Microbiome modulator composition for enhancing the immune system

Abstract

The invention relates to a microbiome modulator composition (MMC) and use thereof for enhancing immune system and for treating cancer and infectious diseases.

Description

[0001] MICROBIOME MODULATOR COMPOSITION FOR ENHANCING THE IMMUNE SYSTEM

[0002] FIELD OF THE INVENTION

[0003] The invention relates to a microbiome modulator composition (MMC) and use thereof for enhancing immune system and for treating cancer and infectious diseases.

[0004] BACKGROUND OF THE INVENTION

[0005] The management of solid tumors has been revolutionized in the past 10 years with the advent of immunotherapy. The latter, takes advantage of the innate and adaptive immunity to generate or take advantage of the immune response directed against cancer. Over the past years, there has been a much better understanding of how the immune system is remodelled by tumors and, in some cases, inhibited.

[0006] In the context of non-small cell lung cancer (NSCLC) or melanoma, immune checkpoint inhibitors have revolutionized patient care and outcome. Patients that had very poor prognosis in the past are now surviving on the long term with excellent quality of life. However, the current studies report that only a fraction of patients respond to therapy and the reasons for this are still under investigation. Some hypotheses rely in the poor immune content of tumors, a condition known as immune desert. A typical tumor considered as an immune desert is the malignant pleural mesothelioma (MPM). MPM is a rare cancer with poor prognosis that originates from mesothelial cells that line the lung chest cavity. MPM is mainly associated with asbestos exposure and shows a latency period of 20-40 years before the cancer develops. Treatment options for Malignant Pleural Mesothelioma (MPM) are limited. For many years, systemic therapies only consisted in platinum and folate antimetabolite compounds for unresectable MPM. Multiple clinical studies conducted on MPM patients with anti-CTLA-4 and anti-PD-l / PD-Ll immune checkpoint inhibitors (ICI) indicate a better tumor response to ICI-based immunotherapy compared to chemotherapy. Indeed, combination therapy using dual immune checkpoint inhibition (DICI) has shown greater efficacy in controlling tumor progression than single-agent treatment or chemotherapy. These studies led the European Society for Medical Oncology (ESMO) to recommend DICI with Nivolumab plus Ipilimumab as a new standard of care for unresectable MPM. However, despite these advancements, some hurdles still need to be overcome before most patients could profit from the full therapeutic potential of this approach. Indeed, less than fifty percent of MPM patients favourably respond to DICI and the percentage of immune adverse events (iAE) remains high, leading in some cases to treatment discontinuation. In addition, the survival among MPM patients is low even with DICI therapy, with a three-year overall survival of 23% when combining Nivolumab plus ipilimumab. Primary or secondary resistance to ICI results from poor tumor immunogenicity, lack of activable anti-tumor T cells in the tumor bulk, impaired function of effector T cells and formation of T-cell memory.

[0007] In other solid cancers such as colorectal cancer, the immune microenvironment was shown to be very favourable for ICI therapy, but other subtypes seem to lack the appropriate microenvironment composition for immunotherapy to act.

[0008] In recent years, the connection between microbiota and immunity has become increasingly apparent, with numerous studies highlighting the relationship between the composition of commensal bacterial population and both innate and adaptive immune responses in both healthy and disease conditions. These findings have opened new perspectives for cancer management, suggesting that modulation of the microbiota could serve as a minimally toxic approach to enhance anti-tumor immunity and improve the response to immunotherapies such as immune checkpoint inhibitor (ICI) therapy. Several past and ongoing studies with faecal microbial transplantation (FMT) from ICI responders to non-responders showed promising results in melanoma, non-small cell lung cancer and colorectal patients. Despite good results obtained with FMT, this approach presents some key limitations such as important infectious risks and demanding screening procedures to identify stool donors. Given these limitations, modulating the gut microbiome through diet supplementation of probiotics or prebiotics appears to be a better option. However, commercially available probiotic supplementation showed mixed results, possibly due to microbiota dysbiosis and, in some cases, reduced infiltration of cytotoxic CD8+ and helper CD4+ T cells. A recent study on melanoma and colon adenocarcinoma mouse models has shown beneficial impact of diet supplementation with mucin and inulin prebiotics on anti-tumor immunity and tumor control through microbiota modulation. Mucin and inulin prebiotics induced a modulation of the expression genes involved in activation of dendritic cells and T cells and gut microbiota was required for the activation of the anti-tumor immune response. Nevertheless, translating the prebiotic dosage to human equivalent based on body weight would require the administration of several hundred grams per day, which presents a challenge for its practical incorporation into the daily diet. Accordingly, there is a need for a next generation microbiome modulator composition that can efficiently enhance human immune system and thereby improve cancer treatment with immune checkpoint inhibitors as well as infectious diseases treatment overcoming the challenges of existing and already studied and available biotic compositions.

[0009] SUMMARY OF THE INVENTION

[0010] An aspect of the present invention provides a microbiome modulator composition (MMC) able to stimulate growths and / or activity of one or more strains of bacteria in the gut selected from Rikenellacaea family and Muribaculaceae family, said composition comprising in wt%:

[0011] • carbohydrates and / or hydrolyzed carbohydrates: 30% to 50%

[0012] • proteins and / or hydrolyzed proteins: 30% to 50%

[0013] • fats: 1% to 4%

[0014] • ash: 1% to 5%

[0015] • nucleotides: 0% to 5%

[0016] • moisture: < 10%

[0017] Another aspect of the present invention provides a pharmaceutical combination comprising the MMC of the invention and one or more cancer immunotherapeutic agent for simultaneous, separate or sequential administration, wherein the cancer immunotherapeutic agent is selected from the group comprising immune checkpoint inhibitor, TCR-T cells, CAR-T cells or combinations thereof.

[0018] Another aspect of the present invention provides a method of enhancing immune system function in a subject comprising administering to the subject an effective amount of the MMC of the invention.

[0019] Another aspect of the present invention provides a method for enhancing the efficacy of a cancer immunotherapy in a subject, the method comprising administering an effective amount of the MMC of the invention before the cancer immunotherapeutic agent, concurrently with the cancer immunotherapeutic agent and after the cancer immunotherapeutic agent, wherein the cancer immunotherapeutic agent is selected from the group comprising immune checkpoint inhibitor, TCR-T cells, CAR-T cells or combinations thereof. Another aspect of the present invention provides a method for treating a solid cancer in a subject in need thereof, said method comprising administering to the subject an effective amount of the MMC of the invention and one or more cancer immunotherapeutic agent, wherein the effective amount of the MMC is 0.01 to 20 mg / kg / day and wherein the cancer immunotherapeutic agent is selected from the group comprising immune checkpoint inhibitor, TCR-T cells, CAR-T cells or combinations thereof.

[0020] Another aspect of the present invention provides a method of preventing and / or treating infectious diseases in a subject comprising administering to the subject an effective amount of the MMC of the invention, wherein the infectious diseases are caused by pathogenic microorganisms selected from the group comprising bacteria, viruses, fungi and parasites.

[0021] BRIEF DESCRIPTION OF THE FIGURES

[0022] Figure 1 shows impact of the microbiome modulator composition (MMC) of the invention on T-cell infiltration and activation. A-C: Representative images of non -treated (NT) and MMC- treated tumours stained for CD3 (red), CD4 or CD8 (green) and GRZB (blue). CD4 (A) or CD8 (B) costained with the lymphocyte marker CD3 (red). Colocalization appears in yellow. Scalebar: 100 pm (C): GRZB (blue) staining and colocalization with CD3 (red) and CD8 (green). Colocalization signal appears in white (arrows). Scalebar: 100 pm. D-F: Quantification of immunofluorescence stainings. Colocalisation area between CD3 and CD4 signal (D), CD3 and CD8 signal (E) normalized by tumor area. F: Quantification of the percentage of Granzyme B+ CD8+ CD3+ lymphocytes. Graphs represent the mean ± SD. -values were calculated by using unpaired t-tests. *p<0.05, **p<0.01.

[0023] Figure 2 shows impact of the MMC of the invention on macrophage proportions in tumors. A- B: Representative images of CD45+ (red) CD68+ cells (green) costained with the activation marker (Ml -like macrophages) CD80 (A) or the M2-like marker CD206 (B) in blue for non- treated (NT) and the MMC treated group. Scalebar: 100 pm. C-F: Quantification of the immunofluorescence images. Quantification of the number of Ml-like macrophages (C) and M2-like macrophages (D) normalized by total amount of macrophages (colocalization CD68 / CD80). (E) Ratio Ml-like / M2-like macrophages. (F). Quantification of total amount of infiltrating macrophages (CD45+CD68+) normalized by tumor area. Graphs represent the mean ± SD. -values were calculated by using unpaired t-tests. *p<0.05, **p<0.01, ***p<0.001. Figure 3 shows impact of the MMC of the invention on tumor growth and animal survival in immunocompetent (A-B) and athymic (T-cell deficient) mice (C-D). A + C: Tumor growth curves for non-treated (NT, grey) and MMC-treated (orange) animals, measured by luminescence in photon per second per square centimetre per steradian (p / s / cm2 / sr). B+D: Kaplan-Meier curves for NT (grey) and the MMC-treated (orange) animals. In immunocompetent animals (B), MMC administration significantly extended overall survival versus non-treated animals (median survival 10 vs 7 days, Hazard ratio (logrank): 0.35 [95% CI 0.14-0.91]). -values was calculated by using log-rank test. **p<0.01. In athymic T-cell deficient animal (D), the positive impact of MMC on tumor control and animal survival was lost in athymic animals (median survival of 7 days for MMC-treated and non-treated animals, Hazard ratio (logrank): 0.87 [95% CI 0.31 -2.41]).

[0024] Figure 4 shows the association between tumor-infiltrating CD8+GRZB+ T-cells, CD4 T-cells and animal survival. A: A positive association was found between CD8+GRZB+ T cells and animal survival with a Spearman correlation coefficient of 0.5965 and an associated p-value of 0.0164. *p<0.05. B: Survival of the animals was not associated with the amount of CD4 T cells found in tumors. Spearman correlation = 0.391.p- value: 0.2435.

