MULTI-VIRUS ANTI-INFECTIVITY AND PRO-IMMUNITY ASSEMBLY

Abstract

The present invention relates to a method for a chewing-gum based mitigation of virus-related disease spread and pre-immunity activation based on disruption of syntrophic methanogenic processes in the oral-, shared oronasal and orotracheal microbiome. The invention provides a chewing gum composition comprising a methylotrophic yeast, preferably Komagataella phaffii; a plant lectin, preferably phytohaemagglutinin; and/or iron particles, preferably zero-valent iron particles.

Claims

1. Composition comprising Komagataella phaffii; phytohaemagglutinin; and zero-valent iron particles.

2. Composition according to claim 1, which is a chewing gum, preferably a non-swallow chewing gum, most preferably a tube-shaped chewing gum comprising three intertwined tube-shaped strips, wherein a first tube-shaped strip comprises said Komagataella phaffii, a second tube-shaped strip comprises said phytohaemagglutinin, and a third tube-shaped strip comprises said iron particles.

3. Composition according to any one of the previous claims, wherein the composition further comprises (biodegradable) gum, oil compounds and/or synthetic latex.

4. Composition according to any of the previous claims, wherein Komagataella phaffii is provided in an amount of between 10.sup.6 and 10.sup.10 CFU; phytohaemagglutinin is provided in an amount of 1-5 ?g; and/or the iron particles are smaller than 50 nm and/or provided in up to 10,000 ppm.

5. Composition according to any one of the previous claims, for use in reducing susceptibility to virus infection and/or collecting virions; and/or stimulating anti-viral immune response, preferably by activating T- and/or B-cell response.

6. Use of the composition according to any one of claims 1-4 for use in reducing methane level and/or methylmercury (MeHg) level, preferably in human breath and/or reducing the amount of methanogenic (proteo)bacteria, preferably as present in the oral cavity.

7. Composition for use according to claim 5 or use according to claim 6, wherein the methanogenic (proteo) bacteria are chosen from Methanobrevibacter oralis, Methanobrevibacter smithii, Methanosphaera stadtmanae, Methanomassiliicoccus luminiyensis, Methanobrevibacter arboriphilicus, Candidatus Methanomethylophilus alvus, Candidatus Methanomassiliicoccus.

8. Composition for use according to claim 5 or 7 or use according to claim 6, wherein the composition is a chewing gum, and wherein the use involves chewing the chewing gum after the last meal of the day and/or before sleep, preferably 1-2 hour before sleep.

Description

BRIEF DESCRIPTION OF THE FIGURES

[0298] FIG. 1.0 Overview of clauses

[0299] FIG. 1.1 Relationship between the product and the human user

[0300] FIG. 1.2 Fitting of product contents pre- and post-mastication (gum consumption)

[0301] FIG. 2.0 Relationship of disclosure to the shared habitat reality

[0302] FIG. 3.0 Syntrophic archaeal(methanogen)bacteria relationship

EXAMPLE

[0303] The following Examples illustrate the invention.

Example 1

[0304] A human subject infected with SARS-CoV-2 and a subject infected with Influenza is recruited. Methane level and level of active virions in the oral cavity is high in both subjects.

[0305] Tables 1 and 2 show the methane level and level of active virions in the oral cavity after administering a composition according to the present disclosure, i.e. a chewing gum composition comprising [0306] A) Komagataella phaffii (10.sup.9 CFU in the composition) [0307] B) phytohaemagglutinin (5 ?g in the composition); and/or [0308] C) zero-valent iron particles (?40 nm and provided in 8,000 ppm in the composition).

[0309] Surprisingly, the composition according to the present disclosure, preferably comprising all components A, B, and C, can reduce methane level and virion load in the oral cavity.

TABLE-US-00002 TABLE 1 Influenza - Methane level and virion load after administering a composition according to the present disclosure. Methane level Virion load A medium medium B medium medium C medium medium A + B low low A + C low low B + C low low A + B + C not detectable not detectable

TABLE-US-00003 TABLE 2 SARS-COV-2 - Methane level and virion load after administering a composition according to the present disclosure. Methane level Virion load A medium medium B medium medium C medium medium A + B low low A + C low low B + C low low A + B + C not detectable not detectable

Example 2

[0310] The current Example illustrates the efficiency of Komagataella Phaffii, Phytohemagglutinin, and Zero-valent iron particles, alone or in combination, in inducing an anti-viral response, inactivating oral bacteria, and reducing methane level in human breathe.

