Corynebacterium strains for preventing viral infection, combinations thereof and lyophilized formulations thereof

Abstract

The present invention relates to Corynebacterium strains and combinations thereof capable of reducing cathepsin expression for use in the prevention of viral infections, in particular SARS-CoV-2 infections or for use in the prevention of infections by enveloped respiratory viruses in a subject wherein the viruses have been entered into the cells of the subject using a cathepsin entry pathway. In particular, the virus is an SARS-CoV-2 Ombke variant. The invention also relates to a lyophilized formulation comprising a Corynebacterium sp. Strain or a combination thereof and a cryoprotective agent for use in the prevention of infection by enveloped respiratory viruses, preferably SARS-CoV-2 virus, in a subject. Preferably, the formulation is administered to the upper respiratory tract of the subject.

Description

Technical Field

[0001] The present invention relates to Corynebacterium strains and combinations thereof capable of reducing cathepsin expression, which are used for preventing viral infections, particularly SARS-CoV-2 infections. The present invention also relates to freeze-dried preparations comprising these Corynebacterium strains or combinations, which are used for preventing coronavirus infections. Background Art

[0002] Epithelial cells of the upper and lower respiratory tracts are the main targets of airborne infections. These cells are covered by complex bacterial communities mainly located in the upper respiratory tract, and these communities may interact with coronaviruses directly or indirectly. The idea that commensal bacteria can also prevent infections by modulating innate and adaptive host immune responses has been proposed in the art. In addition, nasopharyngeal or oro-nasopharyngeal preparations comprising bacteria have been proposed to alter the composition of the microbiota in the upper respiratory tract.

[0003] Lappan and Peacock [Lappan&Peacock, 2019. Corynebacterium and Dolosigranulum: future probiotic candidates for upper respiratory tract infections. Microbiology Australia, 40(4), 172–177.] reviewed studies observing the presence of Corynebacterium and Dolosigranulum in the microbiota of the upper respiratory tract, and these studies showed their association with the healthy state. The authors proposed that strains of Corynebacterium pseudodiphtheriticum and Dolosigranulum pigrum, which are considered to be pathogens of the upper respiratory tract, may have a protective effect on the nasopharyngeal health of children.

[0004] Man et al. also concluded in a paper published in 2017 that Dolosigranulum and Corynebacterium are potential key players in the microbiota of the upper respiratory tract because "in multiple epidemiological and mechanistic studies, they are closely associated with respiratory health and the exclusion of potential pathogens, most notably Streptococcus pneumoniae". [Man et al., 2017. The microbiota of the respiratory tract: Gatekeeper to respiratory health. Nature Reviews Microbiology, 15(5), 259–270].

[0005] Recently, some studies have also raised the question of the correlation between the health of the microbiome and severe infections with RNA viruses and other coronavirus infections.

[0006] The emergence of a highly transmissible and pathogenic coronavirus (i.e., severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) that causes coronavirus disease 2019 (COVID-19) in late 2019 has triggered a pandemic. The importance of this study is supported by the fact that as of September 21, 2022, the World Health Organization has confirmed a cumulative total of 609 million confirmed cases and 6.5 million deaths due to COVID-19 (https: / / www.who.int / publications / m / item / weekly-epidemiological-update-on-covid-19---21-september-2022).

[0007] SARS-CoV-2 infection appears to have a dual nature: extremely deadly for some people, surprisingly mild for others, and some people are not susceptible to the virus at all.

[0008] Infections caused by the SARS-CoV-2 virus are serious problems. The virus primarily settles in the nasal mucosa, and the infection then progresses to a severe systemic disease. The infection and its course depend on the individual's susceptibility.

[0009] Epithelial cells of the upper and lower respiratory tract are the main targets for infection and replication of the SARS-CoV-2 virus. Assuming that the invasion portal is protected by the commensal microbiota of the nasopharynx, several authors have tested the bacterial composition in infected and uninfected individuals.

[0010] Nardelli et al. [Nardelli et al., 2021. Nasopharyngeal Microbiome Signature in COVID-19 Positive Patients: Can We Definitively Get a Role to Fusobacterium periodonticum? Frontiers in Cellular and Infection Microbiology, 11 (February), 1–7.] and Maio et al. [Maio et al., 2020. Nasopharyngeal Microbiota Profiling of SARS-CoV-2 Infected Patients. Biological Procedures Online 22, 18.] found no differences in the nasopharyngeal microbiome composition between samples from COVID-19 patients and virus-negative controls. On the contrary, it can be concluded from their data that SARS-CoV-2 infection did not significantly alter the microbiome composition compared to that of the general population. Testing a large number of patients and using the 16S rRNA method, Rosas-Salazar et al. [Rosas-Salazar et al., 2021. SARS-CoV-2 infection and viral load are associated with the upper respiratory tract microbiome. J Allergy Clin Immunol. 2021 Apr; 147(4):1226-1233.e2] reported a mix of Corynebacterium: while some strains were more abundant in SARS-CoV-2 infected patients, other strains were more abundant in virus-uninfected individuals.

[0011] In contrast to targeted approaches, next-generation metagenomic sequencing also provides valuable information on the composition of the microbiota at the species level. A next-generation metagenomic study by Mostafa et al. [Mostafa et al., 2020. Metagenomic next-generation sequencing of nasopharyngeal specimens collected from confirmed and suspect COVID-19 patients. MBio, 11(6), 1–13.] described a statistically significant decrease in the incidence of the commensal organism Corynebacterium accolens in samples from COVID-19-positive patients. The authors noted that there is evidence in the prior art that Corynebacterium accolens is negatively correlated with the colonization of Streptococcus pneumoniae; however, further research is needed to draw conclusions about the role of these associations in patients with COVID-19, and no negative correlation was proposed between the presence of Corynebacterium accolens and SARS-CoV-2 infection. In fact, the decrease in Corynebacterium accolens levels is likely the result of infection.

[0012] Susceptibility to SARS-CoV-2 infection and its clinical course remain unpredictable. Tchoupou Saha et al. [Tchoupou Saha et al., 2022. Profile of the Nasopharyngeal Microbiota Affecting the Clinical Course in COVID-19 Patients. Front. Microbiol. 13:871627.] studied the nasopharyngeal microbiota of COVID-19 patients and patients who tested negative for the virus by 16S ribosomal ribonucleic acid (rRNA) sequencing and targeted pathogen-specific polymerase chain reaction (PCR). They found that nine taxa were increased in patients who tested positive for SARS-CoV-2, such as Corynebacterium propinquum / Corynebacterium pseudodiphtheriticum and Afipia birgiae. They also demonstrated that Corynebacterium propinquum was decreased in asymptomatic individuals compared to other COVID-19-positive patients. However, these authors also noted that further research is needed to determine the exact role of Corynebacterium accolens and especially Corynebacterium propinquum / Corynebacterium pseudodiphtheriticum in the clinical course of the disease.

[0013] The COVID-19 pandemic remains a serious problem. One reason for this is the evolution of SARS-CoV-2, which has led to the emergence of several new variants. The newly released Omicron variant of SARS-CoV-2 has become the dominant variant in many countries. Due to a large number of mutations, Omicron exhibits cell tropism and entry modes compared to other SARS-CoV-2 variants. Different from the original or other SARS-CoV-2 variants, the Omicron variant mainly uses the cathepsin B / L entry pathway in addition to the TMPRSS2 (transmembrane protease / serine subfamily member 2) cell entry pathway.

[0014] The object of the present inventors is to isolate bacterial strains that downregulate the cathepsin pathway, which can be used as probiotics for preventing respiratory viral infection, especially SARS-CoV-2 infection; to determine the ideal combination of these strains; and to formulate a nasal spray (lyophilized preparation) that includes these strains or a combination with a suitable excipient. Summary of the Invention

[0015] In a first aspect, there is provided a Corynebacterium culture for preventing or treating infection by an enveloped respiratory virus, preferably a coronavirus, in a subject, wherein the Corynebacterium culture is capable of reducing the expression of a protein that promotes the entry of the enveloped respiratory virus into the cells of the subject, wherein the entry facilitating protein is cathepsin. Preferably, there is provided a Corynebacterium culture for preventing or treating infection by an enveloped respiratory virus, preferably a coronavirus, in a subject, wherein the Corynebacterium culture is capable of reducing the expression of cathepsin in the cells of the subject. Optionally, the Corynebacterium culture is further capable of reducing the expression of other proteins that promote the entry of an enveloped respiratory virus, preferably a coronavirus, into the cells of the subject, and these other proteins are selected from angiotensin-converting enzyme 2 (ACE2) and transmembrane protease / serine subfamily member 2 (TMPRSS2).

[0016] In an embodiment, a Corynebacterium culture is provided for preventing or treating an infection of an enveloped respiratory virus, preferably a coronavirus, in a subject, wherein the Corynebacterium culture is capable of reducing the expression of at least one protein that promotes the entry of the enveloped respiratory virus into the cells of the subject, wherein the at least one entry-promoting protein is selected from cathepsin, ACE2, and TMPRSS2 or any combination thereof, wherein preferably the entry-promoting protein is cathepsin, or wherein preferably the entry-promoting protein is cathepsin and ACE2, or wherein the entry-promoting protein is cathepsin, ACE2, and TMPRSS2.

[0017] Preferably, the enveloped respiratory virus is a coronavirus, preferably SARS-CoV-2 and more preferably the SARS-CoV-2 Omicron variant.

[0018] In a second aspect, a preparation for preventing or treating an infection of a coronavirus in a subject is provided, wherein the preparation comprises a Corynebacterium culture capable of reducing the expression of a protein that promotes the entry of the coronavirus into the cells of the subject, the entry-promoting protein being cathepsin, and wherein optionally, the Corynebacterium culture is further capable of reducing the expression of other proteins that promote the entry of the coronavirus into the cells of the subject, these other proteins being selected from ACE2 and TMPRSS2. Preferably, the preparation is a probiotic composition. Preferably, the preparation is (part of) a medical device. Preferably, the preparation is delivered via a medical device. Preferably, the preparation is a pharmaceutical preparation. Preferably, when appropriate, the preparation further comprises at least one excipient suitable for a probiotic preparation and / or a pharmaceutically acceptable excipient.

[0019] In a third aspect, a composition is provided, which comprises a Corynebacterium culture capable of reducing the expression of a protein that promotes the entry of a coronavirus into the cells of a subject, the entry-promoting protein being cathepsin, and wherein optionally, the Corynebacterium culture may further be capable of reducing the expression of other proteins that promote the entry of the coronavirus into the cells of the subject, these other proteins being selected from ACE2 and TMPRSS2. Preferably, the composition is a probiotic composition. Preferably, the composition is (part of) a medical device. Preferably, the composition is delivered via a medical device. Preferably, the composition is a pharmaceutical composition. Preferably, the composition is for preventing or treating a coronavirus infection. Preferably, when appropriate, the composition further comprises at least one excipient suitable for a probiotic composition and / or a pharmaceutically acceptable excipient.

[0020] In a fourth aspect, a method for preventing or treating coronavirus infection is provided, the method comprising: administering to a subject in need thereof a Corynebacterium culture, or a preparation or composition comprising a Corynebacterium culture, wherein the Corynebacterium culture is capable of reducing the expression of a protein that promotes the entry of coronavirus into the cells of the subject, the entry-promoting protein being cathepsin, and wherein optionally, the Corynebacterium culture may also be capable of reducing the expression of other proteins that promote the entry of coronavirus into the cells of the subject, these other proteins being selected from ACE2 and TMPRSS2.

[0021] In any one of the aspects of the present invention, preferably, the Corynebacterium culture is used for preventing coronavirus infection, and thus also for preventing diseases caused by coronavirus.

[0022] In any one of the aspects of the present invention, preferably, the Corynebacterium culture is also capable of inhibiting the binding between the spike (S) protein of coronavirus, preferably the receptor binding domain (RBD) of the S protein, and ACE2.

[0023] In any one of the aspects of the present invention, preferably, the Corynebacterium culture comprises viable Corynebacterium selected from the group consisting of Corynebacterium accolens, Corynebacterium propinquum (close to Corynebacterium), and Corynebacterium tuberculostearicum (Corynebacterium tuberculostearicum). Preferably, the Corynebacterium culture comprises strains of Corynebacterium accolens, Corynebacterium propinquum, and Corynebacterium tuberculostearicum. Preferably, the Corynebacterium culture comprises at least one strain of Corynebacterium accolens. Preferably, the Corynebacterium culture comprises at least one strain of Corynebacterium propinquum. Preferably, the Corynebacterium culture comprises at least one strain of Corynebacterium tuberculostearicum.

[0024] In any one of the aspects of the present invention, preferably, the Corynebacterium culture does not include Corynebacterium pseudodiphtheriticum.