[0025] Figure 5 shows the impact of MMC on gut microbiome community composition and diversity. A: Principle Component Analysis (PCoA) depicting the differences in community composition between non-treated controls (NT - grey) and MMC-treated (orange) subjects. Distance matrixes were calculated using Weighted UniFrac. The community composition of the MMC- treated subject is altered compared to non-treated mice. MMC-treated subjects have a significantly different microbial community composition in the [a] colon (P = 0.0012), [b] feces (p = 0.2946) and [c] ileum (p = 0.017) compared to the non-treated controls. MMC-treated subjects have a decrease in beta-diversity (relative abundance between communities) in the colon, as evidenced by the decrease in dispersion. Permutational Multivariate Analysis of Variance (i.e., ADONIS) was performed to followed by the Tukey’s Honest Test. -values indicated the significance adjusted for variance variations between groups. B: Alpha diversity analyses of untreated controls (NT - Grey) and MMC-treated (Orange) subjects across all sampling locations (i.e., ileum, colon and feces), [a] Shannon diversity is significantly changed in the feces and ileum, [b] Faith’s phylodiversity is significantly reduces in fecal samples of MMC-treated subjects. Significant results for Wilcoxon test are shown. *p < 0.05. Figure 6 shows the impact of the MMC on the relative abundance of specific gut microbiota taxa. A: [a] Colon, [b] Feces and [c] Ileum. Top 12 most abundant genera shown in the non- treated (NT) and MMC groups. MMC supplementation has a clear effect on relative ratios of the selected genera. B: Relative abundance per sampling site (i.e., colon, feces and ileum) for non-treated (“NT” - grey) and MMC-treated (orange), showing the selective enrichment of specific taxa by MMC supplementation. Analyses were performed on [a] Alistipes, [b] the Rikenellaceae RC9 gut group, [c] Muribaculaceae (FAMILY), [d] Muci spirillum, [e] the Lachnospiraceae NK4A136 group and [f] Lachnospiraceae (FAMILY). MMC-treated colon samples show the largest change compared to the non-treated controls. Statistically significant Wilcoxon Test are shown: *p<0.05, **p<0.01, ***p<0.001. C: Venn Diagram indicating the number of genera found in at least one subject in non-treated (“Control” - grey) and MMC- treated (orange) groups across all sample types. Of the total 170 genera identified, 108 were present in all groups. No unique taxa were found in the MMC-treated group, indicating that MMC may exert a selection pressure on the microbial population.

[0026] Figure 7 shows the association between fecal short-chain fatty acid (SCFA) and level of tumor- infiltrating CD8+GRZB+ T-cells. A: Level of SCFA in fecal samples of untreated controls (NT) and MMC-treated mice. Graphs show the mean ± SD. -values were calculated by using unpaired t-tests or unpaired t-tests with Welch’s correction (for propionic acid) to correct for unequal variances. B: Linear correlation of SCFA concentrations (pmol / g) with CD8+GRZB+ T-cells into tumours (% of Granzyme B+ CD8+ T-cells). Non-treated controls are depicted in grey and the MMC-treated group is depicted in orange. The supplementation of MMC alters the relationship between SCFA levels in feces and presence of Granzyme B+ CD8+ T-cells in tumours. In the MMC-treated group, higher levels of Granzyme B+ CD8+ T-cells coincide with higher levels of fecal SCFAs. [a] 2-Methylbuteric acid, [b] Isobutyric acid, and [c] isovaleric acid show strong correlations with R2values of 0.97, 0.91, and 0.88 respectively. Butyric acid [d] shows strong correlations with R2values of 0.91 with the level of CD8+ in tumors. All correlations shown are statistically significant (*p< 0.05).

[0027] Figure 8 shows the impact of MMC on PD-1 and CTLA-4 immune checkpoint molecules expression in CD8+ cells. A-B: Representative images of non-treated (NT in grey) and the MMC-treated (orange) tumours stained for CD8 (red) and PD-1 or CTLA-4 (green). [A] CD8 / PD-1 and [B] CD8 / CTLA-4 double staining. Colocalization appears in yellow. Scalebar: 100 pm. C-D: Quantification of immunofluorescence stainings. [C]: Colocalisation area between CD8 and PD-1 signal and [D] CD8 and CTLA-4 signal normalized by tumor area. Graphs represent the mean ± SD. -values were calculated by using unpaired t-tests. *p<0.05.

[0028] Figure 9 shows size exclusion chromatogram for the MMC. A: RI detector, B: UV detector.

[0029] DETAILED DESCRIPTION OF THE INVENTION

[0030] All publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety. The publications and applications discussed herein are provided solely for their disclosure prior to the filing date of the present application. Nothing herein is to be construed as an admission that the present invention is not entitled to antedate such publication by virtue of prior invention. In addition, the materials, methods, and examples are illustrative only and are not intended to be limiting.

[0031] In the case of conflict, the present specification, including definitions, will control. Unless defined otherwise, all technical and scientific terms used herein have the same meaning as is commonly understood by one of skill in art to which the subject matter herein belongs. As used herein, the following definitions are supplied in order to facilitate the understanding of the present invention.

[0032] The term “comprise” is generally used in the sense of include, permitting the presence of one or more features or components. Also as used in the specification and claims, the language "comprising" can include analogous embodiments described in terms of "consisting of “ and / or "consisting essentially of’. The terms "including," "comprising," or "having," and variations thereof, is meant to encompass the elements listed thereafter and equivalents thereof as well as additional elements. Embodiments recited as "including", "comprising" or "having" certain elements are also contemplated as "consisting essentially of' and "consisting of' those certain elements.

[0033] As used in the specification and claims, the singular form "a", "an" and "the" include plural references unless the context clearly dictates otherwise.

[0034] As used in the specification and claims, the term "and / or" used in a phrase such as "A and / or B" herein is intended to include "A and B", "A or B", "A", and "B". As used herein the terms "subject" and “patient” are well -recognized in the art, and, are used herein to refer to a mammal, and most preferably a human. These terms do not denote a particular age or sex. Thus, adult and newborn subjects or patients, whether male or female, are intended to be covered. The subject or patient may be healthy, or may be suffering from a cancer or an infectious disease, at any developmental stage. In some embodiments, the subject or the patient has undergone a cancer immunotherapy or infectious disease therapy. In some embodiments, the subject or the patient is a subject in need of treatment or a subject having a cancer or being infected or having an infectious disease, who is likely to benefit from a treatment of the present invention.

[0035] The term "effective amount" or "therapeutically effective amount" means a sufficient amount of the specified component / composition to have the specified properties under the specified conditions. For example, an effective amount of a MMC means an amount sufficient to cause a desired increase in the metabolite level and / or bacterial counts of one or more selected microorganisms in vitro and / or in vivo and thereby enhancing the immune system.

[0036] The term "microbiome modulator" means any substance or combination of substances that can be utilized as a nutrient, a stimulant and conferring a regulatory and modulatory effect on selected microorganism (e.g., beneficial microorganisms) or group, family or phyla of microorganisms, can induce the growth and / or activity of a selected microorganism, can induce the replication of a selected microorganism, can be utilized as an energy source by a selected microorganism, and / or can be utilized by a selected microorganism for the production of biomolecules (i.e. RNA, DNA, and proteins). In other cases, the microbiome modulator can suppress the growth and even eliminate the presence of certain microorganisms, group, family or phyla of microorganisms. Non-limiting examples of microbiome modulators include mucopolysaccharides, oligosaccharides such as fructooligosaccharides ("FOS"), polysaccharides, amino acids, vitamins, nutrient precursors, harvested metabolic products of biological organisms, lipids, proteins and peptides or isolated fragments of such naturally occurring substances, including naturally occurring linked combinations thereof.

[0037] As used herein, the terms "immune checkpoint inhibitors" (ICIs), "checkpoint inhibitors," and the like refer to compounds that inhibit the activity of negative regulators of the immune response. Immune system checkpoints, or immune checkpoints, are regulatory molecules involved in the control of the duration and amplitude of physiological immune responses as well as maintenance of self-tolerance to minimize collateral tissue damage. Immune checkpoint signalling requires ligand-receptor interaction to send, via receptors, co-stimulatory (positive immune checkpoints) or co-inhibitory (negative immune checkpoints) signals. ICI are molecules disrupting immunosuppressive interaction between immune checkpoint ligand and receptors, reinvigorating thus innate and adaptive immunity to fight cancers. Negative immune checkpoints include, but are not limited to cytotoxic T-lymphocyte-associated protein 4 (CTLA-4), programmed cell death- ligand 1 (PD-L1), programmed cell death- ligand 2 (PD- L2), programmed cell death 1 (PD-1), Lymphocyte-activation gene 3 (LAG-3), T cell immunoreceptor with Ig and ITIM domains (TIGIT), V-domain immunoglobulin suppressor of T-cell activation (VISTA), CD276, and V-set domain containing T cell activation inhibitor 1 (VTCN1). As such, ICI include antagonists of, for example, immune checkpoints such as CTLA-4, PD-1, or PD-L1. For example, antibodies that bind to CTLA-4, PD-1, or PD-L1 and antagonize their function are ICI. Moreover, any molecule (e.g., peptide, nucleic acid, small molecule, etc.) that inhibits the inhibitory function of an immune checkpoint is an ICI.

[0038] The term "hydrolysis" as used within the present disclosure refers to depolymerization of a polymer by a hydrolysis reaction. Hydrolysis reaction is to be understood as the cleavage of chemical bonds in the presence of water. One way to perform hydrolysis technically is to contact the yeast with ionic polymers disclosed herein in the presence of water.

[0039] An aspect of the present invention provides a method for yeast hydrolysis into a microbiome modulator composition (MMC) with excellent properties for enhancing immune system, and especially enhancing anticancer immunity and immunity against infectious diseases.

[0040] In an embodiment, the present invention provides a method for producing a microbiome modulator composition (MMC) from yeast, the method comprising the steps of: a) providing the yeast; b) optionally pre-treating the yeast, wherein the optional pre-treatment is hot water pre- treatment at about 90°C and / or milling; c) contacting the yeast with a catalyst in the presence of water and / or an organic solvent to form a reaction mixture, wherein the catalyst is an ionic polymer or a combination of ionic polymers, the ionic polymer network, a solid-supported ionic polymers and / or a polymer membrane incorporating ionic polymers; d) heating the reaction mixture between 100°C to 170°C and degrading the yeast in the reaction mixture to produce a liquid phase and a solid phase, wherein the liquid phase includes the MMC, and the solid phase includes residual components; e) cooling the reaction mixture to room temperature; f) isolating at least a portion of the liquid phase from the solid phase; g) optionally adding one or more compounds selected from bentonite, charcoal, zeolite, amorphous silica and / or ion exchange resin to the isolated liquid phase and filtering it / them out; and h) recovering the MMC from the isolated liquid phase.

