of zeolite and at least one member species of Lactobacillus thus provides a unique, synergistic, and effective treatment for Helicobacter pylori, a novel non-antibiotic treatment that can be used to effectively eradicate Helicobacter pylori infection, urease-positive bacteria, and ammonium in the gastrointestinal tract.

[0252] Initial modeling suggests that compositions of the present invention, including zeolite (especially when the zeolite is activated clinoptilolite) and at least one species of Lactobacillus (e.g., Lactobacillus reuteri), may more rapidly eliminate Helicobacter pylori than zeolite alone. Initial modeling suggests that compositions of the present invention, including zeolite and at least one species of Lactobacillus, may more completely eliminate Helicobacter pylori from a patient's gastrointestinal tract than zeolite alone.

[0253] It is believed that at least a portion of the composition is substantially insoluble in water at body temperature, and presumably at least a significant portion, or perhaps all, of the zeolite component is insoluble, thereby aiding in clearance.

[0254] The disclosure illustratively described herein may suitably be practiced in the absence of one or more elements or limitations not specifically disclosed herein. Thus, for example, terms such as "comprising," "including," and "containing" are to be read expansively and without limitation. Furthermore, the terms and expressions used herein are used as terms of description and not of limitation, and the use of such terms and expressions is not intended to exclude any equivalents of the shown and described features or portions thereof, recognizing that various modifications are possible within the scope of the claimed disclosure.

Claims

1. An antibacterial composition comprising a zeolite and at least one member of the genus Lactobacillus for use in treating a condition associated with infection with pathogenic urease-active bacteria in the gastrointestinal tract of a vertebrate.

2. An antibacterial composition comprising a zeolite and at least one member of the genus Lactobacillus for use in treating a condition associated with Helicobacter in a vertebrate.

3. 3. The antibacterial composition according to claim 1, wherein the zeolite is a naturally occurring zeolite or a synthetic zeolite.

4. 4. The antimicrobial composition of claim 1, wherein the zeolite is zeolite clinoptilolite (ZC).

5. 5. An antimicrobial composition according to any one of claims 1 to 4, wherein the zeolite is the activated zeolite clinoptilolite (aZC), preferably the doubly activated zeolite clinoptilolite (aZC).

6. At least one member species of the genus Lactobacillus is selected from the group consisting of Lactobacillus reuteri, Lactobacillus delbrueckii, Lactobacillus rhamnosus, Lactobacillus casei, Lactobacillus acidophilus, Lacticaseibacillus paracasei, Lacticaseibacillus plantarum, Lactiplantibacillus spp. plantarum, Levilactobacillus brevis, Ligilactobacillus salivarius, Limosilactobacillus fermentum, Lactobacillus bulgaricus, Lactobacillus crispatus, Lactobacillus helveticus, and Lactobacillus johnsonii.

6. The antimicrobial composition of claim 1, wherein the antimicrobial composition is selected from the group consisting of:

7. 7. The antibacterial composition of claim 6, wherein at least one member of the genus Lactobacillus is Lactobacillus reuteri.

8. The antibacterial composition according to claim 6 or 7, wherein the Lactobacillus reuteri is Lactobacillus reuteri DSM17648 or Lactobacillus reuteri UBLRu-87.

9. The antimicrobial composition of claim 1 , wherein the vertebrate is administered a therapeutically effective amount of the antimicrobial composition.

10. 10. The antimicrobial composition of claim 9, wherein said therapeutically effective amount is achieved by a dosing regimen of said zeolite and at least one member species of said Lactobacillus genus.

11. 11. The antimicrobial composition of claim 10, wherein the regimen comprises one or more administrations of the zeolite and at least one member species of the Lactobacillus genus.

12. 12. The antibacterial composition of claim 1, wherein at least one member of the Lactobacillus genus is a live bacterium.

13. 13. The antimicrobial composition of any one of claims 1 to 12, further comprising a bioactive substance or mineral, such as calcium carbonate, magnesium carbonate, vitamins, such as vitamin D and vitamin E, pharmaceutically acceptable carriers, additives, and adjuvants, such as antimicrobial chitins, such as chitosan, α-ketoglutarate, citrate, and lactate.

14. 14. The antibacterial composition of any one of claims 9 to 13, wherein the administration regimen of the zeolite and the at least one member species of the Lactobacillus genus is suitable for oral or intragastric administration.