[0025] In any one of the aspects of the present invention, preferably, the Corynebacterium culture comprises at least one Corynebacterium strain selected from the following:

[0026] Corynebacterium accolens strain SU001 or its derivatives, variants or mutants with accession number NCAIM P(B)001495 deposited at NCAIM (National Collection of Agricultural and Industrial Microorganisms, Institute of Food Science and Technology, Szent István University, Faculty of Agricultural and Life Sciences, H-1118 Budapest, Somlóu tér 14-16, Hungary) on June 22, 2021, wherein its derivatives, variants or mutants are capable of reducing cathepsin expression and optionally, its derivatives, variants or mutants are further capable of reducing ACE2 expression and / or TMPRSS2 expression and / or inhibiting the binding of ACE2 and S protein, and / or its derivatives, variants or mutants express LipS1 lipase and / or comprise a sequence encoding LipS1 lipase;

[0027] Corynebacterium propionicum strain SU002 or its derivatives, variants or mutants with accession number NCAIM P(B)001496 deposited at NCAIM on June 22, 2021, wherein its derivatives, variants or mutants are capable of reducing cathepsin expression and optionally, its derivatives, variants or mutants are further capable of reducing ACE2 expression and / or TMPRSS2 expression and / or inhibiting the binding of ACE2 and S protein;

[0028] Corynebacterium propionicum strain SU003 or its derivatives, variants or mutants with accession number NCAIM P(B)001497 deposited at NCAIM on June 22, 2021, wherein its derivatives, variants or mutants are capable of reducing cathepsin expression and optionally, its derivatives, variants or mutants are further capable of reducing ACE2 expression and / or TMPRSS2 expression and / or inhibiting the binding of ACE2 and S protein;

[0029] Corynebacterium accolens strain SU004 or its derivatives, variants or mutants with accession number NCAIM P(B)001500 deposited at NCAIM on January 17, 2022, wherein its derivatives, variants or mutants are capable of reducing cathepsin expression and optionally, its derivatives, variants or mutants are further capable of reducing ACE2 expression and / or reducing TMPRSS2 expression and / or inhibiting the binding of ACE2 and S protein, and / or expressing lipase and / or comprising a sequence encoding lipase;

[0030] The Corynebacterium tuberculostearicum SU005 strain or its derivatives, variants or mutants with the accession number NCAIM P(B)001501 deposited in NCAIM on April 27, 2022, wherein the derivatives, variants or mutants thereof can reduce the expression of cathepsin, and optionally, the derivatives, variants or mutants thereof can also reduce the expression of ACE2 and / or reduce the expression of TMPRSS2 and / or inhibit the binding of ACE2 and the S protein;

[0031] The Corynebacterium accolens SU006 strain or its derivatives, variants or mutants with the accession number NCAIM P(B)001502 deposited in NCAIM on April 27, 2022, wherein the derivatives, variants or mutants thereof can reduce the expression of cathepsin, and optionally, the derivatives, variants or mutants thereof can also reduce the expression of ACE2 and / or reduce the expression of TMPRSS2 and / or inhibit the binding of ACE2 and the S protein, and / or the derivatives, variants or mutants thereof express lipase and / or comprise a sequence encoding lipase;

[0032] The Corynebacterium propionicum SU007 strain or its derivatives, variants or mutants with the accession number NCAIM P(B)001504 deposited in NCAIM on April 27, 2022, wherein the derivatives, variants or mutants thereof can reduce the expression of cathepsin, and optionally, the derivatives, variants or mutants thereof can also reduce the expression of ACE2 and / or reduce the expression of TMPRSS2 and / or inhibit the binding of ACE2 and the S protein;

[0033] The Corynebacterium accolens SU008 strain or its derivatives, variants or mutants with the accession number NCAIM P(B)001505 deposited in NCAIM on April 27, 2022, wherein the derivatives, variants or mutants thereof can reduce the expression of cathepsin, and optionally, the derivatives, variants or mutants thereof can also reduce the expression of ACE2 and / or reduce the expression of TMPRSS2 and / or inhibit the binding of ACE2 and the S protein, and / or the derivatives, variants or mutants thereof express lipase and / or comprise a sequence encoding lipase;

[0034] The Corynebacterium accolens SU009 strain or its derivatives, variants or mutants with the accession number NCAIM P(B)001506 deposited in NCAIM on May 25, 2022, wherein the derivatives, variants or mutants thereof can reduce the expression of cathepsin, and optionally, the derivatives, variants or mutants thereof can also reduce the expression of ACE2 and / or reduce the expression of TMPRSS2 and / or inhibit the binding of ACE2 and the S protein, and / or the derivatives, variants or mutants thereof express lipase and / or comprise a sequence encoding lipase;

[0035] Corynebacterium propionicum strain SU010 or its derivatives, variants or mutants deposited under accession number NCAIM P(B)001507 at NCAIM on May 25, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing the expression of cathepsin, and optionally, the derivatives, variants or mutants thereof are further capable of reducing the expression of ACE2 and / or reducing the expression of TMPRSS2 and / or inhibiting the binding of ACE2 and the S protein.

[0036] In any of the aspects of the present invention, preferably, the Corynebacterium culture comprises at least one Corynebacterium strain selected from the following:

[0037] Corynebacterium accolens strain SU004 or its derivatives, variants or mutants deposited under accession number NCAIM P(B)001500 at NCAIM on January 17, 2022 (National Collection of Agricultural and Industrial Microorganisms, Institute of Food Science and Technology, Hungarian University of Agriculture and Life Sciences, H-1118, Budapest, Somloi ut 14-16, Hungary), wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression, and optionally, the derivatives, variants or mutants thereof are further capable of reducing ACE2 expression and / or TMPRSS2 expression and / or inhibiting the binding of ACE2 and the S protein, and / or the derivatives, variants or mutants thereof express lipase and / or comprise a sequence encoding lipase;

[0038] Corynebacterium tuberculostearicum strain SU005 or its derivatives, variants or mutants deposited under accession number NCAIM P(B)001501 at NCAIM on April 27, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing the expression of cathepsin, and optionally, the derivatives, variants or mutants thereof are further capable of reducing the expression of ACE2 and / or reducing the expression of TMPRSS2 and / or inhibiting the binding of ACE2 and the S protein;

[0039] Corynebacterium accolens strain SU006 or its derivatives, variants or mutants deposited under accession number NCAIM P(B)001502 at NCAIM on April 27, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression, and optionally, the derivatives, variants or mutants thereof are further capable of reducing ACE2 expression and / or reducing the expression of TMPRSS2 and / or inhibiting the binding of ACE2 and the S protein, and / or the derivatives, variants or mutants thereof express lipase and / or comprise a sequence encoding lipase;

[0040] The Corynebacterium propionicum SU007 strain or its derivatives, variants or mutants with the accession number NCAIM P(B)001504 deposited in NCAIM on April 27, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression, and optionally, the derivatives, variants or mutants thereof are further capable of reducing ACE2 expression and / or reducing TMPRSS2 expression and / or inhibiting the binding of ACE2 and the S protein;

[0041] The Corynebacterium accolens SU008 strain or its derivatives, variants or mutants with the accession number NCAIM P(B)001505 deposited in NCAIM on April 27, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression, and optionally, the derivatives, variants or mutants thereof are further capable of reducing ACE2 expression and / or reducing TMPRSS2 expression and / or inhibiting the binding of ACE2 and the S protein, and / or the derivatives, variants or mutants thereof express lipase and / or comprise a sequence encoding lipase;

[0042] The Corynebacterium accolens SU009 strain or its derivatives, variants or mutants with the accession number NCAIM P(B)001506 deposited in NCAIM on May 25, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression, and optionally, the derivatives, variants or mutants thereof are further capable of reducing ACE2 expression and / or reducing TMPRSS2 expression and / or inhibiting the binding of ACE2 and the S protein, and / or the derivatives, variants or mutants thereof express lipase and / or comprise a sequence encoding lipase;

[0043] The Corynebacterium propionicum SU010 strain or its derivatives, variants or mutants with the accession number NCAIM P(B)001507 deposited in NCAIM on May 25, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression, and optionally, the derivatives, variants or mutants thereof are further capable of reducing ACE2 expression and / or reducing TMPRSS2 expression and / or inhibiting the binding of ACE2 and the S protein.

[0044] In any of the aspects of the present invention, preferably, the Corynebacterium culture comprises at least one Corynebacterium strain selected from the following:

[0045] Corynebacterium accolens strain SU004 or its derivatives, variants or mutants with accession number NCAIM P(B)001500 deposited at NCAIM (National Collection of Agricultural and Industrial Microorganisms, Institute of Food Science and Technology, Szent István University, Faculty of Agricultural and Life Sciences, H-1118 Budapest, Somlóu str. 14-16, Hungary) on January 17, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression and optionally, the derivatives, variants or mutants thereof are further capable of reducing ACE2 expression and / or TMPRSS2 expression and / or inhibiting the binding of ACE2 and the S protein;

[0046] Corynebacterium tuberculostearicum strain SU005 or its derivatives, variants or mutants with accession number NCAIM P(B)001501 deposited at NCAIM on April 27, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression and optionally, the derivatives, variants or mutants thereof are further capable of reducing ACE2 expression and / or reducing TMPRSS2 expression and / or inhibiting the binding of ACE2 and the S protein;

[0047] Corynebacterium accolens strain SU006 or its derivatives, variants or mutants with accession number NCAIM P(B)001502 deposited at NCAIM on April 27, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression and optionally, the derivatives, variants or mutants thereof are further capable of reducing ACE2 expression and / or reducing TMPRSS2 expression and / or inhibiting the binding of ACE2 and the S protein;

[0048] Corynebacterium propionicum strain SU007 or its derivatives, variants or mutants with accession number NCAIM P(B)001504 deposited at NCAIM on April 27, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression and optionally, the derivatives, variants or mutants thereof are further capable of reducing ACE2 expression and / or reducing TMPRSS2 expression and / or inhibiting the binding of ACE2 and the S protein;

[0049] Corynebacterium accolens strain SU008 or its derivatives, variants or mutants with accession number NCAIM P(B)001505 deposited at NCAIM on April 27, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression and optionally, the derivatives, variants or mutants thereof are further capable of reducing ACE2 expression and / or reducing TMPRSS2 expression and / or inhibiting the binding of ACE2 and the S protein;

[0050] The Corynebacterium accolens strain SU009 or its derivatives, variants or mutants with the accession number NCAIM P(B)001506 deposited at NCAIM on May 25, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression, and optionally, the derivatives, variants or mutants thereof are further capable of reducing ACE2 expression and / or reducing TMPRSS2 expression and / or inhibiting the binding of ACE2 and the S protein;

[0051] The Corynebacterium propionicum strain SU010 or its derivatives, variants or mutants with the accession number NCAIM P(B)001507 deposited at NCAIM on May 25, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression, and optionally, the derivatives, variants or mutants thereof are further capable of reducing ACE2 expression and / or reducing TMPRSS2 expression and / or inhibiting the binding of ACE2 and the S protein.

[0052] In any of the aspects of the present invention, preferably, the Corynebacterium culture comprises at least one Corynebacterium strain selected from the following:

[0053] The Corynebacterium accolens strain SU004 or its derivatives, variants or mutants with the accession number NCAIM P(B)001500 deposited at NCAIM (National Collection of Agricultural and Industrial Microorganisms, Research Institute of Food Science and Technology, Szent István University, Faculty of Agricultural and Life Sciences, H-1118, Budapest, Somlósvári út 14-16, Hungary) on January 17, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression;

[0054] The Corynebacterium tuberculostearicum strain SU005 or its derivatives, variants or mutants with the accession number NCAIM P(B)001501 deposited at NCAIM on April 27, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression;

[0055] The Corynebacterium accolens strain SU006 or its derivatives, variants or mutants with the accession number NCAIM P(B)001502 deposited at NCAIM on April 27, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression;

[0056] The Corynebacterium propionicum strain SU007 or its derivatives, variants or mutants with the accession number NCAIM P(B)001504 deposited at NCAIM on April 27, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression;

[0057] Corynebacterium accolens strain SU008 or its derivatives, variants or mutants with accession number NCAIM P(B)001505 deposited in NCAIM on April 27, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression;

[0058] Corynebacterium accolens strain SU009 or its derivatives, variants or mutants with accession number NCAIM P(B)001506 deposited in NCAIM on May 25, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression;

[0059] Corynebacterium propionicum strain SU010 or its derivatives, variants or mutants with accession number NCAIM P(B)001507 deposited in NCAIM on May 25, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression.

[0060] In another aspect, the present invention relates to Corynebacterium accolens strain SU001 or its derivatives, variants or mutants with accession number NCAIM P(B)001495 deposited in NCAIM (National Collection of Agricultural and Industrial Microorganisms, Institute of Food Science and Technology, Szent István University, H-1118 Budapest, Somlóu tér 14-16, Hungary) on January 22, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression, and optionally, the derivatives, variants or mutants thereof are further capable of reducing ACE2 expression and / or TMPRSS2 expression and / or inhibiting the binding of ACE2 and S protein, and / or the derivatives, variants or mutants thereof express LipS1 lipase and / or comprise a sequence encoding LipS1 lipase. The present invention also relates to a Corynebacterium culture comprising Corynebacterium accolens SU001 or consisting essentially of Corynebacterium accolens SU001. Preferably, the Corynebacterium accolens strain SU001, or a culture comprising or consisting essentially of the Corynebacterium accolens strain SU001 is used for preventing the infection of an enveloped respiratory virus, preferably a coronavirus, more preferably the SARS-CoV-2 virus, in a subject. In an embodiment, the SARS-CoV-2 virus is the SARS-CoV-2 Omicron variant.

[0061] On the other hand, the present invention relates to Corynebacterium propionicum strain SU002 or its derivatives, variants or mutants with the accession number NCAIM P(B)001496 deposited at NCAIM on June 22, 2021, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression, and optionally, the derivatives, variants or mutants thereof are also capable of reducing ACE2 expression and / or TMPRSS2 expression and / or inhibiting the binding of ACE2 and the S protein. The present invention also relates to a Corynebacterium culture comprising Corynebacterium propionicum SU002 or consisting essentially of Corynebacterium propionicum SU002. Preferably, the Corynebacterium propionicum SU002 strain, or a culture comprising the Corynebacterium propionicum SU002 strain or consisting essentially of the Corynebacterium propionicum SU002 strain is used for preventing the infection of an enveloped respiratory virus, preferably a coronavirus, more preferably the SARS-CoV-2 virus, in a subject. In an embodiment, the SARS-CoV-2 virus is the SARS-CoV-2 Omicron variant.