[0041] In some embodiments of the method of the invention, the yeast is any suitable yeast for human consumption, such as brewer’s yeast or baker's yeast, In the preferred embodiments, the yeast is Saccharomyces cerevisiae.

[0042] In one embodiment, the step c) contacting the yeast with a catalyst to form a reaction mixture consists in adding water and / or an appropriate organic solvent and an effective amount of the catalyst to the yeast to form a reaction mixture, wherein the catalyst is an ionic polymer of the invention or a combination of ionic polymers of the invention, the ionic polymer network of the invention, a membrane incorporating ionic polymers of the invention and / or a solid- supported ionic polymers of the invention; and degrading step d) consists in heating the reaction mixture of step c) during appropriate time and subsequently cooling the temperature (typically to 80-90 °C).

[0043] In some embodiments of the method for producing the MMC from the yeast, the method further comprises applying a pressure of N2 or CO2 during the degrading step d). The pressure may range from 10 bar to 300 bar, preferably from 10 to 150 bar.

[0044] In some embodiments of the method for producing the MMC from the yeast, the step g) is performed, i.e. it is not optional.

[0045] Optionally, prior to contacting the yeast with a catalyst (step c)), determining lipids, proteins and / or carbohydrates contents in the yeast is carried out. Lipids, proteins and / or carbohydrates contents are determined in the yeast based on the standard methods. Lipids can be determined / extracted using Folch method (Folch J, Lees M, Stanley, GHS, 1957, 226, 497-509) involving a mixture of methanol, chloroform and water (2: 1 :0.8, v / v / v), and phase separation afterwards. Determination of carbohydrates is performed according, for example, NREL protocol for “Determination of Structural Carbohydrates and Lignin in Biomass”. For example, 1 ml of 72 % sulfuric acid was added to 100 mg of the yeast. The slurry was stirred for 1 h at 30 °C, followed by addition of 28 ml of deionized water. Mixture was autoclaved at 120 C for 1 h, cooled to room temperature and was used for sugar analysis by HPLC and acid-soluble lignin determination using UV-spectrophotometry at 205 nm wavelength. The same hydrolysate was used for proteins analysis according to the Bradford protein assay. The residue from acid hydrolysis was washed with 100 mL of water and then dried at 105 °C to determine Klason lignin.

[0046] The optional pre-treatment of the yeast, used in the methods described herein, uses one or more methods selected from the group consisting of washing, solvent-extraction, solvent-swelling, comminution, milling, steam pre-treatment, explosive steam pre-treatment, dilute acid pre- treatment, hot water pre-treatment (at about 90°C), alkaline pre-treatment, lime pre-treatment, wet oxidation, wet explosion, ammonia fibre explosion, organosolvent pre-treatment, biological pre-treatment, ammonia percolation, ultrasound, electroporation, microwave, supercritical CO2, supercritical H2O, ozone, and gamma irradiation. The optional pre-treatment of the yeast includes for example the milling of the yeast. To overcome the obstacle of the reaction rate being limited by the surface reaction and mass transfer, a pre-treatment processes of the yeast via ball milling, which leads to cell wall disruption and an increase in the specific surface area of biomass, is highly recommended. Depending on performed mechanical ball milling of the yeast there is a decrease in structural particle size, reduction of the degree of polymerization of mannoprotein and b-glucan.

[0047] In an embodiment of the method for producing a MMC of the invention, the optional pre- treating of the yeast (step b) consists in the hot water pre-treatment of the yeast at about 90°C during 30 minutes to 2 hours, preferably 1 hour, followed by cooling to room temperature (20°C-25°C), and filtration in order to obtain a solid phase (i.e. pre-treated yeast) to be used in step c).

[0048] Some ionic polymers used in the method of the invention for producing the MMC from yeast consists of anions and a polymeric backbone containing cations as disclosed in WO 2019 / 058270 Al incorporated by reference in its entirety. Specifically, the ionic polymer (IP) used in the method of the invention for producing the MMC from the yeast consists of a monomer of formula I formula I or consists of a first monomer of formula I formula I and at least one second monomer selected from the group consisting of formula IV formula V formula VI wherein n and m are independently selected from 1, 2, 3, 4, 5, 6; preferably n and m are independently selected from 1, 2, 3; most preferably n is 2 and m is 1 or 2. z and w are independently selected from 0, 1, 2, 3; preferably z and w are independently selected from 0 and 1 ; most preferably z and w are 0 or 1.

[0049] Zi, Z2 and Z3 are cations each independently selected from the group comprising: preferably Zi, Z2 and Z3 are cations each independently selected from the group comprising: most preferably Zi, Z2 and Z3 are cations each independently selected from the group comprising: R1, R2, R3, R4, R5, R6 and R7 are each independently selected from the group comprising a bond, H, C1-C6alkyl, C1-C6allyl, -CH2-(CH2)p-O-(CH2)q-CH3, C1-C6alkoxy, C1- C6alkoxyalkyl, benzyl, -SO3H, -(CH2)q-SO3H, provided that two of R1, R2, R3, R4, R5, R6 and R7 are each a bond; preferably R1, R2, R3, R4, R5, R6 and R7 are each independently selected from the group comprising a bond, H, C1-C6alkyl, provided that two of R1, R2, R3, R4, R5, R6 and R7 are each a bond; most preferably R1, R2, R3, R4, R5, R6 and R7 are each independently selected from the group comprising a bond and H, provided that two of R1, R2, R3, R4, R5, R6 and R7 are each a bond; p and q are independently selected from 0, 1, 2, 3, 4, 5, 6; L is an optional (present or not present) linker and each occurrence of L, if present, is independently selected from H, substituted or unsubstituted C1-C20alkylene, C1-C20alkenylene, C1-C20 alkynylene and substituted or unsubstituted C5-C10 aryl, wherein the substituents are selected from the group comprising H, –SO3H, –COOH, –[P(=O)(OH)2], –O–SO3H, –O– COOH, –O–[P(=O)(OH)2], preferably L is absent; A is an optional (present or not present) acidic group and each occurrence of A, if present, is independently selected from the group comprising H, –SO3H, –COOH, – [P(=O)(OH)2], –O–SO3H, –O–COOH, –O–[P(=O)(OH)2], -CH2-COOH, provided that when z and w are 0, A is present in formula IV; preferably each occurrence of A, if present, is independently selected from the group comprising –SO3H, –COOH, –O–COOH, -CH2-COOH, provided that when z and w are 0, A is present in formula IV; most preferably A is absent or occurrence of A, if present, is independently selected from the group comprising –COOH, - CH2-COOH, provided that when z and w are 0, A is present in formula IV; X- is selected from the group comprising F−, Cl−, Br−, I−, ClO4−, BF4−, PF6−, AsF6−, SbF6−, NO2−, NO3−, HSO4−, SO42−, PO43−, HPO42−, CF3CO2−, CF3CO3−, CO32−, CF3SO3-, C1-C6 carboxylate, CN-, SCN-, OCN-, CNO-, N3-, tosylate, mesylate, trifluoromethanesulfonate, trifluoroethane sulfonate, di-trifluoromethanesulfonyl amino, docusate, xylenesulfonate; preferably X- is selected from the group comprising F−, Cl−, HSO4−, SO42−, PO43−, HPO42−, CF3CO2−, CF3CO3−, CF3SO3-; most preferably X- is selected from the group comprising Cl−, HSO4−, SO42−, CF3SO3-. Ra is C1-C24alkyl; Rb and Rc are each independently selected from the group comprising H and CH3or absent, preferably Rc is absent; Rd is C1-C24alkylene or C1-C24alkyl, optionally substituted by C1-C24alkyl, preferably Rd is C1-C2alkylene or C1-C2alkyl or C1-C3alkyl; Re and Rf are each independently C1-C24 alkyl, preferably CH3; Y is N or O, provided that when Y is O, Rc is absent; R is selected from the group comprising C1-C24alkyl and C5-C10aryl or is absent; preferably R is CH3; In the context of the present invention, the feature "A is an optional acidic group" means that A is a group, preferably an acidic group, that is optional, i.e that A is present or absent. In some embodiments of the method, the second monomer of formula VI of the ionic polymer (IP) is In some embodiments of the method, the second monomer of formula VI of the ionic polymer (IP) is In some embodiments of the method, the (first) monomer of formula I of the ionic polymer (IP) is formula I In some embodiments of the ionic polymer of the method of the present invention, Z1and Z2are same (identical). In other embodiments, Z1and Z2are different. In some embodiments of the ionic polymer of the method of the present invention, when Z1 and Z2is , wherein R2 and R5 are bonds and R1, R3 and R4 are H, n is not 4. In other embodiments of the ionic polymer of the present invention, when Z1and Z2is , wherein R2 and R5 are bonds and n is 4, at least one of R1, R3 and R4 is not H. In some preferred embodiments of the ionic polymer of the method of the present invention, C1-C6carboxylate are selected from the group comprising formate, acetate, propionate, butyrate, hexanoate, maleate, fumarate, oxalate, lactate, pyruvate. The ratio between different monomers in the ionic polymers of the invention that comprises the first monomer and the second monomers can be any suitable ratio and may vary depending on the yeast to be processed. In some embodiments, the first and the second monomers are present in ratio 1 : 1 or 4: 1. In some other embodiments, the ratio between the first and the second monomers used in the methods described herein is ranging from 4: 1 to 1 :4. According to some embodiments of the method of the present invention, monomers according to formula I are selected from the group comprising

[0050]

[0051] According to further embodiments of the method of the present invention, monomers according to formula I are selected from or

[0052] According to some embodiments of the method of the present invention, monomer according to formula II is According to some embodiments, the method of the present invention provides ionic polymers selected from the group comprising

[0053]

[0054] x and y are integers each independently selected within the range 1 to 1000; preferably 1 to 500 or 1 to 200; more preferably 1 to 100 or 1 to 50;

[0055] According to some embodiments, the method of the present invention provides ionic polymers selected from the group comprising x and y are integers each independently selected within the range 1 to 1000; preferably 1 to 500 or 1 to 200; more preferably 1 to 100 or 1 to 50;

[0056] According to an embodiment of the method of the invention, the ionic polymer is

[0057] According to another embodiment of the method of the invention, the ionic polymer (IP-A5) consists of the monomer of formula (I) and the monomer of formula VI as follows: formula (I) formula (VI)

[0058] According to another embodiment of the method of the invention, the ionic polymer (IP-A6) consists of the monomer of formula (I) and the monomer of formula (VI) as follows: formula (I) formula (VI)

[0059] In some embodiments of the method for producing the MMC, the organic solvent is selected from the group comprising alcohol (such as methanol, ethanol, butanol, ethylene glycol, etc., preferably ethanol), ether (such as dimethoxyethane, diglyme, butyl methyl ether, etc.), ketone (such as methyl isobutyl ketone, V-methyl-2-pyrrolidone, etc.), eutectic solvent (such as glycerol, choline chloride, octanoic acid, tetrabutylammonium chloride, poly (ethylene glycol), choline chloride, lactic acid, glycine).