15. 15. The antibacterial composition of any one of claims 1 to 14, comprising activated zeolite clinoptilolite (aZC), Lactobacillus reuteri DSM 17648 or Lactobacillus reuteri UBL Ru-87, calcium carbonate and magnesium carbonate.

16. 16. The antimicrobial composition of any one of claims 1 to 15, wherein the condition is associated with Helicobacter, Staphylococcus, Proteus, Klebsiella, and Mycobacterium.

17. 17. The antibacterial composition of any one of claims 1 to 16, wherein the condition is associated with Helicobacter pylori.

18. 18. The antimicrobial composition of claim 1, wherein the vertebrate is a human.

19. 19. The antimicrobial composition of any one of claims 1 to 18, wherein the antimicrobial composition results in a reduction of one or more markers.

20. 20. The antimicrobial composition of claim 19, wherein the marker is a predictive marker selected from the group consisting of urea, gastric ammonia, acid reflux, bloating, and urease activity.

21. 21. The antimicrobial composition of claim 19 or claim 20, wherein the predictive marker is gastric ammonia.

22. 22. The antimicrobial composition of claim 21, wherein the antimicrobial composition results in a reduction of intragastric ammonia levels in a vertebrate infected with Helicobacter pylori.

23. 23. The antimicrobial composition of any one of claims 20 to 22, wherein the reduction in gastric ammonia is at least 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 100% or more compared to placebo.

24. 23. The antimicrobial composition of any one of claims 20 to 22, wherein the reduction in gastric ammonia is at least 0.1, 0.25, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, 5, 5.5, 6, 6.5, 7, 7.5, 8, 8.5, 9, 9.5, or 10 times or more compared to placebo.

25. 23. The antibacterial composition of any one of claims 20 to 22, wherein the reduction in the predictive marker is statistically significant compared to placebo.

26. 26. The antibacterial composition of any one of claims 1 to 25, wherein the antibacterial composition results in a statistically significant clinical improvement in humans infected with one or more of Helicobacter, Staphylococcus, Proteus, Klebsiella, and Mycobacterium.

27. 24. The antibacterial composition of claim 23, wherein the reduction in gastric ammonia marker levels in humans infected with Helicobacter pylori is statistically significant.

28. 28. The antibacterial composition of any one of claims 1 to 27, wherein the antibacterial composition produces a statistically significant clinical improvement in humans infected with Helicobacter pylori.

29. 29. The antimicrobial composition of any one of claims 1 to 28, wherein the antimicrobial composition is substantially insoluble in water.

30. 30. Use of the antibacterial composition of any one of claims 1 to 29 for the manufacture of a pharmaceutical formulation, for single, combined or simultaneous administration, designed for the treatment, alleviation or prevention of GIT diseases or disorders, gastritis, gastric ulcers, duodenal ulcers, gastric cancer, duodenal cancer in vertebrates, including mammals and birds, in need thereof.

31. 31. The use of claim 30, wherein the pharmaceutical formulation is in a form suitable for administering a dose of the antibacterial composition in a single dose or as divided doses or subdoses administered at appropriate intervals per day.

32. 32. The use of claim 31 , wherein the pharmaceutical formulation is administered as one, two, three, four or more doses or subdoses per day or administration.

33. The pharmaceutical preparation comprises 10 ７ ~10 ９ 33. The use of any one of claims 30 to 32, comprising CFU / g of Lactobacillus reuteri DSM 17648 or Lactobacillus reuteri UBL Ru-87, and a zeolite, wherein the zeolite is present in an amount of at least 50 mg / gram of formulation.

34. 34. The use of claim 33, wherein the pharmaceutical formulation further comprises a mineral salt such as calcium carbonate and / or magnesium carbonate, the calcium carbonate and magnesium carbonate each being present in an amount of at least 0.5 mg per gram of formulation.

35. 35. The use of any one of claims 30 to 34, wherein the dose of the antimicrobial composition is formulated as a capsule, a lyophilized formulation, a liquid, a pill, a powder, or a gel.

36. 36. The use according to any one of claims 30 to 35, wherein the treatment results in complete remission of a disease or disorder caused by or associated with infection with Helicobacter.

37. 36. The use of any one of claims 30 to 35, wherein the treatment results in complete remission of a disease or disorder caused by or associated with any one or more of a Staphylococcus infection, a Proteus infection, a Klebsiella infection, or a Mycobacterium infection.

38. 30. An antibacterial composition according to any one of claims 1 to 29 for use in the reduction of ammonia in the gastrointestinal tract, preferably in the reduction of ammonia concentrations in gastric juices.

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