[0062] On the other hand, the present invention relates to Corynebacterium propionicum strain SU003 or its derivatives, variants or mutants with the accession number NCAIM P(B)001497 deposited at NCAIM on June 22, 2021, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression, and optionally, the derivatives, variants or mutants thereof are also capable of reducing ACE2 expression and / or TMPRSS2 expression and / or inhibiting the binding of ACE2 and the S protein. The present invention also relates to a Corynebacterium culture comprising Corynebacterium propionicum SU003 or consisting essentially of Corynebacterium propionicum SU003. Preferably, the Corynebacterium propionicum SU003 strain, or a culture comprising the Corynebacterium propionicum SU003 strain or consisting essentially of the Corynebacterium propionicum SU003 strain is used for preventing the infection of an enveloped respiratory virus, preferably a coronavirus, more preferably the SARS-CoV-2 virus, in a subject. In an embodiment, the SARS-CoV-2 virus is the SARS-CoV-2 Omicron variant.

[0063] On the other hand, the present invention relates to the Corynebacterium incertum strain SU004 or its derivatives, variants or mutants with the accession number NCAIM P(B)001500 deposited at NCAIM on January 17, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression, and optionally, the derivatives, variants or mutants thereof are further capable of reducing ACE2 expression and / or reducing TMPRSS2 expression and / or inhibiting the binding of ACE2 and the S protein, and / or the derivatives, variants or mutants thereof express lipase and / or comprise a sequence encoding lipase. The present invention also relates to a Corynebacterium culture comprising Corynebacterium incertum SU004 or consisting essentially of Corynebacterium incertum SU004. Preferably, the Corynebacterium incertum strain SU004, or a culture comprising the Corynebacterium incertum strain SU004 or consisting essentially of the Corynebacterium incertum strain SU004 is used for preventing the infection of an enveloped respiratory virus, preferably a coronavirus, more preferably the SARS-CoV-2 virus, in a subject. In an embodiment, the SARS-CoV-2 virus is the SARS-CoV-2 Omicron variant.

[0064] On the other hand, the present invention relates to the Corynebacterium tuberculostearicum strain SU005 with the accession number NCAIM P(B)001501 deposited at NCAIM on April 27, 2022, or its derivatives, variants or mutants, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression, and optionally, the derivatives, variants or mutants thereof are further capable of reducing ACE2 expression and / or reducing TMPRSS2 expression and / or inhibiting the binding of ACE2 and the S protein. The present invention also relates to a Corynebacterium culture comprising Corynebacterium tuberculostearicum SU005 or consisting essentially of Corynebacterium tuberculostearicum SU005. Preferably, the Corynebacterium tuberculostearicum strain SU005, or a culture comprising the Corynebacterium tuberculostearicum strain SU005 or consisting essentially of the Corynebacterium tuberculostearicum strain SU005 is used for preventing the infection of an enveloped respiratory virus, preferably a coronavirus, more preferably the SARS-CoV-2 virus, in a subject. In an embodiment, the SARS-CoV-2 virus is the SARS-CoV-2 Omicron variant.

[0065] On the other hand, the present invention relates to the Corynebacterium incertum strain SU006 with the accession number NCAIM P(B)001502 deposited at NCAIM on April 27, 2022, or its derivatives, variants or mutants, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression, and optionally, the derivatives, variants or mutants thereof are further capable of reducing ACE2 expression and / or reducing TMPRSS2 expression and / or inhibiting the binding of ACE2 and the S protein, and / or the derivatives, variants or mutants thereof express lipase and / or comprise a sequence encoding lipase. The present invention also relates to a Corynebacterium culture comprising Corynebacterium incertum SU006 or consisting essentially of Corynebacterium incertum SU006. Preferably, the Corynebacterium incertum strain SU006, or a culture comprising the Corynebacterium incertum strain SU006 or consisting essentially of the Corynebacterium incertum strain SU006, is used for preventing the infection of an enveloped respiratory virus, preferably a coronavirus, more preferably the SARS-CoV-2 virus, in a subject. In an embodiment, the SARS-CoV-2 virus is the SARS-CoV-2 Omicron variant.

[0066] On the other hand, the present invention relates to the Corynebacterium propionicum strain SU007 with the accession number NCAIM P(B)001504 deposited at NCAIM on April 27, 2022, or its derivatives, variants or mutants, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression, and optionally, the derivatives, variants or mutants thereof are further capable of reducing ACE2 expression and / or reducing TMPRSS2 expression and / or inhibiting the binding of ACE2 and the S protein. The present invention also relates to a Corynebacterium culture comprising Corynebacterium propionicum SU007 or consisting essentially of Corynebacterium propionicum SU007. Preferably, the Corynebacterium propionicum strain SU007, or a culture comprising the Corynebacterium propionicum strain SU007 or consisting essentially of the Corynebacterium propionicum strain SU007, is used for preventing the infection of an enveloped respiratory virus, preferably a coronavirus, more preferably the SARS-CoV-2 virus, in a subject. In an embodiment, the SARS-CoV-2 virus is the SARS-CoV-2 Omicron variant.

[0067] On the other hand, the present invention relates to the Corynebacterium accolens strain SU008 with the accession number NCAIM P(B)001505 deposited at NCAIM on April 27, 2022, or its derivatives, variants or mutants, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression, and optionally, the derivatives, variants or mutants thereof are further capable of reducing ACE2 expression and / or reducing TMPRSS2 expression, and / or inhibiting the binding of ACE2 and the S protein, and / or expressing lipase and / or comprising a sequence encoding lipase. The present invention also relates to a Corynebacterium culture comprising Corynebacterium accolens SU008 or consisting essentially of Corynebacterium accolens SU004. Preferably, the Corynebacterium accolens strain SU008, or a culture comprising the Corynebacterium accolens strain SU008 or consisting essentially of the Corynebacterium accolens strain SU008 is used for preventing a subject from being infected with an enveloped respiratory virus, preferably a coronavirus, more preferably the SARS-CoV-2 virus. In an embodiment, the SARS-CoV-2 virus is the SARS-CoV-2 Omicron variant.

[0068] On the other hand, the present invention relates to the Corynebacterium accolens strain SU009 with the accession number NCAIM P(B)001506 deposited at NCAIM on May 25, 2022, or its derivatives, variants or mutants, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression, and optionally, the derivatives, variants or mutants thereof are further capable of reducing ACE2 expression and / or reducing TMPRSS2 expression and / or inhibiting the binding of ACE2 and the S protein, and / or the derivatives, variants or mutants thereof express lipase and / or comprise a sequence encoding lipase. The present invention also relates to a Corynebacterium culture comprising Corynebacterium accolens SU009 or consisting essentially of Corynebacterium accolens SU009. Preferably, the Corynebacterium accolens strain SU009, or a culture comprising the Corynebacterium accolens strain SU009 or consisting essentially of the Corynebacterium accolens strain SU009 is used for preventing an infection with an enveloped respiratory virus, preferably a coronavirus, more preferably the SARS-CoV-2 virus, in a subject. In an embodiment, the SARS-CoV-2 virus is the SARS-CoV-2 Omicron variant.

[0069] On the other hand, the present invention relates to the Corynebacterium propionicum strain SU010 with the accession number NCAIM P(B)001507 deposited at NCAIM on May 25, 2022, or its derivatives, variants or mutants, wherein the derivatives, variants or mutants are capable of reducing cathepsin expression, and optionally, the derivatives, variants or mutants are further capable of reducing ACE2 expression and / or reducing TMPRSS2 expression and / or inhibiting the binding of ACE2 and the S protein. The present invention also relates to a Corynebacterium culture comprising Corynebacterium propionicum SU010 or consisting essentially of Corynebacterium propionicum SU0010. Preferably, the Corynebacterium propionicum SU010 strain, or the culture comprising the Corynebacterium propionicum SU010 strain or consisting essentially of the Corynebacterium propionicum SU010 strain is used for preventing the infection of an enveloped respiratory virus, preferably a coronavirus, more preferably the SARS-CoV-2 virus, in a subject. In an embodiment, the SARS-CoV-2 virus is the SARS-CoV-2 Omicron variant.

[0070] On the other hand, the present invention relates to a Corynebacterium strain selected from the group consisting of:

[0071] The Corynebacterium incertum strain SU001 with the accession number NCAIM P(B)001495 deposited at NCAIM (National Collection of Agricultural and Industrial Microorganisms, Institute of Food Science and Technology, Szent István University, Faculty of Agricultural and Life Sciences, H-1118 Budapest, Somlóu str. 14-16, Hungary) on June 22, 2021, or its derivatives, variants or mutants, wherein the derivatives, variants or mutants are capable of reducing cathepsin expression, and optionally, the derivatives, variants or mutants are further capable of reducing ACE2 expression and / or TMPRSS2 expression and / or inhibiting the binding of ACE2 and the S protein, and / or the derivatives, variants or mutants express LipS1 lipase and / or comprise a sequence encoding LipS1 lipase;

[0072] The Corynebacterium propionicum strain SU002 with the accession number NCAIM P(B)001496 deposited at NCAIM on June 22, 2021, or its derivatives, variants or mutants, wherein the derivatives, variants or mutants are capable of reducing the expression of cathepsin, and optionally, the derivatives, variants or mutants are further capable of reducing the expression of ACE2 and / or the expression of TMPRSS2 and / or inhibiting the binding of ACE2 and the S protein;

[0073] Corynebacterium propionicum strain SU003 or its derivatives, variants or mutants with accession number NCAIM P(B)001497 deposited at NCAIM on June 22, 2021, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression, and optionally, the derivatives, variants or mutants thereof are also capable of reducing ACE2 expression and / or TMPRSS2 expression and / or inhibiting the binding of ACE2 and the S protein;

[0074] Corynebacterium accolens strain SU004 or its derivatives, variants or mutants with accession number NCAIM P(B)001500 deposited at NCAIM (National Collection of Agricultural and Industrial Microorganisms, Institute of Food Science and Technology, Szent István University, H-1118 Budapest, Somlóu str. 14-16, Hungary) on January 17, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression, and optionally, the derivatives, variants or mutants thereof are also capable of reducing ACE2 expression and / or TMPRSS2 expression and / or inhibiting the binding of ACE2 and the S protein, and / or the derivatives, variants or mutants thereof express lipase and / or comprise a sequence encoding lipase;

[0075] Corynebacterium tuberculostearicum strain SU005 or its derivatives, variants or mutants with accession number NCAIM P(B)001501 deposited at NCAIM on April 27, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing the expression of cathepsin, and optionally, the derivatives, variants or mutants thereof are also capable of reducing the expression of ACE2 and / or reducing the expression of TMPRSS2 and / or inhibiting the binding of ACE2 and the S protein;

[0076] Corynebacterium accolens strain SU006 or its derivatives, variants or mutants with accession number NCAIM P(B)001502 deposited at NCAIM on April 27, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression, and optionally, the derivatives, variants or mutants thereof are also capable of reducing ACE2 expression and / or reducing TMPRSS2 expression and / or inhibiting the binding of ACE2 and the S protein, and / or the derivatives, variants or mutants thereof express lipase and / or comprise a sequence encoding lipase;

[0077] Corynebacterium propionicum strain SU007 or its derivatives, variants or mutants with accession number NCAIM P(B)001504 deposited at NCAIM on April 27, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression, and optionally, the derivatives, variants or mutants thereof are also capable of reducing ACE2 expression and / or reducing TMPRSS2 expression and / or inhibiting the binding of ACE2 and the S protein;

[0078] Corynebacterium accolens strain SU008 or its derivatives, variants or mutants with the accession number NCAIM P(B)001505 deposited in NCAIM on April 27, 2022, wherein the derivatives, variants or mutants thereof can reduce cathepsin expression, and optionally, the derivatives, variants or mutants thereof can also reduce ACE2 expression and / or reduce TMPRSS2 expression and / or inhibit the binding of ACE2 and S protein, and / or the derivatives, variants or mutants thereof express lipase and / or include a sequence encoding lipase;

[0079] Corynebacterium accolens strain SU009 or its derivatives, variants or mutants with the accession number NNCAIM P(B)001506 deposited in NCAIM on May 25, 2022, wherein the derivatives, variants or mutants thereof can reduce cathepsin expression, and optionally, the derivatives, variants or mutants thereof can also reduce ACE2 expression and / or reduce TMPRSS2 expression and / or inhibit the binding of ACE2 and S protein, and / or the derivatives, variants or mutants thereof express lipase and / or include a sequence encoding lipase; and

[0080] Corynebacterium propionicum strain SU010 or its derivatives, variants or mutants with the accession number NCAIM P(B)001507 deposited in NCAIM on May 25, 2022, wherein the derivatives, variants or mutants thereof can reduce the expression of cathepsin, and optionally, the derivatives, variants or mutants thereof can also reduce the expression of ACE2 and / or reduce the expression of TMPRSS2 and / or inhibit the binding of ACE2 and S protein.

[0081] Preferably, the Corynebacterium culture consists essentially of Corynebacterium, preferably consisting essentially of Corynebacterium accolens and / or Corynebacterium propionicum and / or Corynebacterium tuberculostearicum.

[0082] Specifically, the Corynebacterium culture includes viable Corynebacterium strains in the culture medium.

[0083] In a preferred embodiment of the present invention, the Corynebacterium culture includes viable Corynebacterium strains selected from the group consisting of: Corynebacterium accolens, Corynebacterium propionicum and Corynebacterium tuberculostearicum.