[0060] In some embodiments of the methods of the present invention, recovering the MMC can be done by any technic known in the art, such as filtration, centrifugation or gravity settling. After the recovering, the MMC can be used in a liquid form or concentrated or dried to a powder form.

[0061] The effective amount of the ionic polymers of the invention or a combination thereof used in the methods described herein can depend on several factors including, for example, the type of the yeast, the amount of the yeast, the content of proteins, carbohydrates and / or lipids in the yeast, the type and number of pre-treatment(s) applied to the yeast, and the reaction conditions (such as temperature and time). An effective amount of the ionic polymer of the invention refers to an amount sufficient to degrade the yeast into the MMC of the invention. In some embodiments, the effective amount of the ionic polymer of the invention is usually 0.005: 1 w / w to 10: 1 w / w, 0.05: 1 w / w to 10: 1 w / w, 0.5: 1 w / w to 10: 1 w / w, 1 : 1 w / w to 1 :5 w / w, preferably 0.1 :1 w / w to 1 :5 w / w, most preferably 0.005: 1 w / w to 0.5: 1 w / w, compared to in the yeast loading.

[0062] The ratio yeast to water used in the methods described herein can depend on several factors, including for example the type of the yeast and the amount of the yeast. In some embodiments, the ratio yeast to water and / or organic solvent (such as alcohol, ether, ketone, eutectic solvent) used in the methods described herein is ranging from 1 : 100 w / v to 1 : 1 w / v, preferably 1 :50 w / v to 1 :10 w / v or preferably 0.5: 10 w / v to 1.5: 10 w / v.

[0063] The preferred temperature profile for the heating used in the methods described herein depends on the yeast starting material being used and also the intended MMC being produced. The heating temperature should preferably be held at a maximum of 170°C, in some embodiments at a maximum of 150°C, 155°C or 160°C. In some embodiments, the heating temperature is between 50°C and 170°C, or between 80°C and 170°C preferably between 100°C to 155°C or between 100°C to 130°C. Preferably, for small-scale applications, the heating is done in a high- pressure autoclave reactor, which after sealing, is heated for appropriate reaction time and temperature.

[0064] In some embodiments, the appropriate reaction time in the methods described herein is for example between 10 minutes and 10 hours, preferably between 0.5 hour and 5 hours, or 0.5 hours and 3 hours, or between 1 hour and 3 hours, depending on the type and amount of the yeast.

[0065] The method for producing the MMC operates at moderate temperatures, typically less than 160°C, whereas the prior art methods need temperatures of more than 170°C. In addition, the method for producing the MMC of the invention provides fewer by-products, which allows easier recovery of the desired products. Another aspect of the present invention provides a microbiome modulator composition (MMC) able to stimulate growths and / or activity of one or more strains of bacteria selected from Rikenellacaea family and Muribaculaceae family, wherein said composition comprising or consisting of in wt%:

[0066] • carbohydrates and / or hydrolyzed carbohydrates: 30% to 50%, preferably 35% to 45% or preferably 40% to 45%;

[0067] • proteins and / or hydrolyzed proteins: 30% to 50%, preferably 35% to 45% or preferably 40% to 45%;

[0068] • fats: 1% to 4%

[0069] • ash: 1% to 5%

[0070] • nucleotides: 0% to 5%, preferably 0.0001% to 5%

[0071] • moisture: < 10%, preferably 0.0001% to 10% or preferably 0.0001% to 5%

[0072] In some embodiments, the bacteria selected from Rikenellacaea family are Rikenellaceae RC9 and Alistipes. In a further embodiment, the MMC inhibits growths and metabolic activity of Micispirillum phylum Deferribacteres.

[0073] According to some embodiments of the MMC of the invention, the hydrolyzed carbohydrates have the degree of polymerisation of less than DP 50.

[0074] According to other embodiments of the MMC of the invention, the hydrolyzed proteins have molecular weight smaller than 100 kDa and consist of or comprise the following amino acids in wt%:

[0075] Asp: 0% to 5%

[0076] Ser: 0% to 5%

[0077] - Glu: 0% to l0%

[0078] Gly: 0% to 4%

[0079] - His: 0% to 3%

[0080] Arg: 0% to 4%

[0081] - Thr: 0% to 4%

[0082] - Ala: 0% to 7%

[0083] - Pro: 0% to 4%

[0084] - GABA: 0% to 2%

[0085] Cystine: 0% to 4% Tyr: 0% to 4%

[0086] - Vai: 0% to 6%

[0087] - Met: 0% to 4

[0088] - Lys: 0% to 7%

[0089] - Lie: 0% to 4%

[0090] - Leu: 0% to 4%

[0091] - Phe: 0% to 5%

[0092] Trp: 0% to 4%.

[0093] According to some embodiment of the MMC of the invention, the hydrolyzed carbohydrates have the distribution of degree of polymerisation between DP2 and DP50.

[0094] According to further embodiments of the MMC of the invention, the hydrolyzed carbohydrates comprise mainly b-glucan and mannan oligo- and polysaccharides.

[0095] In an embodiment, the MMC of the invention comprises or consists of (in wt %):

[0096] • 41% carbohydrates and / or hydrolysed carbohydrates

[0097] • 45% proteins and / or hydrolysed proteins

[0098] • 2.5% fats

[0099] • 3.5% ash

[0100] • 8% moisture

[0101] In another embodiment, the MMC of the invention comprises or consists of (in wt %):

[0102] • 41.6% carbohydrates and / or hydrolysed carbohydrates

[0103] • 42% proteins and / or hydrolysed proteins

[0104] • 2.5% fats

[0105] • 3.5% ash

[0106] • 8% moisture

[0107] In further embodiments, the MMC of the invention is derived from a yeast. In some embodiments, the yeast is any suitable yeast for human consumption, such as brewer’s yeast or baker's yeast, In the preferred embodiments, the yeast is Saccharomyces cerevisiae. In another embodiment, the MMC of the invention is able to inhibit growths and / or activity of one or more strains of bacteria shown in Table 2. In an embodiment, the MMC inhibits growths and metabolic activity of Mi cispirillum phylum Deferribacteres.

[0108] In some embodiments of the MMC, carbohydrates and hydrolysed carbohydrates are at least glucose and mannose.

[0109] In some embodiments of the MMC, the distribution of degree of polymerisation (DP) of said carbohydrates and hydrolysed carbohydrates is 2-50.

[0110] In various embodiments, the MMC of the invention includes primarily carbohydrates and proteins and hydrolysed forms thereof. Such a carbohydrate compound can be a bio-based compound. A benefit of a carbohydrate compound can be improving the balance of a body area commensal microbiota, by providing an energy source to promote beneficial microbial growth in the body area. In some embodiments, the at least one carbohydrates or hydrolysed carbohydrates includes at least glucose and mannose. In other embodiments, carbohydrate or hydrolysed carbohydrate can be further an inulin, an alpha-glucan oligosaccharide, a fructooligosaccharide, an isomaltooligosaccharide, an xylooligosaccharide, an arabinoxylo- oligosaccharide, a beta-glucan, a transgalactooligosaccharide, mannan-oligosaccharide, lactulose, xylitol, lactitol, trehalose, or combinations thereof. The MMC of the invention can further include phenolics (such as ferulic acid).

[0111] Another embodiment of the present invention provides the microbiome modulator composition (MMC) of the invention obtained by the method of the invention for producing the microbiome modulator composition (MMC).

[0112] The production (preparation) of the microbiome composition (MMC) of the invention is not limited to the method of the invention for producing the MMC. According to other embodiments, the microbiome modulator composition (MMC) of the invention can be produced, obtained or manufactured by chemical synthesis, fermentation and manufacturing processes known in the art. For example, the MMC of the invention may be formulated as nutraceutical, nutritional or pharmaceutical compositions. These formulations comprise effective amounts of the essential ingredients of the MMC of the invention, such as carbohydrates, proteins and hydrolysed forms thereof. The different formulations can be prepared according to standard rules and proceeding established in the corresponding art.

[0113] The term "nutraceutical” means a pharmaceutical -grade standardized nutrient. The term "pharmaceutical" in the present content means a pharmaceutical grade compound prescribed as medicament to treat a disease. The term "nutrient" means in the present context substance that provides nourishment essential for the maintenance of life of a human. The term "nutritional" in the present context means that the composition is for the dietary supplementation of a human individual. The term "dietary supplement" means a product taken by mouth that contains a dietary ingredient, e.g. a nutrient, intended to supplement the diet.

[0114] In one embodiment, the MMC for use and methods in accordance with the present invention are applicable to humans or more generally to animals. In an embodiment, administration of the MMC of the invention is oral and therefore the present invention provides an oral MMC.

[0115] For example, the MMC of the invention can be formulated in the form of freeze-dried power, tablet, capsules, pills, suspension, lozenge, emulsion, liquid preparations, gel, syrup, cream, ointment, etc. In a preferred embodiment of the invention, the MMC of the invention is a liquid preparation, the most preferably an aqueous preparation.

[0116] While it is possible to administer the MMC of the invention alone, it is also possible to administer on or in a support (for example a carrier) as part of a product, in particular as a component of a food product, a drink product, a dietary supplement, medicament, medical food, nutraceutical or a pharmaceutical formulation / combination. These products typically contain additional components, acceptable excipients, carriers or adequate additives well known to those skilled in the art. The term "acceptable excipients and carriers" as used herein pertains to those that are compatible with the other ingredients in the formulation and biologically acceptable.