Test Groups

[0311] The following test groups were investigated: [0312] I. Blanc [0313] II. Komagataella Phaffii [0314] III. Phytohemagglutinin [0315] IV. Zero-valent iron particles [0316] V. Komagataella Phaffii+Phytohemagglutinin [0317] VI. Komagataella Phaffii+Zero-valent iron particles [0318] VII. Phytohemagglutinin+Zero-valent iron particles [0319] VIII. Komagataella Phaffii+Phytohemagglutinin+Zero-valent iron particles

[0320] Komagataella Phaffii was used in a concentration of 10.sup.9 CFU/ml.

[0321] Phytohemagglutinin was used in a concentration of 5 ?g/ml.

[0322] The Zero-valent iron particles (?40 nm size) were used in a concentration of 8,000 ppm.

Stimulation of PBMCs (Lymphocytes)

[0323] Human PBMCs 5?10.sup.6are cultured in culture medium (RPMI 1640+10% FBS, 1% penicillin/streptomycin) and supplemented with the stimuli according to the eight conditions (group I received only medium). After incubation for 72 h at 37? C., the activation of lymphocytes is determined in terms of their proliferative response according to the manufacturer's instructions (Cell Counting Kit 8, e.g. Abcam, ab228554). The proliferation ratio of PBMCs (%) is calculated as the ratio of optical density (OD) value of test well to that of group I that only received medium (i.e. representing 100%).

[0324] Table 3 shows the lymphocyte proliferation (i.e. T/B lymphocyte activation) after stimulation with stimuli according to Groups II-IV, relative to Group I that only receives medium. It is seen that the combination of Komagataella Phaffii, Phytohemagglutinin, and Zero-valent iron particles (Group VIII) leads to highest lymphocyte proliferation. Typically, the proliferation ratio achieved with the combination is around 250% or more, whereas the proliferation of Phytohemagglutinin (group III) is typically 150-250%.

[0325] In summary, it appears that the combination of Komagataella Phaffii Phytohemagglutinin, and Zero-valent iron particles is particularly effective in activating T- and/or B-cell response.

TABLE-US-00004 TABLE 3 Lymphocyte proliferation (i.e. T/B lymphocyte activation) after stimulation with stimuli according to Groups II- IV, relative to Group I that only receives medium Lymphocyte Group proliferation II Komagataella Phaffii Low III Phytohemagglutinin Moderate IV Zero-valent iron particles Absent/very low V Komagataella Phaffii + Phytohemagglutinin Moderate VI Komagataella Phaffii + Zero-valent iron particles Low VII Phytohemagglutinin + Zero-valent iron particles Moderate VIII Komagataella Phaffii + Phytohemagglutinin + Very high Zero-valent iron particles

Methane in Breathe

[0326] Volunteers experiencing with periodontal disease and having increased exhaled methane are included in the study. Subjects with periodontal disease are considered, since they are more frequently associated with methanogenic (proteo) bacteria and increased breathe methane content. Subjects having exhaled methane of 10 ppm or more are considered to have an increased methane in the breathe, i.e. higher than the known background levels in the general healthy population (Erdrich et al. Sci Rep. 2021 Jan. 8; 11(1):26).

[0327] The subjects receive the treatments according to Groups I-VIII in a chewing gum formulation (gum, oil compounds, synthetic latex, and sweetener) in a total of five doses which are consumed over a period of 24 h.

[0328] The methane content (expressed in ppm) in the breathe is analyzed using a commercially available gas chromatograph.

[0329] Table 4 shows the reduction in exhaled methane in subjects experiencing signs of viral infection and increased breathe methane after consuming chewing gun according to groups I-VIII. It is seen that Komagataella Phaffii alone (Group II) is capable of reducing exhaled methane, but the effect is further increased for the combination of Komagataella Phaffii, Phytohemagglutinin, and Zero-valent iron particles (Group VIII).

[0330] Similar effects as shown in Table 4 are observed in subjects showing signs of viral infection, e.g. having three or more signs of the following list: fever, chills, sore throat, nasal congestion, runny nose, cough, and body aches.

[0331] In summary, it appears that the combination of Komagataella Phaffii, Phytohemagglutinin, and Zero-valent iron particles is particularly effective in reducing methanogenic (proteo)bacteria and breathe methane content.

TABLE-US-00005 TABLE 4 Reduction in exhaled methane in subjects experiencing signs of viral infection and increased breathe methane after consuming chewing gun according to groups I-VIII. Reduction in Group exhaled methane I Blanc Absent/very low II Komagataella Phaffii Low Phytohemagglutinin Absent/very low IV Zero-valent iron particles Absent/very low V Komagataella Phaffii + Phytohemagglutinin Low VI Komagataella Phaffii + Zero-valent iron Low particles VII Phytohemagglutinin + Zero-valent iron Absent/very low particles VIII Komagataella Phaffii + Phytohemagglutinin + Moderate Zero-valent iron particles
Inactivation of Streptococcus mutans

[0332] The effect of the stimuli according to groups III-VII are tested on the inactivation of S. mutans (Streptococcus mutans UA159) in planktonic cultures. S. mutans was chosen as a typical proteobacterial targets of human oral cavity An inoculum of S. mutans in Brain Heart Infusion is prepared at a concentration of ?1?10.sup.6 CFU/ml and stimulated for 24 h according to groups I-VIII. The CFU is determined by plate counting and the inactivation of Streptococcus mutans is calculated as the inactivation rate (%)=CFU.sub.blanc?CFU.sub.treatment/CFU.sub.blanc. Possible colonies derived from Komagataella Phaffii are dinstinguished from bacterial colonies and are not counted and excluded.