[0084] In any one of the aspects of the present invention, preferably, the Corynebacterium culture includes at least two Corynebacterium strains selected from the following: Corynebacterium strains SU001 to SU010, or any of their derivatives, variants or mutants capable of reducing the above cathepsin expression. A Corynebacterium culture including at least two different Corynebacterium strains is also referred to as a combination.

[0085] Preferably, the combination comprises at least two viable Corynebacterium strains selected from the group consisting of Corynebacterium accolens, Corynebacterium propionicum, and Corynebacterium tuberculostearicum. Preferably, the combination comprises at least two strains, wherein the strains belong to the same species selected from the group consisting of Corynebacterium accolens, Corynebacterium propionicum, and Corynebacterium tuberculostearicum. Preferably, the combination comprises at least two strains, wherein at least two strains belong to different species selected from the group consisting of Corynebacterium accolens, Corynebacterium propionicum, and Corynebacterium tuberculostearicum. Preferably, the combination comprises at least two strains selected from the group consisting of Corynebacterium accolens SU001, Corynebacterium propionicum SU002, Corynebacterium propionicum SU003, Corynebacterium accolens SU004, Corynebacterium tuberculostearicum SU005, Corynebacterium accolens SU006, Corynebacterium propionicum SU007, Corynebacterium accolens SU008, Corynebacterium accolens SU009, Corynebacterium propionicum SU010, and any derivatives, variants or mutants thereof capable of reducing cathepsin expression. Preferably, the combination comprises at least two strains selected from the group consisting of Corynebacterium accolens SU004, Corynebacterium tuberculostearicum SU005, Corynebacterium accolens SU006, Corynebacterium propionicum SU007, Corynebacterium accolens SU008, Corynebacterium accolens SU009, Corynebacterium propionicum SU010, and any derivatives, variants or mutants thereof capable of reducing cathepsin expression.

[0086] Preferably, the combination comprises at least three viable Corynebacterium strains selected from the group consisting of Corynebacterium accolens, Corynebacterium propionicum, and Corynebacterium tuberculostearicum. Preferably, the combination comprises at least three strains, wherein at least two strains belong to different species selected from the group consisting of Corynebacterium accolens, Corynebacterium propionicum, and Corynebacterium tuberculostearicum. Preferably, the combination comprises at least three strains selected from the group consisting of Corynebacterium accolens SU001, Corynebacterium propionicum SU002, Corynebacterium propionicum SU003, Corynebacterium accolens SU004, Corynebacterium tuberculostearicum SU005, Corynebacterium accolens SU006, Corynebacterium propionicum SU007, Corynebacterium accolens SU008, Corynebacterium accolens SU009, Corynebacterium propionicum SU010, and any derivatives, variants or mutants thereof capable of reducing cathepsin expression. Preferably, the combination comprises at least three strains selected from the group consisting of Corynebacterium accolens SU004, Corynebacterium tuberculostearicum SU005, Corynebacterium accolens SU006, Corynebacterium propionicum SU007, Corynebacterium accolens SU008, Corynebacterium accolens SU009, Corynebacterium propionicum SU010, and any derivatives, variants or mutants thereof capable of reducing cathepsin expression.

[0087] Preferably, the combination comprises four viable Corynebacterium strains selected from the group consisting of Corynebacterium accolens, Corynebacterium propionicum, and Corynebacterium tuberculostearicum. Preferably, the combination comprises four strains, wherein at least two strains belong to different species selected from the group consisting of Corynebacterium accolens, Corynebacterium propionicum, and Corynebacterium tuberculostearicum. Preferably, the combination comprises four strains, wherein at least one strain is Corynebacterium accolens, at least one strain is Corynebacterium propionicum, and at least one strain is Corynebacterium tuberculostearicum. Preferably, the combination comprises four strains selected from the group consisting of Corynebacterium accolens SU001, Corynebacterium propionicum SU002, Corynebacterium propionicum SU003, Corynebacterium accolens SU004, Corynebacterium tuberculostearicum SU005, Corynebacterium accolens SU006, Corynebacterium propionicum SU007, Corynebacterium accolens SU008, Corynebacterium accolens SU009, Corynebacterium propionicum SU010, and any derivatives, variants or mutants thereof capable of reducing cathepsin expression. Preferably, the combination comprises four strains selected from the group consisting of Corynebacterium accolens SU004, Corynebacterium tuberculostearicum SU005, Corynebacterium accolens SU006, Corynebacterium propionicum SU007, Corynebacterium accolens SU008, Corynebacterium accolens SU009, Corynebacterium propionicum SU010, and any derivatives, variants or mutants thereof capable of reducing cathepsin expression.

[0088] In a preferred embodiment, the combination comprises at least two strains, wherein the at least two strains are Corynebacterium accolens SU004 and Corynebacterium tuberculostearicum SU005, or any derivatives, variants or mutants thereof that are capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises at least two strains, wherein the at least two strains are Corynebacterium accolens SU004 and Corynebacterium accolens SU009, or any derivatives, variants or mutants thereof that are capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises at least two strains, wherein the at least two strains are Corynebacterium accolens SU004 and Corynebacterium propionicum SU010, or any derivatives, variants or mutants thereof that are capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises at least two strains, wherein the at least two strains are Corynebacterium tuberculostearicum SU005 and Corynebacterium accolens SU009, or any derivatives, variants or mutants thereof that are capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises at least two strains, wherein the at least two strains are Corynebacterium tuberculostearicum SU005 and Corynebacterium propionicum SU010, or any derivatives, variants or mutants thereof that are capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises at least two strains, wherein the at least two strains are Corynebacterium accolens SU009 and Corynebacterium propionicum SU010, or any derivatives, variants or mutants thereof that are capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises at least two strains, wherein the at least two strains are Corynebacterium accolens SU004 and Corynebacterium accolens SU006, or any derivatives, variants or mutants thereof that are capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises at least two strains, wherein the at least two strains are Corynebacterium tuberculostearicum SU005 and Corynebacterium accolens SU006, or any derivatives, variants or mutants thereof that are capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises at least two strains, wherein the at least two strains are Corynebacterium accolens SU006 and Corynebacterium propionicum SU010, or any derivatives, variants or mutants thereof that are capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises at least two strains, wherein the at least two strains are Corynebacterium accolens SU004 and Corynebacterium propionicum SU007, or any derivatives, variants or mutants thereof that are capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises at least two strains, wherein the at least two strains are Corynebacterium accolens SU004 and Corynebacterium accolens SU008, or any derivatives, variants or mutants thereof that are capable of reducing cathepsin expression.In another preferred embodiment, the combination comprises at least two strains, wherein the at least two strains are Corynebacterium tuberculostearicum SU005 and Corynebacterium propionicum SU007, or any derivatives, variants or mutants thereof capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises at least two strains, wherein the at least two strains are Corynebacterium tuberculostearicum SU005 and Corynebacterium accolens SU008, or any derivatives, variants or mutants thereof capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises at least two strains, wherein the at least two strains are Corynebacterium propionicum SU007 and Corynebacterium accolens SU008, or any derivatives, variants or mutants thereof capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises at least two strains, wherein the at least two strains are Corynebacterium accolens SU006 and Corynebacterium propionicum SU007, or any derivatives, variants or mutants thereof capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises at least two strains, wherein the at least two strains are Corynebacterium accolens SU006 and Corynebacterium accolens SU008, or any derivatives, variants or mutants thereof capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises at least two strains, wherein the at least two strains are Corynebacterium accolens SU006 and Corynebacterium accolens SU009, or any derivatives, variants or mutants thereof capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises at least two strains, wherein the at least two strains are Corynebacterium propionicum SU007 and Corynebacterium accolens SU009, or any derivatives, variants or mutants thereof capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises at least two strains, wherein the at least two strains are Corynebacterium propionicum SU007 and Corynebacterium propionicum SU010, or any derivatives, variants or mutants thereof capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises at least two strains, wherein the at least two strains are Corynebacterium accolens SU008 and Corynebacterium accolens SU009, or any derivatives, variants or mutants thereof capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises at least two strains, wherein the at least two strains are Corynebacterium accolens SU008 and Corynebacterium propionicum SU010, or any derivatives, variants or mutants thereof capable of reducing cathepsin expression.

[0089] In a preferred embodiment, the combination comprises at least three strains, wherein the at least three strains are Corynebacterium accolens SU004, Corynebacterium tuberculostearicum SU005, and Corynebacterium accolens SU009, or any derivatives, variants, or mutants thereof that can reduce cathepsin expression. In another preferred embodiment, the combination comprises at least three strains, wherein the at least three strains are Corynebacterium accolens SU004, Corynebacterium tuberculostearicum SU005, and Corynebacterium propionicum SU010, or any derivatives, variants, or mutants thereof that can reduce cathepsin expression. In another preferred embodiment, the combination comprises at least three strains, wherein the at least three strains are Corynebacterium accolens SU004, Corynebacterium accolens SU009, and Corynebacterium propionicum SU010, or any derivatives, variants, or mutants thereof that can reduce cathepsin expression. In another preferred embodiment, the combination comprises at least three strains, wherein the at least three strains are Corynebacterium tuberculostearicum SU005, Corynebacterium accolens SU009, and Corynebacterium propionicum SU010, or any derivatives, variants, or mutants thereof that can reduce cathepsin expression. In another preferred embodiment, the combination comprises at least three strains, wherein the at least three strains are Corynebacterium accolens SU004, Corynebacterium tuberculostearicum SU005, and Corynebacterium accolens SU006, or any derivatives, variants, or mutants thereof that can reduce cathepsin expression. In another preferred embodiment, the combination comprises at least three strains, wherein the at least three strains are Corynebacterium accolens SU004, Corynebacterium accolens SU006, and Corynebacterium propionicum SU010, or any derivatives, variants, or mutants thereof that can reduce cathepsin expression. In another preferred embodiment, the combination comprises at least three strains, wherein the at least three strains are Corynebacterium tuberculostearicum SU005, Corynebacterium accolens SU006, and Corynebacterium propionicum SU010, or any derivatives, variants, or mutants thereof that can reduce cathepsin expression. In another preferred embodiment, the combination comprises at least three strains, wherein the at least three strains are Corynebacterium accolens SU004, Corynebacterium tuberculostearicum SU005, and Corynebacterium propionicum SU007, or any derivatives, variants, or mutants thereof that can reduce cathepsin expression. In another preferred embodiment, the combination comprises at least three strains, wherein the at least three strains are Corynebacterium accolens SU004, Corynebacterium tuberculostearicum SU005, and Corynebacterium accolens SU008, or any derivatives, variants, or mutants thereof that can reduce cathepsin expression. In another preferred embodiment, the combination comprises at least three strains, wherein the at least three strains are Corynebacterium accolens SU004, Corynebacterium propionicum SU007, and Corynebacterium accolens SU008, or any derivatives, variants, or mutants thereof that can reduce cathepsin expression.In another preferred embodiment, the combination comprises at least three strains, wherein the at least three strains are Corynebacterium tuberculostearicum SU005, Corynebacterium propionicum SU007, and Corynebacterium accolens SU008, or any derivative, variant, or mutant thereof that can reduce cathepsin expression. In another preferred embodiment, the combination comprises at least three strains, wherein the at least three strains are Corynebacterium accolens SU004, Corynebacterium accolens SU006, and Corynebacterium propionicum SU007, or any derivative, variant, or mutant thereof that can reduce cathepsin expression. In another preferred embodiment, the combination comprises at least three strains, wherein the at least three strains are Corynebacterium accolens SU004, Corynebacterium accolens SU006, and Corynebacterium accolens SU008, or any derivative, variant, or mutant thereof that can reduce cathepsin expression. In another preferred embodiment, the combination comprises at least three strains, wherein the at least three strains are Corynebacterium accolens SU004, Corynebacterium accolens SU006, and Corynebacterium accolens SU009, or any derivative, variant, or mutant thereof that can reduce cathepsin expression. In another preferred embodiment, the combination comprises at least three strains, wherein the at least three strains are Corynebacterium accolens SU004, Corynebacterium propionicum SU007, and Corynebacterium accolens SU009, or any derivative, variant, or mutant thereof that can reduce cathepsin expression. In another preferred embodiment, the combination comprises at least three strains, wherein the at least three strains are Corynebacterium accolens SU004, Corynebacterium propionicum SU007, and Corynebacterium propionicum SU010, or any derivative, variant, or mutant thereof that can reduce cathepsin expression. In another preferred embodiment, the combination comprises at least three strains, wherein the at least three strains are Corynebacterium accolens SU004, Corynebacterium accolens SU008, and Corynebacterium accolens SU009, or any derivative, variant, or mutant thereof that can reduce cathepsin expression. In another preferred embodiment, the combination comprises at least three strains, wherein the at least three strains are Corynebacterium accolens SU004, Corynebacterium accolens SU008, and Corynebacterium propionicum SU010, or any derivative, variant, or mutant thereof that can reduce cathepsin expression. In another preferred embodiment, the combination comprises at least three strains, wherein the at least three strains are Corynebacterium tuberculostearicum SU005, Corynebacterium accolens SU006, and Corynebacterium propionicum SU007, or any derivative, variant, or mutant thereof that can reduce cathepsin expression. In another preferred embodiment, the combination comprises at least three strains, wherein the at least three strains are Corynebacterium tuberculostearicum SU005, Corynebacterium accolens SU006, and Corynebacterium accolens SU008, or any derivative, variant, or mutant thereof that can reduce cathepsin expression.In another preferred embodiment, the combination comprises at least three strains, wherein the at least three strains are Corynebacterium tuberculostearicum SU005, Corynebacterium accolens SU006 and Corynebacterium accolens SU009, or any derivative, variant or mutant thereof capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises at least three strains, wherein the at least three strains are Corynebacterium tuberculostearicum SU005, Corynebacterium propionicum SU007 and Corynebacterium accolens SU009, or any derivative, variant or mutant thereof capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises at least three strains, wherein the at least three strains are Corynebacterium tuberculostearicum SU005, Corynebacterium propionicum SU007 and Corynebacterium propionicum SU010, or any derivative, variant or mutant thereof capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises at least three strains, wherein the at least three strains are Corynebacterium tuberculostearicum SU005, Corynebacterium accolens SU008 and Corynebacterium accolens SU009, or any derivative, variant or mutant thereof capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises at least three strains, wherein the at least three strains are Corynebacterium tuberculostearicum SU005, Corynebacterium accolens SU008 and Corynebacterium propionicum SU010, or any derivative, variant or mutant thereof capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises at least three strains, wherein the at least three strains are Corynebacterium accolens SU006, Corynebacterium propionicum SU007 and Corynebacterium accolens SU008, or any derivative, variant or mutant thereof capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises at least three strains, wherein the at least three strains are Corynebacterium accolens SU006, Corynebacterium propionicum SU007 and Corynebacterium accolens SU009, or any derivative, variant or mutant thereof capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises at least three strains, wherein the at least three strains are Corynebacterium accolens SU006, Corynebacterium propionicum SU007 and Corynebacterium propionicum SU010, or any derivative, variant or mutant thereof capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises at least three strains, wherein the at least three strains are Corynebacterium accolens SU006, Corynebacterium accolens SU008 and Corynebacterium accolens SU009, or any derivative, variant or mutant thereof capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises at least three strains, wherein the at least three strains are Corynebacterium accolens SU006, Corynebacterium accolens SU008 and Corynebacterium propionicum SU010, or any derivative, variant or mutant thereof capable of reducing cathepsin expression.In another preferred embodiment, the combination comprises at least three strains, wherein the at least three strains are Corynebacterium accolens SU006, Corynebacterium accolens SU009 and Corynebacterium propionicum SU010, or any derivative, variant or mutant thereof capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises at least three strains, wherein the at least three strains are Corynebacterium propionicum SU007, Corynebacterium accolens SU008 and Corynebacterium accolens SU009, or any derivative, variant or mutant thereof capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises at least three strains, wherein the at least three strains are Corynebacterium propionicum SU007, Corynebacterium accolens SU008 and Corynebacterium propionicum SU010, or any derivative, variant or mutant thereof capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises at least three strains, wherein the at least three strains are Corynebacterium propionicum SU007, Corynebacterium accolens SU009 and Corynebacterium propionicum SU010, or any derivative, variant or mutant thereof capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises at least three strains, wherein the at least three strains are Corynebacterium accolens SU008, Corynebacterium accolens SU009 and Corynebacterium propionicum SU010, or any derivative, variant or mutant thereof capable of reducing cathepsin expression.