[0117] The MMC of the invention can be also used as an ingredient in food products such as milk products, yogurt, curd, cheese (for example quark, cream, processed, soft and hard), fermented milk, milk powder, milk based fermented product, ice-cream, a fermented cereal based product, milk-based powder (such as for example a whey concentrate), a beverage, a dressing, meat products (e.g. liver paste, frankfurter and salami sausages or meat spreads), spreads, fillings, frostings, chocolate, confectionery (e.g. caramel, candy, fondants or toffee), baked goods (cakes, pastries), sauces and soups, fruit juices or coffee whiteners.

[0118] Another aspect of the present invention provides a pharmaceutical combination of the MMC of the invention and one or more cancer immunotherapeutic agent for simultaneous, separate or sequential administration, wherein the cancer immunotherapeutic agent is selected from the group comprising immune checkpoint inhibitor, TCR-T cells, CAR-T cells or combinations thereof.

[0119] In some embodiments of the pharmaceutical combination of the invention, the immune checkpoint inhibitor is selected from the group comprising anti-PDl, anti-PDLl, anti-CTLA4, TIGIT receptor inhibitors, anti-LAG3, anti-TIM-3, anti-TIM-4 and anti-VISTA.

[0120] In other embodiments of the pharmaceutical combination of the invention, the immune checkpoint inhibitor is selected from the group consisting of nivolumab, pembrolizumab, ipilimumab, atezolizumab, jemperli, pidilizumab, durvalumab, lambrolizumab and avelumab. or any combinations thereof.

[0121] In an embodiment, the pharmaceutical combination of the invention comprises: a first composition comprising or consisting of the MMC of the invention, wherein the first composition is capable of increasing the efficacy of the cancer immunotherapeutic agent; and a second composition comprising a therapeutically effective amount of one or more cancer immunotherapeutic agent, wherein the cancer immunotherapeutic agent is selected from the group comprising immune checkpoint inhibitor, TCR-T cells, CAR-T cells or combinations thereof. In one embodiment, the first and second compositions are provided in a single formulation, are provided concurrently, or are provided separately and / or sequentially.

[0122] The MMC of the invention for use with the invention can be provided in a variety of dosage forms. For example, freeze-dried power, tablet, capsules, pills, suspension, lozenge, emulsion, liquid preparations, gel, syrup, cream, ointment, may be used to provide the MMC of the invention to a patient in need of therapy for cancer with the cancer immunotherapeutic agent(s). The cancer immunotherapeutic agent(s) and the MMC of the invention can be provided concurrently, in the same dosage form, simultaneously (at the same time) thorough the same or different routes, sequentially (one after the other), or separately in location and / or time. In the present disclosure, the inventors show for the first time the beneficial impact of the MMC of the invention obtained from hydrolysis of the yeast on antitumor immunity in a syngeneic MPM mouse model. Based on an estimation of daily consumption of water per mouse, only a few micrograms of the MMC of the invention per day, incorporated in the drinking water, were sufficient to induce a tumour growth delay and improve survival although no effects on tumour engraftment were reported. Importantly, no adverse effects of the administration of the MMC of the invention were observed in mice, including body weight and stool appearance.

[0123] The MMC of the invention demonstrated an enhancement in the activation and infiltration of CD8+ T-cells into tumours. The recruitment of cytotoxic lymphocytes into tumours appeared to be one of the requirements for the tumour control observed in response to the MMC of the invention, as no significant impact on tumour-infiltrating CD4 T cells was reported and the beneficial effects of MMC on tumour growth and animal survival was completely lost / abolished in T-cell deficient animals. In addition to T cell modulation, macrophages infiltration in the tumour of MMC group was significantly increased. The immunostaining analyses did not reveal any apparent alterations in the ratio between Ml -like and M2 -like macrophages, respectively localized by CD80 and CD206 markers.

[0124] The administration of the MMC of the invention shows a significant modification of the microbial community at the three sites of investigation, including ileum, faeces, and most prominently in the colon. The administration of the MMC of the invention also shows a change in the metabolomic composition of the gut and a link between short chain fatty acids but also 2-Methylbuteric acid, Isobutyric acid, Butyric acid and Isovaleric acid and CD8+GRZB+ tumour infiltrating lymphocytes.

[0125] The MMC of the invention has the capacity to significantly change the tumour immune microenvironment and thus improve tumour control. Such effect of the MMC of the invention is valuable in combination with cancer immunotherapy, such as immune checkpoint inhibition, in patients bearing solid tumours.

[0126] The MMC of the invention triggers the sequence of metabolic reactions which amplifies the effect of the product and makes it more pronounced than usual prebiotics. In some embodiments, the MMC of the invention changes the gut microbiota to specific metabolic creating strains which are strongly associated to the infiltration of active CD8 granzyme B positive T cells (cytotoxic T cells). This step is important for example for better tumour control and improves the likelihood and depth of responsiveness to any form of immunotherapy.

[0127] Another aspect of the present invention provides a method for stimulating growths and / or activity of one or more strains of bacteria selected from Rikenellacaea family and Muribaculaceae family in a subject, the method comprising administering to the subject an effective amount of the MMC of the invention.

[0128] According to an embodiment of the method for stimulating growths and / or activity of one or more strains of bacteria selected from Rikenellacaea family and Muribaculaceae family in a subject, the effective amount of the MMC of the invention is 0.01 to 20 mg / kg / day, preferably 0.01 to 5 mg / kg / day or 0.4 to 2 mg / kg / day, preferably 0.4 to 1 mg / kg / day or 0.8 to 1.2 mg / kg / day or 0.8 to 1.4 mg / kg / day or 5 to 10 mg / kg / day or 10 to 20 mg / kg / day, in one dose or in several doses, such as two, three, four or more doses, preferably one or two doses.

[0129] In an embodiment, the method for stimulating growths and / or activity of one or more strains of bacteria selected from Rikenellacaea family and Muribaculaceae family in a subject, includes inhibiting growths and metabolic activity of Mi cispirillum phylum Deferribacteres.

[0130] Another aspect of the present invention provides a method of enhancing immune system function in a subject comprising administering to the subject an effective amount of the MMC of the invention. In a specific embodiment, the MMC of the invention stimulates immune response against a solid cancer.

[0131] According to an embodiment, the present invention provides the MMC of the invention for use in a method of enhancing immune system function in a subject. In a specific embodiment, the MMC of the invention stimulates immune response against a solid cancer.

[0132] According to another embodiment, the present invention provides use of the MMC of the invention for the manufacture of a medicament use for enhancing immune system function in a subject. In a specific embodiment, the MMC of the invention stimulates immune response against a solid cancer. As used herein, the term "immune system function" is intended to include any function of the immune system, e.g., functions associated with non-specific (innate) immunity as well as functions associated with specific (adaptive) immunity.

[0133] According to an embodiment of the method of enhancing immune system function of the invention, the effective amount of the MMC of the invention is 0.01 to 20 mg / kg / day, preferably 0.01 to 5 mg / kg / day or 0.4 to 2 mg / kg / day, preferably 0.4 to 1 mg / kg / day or 0.8 to 1.2 mg / kg / day or 0.8 to 1.4 mg / kg / day or 5 to 10 mg / kg / day or 10 to 20 mg / kg / day, in one dose or in several doses, such as two, three, four or more doses, preferably one or two doses.

[0134] Another aspect of the present invention provides a method for enhancing the efficacy of a cancer immunotherapy in a subject, the method comprising administering an effective amount of the MMC of the invention before the cancer immunotherapeutic agent, concurrently with the cancer immunotherapeutic agent and after the cancer immunotherapeutic agent, wherein the cancer immunotherapeutic agent is selected from the group comprising immune checkpoint inhibitor, TCR-T cells, CAR-T cells or combinations thereof.

[0135] According to an embodiment, the present invention provides the MMC of the invention for use in a method for enhancing the efficacy of a cancer immunotherapy in a subject, the method comprising administering an effective amount of the MMC of the invention before the cancer immunotherapeutic agent, concurrently with the cancer immunotherapeutic agent and after the cancer immunotherapeutic agent, wherein the cancer immunotherapeutic agent is selected from the group comprising immune checkpoint inhibitor, TCR-T cells, CAR-T cells or combinations thereof.

[0136] According to an embodiment, the present invention provides use of the MMC of the invention for the manufacture of a medicament for the method of enhancing the efficacy of a cancer immunotherapy in a subject, the method comprising administering an effective amount of the MMC of the invention before the cancer immunotherapeutic agent, concurrently with the cancer immunotherapeutic agent and after the cancer immunotherapeutic agent, wherein the cancer immunotherapeutic agent is selected from the group comprising immune checkpoint inhibitor, TCR-T cells, CAR-T cells or combinations thereof. In an embodiment of the method for enhancing the efficacy of a cancer immunotherapy of the invention, administering the MMC of the invention is during at least one week before the cancer immunotherapeutic agent, concurrently with the cancer immunotherapeutic agent and during at least one week after the cancer immunotherapeutic agent.

[0137] According to another embodiment of the method for enhancing the efficacy of a cancer immunotherapy of the invention, the immune checkpoint inhibitor is selected from the group comprising comprising anti-PDl, anti-PDLl, anti-CTLA4, TIGIT receptor inhibitors, anti- LAG3, anti-TIM-3, anti-TIM-4 and anti-VISTA.

[0138] According to another embodiment of the method for enhancing the efficacy of a cancer immunotherapy of the invention, the immune checkpoint inhibitor is selected from the group comprising nivolumab, pembrolizumab, ipilimumab, atezolizumab, jemperli, pidilizumab, durvalumab, lambrolizumab and avelumab or any combination thereof.

[0139] According to another embodiment of the method for enhancing the efficacy of a cancer immunotherapy of the invention, the effective amount of the MMC of the invention is 0.01 to 20 mg / kg / day, preferably 0.01 to 5 mg / kg / day or 0.4 to 2 mg / kg / day, preferably 0.4 to 1 mg / kg / day or 0.8 to 1.2 mg / kg / day or 0.8 to 1.4 mg / kg / day or 5 to 10 mg / kg / day or 10 to 20 mg / kg / day, in one dose or in several doses, such as two, three, four or more doses, preferably one or two doses.

[0140] In some embodiments, the cancer immunotherapy is for treating a solid cancer. According to a preferred embodiment, the solid cancer is selected from malignant pleural mesothelioma, melanoma, NSCLC / SCLC, ovarian cancer, cervical cancer, gastro-intestinal cancer. In another preferred embodiment, the solid cancer is malignant pleural mesothelioma.