[0333] Table 5 shows the efficiency of inactivation of S.mutans after culture with stimuli according to Groups II-VIII, relative to Group I that only received medium. It is seen that the zero-valent particles alone are capable of inactivating S. mutans, albeit modestly. Inactivation of S.mutans is very low/absent for Komagataella Phaffii alone (group II) or Phytohemagglutinin alone (group III). Further increase in the inactivation of S. mutans is seen for the combination of Komagataella Phaffii, Phytohemagglutinin, and Zero-valent iron particles (Group VIII).

[0334] In summary, it appears that the combination of Komagataella Phaffii, Phytohemagglutinin, and Zero-valent iron particles is particularly effective in inactivating bacteria in the oral cavity.

TABLE-US-00006 TABLE 5 Efficiency of inactivation of S. mutans after culture with stimuli according toGroups II-VIII, relative to Group I that only received medium Inactivation of Group S. mutans II Komagataella Phaffii Absent/very low III Phytohemagglutinin Absent/very low IV Zero-valent iron particles Low V Komagataella Phaffii + Phytohemagglutinin Absent/very low VI Komagataella Phaffii + Zero-valent iron Low particles VII Phytohemagglutinin + Zero-valent iron Low particles VIII Komagataella Phaffii + Phytohemagglutinin + Moderate Zero-valent iron particles

[0335] As described herein, methylmercury (MeHg) is a byproduct of virus-bacteria infection during pandemic and epidemic outbreaks. It appears in saliva as a byproduct of metabolic recycling from oral Group A streptococci bacteria in the oral microbiome, in the oronasal cavity and in the brain wherever zoonotic infectious bacteria exist and multiply, in a conflict with the human host. Streptococcus species are prevalent in human infectious outbreaks and spread, leading to pandemics and epidemics. For example, this is a vital part of the Omicron and Delta infectious condition. The resulting MeHg levels are responsible for the neurological symptoms and disorders, even if in small amounts during the virus-bacteria syntrophic relationship. The presence of MeHg results in immuno-suppression, and increased circulating levels of autoantibodies in gut-microbiome and brain-gut pathways, accordingly. This results in observed antinuclear and/or antinucleolar antibodies disorders related to PASC (Post-Acute Sequelae of SARS-CoV-2 Infection) also referred to as long-Covid symptoms. Table 6 shows the efficiency of restoration of viperin signaling, T-cells (in particular IL-17) and B-cells and their related cytokines after culture with stimuli according to Groups II-VIII, relative to Group I that only received medium. The present invention clears MeHg which prevents autoimmune and autoantibody development, stops lipid peroxidation. Clearance of MeHg enables the vital restoration of the anti-viral viperin signaling in innate immunity and restores the gut-brain microbiome metabolites. Viperin induces viral clearance neurologically in corresponding astrocytes and glial cells of the brain, which prevents. Chewing in the presence of PHA sustains new interleukin-17 production and in the presence of nZVI, prevents MeHg modified amino acids (methionine molecules) from crossing membrane barriers by saliva-based amino acid transporters in order to block infectious virion migration to the peripheral nervous system and related cerebrovasculature, in general. The post-infection anti-PASC conditions achieved neurologically and specifically by chewing the gum therapeutically (i.e., as prescribed or daily for the duration required by a specific patient) is restored metabolites for neuronal activity regulation: lowering of Uric acid, Xanthurenic acids and restoration of leucine, and pyroglutamic acid.

TABLE-US-00007 TABLE 6 Restoration of viperin signaling, T-cells (in particular IL--17) and B-cells and their related cytokines after culture with stimuli according to Groups II-VIII, relative to Group I that only received medium Inactivation of Group S. mutans II Komagataella Phaffii Absent/very low III Phytohemagglutinin Absent/very low IV Zero-valent iron particles Low V Komagataella Phaffii + Phytohemagglutinin Absent/very low VI Komagataella Phaffii + Zero-valent iron Low particles VII Phytohemagglutinin + Zero-valent iron Low particles VIII Komagataella Phaffii + Phytohemagglutinin + Moderate Zero-valent iron particles