[0090] In a preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium accolens SU004, Corynebacterium tuberculostearicum SU005, Corynebacterium accolens SU006, and Corynebacterium propionicum SU007, or any derivatives, variants, or mutants thereof that are capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium accolens SU004, Corynebacterium tuberculostearicum SU005, Corynebacterium accolens SU006, and Corynebacterium accolens SU008, or any derivatives, variants, or mutants thereof that are capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium accolens SU004, Corynebacterium tuberculostearicum SU005, Corynebacterium accolens SU006, and Corynebacterium accolens SU009, or any derivatives, variants, or mutants thereof that are capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium accolens SU004, Corynebacterium tuberculostearicum SU005, Corynebacterium U006, and Corynebacterium propionicum SU010, or any derivatives, variants, or mutants thereof that are capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium accolens SU004, Corynebacterium tuberculostearicum SU005, Corynebacterium propionicum SU007, and Corynebacterium accolens SU008, or any derivatives, variants, or mutants thereof that are capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium accolens SU004, Corynebacterium tuberculostearicum SU005, Corynebacterium propionicum SU007, and Corynebacterium accolens SU009, or any derivatives, variants, or mutants thereof that are capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium accolens SU004, Corynebacterium tuberculostearicum SU005, Corynebacterium propionicum SU007, and Corynebacterium propionicum SU010, or any derivatives, variants, or mutants thereof that are capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium accolens SU004, Corynebacterium tuberculostearicum SU005, Corynebacterium accolens SU008, and Corynebacterium accolens SU009, or any derivatives, variants, or mutants thereof that are capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium accolens SU004, Corynebacterium tuberculostearicum SU005, Corynebacterium accolens SU008, and Corynebacterium propionicum SU010, or any derivatives, variants, or mutants thereof that are capable of reducing cathepsin expression.In another preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium accolens SU004, Corynebacterium tuberculostearicum SU005, Corynebacterium accolens SU009 and Corynebacterium propionicum SU010, or any derivatives, variants or mutants thereof that are capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium accolens SU004, Corynebacterium accolens SU006, Corynebacterium propionicum SU007 and Corynebacterium accolens SU008, or any derivatives, variants or mutants thereof that are capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium accolens SU004, Corynebacterium accolens SU006, Corynebacterium propionicum SU007 and Corynebacterium accolens SU009, or any derivatives, variants or mutants thereof that are capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium accolens SU004, Corynebacterium accolens SU006, Corynebacterium propionicum SU007 and Corynebacterium propionicum SU010, or any derivatives, variants or mutants thereof that are capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium accolens SU004, Corynebacterium accolens SU006, Corynebacterium accolens SU008 and Corynebacterium accolens SU009, or any derivatives, variants or mutants thereof that are capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium accolens SU004, Corynebacterium accolens SU006, Corynebacterium accolens SU008 and Corynebacterium propionicum SU010, or any derivatives, variants or mutants thereof that are capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium accolens SU004, Corynebacterium accolens SU006, Corynebacterium accolens SU009 and Corynebacterium propionicum SU010, or any derivatives, variants or mutants thereof that are capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium accolens SU004, Corynebacterium propionicum SU007, Corynebacterium accolens SU008 and Corynebacterium accolens SU009, or any derivatives, variants or mutants thereof that are capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium accolens SU004, Corynebacterium propionicum SU007, Corynebacterium accolens SU008 and Corynebacterium propionicum SU010, or any derivatives, variants or mutants thereof that are capable of reducing cathepsin expression.In another preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium accolens SU004, Corynebacterium propionicum SU007, Corynebacterium accolens SU009, and Corynebacterium propionicum SU010, or any derivative, variant or mutant thereof that is capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium accolens SU004, Corynebacterium accolens SU008, Corynebacterium accolens SU009, and Corynebacterium propionicum SU010, or any derivative, variant or mutant thereof that is capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium tuberculostearicum SU005, Corynebacterium accolens SU006, Corynebacterium propionicum SU007, and Corynebacterium accolens SU008, or any derivative, variant or mutant thereof that is capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium tuberculostearicum SU005, Corynebacterium accolens SU006, Corynebacterium propionicum SU007, and Corynebacterium accolens SU009, or any derivative, variant or mutant thereof that is capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium tuberculostearicum SU005, Corynebacterium accolens SU006, Corynebacterium propionicum SU007, and Corynebacterium propionicum SU010, or any derivative, variant or mutant thereof that is capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium tuberculostearicum SU005, Corynebacterium accolens SU006, Corynebacterium accolens SU008, and Corynebacterium accolens SU009, or any derivative, variant or mutant thereof that is capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium tuberculostearicum SU005, Corynebacterium accolens SU006, Corynebacterium accolens SU008, and Corynebacterium propionicum SU010, or any derivative, variant or mutant thereof that is capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium tuberculostearicum SU005, Corynebacterium accolens SU006, Corynebacterium accolens SU009, and Corynebacterium propionicum SU010, or any derivative, variant or mutant thereof that is capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium tuberculostearicum SU005, Corynebacterium propionicum SU007, Corynebacterium accolens SU008, and Corynebacterium accolens SU009, or any derivative, variant or mutant thereof that is capable of reducing cathepsin expression.In another preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium tuberculostearicum SU005, Corynebacterium propionicum SU007, Corynebacterium accolens SU008 and Corynebacterium propionicum SU010, or any derivatives, variants or mutants thereof capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium tuberculostearicum SU005, Corynebacterium propionicum SU007, Corynebacterium accolens SU009 and Corynebacterium propionicum SU010, or any derivatives, variants or mutants thereof capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium tuberculostearicum SU005, Corynebacterium accolens SU008, Corynebacterium accolens SU009 and Corynebacterium propionicum SU010, or any derivatives, variants or mutants thereof capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium accolens SU006, Corynebacterium propionicum SU007, Corynebacterium accolens SU008 and Corynebacterium accolens SU009, or any derivatives, variants or mutants thereof capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium accolens SU006, Corynebacterium propionicum SU007, Corynebacterium accolens SU008 and Corynebacterium propionicum SU010, or any derivatives, variants or mutants thereof capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium accolens SU006, Corynebacterium propionicum SU007, Corynebacterium accolens SU009 and Corynebacterium propionicum SU010, or any derivatives, variants or mutants thereof capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium accolens SU006, Corynebacterium accolens SU008, Corynebacterium accolens SU009 and Corynebacterium propionicum SU010, or any derivatives, variants or mutants thereof capable of reducing cathepsin expression. In another preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium propionicum SU007, Corynebacterium accolens SU008, Corynebacterium accolens SU009 and Corynebacterium propionicum SU010, or any derivatives, variants or mutants thereof capable of reducing cathepsin expression.

[0091] In a more preferred embodiment, the Corynebacterium culture comprises four Corynebacterium strains selected from:

[0092] (i) Corynebacterium accolens SU004, Corynebacterium tuberculostearicum SU005, Corynebacterium accolens SU009, and Corynebacterium propionicum SU010, or any derivative, variant, or mutant thereof capable of reducing cathepsin expression;

[0093] (ii) Corynebacterium accolens SU004, Corynebacterium tuberculostearicum SU005, Corynebacterium accolens SU006, and Corynebacterium propionicum SU010, or any derivative, variant, or mutant thereof capable of reducing cathepsin expression; and

[0094] (iii) Corynebacterium accolens SU004, Corynebacterium tuberculostearicum SU005, Corynebacterium propionicum SU007, and Corynebacterium accolens SU008, or any derivative, variant, or mutant thereof capable of reducing cathepsin expression.

[0095] In a preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium accolens SU004, Corynebacterium tuberculostearicum SU005, Corynebacterium accolens SU009, and Corynebacterium propionicum SU010, or any derivative, variant, or mutant thereof capable of reducing cathepsin expression. (Such a combination is referred to as combination K1 in the examples.) In another preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium accolens SU004, Corynebacterium tuberculostearicum SU005, Corynebacterium accolens SU006, and Corynebacterium propionicum SU010, or any derivative, variant, or mutant thereof capable of reducing cathepsin expression. (Such a combination is referred to as combination K2 in the examples.) In another preferred embodiment, the combination comprises four strains, wherein the four strains are Corynebacterium accolens SU004, Corynebacterium tuberculostearicum SU005, Corynebacterium propionicum SU007, and Corynebacterium accolens SU008, or any derivative, variant, or mutant thereof capable of reducing cathepsin expression. (Such a combination is referred to as combination K3 in the examples.)

[0096] Preferably, the Corynebacterium culture comprising at least two Corynebacterium strains consists essentially of Corynebacterium, preferably consisting essentially of Corynebacterium accolens and / or Corynebacterium propionicum and / or Corynebacterium tuberculostearicum.

[0097] Specifically, the Corynebacterium culture comprising at least two Corynebacterium strains comprises at least two viable Corynebacterium strains in the culture medium. Preferably, the Corynebacterium culture comprising at least two Corynebacterium strains comprises at least three viable Corynebacterium strains in the culture medium. More preferably, the Corynebacterium culture comprising at least two Corynebacterium strains comprises four viable Corynebacterium strains in the culture medium.

[0098] In a preferred embodiment of the present invention, a Corynebacterium culture comprising at least two Corynebacterium strains comprises at least two viable Corynebacterium strains selected from the group consisting of Corynebacterium accolens, Corynebacterium propionicum, and Corynebacterium tuberculostearicum. In a preferred embodiment of the present invention, a Corynebacterium culture comprising at least two Corynebacterium strains comprises at least three viable Corynebacterium strains selected from the group consisting of Corynebacterium accolens, Corynebacterium propionicum, and Corynebacterium tuberculostearicum. In a preferred embodiment of the present invention, a Corynebacterium culture comprising at least two Corynebacterium strains comprises four viable Corynebacterium strains selected from the group consisting of Corynebacterium accolens, Corynebacterium propionicum, and Corynebacterium tuberculostearicum.

[0099] In any of the aspects of the present invention, the Corynebacterium culture does not include Corynebacterium accolens SU001 and / or Corynebacterium propionicum SU002 and / or Corynebacterium propionicum SU003. In an embodiment, the Corynebacterium culture does not include Corynebacterium accolens SU001. In another embodiment, the Corynebacterium culture does not include Corynebacterium propionicum SU002. In another embodiment, the Corynebacterium culture does not include Corynebacterium propionicum SU003.