[0141] Another aspect of the present invention provides a method for treating a solid cancer in a subj ect in need thereof, said method comprising administering to the subject an effective amount of the MMC of the invention and one or more cancer immunotherapeutic agent, wherein the effective amount of the MMC is 0.01 to 20 mg / kg / day, preferably 0.01 to 5 mg / kg / day or 0.4 to 2 mg / kg / day, preferably 0.4 to 1 mg / kg / day or 0.8 to 1.2 mg / kg / day or 0.8 to 1.4 mg / kg / day or 5 to 10 mg / kg / day or 10 to 20 mg / kg / day, and wherein the cancer immunotherapeutic agent is selected from the group comprising immune checkpoint inhibitor, TCR-T cells, CAR-T cells or combinations thereof.

[0142] According to an embodiment, the present invention provides the MMC of the invention for use in a method for treating a solid cancer in a subject in need thereof, said method comprising administering to the subject an effective amount of the MMC of the invention and one or more cancer immunotherapeutic agent, wherein the effective amount of the MMC is 0.01 to 20 mg / kg / day, preferably 0.01 to 5 mg / kg / day or 0.4 to 2 mg / kg / day, preferably 0.4 to 1 mg / kg / day or 0.8 to 1.2 mg / kg / day or 0.8 to 1.4 mg / kg / day or 5 to 10 mg / kg / day or 10 to 20 mg / kg / day, and wherein the cancer immunotherapeutic agent is selected from the group comprising immune checkpoint inhibitor, TCR-T cells, CAR-T cells or combinations thereof.

[0143] According to another embodiment, the present invention provides use of the MMC of the invention for the manufacture of a medicament for a method of treating a solid cancer in a subject in need thereof, said method comprising administering to the subject an effective amount of the MMC of the invention and one or more cancer immunotherapeutic agent, wherein the effective amount of the MMC is 0.01 to 20 mg / kg / day, preferably 0.01 to 5 mg / kg / day or 0.4 to 2 mg / kg / day, preferably 0.4 to 1 mg / kg / day or 0.8 to 1.2 mg / kg / day or 0.8 to 1.4 mg / kg / day or 5 to 10 mg / kg / day or 10 to 20 mg / kg / day, and wherein the cancer immunotherapeutic agent is selected from the group comprising immune checkpoint inhibitor, TCR-T cells, CAR-T cells or combinations thereof.

[0144] According to an embodiment of the method for treating a solid cancer of the invention, the MMC stimulates the enrichment and metabolomic activity of one or more strains of bacteria in the gut selected from Rikenellacaea and Muribaculaceae families. In some embodiments, the bacteria selected from Rikenellacaea family are Rikenellaceae RC9 and Alistipes. In a further embodiment, the MMC inhibits growths and metabolic activity of Micispirillum phylum Deferribacteres.

[0145] In another embodiment, the MMC of the invention inhibits growths and metabolomic activity of one or more strains of bacteria shown in Table 2. In an embodiment, the MMC inhibits growths and metabolic activity of Micispirillum phylum Deferribacteres. According to an embodiment of the method for treating a solid cancer of the invention, the solid cancer is a cancer sensitive to immunotherapies. According to a preferred embodiment, the solid cancer is selected from malignant pleural mesothelioma, melanoma, NSCLC / SCLC, ovarian cancer, cervical cancer, gastro-intestinal cancer. In another preferred embodiment, the solid cancer is malignant pleural mesothelioma.

[0146] According to another embodiment of the method for treating a solid cancer of the invention, the immune checkpoint inhibitor selected from the group comprising anti-PDl, anti-PDLl, anti- CTLA4, TIGIT receptor inhibitor, anti-LAG3, anti-TIM-3, anti-TIM-4 and anti-VISTA.

[0147] According to another embodiment of the method for treating a solid cancer of the invention, the immune checkpoint inhibitor selected from the group comprising nivolumab, pembrolizumab, ipilimumab, atezolizumab, jemperli, pidilizumab, durvalumab, lambrolizumab and avelumab or any combinations thereof.

[0148] Cancer immunotherapies are usually given every 3 weeks (intervals may range between every two weeks to every four weeks). Cancer immunotherapies are given for a maximum of two years.

[0149] According to the present invention, the MMC of the invention is administered before the cancer immunotherapeutic agent, concurrently with the cancer immunotherapeutic agent and after the cancer immunotherapeutic agent. Since the cancer immunotherapy is usually given every 3 weeks, typically the MMC of the invention is administered in between two sessions of cancer immunotherapy.

[0150] The term “before the cancer immunotherapeutic agent” may refer to any point in time after a subject has been selected to undergo the cancer immunotherapy but prior to the commencement of administration of the cancer immunotherapeutic agent.

[0151] The term “undergoing cancer immunotherapy” is defined as encompassing a subject who has been selected to undergo immunotherapy in the future but is yet to begin immunotherapy, and / or a subject concurrently undergoing immunotherapy and / or a subject who has undergone immunotherapy. It is understood that immunotherapy encompasses the administration of the cancer immunotherapeutic agent to the patient, either directly or indirectly. According to some embodiments of the present invention, the MMC of the invention may be administered at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 days, at least 8 days, at least 9 days, at least 10 days, at least, 11 days, at least 12 days, at least 13 days, at least 14 days before administration of the cancer immunotherapeutic agent. The MMC of the invention may be administered 1 to 14 days, 5 to 30 days, 10 to 30 days, 15 to 30 days, 20 to 30 days, or 25 to 30 days before administration of the cancer immunotherapeutic agent.

[0152] The term “concurrent to cancer immunotherapy” may refer to any point in time while the cancer immunotherapy is being administered through an individual treatment or treatment programme over time, or concurrently as the cancer immunotherapeutic agent is administered. As used herein, “concurrent” is used to mean administration of the MMC of the invention and the cancer immunotherapeutic agent by simultaneous, sequential or separate means.

[0153] As used herein, “simultaneous” is used to mean that the MMC of the invention and the cancer immunotherapeutic agent are administered at an overlapping or the same time. As used herein, “sequential” is used to mean that the MMC of the invention and the cancer immunotherapeutic agent are not administered simultaneously, but one after the other. In contrast to “sequentially”, “separately” is used herein to mean that the gap between administering one agent and the other is significant i.e. the first administered agent may no longer be present in the bloodstream in a therapeutically effective amount when the second agent is administered. Thus, administration “sequentially” may permit the MMC of the invention to be administered within 5 minutes, 10 minutes or a matter of hours before or after the cancer immunotherapeutic agent administration provided the circulatory half-life of the first administered agent is such that they are both concurrently present in therapeutically effective amounts. The time delay between administration of the MMC of the invention and the cancer immunotherapeutic agent will vary depending on the exact nature of the cancer immunotherapeutic agent, the interaction there between, and their respective half-lives.

[0154] The term “after cancer immunotherapy” may refer to any point in time after the administration of any individual cancer immunotherapeutic agent or treatment programme has completed. According to some embodiments of the present invention, the MMC of the invention may be administered during 1 day, 2 days, 3 days, 4 days, 5 days, 6 days, 7 days, 8 days, 9 days, 10 days, 11 days, 12, days, 13 days, 14 days, 15 days, 16 days, 17 days, 18 days, 19 days 20 days, 21 days, 22 days, 23 days, 24 days, 25 days, 26 days, 27 days, 28 days, 29 days, 30 days after administration of the cancer immunotherapeutic agent. The MMC of the invention may be administered during 1 to 14 days, 1 to 28 days, 1 to 30 days, 5 to 30 days, 7 to 14 days, 7 to 28 days, 10 to 30 days, 15 to 30 days, 20 to 30 days, 21 to 28 days, or 25 to 30 days after administration of the cancer immunotherapeutic agent. Since the cancer immunotherapy is usually given every 3 weeks (21 days), typically the MMC of the invention is administered in between two sessions of cancer immunotherapy.

[0155] In one embodiment, the MMC of the invention may be administered once per day. In another embodiment the MMC of the invention may be administered more than once, for example two, three, four, five, six, seven, eight, nine, ten or more times per day.

[0156] The administration of the MMC of the invention was associated with a better tumour control and patient survival with limited toxicity. This opens new perspectives for cancer management, suggesting that modulation of the microbiota could serve as a minimally toxic approach to enhance anti-tumour immunity and improve the response to immune checkpoint inhibitor (ICI) therapy.

[0157] Another aspect of the present invention provides a method of preventing and / or treating infectious diseases in a subject, the method comprising administering to the subject an effective amount of the MMC of the invention, wherein the infectious diseases are caused by pathogenic microorganisms selected from the group comprising bacteria, viruses, fungi and parasites., preferably the infectious diseases are caused by bacteria. Indeed, the MMC of the invention has the immunostimulant effect and can thus boost the immune system and enhance adaptive immune response against foreign bodies, such as pathogens, causing infection.

[0158] According to an embodiment of the method of preventing and / or treating infectious diseases, the method further comprises administering to the subject one or more additional active agents selected from the group comprising antibiotics, anti-bacterial agents, antifungal agents, antiparasitic agents, immunomodulating drugs, anti-viral agents and combinations thereof. Those skilled in the art will appreciate that the invention described herein is susceptible to variations and modifications other than those specifically described. It is to be understood that the invention includes all such variations and modifications without departing from the spirit or essential characteristics thereof. The invention also includes all of the steps, features, compositions and compounds referred to or indicated in this specification, individually or collectively, and any and all combinations or any two or more of said steps or features. The present disclosure is therefore to be considered as in all aspects illustrated and not restrictive, the scope of the invention being indicated by the appended claims, and all changes which come within the meaning and range of equivalency are intended to be embraced therein.

[0159] The foregoing description will be more fully understood with reference to the following Examples. Such Examples, are, however, exemplary of methods of practising the present invention and are not intended to limit the application and the scope of the invention.

[0160] EXAMPLES

[0161] Cell lines and culture conditions

[0162] Biphasic MPM cells AB 12 (RRID:CVCL_4405) isolated from ascites of asbestos-exposed wild-type BALB / c mice were transduced with a luciferase reporter gene to generate AB12-luc cells. This allowed monitoring of tumor growth in the orthotopic site using bioluminescence imaging. Once inoculated in mice, these cells cause the development of a biphasic MPM with most cells presenting a sarcomatoid morphology. Cells were grown in RPMI 1640 medium supplemented with 10% Fetal Bovine Serum (FBS) and 5 pg / ml puromycin and passaged at 80% of confluency.

[0163] Animals and tumor model

[0164] Housing and treatment assignation: Animal experiments were initiated on ten to twelve-week- old BALB / c or BALB / c nude mice imported from Charles River (50-50 male-to-female ratio). All animals were kept in a specific pathogen free environment, which included filtered air, sterilized food, water, bedding, and cages. The animals were acclimated for at least one week prior to the beginning of experiments and all experiments were conducted in accordance with the Animal Welfare Act and the National Institutes of Health “Guidelines for the Care and Use of Laboratory Animals” and approved by the Committee for Animal Experiment for the Canton Vaud, Switzerland (authorization VD3345). After the acclimatation period, animals were randomly assigned to untreated (NT) or MMC groups with a sex ratio of 1 : 1. The MMC was administered through drinking water two weeks before cancer cells inoculation and kept along the course of the experiment. Drinking water was replaced every week.

[0165] Orthotopic tumor model

[0166] Mice were anaesthetized with a mix of ketamin / xylazin (80 / 10 mg / kg) and placed in a supine position. 25x 104 AB12-luc cells resuspended in 50pl of DMEM without serum were injected through the fourth intercostal space using a 29-gauge needle inserted about 5mm into the left pleural cavity. Tumor growth was recorded using bioluminescence imaging. Tumor growth curves were started when the tumors reached a volume associated to a photon flux > 107 which correspond to the beginning of the tumor exponential growth phase. After tumor cell inoculation, animals were monitored daily and euthanized if they presented a weight loss of more than 15% compared to the start of the experiment, rapid respiration, or strong decreased activity.

[0167] Bioluminescence imaging

[0168] Tumor sizes were assessed by bioluminescence measurement using the In-Vivo Imaging System (IVIS) Lumina S5 every three days. 100 pl of VivoGloTM Luciferin (15 mg / ml, Promega) were injected intraperitoneally (i.p.) 15 minutes prior imaging and animals were kept under isoflurane anesthesia during the entire course of monitoring. The tumor sites showing bioluminescence signals were identified as the regions of interest (ROI) and the total photon counts were quantified using the Living Image® software.

[0169] Immunofluorescence staining of tumor sections

[0170] At the end of tumor growth assessment, mice were euthanized by i.p. injection of pentobarbital (150 mg / kg) and intracardiac perfusion with NaCl 0.9% was performed. Pleural tumors were collected, embedded in OCT, cut into 8 pm sections and mounted on glass slides. Tumor sections were then fixed at -20°C in methanol for 10 minutes, washed three times in PBS and mounted on Shandon coverplate system. Non-specific reactivity was blocked by incubating sections for 1 hour with blocking buffer (5% normal donkey serum, 0.1% bovine serum albumin, 0.3% triton in PBS). Samples were then incubated overnight at 4°C in blocking buffer containing primary antibodies, washed three times with PBS, and incubated for 1 hour at room temperature with secondary antibodies in blocking buffer. At the end of the procedure, sections were washed in PBS and mounted in Fluoromount-G™ mounting medium. Primary and secondary antibodies used are recapitulated in the table below (Table 1).

[0171] Table 1: Antibodies for immunofluorescence staining of tumor sections

[0172] Image analysis

[0173] Tumor sections were scanned with a Zeiss Axioscan Z1 slide scanner at lOx magnification. Image analysis was performed using ImageJ software. The following threshold values were used for the different staining: CD3: 65-150, CD4: 60-120, CD8: 50-120, CD45: 40-150, CD68: 20-180, CD206: 70-240, CD80: 10-70.

[0174] Colocalization data between CD4 / CD3, CD8 / CD3 and CD45 / CD68 were obtained using the RG2B colocalization plugin. Sequential colocalizations were performed for Granzyme B, CD206 and CD80. Colocalization data were normalized to tumor area.

[0175] GC-MS analysis of Short Chain Fatty Acids

[0176] Short chain fatty acids from C2 to C7 including hydroxylated and methylated derivatives were separated in a DB-FFAP column (30 m x 0.25 mm i.d. x 0.25 pm) (Agilent Technologies) and analyzed by single quadrupole mass spectrometer operating in SIM mode (El 70 eV) using a 2 ms dwell time (Agilent Technologies, data not shown). Chromatographic conditions were set as follows: carrier gas He 1.2 ml / min at constant flow. Initial temperature was set at 40 °C, the temperature increment was 10 °C / min along the gradient until it reached 200 °C, and afterwards 40°C / min until 240°C (over 3 min).

[0177] Feces samples (10 mg) were homogenized with 1 mL of water. An aliquot (100 μL) of homogenate was extracted with 140 μL of methyl -tert-butyl ether (MTBE) spiked with the internal standard solution as previously described by Lotti et al. Samples were centrifuged for 15 min at 15000 rpm (4°C) and 1.5 μL of supernatant was injected in spitless mode (240 °C) into the GC-MS system.

[0178] Bacterial DNA extraction

[0179] One fecal pellet, colon or ileum (without luminal content) from each mouse was collected into a sterile 1.5-ml Biopure tube (Eppendorf), placed on dry ice, and stored at -80 °C until further processing. Total bacterial DNA was isolated using the QiaAMP Fast DNA Stool Mini Kit (QIAGEN) according to the manufacturer’s instructions. DNA was eluted with 200 pl of AE buffer (provided with the kit). DNA was stored at 4 °C until being used for the PCR.

[0180] 16S rRNA gene library preparation and amplicon sequencing

[0181] Amplicon sequencing targeted the V1-V2 region of the 16S rRNA gene with primers F-27 and R-338 (data not shown). Amplification was performed using the Accuprime Taq DNA Polymerase High Fidelity kit (Invitrogen, Waltham, MA). No-template PCR reaction controls (N = 2) were included. Libraries were loaded onto an Illumina MiSeq using pairwise chemistry, generating 250 x 2 read lengths (Lausanne Genomic technologies facility, University of Lausanne, Switzerland).

[0182] Bioinformatics and statistical analysis

[0183] All analyses were performed in R version 4.1.0. Demultiplexing, removal of chimeric and short reads, singlebase resolution of reads into amplicon sequence variants (ASVs) using the Divisive Amplicon Denoising Algorithm 2 (DADA2) algorithm, and taxonomic annotation using the SILVA database were performed using a dedicated pipeline available at https: / / github.com / chuvpne / dada2-pipeline with default parameters. Taxonomic classification and exact sequence matching were performed using the SILVA database (version 123). ASV filtering, normalization, ordination and diversity analyses were performed using the phyloseq R package (version 1.42.0) and visualized using the ggplot2 R package (version 3.4.0). Unclassified ASVs at the genus level were removed and filtered based on prevalence (25% of total samples) and counts (15 000 reads minimum). The ASV count table then was normalized using total sum scaling, where each ASV count is divided by the total read count for each sample. PCoA was performed using Bray-Curtis distance matrix calculated using the vegan R package (version 2.6.4).

[0184] Statistical analysis

[0185] Statistical analyses were performed using GraphPad Prism version 9.1.0 for Windows (GraphPad Software, Inc). Survival curves were compared using log-rank test and a difference in survival distribution was assumed when p < 0.05. For immunostainings comparisons, a two- tailed Student’s t-test was applied to assess differences in the distribution of untreated versus MMC-treated samples. All data are expressed as mean ± SD. P < 0.05 was considered significant.

[0186] Results

[0187] Tumor immune microenvironment in MMC treated mice.

[0188] The MMC used in the study comprises (in wt %):

[0189] • 41,6% carbohydrates and / or hydrolysed carbohydrates

[0190] • 42% proteins and / or hydrolysed proteins

[0191] • 2.5% fats

[0192] • 3.5% ash

[0193] • 8% moisture

[0194] The impact on the composition of the immune microenvironment associated to the MMC of the invention intake in the syngeneic orthotopic MPM mouse model was first assessed. It was particularly looked at the effects on T lymphocytes due to their crucial role in the anti-tumor response and macrophages because of their predominance in MPM tumors. Immunostainings of tumor sections revealed a significant increase of CD3+CD8+ T-cells in tumor whereas CD3+CD4+ T-cells level was not affected (Fig. lA, B, D and E). Of interest, an increased expression of the granule protein Granzyme B (GRZB) in the infiltrating CD8+ T-cells upon MMC administration was observed, indicating cytotoxic activity (Fig 1 C and F). The total amount of macrophages (CD45+CD68+) was also increased in the MMC group, however the ratio between Ml -like (CD45+CD68+CD80+) and M2-like (CD45+CD68+CD206+) macrophages was unchanged (Fig 2). Delayed tumor progression and extended survival in immunocompetent but not in athymic mice (T-cell deficient) supplemented with MMC.

[0195] To assess the impact of MMC-related immunomodulatory effects on tumor control, tumor growths in MMC-treated versus untreated immunocompetant animals were compared. As shown in Figure 3 A, tumor growths were significantly decreased in the MMC group compared to controls. The delay in tumor growth correlated with a significant enhancement in survival time with median survival of 10 versus 7 days for MMC compared to untreated group (Hazard ratio (logrank): 0.35 [95% CI 0.14-0.91]) (Fig 3B). To assess the contribution of T-cells in the observed response, the experiment was repeated in athymic mice of the same background who lack T cells. In T cell deficient mice, MMC had no impact on tumor growth and mouse survival (Fig 3C and 3D). In addition, a positive correlation between the amount of CD8+GRZB+T cells in tumors and survival of the animals was found with a Spearman coefficient of 0.5965 (p- value: 0.0164) (Fig 4A) whereas no significant association was found between CD4+T cells infiltration and survival (Fig 4B).

[0196] MMC promotes a specific gut microbiome composition dependent on the sampling site.

[0197] The impact of the MMC of the invention supplementation on microbiome composition in the ileum, colon, and feces was assessed following 16S rRNA amplicon sequencing. Principal coordinate analysis of community composition in the three different sampling sites revealed that the microbiome in the colon and Ileum of the MMC of the invention treated mice were statistically different when compared to non-treated animals (Fig 5A). The most pronounced changes in community composition occurred in the colon (Fig 5A, a). Interestingly, when assessing Shannon diversity significant decreases were observed in the fecal and Ileum samples of the MMC of the invention treated group (Fig 5B, a); whilst Faith’s phylodiversity was only significantly decreased in in feces (Fig 5B, b).