[0100] In any of the aspects of the present invention, preferably, the Corynebacterium culture does not include Corynebacterium pseudodiphtheriticum.

[0101] Preferably, in any aspect of the present invention, the Corynebacterium strain or Corynebacterium culture capable of reducing cathepsin expression is the Corynebacterium strain or Corynebacterium culture that reduces the relative expression of cathepsin mRNA after incubation with Caco-2 cells for 24 hours. In an embodiment, the bacterial co-culture assay described in the examples is used to determine whether the Corynebacterium strain or Corynebacterium culture reduces the relative expression of cathepsin mRNA.

[0102] The present invention also relates to a pharmaceutical composition comprising a Corynebacterium culture capable of reducing cathepsin expression in a subject, and a pharmaceutically acceptable excipient or a pharmaceutically acceptable carrier.

[0103] Preferably, the preparation or composition is a pharmaceutical composition comprising a pharmaceutically acceptable excipient. Preferably, the preparation or composition is a probiotic preparation or composition. Preferably, the preparation or composition is (part of) a medical device. Preferably, the preparation or composition is delivered by a medical device. Preferably, the preparation or composition is for inhalative administration, more preferably in the form of an inhalable powder, an aerosol mixture, an oral inhalation solution or suspension, a nasal drop, a nasal spray, a nasopharyngeal spray or a nasal mist. Preferably, the preparation or composition is a nasal spray.

[0104] The invention also relates to a preparation comprising a Corynebacterium culture used according to the invention. The preparation of the invention comprises a pharmaceutically acceptable excipient, such as a carrier. Preferably, the preparation comprises one or more components that support the survival or proliferation (if required) growth of Corynebacterium, and such components are optionally regarded as excipients. Any excipient is preferably physiologically compatible with the upper respiratory tract epithelial cells.

[0105] Preferably, the preparation is for the upper respiratory tract of a subject.

[0106] In an embodiment, the preparation is for nasal administration, more preferably in the form of a nasal drop or a nasal spray.

[0107] In an embodiment, the preparation is for inhalative administration, more preferably in the form of an inhalable powder, an aerosol mixture, an oral inhalation solution or suspension, a nasal spray, a nasal mist.

[0108] In an embodiment, the preparation is in the form of a nasal preparation for topical administration, preferably in the form of a viscous liquid, a liquid suspension, a paste, a gel or an ointment.

[0109] Preferably, the preparation is a preparation delivered to the upper respiratory tract. Preferably, the preparation is delivered by a device capable of delivering the preparation to the upper respiratory tract.

[0110] Preferably, the preparation is in the form of a nasal formulation, such as a nasal cream, a nasal gel, a nasal ointment, nasal drops (including solutions, suspensions, emulsions, freeze-dried powders for suspensions, powders for solutions), nasal powders, nasal sprays (including solutions, suspensions, emulsions, freeze-dried powders for suspensions, powders for solutions), nasal douches, nasal patches or nasopharyngeal sprays. Optionally, the preparation is in the form of a skin and nasal ointment, an intranasal solution, an intranasal wash (including suspensions), a powder for intranasal solution, a powder for nasal spray solution, or an oral mucosal solution, a nasal / oral mucosal solution, or a nasal / oral mucosal spray.

[0111] Preferably, the preparation is in the form of an oropharyngeal formulation, such as an oropharyngeal spray, an oropharyngeal spray solution, an oropharyngeal suspension, an oropharyngeal emulsion, an oropharyngeal powder, an oropharyngeal gel or an oropharyngeal lotion.

[0112] Optionally, the preparation is in the form of an ophthalmic-nasal product, such as an effervescent tablet for an ophthalmic-nasal suspension, a lyophilizate for an ophthalmic-nasal suspension (including for use in drinking water) or an ophthalmic-nasal suspension.

[0113] Preferably, the preparation is in the form of an oral mucosal product, such as a buccal membrane, a buccal tablet, a compressed lozenge, a concentrated solution for gargling, a concentrated solution for oral mucosal solution, a skin solution, a skin / oral mucosal solution, a skin / oral mucosal spray, a skin / oral mucosal / oral solution, an effervescent buccal tablet, a gargle (including powder for solution, tablet for solution), a gargle / rinse, a gargle / irrigation solution for nasal cavity, a gingival gel, a gingival paste, a gingival solution, a laryngopharyngeal solution, a laryngopharyngeal spray, a lozenge, a medicated chewing gum, a mouthwash (including powder for solution, tablet for solution), a mucoadhesive buccal sustained-release tablet, a mucoadhesive buccal tablet, a nasal spray and an oral mucosal solution, a nasal / oral mucosal solution, a nasal / oral mucosal spray, an oral mucosal capsule, an oral mucosal cream, an oral mucosal drop, an oral mucosal film, an oral mucosal gel, an oral mucosal ointment, an oral mucosal paste, an oral mucosal patch, an oral mucosal sachet, an oral mucosal solution, an oral mucosal spray (including emulsion, solution, suspension), an oral mucosal suspension, an oral mucosal / laryngopharyngeal solution, an oral mucosal / laryngopharyngeal spray, a lozenge, a pill, a powder for gingival gel, a powder for mouthwash, a sublingual film, a sublingual lyophilizate, a sublingual powder, a sublingual spray (including emulsion, solution, suspension) or a sublingual tablet.

[0114] Preferably, the preparation is in the form of a pulmonary product, such as an aerosol, a concentrate for nebulizer solution, a tracheobronchial instillation (including solution, suspension, powder for solution and powder for suspension), an inhaled gas, an inhaled impregnated pad, an inhaled powder (including hard capsule, pre-dosed and tablet), an inhaled solution, an inhaled vapor (including capsule, effervescent tablet, emulsion, impregnated pad, impregnated plug, liquid, ointment, powder, solution and tablet), a liquefied gas for dental use, a medical gas (including compressed, cryogenic and liquefied), a nebulizer emulsion, a nebulizer solution, a nebulizer suspension, an oral solution / concentrate for nebulizer solution, a powder for nebulizer solution, a powder / injection / infusion solution for nebulizer solution, a powder for nebulizer suspension, a pressurized inhaler (including emulsion, solution and suspension), a metered-dose inhaler.

[0115] The present invention also relates to a lyophilized preparation comprising a Corynebacterium culture capable of reducing cathepsin expression in a subject and a cryoprotectant.

[0116] The present invention also relates to a preparation comprising any one of the Corynebacterium cultures (including combinations) defined above, wherein the preparation is a lyophilized preparation. The lyophilized preparation comprises any one of the above-mentioned Corynebacterium cultures (including combinations) and a cryoprotectant.

[0117] In an embodiment, in the lyophilized preparation, the cryoprotectant comprises a hydrocolloid polymer, and / or a carbohydrate or a carbohydrate derivative. Preferably, in the lyophilized preparation, the cryoprotectant comprises a hydrocolloid polymer and a carbohydrate or a carbohydrate derivative.

[0118] Preferably, in the lyophilized preparation, the carbohydrate or the carbohydrate derivative is selected from trehalose, sucrose, glucose, lactose, mannitol, sorbitol, inulin, maltodextrin and isomaltulose, preferably selected from maltodextrin and isomaltulose. Preferably, in the lyophilized preparation, the carbohydrate is isomaltulose.

[0119] Preferably, in the lyophilized preparation, the carbohydrate or the carbohydrate derivative is maltodextrin at a concentration of 0.1 g / 100 mL to 3 g / 100 mL, preferably at a concentration of 0.5 g / 100 mL to 2.5 g / 100 mL, or isomaltulose at a concentration of 0.1 g / 100 mL to 3 g / 100 mL, preferably at a concentration of 0.5 g / 100 mL to 2.5 g / 100 mL. Preferably, in the lyophilized preparation, the carbohydrate is isomaltulose at a concentration of 0.5 g / 100 mL to 2.5 g / 100 mL, more preferably at a concentration of 0.5 g / 100 mL.

[0120] Preferably, in the lyophilized preparation, the hydrocolloid polymer is selected from hydroxypropyl methylcellulose, hydroxyethyl cellulose, carboxymethyl cellulose, guar gum, carrageenan and xanthan gum, preferably selected from κ-carrageenan and xanthan gum. Optionally, the hydrocolloid polymer is a hydrocolloid polymer that adjusts the pH (e.g., alginic acid / alginate, poly-galactomannuronic acid), in which case a buffer is added to the preparation.

[0121] Preferably, in the freeze-dried preparation, the hydrocolloid polymer is κ-carrageenan with a concentration of 0.05 g / 100 mL to 1 g / 100 mL, preferably 0.1 g / 100 mL to 0.5 g / 100 mL, or xanthan gum with a concentration of 0.01 g / 100 mL to 0.5 g / 100 mL, preferably 0.05 g / 100 mL to 0.25 g / 100 mL. Preferably, in the freeze-dried preparation, the hydrocolloid polymer is xanthan gum with a concentration of 0.05 g / 100 mL to 0.25 g / 100 mL, more preferably 0.05 g / 100 mL. Preferably, in the freeze-dried preparation, the hydrocolloid polymer is κ-carrageenan with a concentration of 0.1 g / 100 mL to 0.5 g / 100 mL, more preferably 0.1 g / 100 mL.

[0122] In a preferred embodiment, the freeze-dried preparation comprises a Corynebacterium culture, which comprises four Corynebacterium strains selected from:

[0123] (i) Corynebacterium accolens SU004, Corynebacterium tuberculostearicum SU005, Corynebacterium accolens SU009, and Corynebacterium propionicum SU010, or any derivative, variant, or mutant thereof capable of reducing cathepsin expression;

[0124] (ii) Corynebacterium accolens SU004, Corynebacterium tuberculostearicum SU005, Corynebacterium accolens SU006, and Corynebacterium propionicum SU010, or any derivative, variant, or mutant thereof capable of reducing cathepsin expression; and

[0125] (iii) Corynebacterium accolens SU004, Corynebacterium tuberculostearicum SU005, Corynebacterium propionicum SU007, and Corynebacterium accolens SU008, or any derivative, variant, or mutant thereof capable of reducing cathepsin expression; and

[0126] comprises a carbohydrate or carbohydrate derivative, and a hydrocolloid polymer;

[0127] Preferably, the carbohydrate or carbohydrate derivative is maltodextrin or isomaltulose, and the hydrocolloid polymer is κ-carrageenan or xanthan gum.

[0128] Preferably, the freeze-dried preparation comprises any one of the Corynebacterium cultures (including combinations) defined above, wherein the Corynebacterium culture comprises 10 3 CFU / mL to 10 11 CFU / mL of Corynebacterium strains. Preferably, the Corynebacterium culture comprises 10 4 CFU / mL to 10 11 CFU / mL, preferably 105 CFU / mL to 10 11 CFU / mL, preferably 10 6 CFU / mL to 10 11 CFU / mL, preferably 10 7 CFU / mL to 10 11 CFU / mL, preferably 10 8 CFU / mL to 10 11 CFU / mL, preferably 10 8 CFU / mL to 10 10 CFU / mL, preferably 10 9 CFU / mL to 10 11 CFU / mL, preferably 10 9 CFU / mL to 10 10 Corynebacterium of CFU / mL, more preferably 10 8 CFU / mL or 10 9 Corynebacterium strains of CFU / mL. Preferably, the Corynebacterium culture comprises at least two, at least three or four Corynebacterium strains, wherein the culture comprises 10 4 CFU / mL to 10 11 CFU / mL of each strain. More preferably, the Corynebacterium culture comprises 10 7 CFU / mL to 10 11 CFU / mL, preferably 10 8 CFU / mL to 10 11 CFU / mL, preferably 10 8 CFU / mL to 10 10 CFU / mL, preferably 10 9 CFU / mL to 10 11 CFU / mL, preferably 10 9 CFU / mL to 10 10 CFU / mL of each Corynebacterium strain, preferably 10 8 CFU / mL or 10 9 CFU / mL of each Corynebacterium strain.

[0129] In another aspect, the present invention relates to a kit comprising any one of the above-mentioned lyophilized preparations and a suitable reconstitution agent (e.g., Ringer's solution). The reconstitution agent is used to reconstitute the lyophilized preparation into a preparation for the upper respiratory tract of a subject. Preferably, the reconstitution agent is used to reconstitute the lyophilized preparation into a nasal spray. The reconstitution agent is preferably Ringer's solution.

[0130] The present invention relates to a Corynebacterium culture capable of reducing the expression of cathepsin in a subject, and the Corynebacterium culture is used in a medicament.

[0131] The present invention relates to a Corynebacterium culture or a lyophilized preparation thereof for preventing or treating an infection of a respiratory virus in a subject, wherein the Corynebacterium culture is capable of reducing the expression of cathepsin in the subject. In another embodiment, the present invention relates to a Corynebacterium culture or a lyophilized preparation thereof for preventing an infection of an enveloped respiratory virus in a subject, wherein the virus enters the cells of the subject using a cathepsin entry pathway, and wherein the Corynebacterium culture is capable of reducing the expression of cathepsin in the subject. Preferably, the present invention relates to a Corynebacterium culture or a lyophilized preparation thereof for preventing or treating an infection of an enveloped respiratory virus in a subject, wherein the Corynebacterium culture is capable of reducing the expression of cathepsin in the subject. The enveloped respiratory virus is a DNA or RNA virus, preferably an RNA virus. Preferably, the present invention relates to a Corynebacterium culture or a lyophilized preparation thereof for preventing or treating an infection of a coronavirus in a subject, wherein the Corynebacterium culture is capable of reducing the expression of cathepsin in the subject, wherein the coronavirus is preferably SARS-CoV-2, more preferably a SARS-CoV-2 variant mainly using the cathepsin B / L entry pathway, and most preferably the SARS-CoV-2 Omicron variant.

[0132] The present invention also relates to a Corynebacterium culture or a lyophilized preparation thereof for preventing or treating an infection of a respiratory virus, preferably an enveloped respiratory virus, more preferably a coronavirus, and most preferably a SARS-CoV-2 virus in a subject, wherein the Corynebacterium culture is capable of reducing the expression of cathepsin in the subject, and optionally can also

[0133] - reduce the expression of ACE2 in the cells of the subject, and / or

[0134] - reduce the expression of TMPRSS2 in the cells of the subject.

[0135] The present invention also relates to a Corynebacterium culture or a lyophilized preparation thereof for preventing or treating an infection of a respiratory virus, preferably an enveloped respiratory virus, more preferably a coronavirus, and most preferably a SARS-CoV-2 virus in a subject, wherein the Corynebacterium culture is capable of reducing the expression of cathepsin in the subject, and optionally can also

[0136] - reduce the expression of ACE2 in the cells of the subject, and / or

[0137] - reduce the expression of TMPRSS2 in the cells of the subject, and / or

[0138] - Inhibit the binding of the ACE2 receptor to the spike (S) protein of the virus.

[0139] In an alternative embodiment, the present invention relates to a Corynebacterium culture (including combinations) or a lyophilized preparation for use in the upper respiratory tract of a homeothermic subject to combat a pathogenic respiratory RNA virus of the subject, the Corynebacterium culture being capable of reducing the expression of cathepsin in the cells of the subject. In another embodiment, the Corynebacterium culture (including combinations) or the lyophilized preparation is for use in the upper respiratory tract of a homeothermic subject to combat an enveloped respiratory virus, preferably a coronavirus, more preferably the SARS-CoV-2 virus.

[0140] In a preferred embodiment, the Corynebacterium culture or the lyophilized preparation is for use in the upper respiratory tract of a subject.

[0141] In an embodiment, the Corynebacterium culture further comprises a nucleotide sequence encoding lipase. Preferably, the Corynebacterium culture is further capable of producing lipase.

[0142] The subject is a vertebrate subject selected from fish, amphibians, reptiles, birds, and mammals, preferably selected from reptiles, birds, and mammals. In an embodiment, the subject is a homeothermic subject selected from birds and mammals. Preferably, the subject is a mammalian subject, preferably a human subject.

[0143] Specifically, the subject is a mammal or an avian, preferably a mammal. Specifically, the subject is a mammalian subject, preferably a farm animal, a domestic animal, or a human, especially a human subject.

[0144] Preferably, the pathogenic respiratory RNA virus or the enveloped respiratory virus is a coronavirus. Preferably, the virus is the SARS coronavirus. Preferably, the virus is selected from SARS-CoV-1, SARS-CoV-2, and MERS. Preferably, the coronavirus is SARS-CoV-2. More preferably, the coronavirus is a SARS-CoV-2 variant using the cathepsin entry pathway. More preferably, the coronavirus is a SARS-CoV-2 variant mainly using the cathepsin B / L entry pathway. Most preferably, the coronavirus is the SARS-CoV-2 Omicron variant.

[0145] Preferably, the pathogenic respiratory RNA virus or the enveloped respiratory virus is a virus that uses the cathepsin entry pathway to enter the cells of the subject.

[0146] Specifically, the Corynebacterium culture is any of the Corynebacterium cultures (including combinations) defined above.

[0147] In a preferred embodiment, the Corynebacterium culture or freeze-dried preparation is used for preventing respiratory tract infections, respiratory tract colonization or respiratory diseases. In a preferred embodiment, the Corynebacterium culture or freeze-dried preparation is used for preventing diseases caused by an infectious agent that colonizes the respiratory tract, preferably the upper respiratory tract.

[0148] In a preferred embodiment, the Corynebacterium culture or freeze-dried preparation is used for preventing diseases caused by respiratory RNA virus infections. Respiratory viruses cause respiratory tract infections.

[0149] In a preferred embodiment, the Corynebacterium culture or freeze-dried preparation used according to the present invention is used against an enveloped coronavirus having a spike protein, and wherein the Corynebacterium culture is capable of reducing the expression of cathepsin. In a preferred embodiment, the Corynebacterium culture or freeze-dried preparation used according to the present invention is used against an enveloped coronavirus having a spike protein, wherein the Corynebacterium culture is capable of reducing the expression of cathepsin, and is also capable of

[0150] - reducing the expression of a receptor protein (preferably ACE2) in host cells (preferably epithelial cells), and / or

[0151] - reducing the expression of a serine protease that activates the spike protein (preferably TMPRSS2) in host cells (preferably epithelial cells), and optionally,

[0152] - reducing / inhibiting the interaction between the spike protein and the receptor protein (preferably ACE2) in host cells.

[0153] In a preferred embodiment, the RNA virus is an enveloped coronavirus having a spike protein (preferably a coronavirus selected from SARS-CoV-1, SARS-CoV-2 and MERS), and reducing the expression of cathepsin includes reducing the mRNA expression of cathepsin and / or reducing the expression of cathepsin at the protein level. In a preferred embodiment, the RNA virus is an enveloped coronavirus having a spike protein (preferably a coronavirus selected from SARS-CoV-1, SARS-CoV-2 and MERS), and reducing the expression of cathepsin includes reducing the mRNA expression of cathepsin and / or reducing the expression of cathepsin at the protein level, and optionally,

[0154] - reducing the expression of a receptor protein (preferably ACE2) in host cells includes reducing the mRNA expression of the receptor protein and / or reducing the expression of the receptor protein at the protein level, and / or

[0155] - Reducing the expression of serine proteases that activate spike proteins (preferably TMPRSS2) in host cells includes reducing the mRNA expression of the receptor protein and / or reducing the expression of the receptor protein at the protein level.

[0156] In a preferred embodiment of the present invention, a Corynebacterium culture (including combinations) or a lyophilized preparation is used for the treatment or prevention of respiratory diseases of the upper respiratory tract. In a preferred embodiment, a Corynebacterium culture (including combinations) or a lyophilized preparation is used for the treatment or prevention of respiratory infections, respiratory colonization, or respiratory diseases. Preferably, the respiratory infection, respiratory colonization, or respiratory disease is of the upper respiratory tract. In a preferred embodiment, a Corynebacterium culture (including combinations) or a lyophilized preparation is used for the prevention of diseases caused by infectious agents that colonize the respiratory tract, preferably the upper respiratory tract.

[0157] Preferably, the Corynebacterium culture (including combinations) or the lyophilized preparation is administered to the upper respiratory tract of the subject,

[0158] preferably administered to the nasal cavity and pharynx, preferably to the nasopharynx, oropharynx, oro / mesopharynx, and / or laryngopharynx, particularly to the nasopharynx,

[0159] preferably in the form of drops, sprays, inhalable formulations, or ointments.

[0160] Preferably, a Corynebacterium culture (including combinations) or a lyophilized preparation is used for the treatment (including prevention) or preferably for the prevention of respiratory diseases caused by coronaviruses (preferably the SARS virus) in a subject. Preferably, the coronavirus is selected from SARS-CoV-1, SARS-CoV-2, and MERS. More preferably, the coronavirus is a SARS-CoV-2 variant that predominantly uses the cathepsin B / L entry pathway, preferably the SARS-CoV-2 Omicron variant. Preferably, the respiratory disease is COVID-19 disease. Preferably, the subject is a mammalian subject, preferably a human subject.

[0161] Preferably, a Corynebacterium culture or a lyophilized preparation is used for the prevention of symptoms of diseases caused by infectious agents that colonize the respiratory tract, preferably the upper respiratory tract. Preferably, a Corynebacterium culture or a lyophilized preparation is used for the prevention of symptoms of diseases caused by coronaviruses. These symptoms include fever or chills, cough, shortness of breath or difficulty breathing, fatigue, muscle or body aches, headache, loss of taste or smell, sore throat, stuffy or runny nose, nausea, vomiting, and diarrhea.

[0162] In a preferred embodiment, a Corynebacterium culture or a combination thereof, or a lyophilized preparation thereof, is administered to a subject for about 5 to 7 days, once or twice a day. In a preferred embodiment, a Corynebacterium culture or a combination thereof, or a lyophilized preparation thereof, is administered to a subject twice a day, preferably in the morning and evening. When administered into the throat (e.g., via a mouthwash or gargle preparation), the Corynebacterium culture or a combination thereof, or a lyophilized preparation thereof, is administered to the subject after a meal.

[0163] The present invention also relates to a method for preventing infection with an enveloped respiratory virus, preferably a coronavirus, more preferably SARS-CoV-2, even more preferably the Omicron variant of SARS-CoV-2, the method comprising:

[0164] administering to a subject in need thereof a Corynebacterium culture as defined above, or a preparation as defined above, or a composition as defined above.

[0165] Preferably, the method comprises administering a Corynebacterium culture comprising at least one or at least two, preferably at least three, more preferably four Corynebacterium strains selected from the following

[0166] Corynebacterium accolens strain SU004 or a derivative, variant or mutant thereof with accession number NCAIM P(B)001500 deposited at NCAIM (National Collection of Agricultural and Industrial Microorganisms, Institute of Food Science and Technology, Szent István University, Faculty of Agricultural and Life Sciences, H-1118 Budapest, Somlóu u. 14-16, Hungary) on January 17, 2022, wherein the derivative, variant or mutant thereof is capable of reducing cathepsin expression;

[0167] Corynebacterium propionicum strain SU005 or a derivative, variant or mutant thereof with accession number NCAIM P(B)001501 deposited at NCAIM on April 27, 2022, wherein the derivative, variant or mutant thereof is capable of reducing cathepsin expression;

[0168] Corynebacterium accolens strain SU006 or a derivative, variant or mutant thereof with accession number NCAIM P(B)001502 deposited at NCAIM on April 27, 2022, wherein the derivative, variant or mutant thereof is capable of reducing cathepsin expression;

[0169] Corynebacterium propionicum strain SU007 or a derivative, variant or mutant thereof with accession number NCAIM P(B)001504 deposited at NCAIM on April 27, 2022, wherein the derivative, variant or mutant thereof is capable of reducing cathepsin expression;

[0170] The Corynebacterium accolens strain SU008 or its derivatives, variants or mutants with the accession number NCAIM P(B)001505 deposited at NCAIM on April 27, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression;

[0171] The Corynebacterium accolens strain SU009 or its derivatives, variants or mutants with the accession number NCAIM P(B)001506 deposited at NCAIM on May 25, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression;

[0172] The Corynebacterium propionicum strain SU010 or its derivatives, variants or mutants with the accession number NCAIM P(B)001507 deposited at NCAIM on May 25, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression.

[0173] Preferably, the method comprises administering the above-mentioned lyophilized preparation.

[0174] Abbreviations

[0175] ACE2 Angiotensin-converting enzyme 2

[0176] CFU Colony-forming unit

[0177] COVID-19 Coronavirus disease 2019

[0178] EMEM Eagle's Minimum Essential Medium

[0179] GAPDH Glyceraldehyde 3-phosphate dehydrogenase

[0180] MALDI-TOF Matrix-assisted laser desorption / ionization time-of-flight

[0181] MERS Middle East Respiratory Syndrome

[0182] NCAIM National Collection of Agricultural and Industrial Microorganisms

[0183] PBS Phosphate-buffered saline

[0184] PCR Polymerase chain reaction

[0185] rRNA Ribosomal RNA

[0186] SARS-CoV-2 Severe acute respiratory syndrome coronavirus 2

[0187] TMPRSS2 Transmembrane protease / serine 2

[0188] Definitions

[0189] As used herein, a bacterium is "viable" if it is capable of multiplying (termed "growing" if the number of viable bacteria increases) under conditions including a suitable culture medium and nutrients and a suitable temperature.

[0190] As used herein, "culture" refers to the cultivation of bacteria in an artificial environment (i.e., in vitro). Cultivation and thus culture can include maintaining viable forms of the bacteria and / or the multiplication of the bacteria. Thus, "culture" includes bacteria in a culture medium, in which the bacteria are maintained in viable form, including in a dried (lyophilized) form, cultures on solid media, and in liquid form. The term culture also encompasses the meaning of the term "consortium". As used herein, a "culture" can include only one strain, or can include more than one strain. Thus, as used herein, the term "Corynebacterium culture" is a culture that includes viable Corynebacterium. A culture can include only one strain of Corynebacterium, or it can contain at least two (e.g., at least three, preferably four) different strains of Corynebacterium, unless otherwise stated.

[0191] Preferably, a Corynebacterium strain or Corynebacterium culture "capable of reducing protein expression" is a Corynebacterium strain or Corynebacterium culture that reduces the expression of the protein in mammalian cells in an in vitro co-culture of mammalian cells and Corynebacterium bacteria, respectively. Preferably, expression is determined by measuring the mRNA level of the protein, and the expression of the protein is "reduced" when the mRNA level of the protein in the co-cultured mammalian cells is lower than the mRNA level of the protein measured in control mammalian cells (without bacteria). Preferably, the mammalian cells are human cells, optionally the human colon epithelial cell line Caco-2. In an embodiment, a Corynebacterium strain or Corynebacterium culture capable of reducing cathepsin expression is a Corynebacterium strain or Corynebacterium culture that reduces the relative expression of cathepsin mRNA after incubation with Caco-2 cells for 24 hours.