[0198] Comparison of community composition based on relative abundance of the top 12 genera showed a differential response based on sample type (Fig 6A). More specifically, the MMC treatment led to an enrichment of Alistipes and Rikenellacaea RC9 gut group (Rikenellacea family) in the colon (Fig 6 B, a-b). In addition, we observed a significant enrichment in Muribaculaceae at the family level following the MMC treatment (Fig 6B, c), whereas a significant reduction was observed in Mucispirillum genus from Deferribacteraceae family (colon), Lachnospiraceae NK4A136 group (feces), and Lachnospiraceae Family (colon and feces) (Fig 6B, d-f). When all sampling locations are considered (i.e., colon, ileum and feces), all genera observed in the MMC samples (MMC treated group) also appeared in the control samples. However, a loss of 62 genera in the MMC samples (MMC treated group) was observed (Fig 6C and table 2)

[0199] Table 2: 62 genera lost in the MMC treated group

[0200]

[0201] Metabolomic changes induced by the MMC of the invention in the gut:

[0202] Metabolite analysis in fecal samples of control and the MMC treated mice showed no differences in SCFA levels between the treatment groups (Fig 7A). However, linear regression analyses show a strong correlation between fecal SCFA levels and and CD8+GRZB+T cell infiltration (Fig. 7B). The MMC -treated mice show a positive and statistically significant correlation between the amount of CD8+GRZB+T cells and of 2-Methylbuteric acid, Isobutyric acid, and Isovaleric acid (Fig. 7B, a-c). Most strikingly, the relationship between the presence of CD8+GRZB+and 2-Methylbuteric acid, Isobuteric acid and Isovaleric was inverted in MMC- treated subjects compared to controls (Fig. 7B, a-c). Furthermore, a correlation between CD8+T cell infiltration and Butyric acid was found (Fig. 7B, d). In particular, the SCFA producing genera Rikenellaceae RC9 gut group and Alistipes (Rikenellacaea family) were both enriched in the colon of MMC-treated mice. In addition, microbiome-derived SCFAs levels in fecal samples of MMC-treated mice were shown to be associated with increased level of cytotoxic T-cells in tumors.

[0203] Impact of the MMC of the invention on immune checkpoints in tumors

[0204] Expression of the immune checkpoints PD-1 and CTLA-4 associated with CD8 cells was assessed by immunofluorescence stainings of tumor sections in order to evaluate the combination potential of the MMC with immune checkpoint inhibitors (ICI) (Fig 8A and B). Quantification of the colocalization signal between PD-1 or CTLA-4 and CD8 signals revealed that the amount of CD8+cells expressing the exhaustion markers PD-1 or CTLA-4 was increased upon MMC treatment (Fig 8 C and D), suggesting a benefit of combination therapy.

[0205] Catalyst and MMC preparations

[0206] Catalyst

[0207] IL of water containing 450 g of Ml and 2.5 ml of propanediol was degassed and added to 2 L of cyclohexane containing 50 ml of Span 80. Afterwards, polymerization initiator (0,25 g) dissolved in 10 ml of degassed water was added to the mixture. The system was subjected to 71 °C under N2. After 10-12 h, another portion of initiator 0,35 g in 10 ml of water was added and heating / stirring under N2 continued for another 12 h. At the end the liquid phase was discarded and obtained solid powder was washed with ethanol and hot water till neutral pH.

[0208] Microbiome Modulator Composition

[0209] The Microbiome Modulator Composition (MMC, EMB008) was provided by Embion Technologies SA (Switzerland) and obtained with Embion’ s proprietary extraction platform by processing yeast material. The MMC is composed of carbohydrates, hydrolysed carbohydrates, proteins and hydrolysed proteins. Importantly the MMC (EMB008) does not include any living organism (probiotic).

[0210] The MMC (EMB008) was provided in the form of a dehydrated powder and was prepared at 31 ppm (7.75 mg of the MMC in 250 ml of drinking water) concentration in the drinking water of mice. The MMC was obtained as followed: 500 g of yeast (dry bases) suspended in 4 L of water together with ionic polymer catalyst “ZPl” were stirred at 155°C for 2 h. After reaction, the mixture was cooled, filtered and the liquid phase was dried using spray drier. The obtained dried powdered product was analysed and used for cancer study.

[0211] The MMC analysis: The composition is composed primarily from carbohydrates and proteins and hydrolysed forms thereof. The total amount of carbohydrates after hydrolysis is 41.6%, of which glucose is 30.6% and mannose is 11%. Most of the carbohydrates are within degree of polymerisation of 2-50. The amount of protein is 42%.

[0212] Size exclusion chromatogram for the MMC is shown in Fig. 9

Claims

CLAIMS1. A microbiome modulator composition (MMC) able to stimulate growths and / or activity of one or more strains of bacteria in the gut selected from Rikenellacaea family and Muribaculaceae family, said composition comprising in wt%:• carbohydrates and / or hydrolyzed carbohydrates: 30% to 50%• proteins and / or hydrolyzed proteins: 30% to 50%• fats: 1% to 4%• ash: 1% to 5%• nucleotides: 0% to 5%• moisture: < 10%2. The microbiome modulator composition (MMC) of claim 1, wherein the hydrolyzed carbohydrates have the degree of polymerization of less than DP 50 and comprise mainly b- glucan and mannan oligo- and polysaccharides, and wherein the hydrolyzed proteins have molecular weight smaller than 100 kDa and consist of the following amino acids in wt%:Asp: 0% to 5%Ser: 0% to 5%- Glu: 0% to l0%Gly: 0% to 4%- His: 0% to 3%Arg: 0% to 4%- Thr: 0% to 4%- Ala: 0% to 7%- Pro: 0% to 4%- GABA: 0% to 2%Cystine: 0% to 4%Tyr: 0% to 4%- Vai: 0% to 6%- Met: 0% to 4- Lys: 0% to 7%- Lie: 0% to 4%- Leu: 0% to 4%- Phe: 0% to 5%Trp: 0% to 4%.

3. A pharmaceutical combination comprising the MMC of claims 1 or 2 and one or more cancer immunotherapeutic agent for simultaneous, separate or sequential administration, wherein the cancer immunotherapeutic agent is selected from the group comprising immune checkpoint inhibitor, TCR-T cells, CAR-T cells or combinations thereof.

4. The pharmaceutical combination of claim 3, wherein the immune checkpoint inhibitor is selected from the group comprising anti-PDl, anti-PDLl, anti-CTLA4, TIGIT receptor inhibitors, anti-LAG3, anti-TIM-3, anti-TIM-4 and anti-VISTA.

5. The pharmaceutical combination of claims 3 or 4, wherein the immune checkpoint inhibitor is selected from the group consisting of nivolumab, pembrolizumab, ipilimumab, atezolizumab, jemperli, pidilizumab, durvalumab, lambrolizumab and avelumab. or any combinations thereof.

6. A method of enhancing immune system function in a subject comprising administering to the subject an effective amount of the MMC of claims 1 or 2.

7. The method of claim 6, wherein the effective amount of the MMC is 0.01 to 20 mg / kg / day.

8. A method for enhancing the efficacy of a cancer immunotherapy in a subj ect, the method comprising administering an effective amount of the MMC of claims 1 or 2 before the cancer immunotherapeutic agent, concurrently with the cancer immunotherapeutic agent and after the cancer immunotherapeutic agent, wherein the cancer immunotherapeutic agent is selected from the group comprising immune checkpoint inhibitor, TCR-T cells, CAR-T cells or combinations thereof.

9. The method of claim 8, wherein administering the MMC of claim 1 before the cancer immunotherapeutic agent is during at least one week before.

10. The method of claim 8 or claim 9, wherein administering the MMC of claims 1 or 2 after the cancer immunotherapeutic agent is during at least one week after.

11. The method of any one of claims 8 to 10, wherein the immune checkpoint inhibitor is selected from the group comprising anti-PDl, anti-PDLl, anti-CTLA4, TIGIT receptor inhibitors, anti-LAG3, anti-TIM-3, anti-TIM-4 and anti-VISTA.

12. The method of any one of claims 8 to 10, wherein the immune checkpoint inhibitor is selected from the group comprising nivolumab, pembrolizumab, ipilimumab, atezolizumab, jemperli, pidilizumab, durvalumab, lambrolizumab and avelumab or any combination thereof.

13. The method of any one of claims 8 to 12, wherein the effective amount of the MMC is 0.01 to 20 mg / kg / day.

14. A method for treating a solid cancer in a subject in need thereof, said method comprising administering to the subject an effective amount of the MMC of claims 1 or 2 and one or more cancer immunotherapeutic agent, wherein the effective amount of the MMC is 0.01 to 20 mg / kg / day and wherein the cancer immunotherapeutic agent is selected from the group comprising immune checkpoint inhibitor, TCR-T cells, CAR-T cells or combinations thereof.

15. The method of claim 14, wherein the administering an effective amount of the MMC is before the cancer immunotherapeutic agent, concurrently with the cancer immunotherapeutic agent and after the cancer immunotherapeutic agent.

16. The method of claim 14 or claim 15, wherein the MMC stimulates the enrichment and metabolomic activity of one or more strains of bacteria in the gut selected from Rikenellacaea and Muribaculaceae families.

17. The method of any one of claims 14 to 16, wherein the solid cancer is a cancer sensitive to immunotherapies.

18. The method of any one of claims 14 to 17, wherein the solid cancer is selected from malignant pleural mesothelioma, melanoma, NSCLC / SCLC, ovarian cancer, cervical cancer, gastro-intestinal cancer.

19. The method of any one of claims 14 to 18, wherein the immune checkpoint inhibitor is selected from the group comprising anti-PDl, anti-PDLl, anti-CTLA4, TIGIT receptor inhibitor, anti-LAG3, anti-TIM-3, anti-TIM-4 and anti-VISTA.

20. The method of any one of claims 14 to 18, wherein the immune checkpoint inhibitor selected from the group comprising nivolumab, pembrolizumab, ipilimumab, atezolizumab, jemperli, pidilizumab, durvalumab, lambrolizumab and avelumab or any combinations thereof.

21. A method of preventing and / or treating infectious diseases in a subject comprising administering to the subject an effective amount of the MMC of claims 1 or 2, wherein the infectious diseases are caused by pathogenic microorganisms selected from the group comprising bacteria, viruses, fungi and parasites.

22. The method of claim 21, further comprising administering to the subject one or more additional active agents selected from the group comprising antibiotics, anti-bacterial agents, antifungal agents, antiparasitic agents, immunomodulating drugs, anti-viral agents and combinations thereof.

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