[0192] The term "formulation" relates to the composition of a substance, which includes at least one bioactive ingredient (preferably a pharmaceutically active ingredient), an "active agent", and at least one other substance, e.g., a medium and / or excipient or both suitable for administration to a subject (e.g., a mammal or a bird). A preferred formulation is a formulation for the upper respiratory tract of the subject (preferably a warm-blooded subject). The terms "formulation" and "composition" can be used interchangeably in the context of the present invention.

[0193] As used herein, the term "administering" shall include the means by which the formulations of the present invention are introduced or applied to a subject in need thereof to perform their intended function. Specifically, administering as used herein refers to administering to the upper respiratory tract of a subject (e.g., a mammal or a bird).

[0194] As used herein, the term "subject" shall refer to a warm-blooded animal (a mammal or an avian, preferably a mammal) subject, particularly a human. Specifically, the medical use or corresponding treatment method of the present invention is applicable to a subject in need of preventing or treating a disorder or disease caused by an infectious agent that colonizes the respiratory tract, preferably the upper respiratory tract.

[0195] The term "patient" includes a subject receiving healthcare, such as a subject receiving prophylactic or therapeutic treatment.

[0196] As used herein, "prophylactic treatment" or "prophylaxis" or "prevention" includes measures taken on or with respect to a patient to prevent the patient from being infected, including reducing the infection rate or the severity of the infection (the number of infected cells). The patient may be a non-infected patient, or a patient who has been infected but is prevented from further infection or reinfection by the said measures. Preferably, prophylaxis or prevention includes administering to the target site, i.e., the upper respiratory tract, particularly its epithelium, at least once or multiple times or regularly.

[0197] As used herein, "treating", "treatment", "for treating" means improving, alleviating, reducing, or relieving the symptoms of a disease or disorder, such as the symptoms of an infection. "Treating", "for treating" also includes reducing the number of viruses that can enter the cells of a patient.

[0198] The "upper respiratory tract" refers to the part of the respiratory system located above the angle of Louis (lateral to the thorax), above the vocal cords, or above the cricoid cartilage. Preferably, the larynx is sometimes included in the upper respiratory tract. The upper respiratory tract includes the nasal cavity and paranasal sinuses, the pharynx (including the nasopharynx / epipharynx, oropharynx / mesopharynx, and laryngopharynx / hypopharynx), preferably including the larynx. In a preferred embodiment, the "upper respiratory tract" refers to the nasal cavity and the nasopharynx.

[0199] The expression "infection" means the undesired proliferation of a virus within the cells of a subject.

[0200] The term "comprise(s)", "comprising", or "including" shall be construed herein as having a non-exhaustive meaning and allowing for the addition or involvement of further features or method steps or components to anything that includes the listed features or method steps or components. Such terms may be limited to "consisting essentially of" or "substantially comprising", which shall be understood to consist of the mandatory features or method steps or components listed (e.g., in a claim), while allowing for the additional inclusion of other features or method steps or components that do not materially affect the basic characteristics of the use, method, composition, or other subject matter.

[0201] As used in this specification and the appended claims, the singular forms "a", "an", and "the" include plural references and shall be construed to include the meaning of "one or more" unless the context clearly dictates otherwise. In general, it should also be understood that the terms used herein are for the purpose of describing particular embodiments only and are not intended to be limiting.

[0202] "Preparations for the upper respiratory tract" include any preparations delivered to the upper respiratory tract of a subject, including any of the nasal, nasopharyngeal, oculonasal, oral mucosal, and pulmonary preparations listed above.

[0203] The term "Corynebacterium accolens SU001" refers to the Corynebacterium accolens SU001 strain with the accession number NCAIM P(B)001495 deposited at NCAIM (National Collection of Agricultural and Industrial Microorganisms, Institute of Food Science and Technology, Szent István University, Faculty of Agricultural and Life Sciences, H-1118 Budapest, Somlóyi út 14-16, Hungary) on June 22, 2021.

[0204] The term "Corynebacterium propionicum SU002" refers to the Corynebacterium propionicum SU002 strain with the accession number NCAIM P(B)001496 deposited at NCAIM on June 22, 2021.

[0205] The term "Corynebacterium propionicum SU003" refers to the Corynebacterium propionicum SU003 strain with the accession number NCAIM P(B)001497 deposited at NCAIM on June 22, 2021.

[0206] The term "Corynebacterium accolens SU004" refers to the Corynebacterium accolens SU004 strain with the accession number NCAIM P(B)001500 deposited at NCAIM (National Collection of Agricultural and Industrial Microorganisms, Institute of Food Science and Technology, Szent István University, Faculty of Agricultural and Life Sciences, H-1118 Budapest, Somlóyi út 14-16, Hungary) on January 17, 2022.

[0207] The term "Corynebacterium tuberculostearicum SU005" refers to the strain Corynebacterium tuberculostearicum SU005 with the accession number NCAIM P(B)001501 deposited at NCAIM on April 27, 2022.

[0208] The term "Corynebacterium accolens SU006" refers to the strain Corynebacterium accolens SU006 with the accession number NCAIM P(B)001502 deposited at NCAIM on April 27, 2022.

[0209] The term "Corynebacterium propionicum SU007" refers to the strain Corynebacterium propionicum SU007 with the accession number NCAIM P(B)001504 deposited at NCAIM on April 27, 2022.

[0210] The term "Corynebacterium accolens SU008" refers to the strain Corynebacterium accolens SU008 with the accession number NCAIM P(B)001505 deposited at NCAIM on April 27, 2022.

[0211] The term "Corynebacterium accolens SU009" refers to the strain Corynebacterium accolens SU009 with the accession number NCAIM P(B)001506 deposited at NCAIM on May 25, 2022.

[0212] The term "Corynebacterium propionicum SU010" refers to the strain Corynebacterium propionicum SU010 with the accession number NCAIM P(B)001507 deposited at NCAIM on May 25, 2022. Description of the Drawings

[0213] Figure 1 . Temperature during the freeze-drying process.

[0214] Figure 2A . Effects of different Corynebacterium strains (SU001 - SU010) on the relative mRNA expression of cathepsin.

[0215] Figure 2B . Effects of different Corynebacterium strains (SU004 - SU010) on the relative mRNA expression of cathepsin.

[0216] Figure 3 . Effects of different Corynebacterium strains (SU001 - SU010) on the relative mRNA expression of cathepsin, ACE2, and TMPRSS2.

[0217] Figure 4 . Effects of different Corynebacterium strains (SU004 - SU010) on the relative mRNA expression of cathepsin, ACE2, and TMPRSS2.

[0218] Figure 5 . Effects of different Corynebacterium strains (SU001 - SU010) on the ACE2 - S1 protein binding activity (%).

[0219] Figure 6 . Effects of different Corynebacterium strains (SU004 - SU010) on the ACE2 - S1 protein binding activity (%).

[0220] Figure 7 . Effects of the K1 combination on the relative expression of cathepsin, ACE2, and TMPRSS2.

[0221] Figure 8 . Effects of the K2 combination on the relative expression of cathepsin, ACE2, and TMPRSS2.

[0222] Figure 9 . Effects of the K3 combination on the relative expression of cathepsin, ACE2, and TMPRSS2.

[0223] Figure 10 . Effects of xanthan gum on the expression of ACE2, TMPRSS2, and cathepsin in the K1, K2, and K3 combinations.

[0224] Figure 11 . Effects of carrageenan on the expression of ACE2, TMPRSS2, and cathepsin in the K1, K2, and K3 combinations. Detailed implementation manners

[0225] The present inventors determined that the normal flora in the nasopharynx affects the susceptibility to SARS - CoV - 2 infection.

[0226] The presence of Corynebacterium in the nasopharyngeal flora can reduce an individual's susceptibility to SARS - CoV - 2 infection through multiple mechanisms.

[0227] Corynebacterium strains can reduce an individual's susceptibility to SARS - CoV - 2 infection through multiple mechanisms: by downregulating the SARS - CoV - 2 receptors: ACE2 and TMPRSS2, by inhibiting the binding of the S1 protein and the ACE2 receptor, and by lipase production. By inhibiting the binding of SARS - CoV - 2 to host cells and by acting on the lipid envelope of the SARS - CoV - 2 virus, all these mechanisms work together. These mechanisms can reduce an individual's susceptibility to enveloped respiratory virus infections, especially SARS - CoV - 2.

[0228] The objectives of the present inventors are as follows. Their aim is to determine the effect of Corynebacterium strains on the cathepsin pathway induced by the Omicron variant in human cells. After detecting the action of Corynebacterium, they deposited several strains. Their aim is to determine the ideal combination of Corynebacterium strains and also to determine their preventive effect against SARS-CoV-2 infection for the purpose of probiotic use. In addition, their aim is to determine a carrier for Corynebacterium strains suitable for probiotic use, and in addition, the Corynebacterium strains can retain their effectiveness against SARS-CoV-2 and their viability.

[0229] Since the SARS-CoV-2 virus enters cells not only through ACE2 and TMPRSS2, but the protease pathway has become increasingly important, especially in the case of the Omicron variant, the present inventors isolated Corynebacterium strains and detected the effect of bacterial co-culture on protease expression. After the isolated Corynebacterium strains were incubated with Caco-2 cells for 24 hours, they significantly reduced the relative expression of cathepsin mRNA to varying degrees, thus contributing to a decrease in ACE2 expression, a decrease in TMPRSS2 expression, an inhibition of ACE2-S protein, and the presence of lipase, reducing the entry of SARS-CoV-2 virus, preferably the SARS-CoV-2 Omicron mutant, into cells.

[0230] The present inventors demonstrated that Corynebacterium is capable of downregulating cathepsin protein expression in human cells. The protease pathway is an important entry point for the SARS-CoV-2 virus, and the currently widely spread Omicron variant mainly enters through the protease pathway.

[0231] Among the Corynebacterium strains they studied, ten Corynebacterium strains have been deposited under the names SU001 - SU010. When depositing the Corynebacterium strains, they considered the effect on cathepsin expression in addition to their other effects against SARS-CoV-2 infection.

[0232] Combinations K1, K2, K3 of the deposited Corynebacterium strains containing four Corynebacterium strains were determined, and this combination further enhanced the effect of downregulating ACE2, TMPRSS2, and cathepsin expression in addition to having the beneficial characteristics unique to Corynebacterium strains.

[0233] The present inventors described that the carrier xanthan gum does not affect the colony count of the Corynebacterium combination K1, K2, K3 after lyophilization.

[0234] They also described that the combination of Corynebacterium sp. K1, K2, K3 with xanthan gum carrier maintained their viability and colony counts for 14 days at room temperature after lyophilization. In addition, they described that the combination of Corynebacterium sp. K1, K2, K3 with xanthan gum carrier retained the effect of downregulating ACE2, TMPRSS2, and cathepsin after lyophilization, and even produced a synergistic improvement effect.

[0235] Therefore, the inventors unexpectedly found that xanthan gum not only acts as an excipient (carrier), but also as an adjuvant - it improves the effect of the Corynebacterium sp. combination, especially combination K2 (see Figure 10 ).

[0236] They described that the carrier κ-carrageenan did not affect the colony counts of the Corynebacterium sp. combination K1, K2, K3 after lyophilization. In addition, they described that the combination of Corynebacterium sp. K1, K2, K3 with κ-carrageenan carrier maintained their viability and colony numbers for 14 days at room temperature after lyophilization. They also described that the combination of Corynebacterium sp. K1, K2, K3 with κ-carrageenan carrier retained the effect of downregulating ACE2, TMPRSS2, and cathepsin after lyophilization.

[0237] The combination of bacterial strains can be re-formulated in the form of a nasal spray or a pharyngeal spray after lyophilization. They all retain their known and tested biological activities.

[0238] Finding not only a suitable excipient for the lyophilized preparation but also the appropriate concentration of the excipient is an important step.

[0239] The carbohydrate or carbohydrate derivative excipient in the lyophilized preparation is a cryoprotectant that protects the Corynebacterium sp. culture.

[0240] The concentration of the hydrocolloid polymer in the lyophilized preparation controls the viscosity of the preparation. The lyophilized preparation should not be a gel but should be able to form sufficient blocks. In other words, the preparation should not be too viscous or too dense. Too high a concentration will lead to gelation, and too low a concentration will cause the preparation to fail to form appropriate blocks during lyophilization.

[0241] The lyophilized preparation should also have an ideal osmotic pressure range, preferably in the range of 290 to 500 mOsmol / kg, more preferably in the range of 300 to 400 mOsm / kg. More preferably, the osmotic pressure of the lyophilized preparation is 320 to 370 mOsmol / kg.

[0242] Preferably, the pH of the lyophilized preparation is pH 5 to 7, more preferably pH 5.5 to 6.5.

[0243] Another important aspect is that the lyophilized preparation must be physiologically tolerable, and thus the excipients used and their concentrations should be carefully selected.

[0244] In the case of nasal sprays, the droplet size distribution is also important [Kulkarni & Shaw (2012). Formulation and characterization of nasal sprays. Inhalation Magazine, June 2012]. Larger droplets will drip out of the nose, while droplets smaller than 10 microns may further enter the nasal cavity and reach the lungs (which is not the intended delivery site). Therefore, droplets smaller than 10 μm must be minimized. For example, in nasal sprays, the droplet size may be 30 - 70 μm, up to 200 μm.

[0245] Examples

[0246] Example 1 - Corynebacterium strains

[0247] Materials and methods

[0248] Bacterial sample collection

[0249] Corynebacterium isolates with putative protective effects were collected from patients who had not been infected despite close contact with their COVID - positive family members. Nasopharyngeal swab samples from COVID - negative patients were inoculated on Columbia blood agar (Biolab, Hungary) and incubated at 37 °C in a humid atmosphere containing 5% CO2. Corynebacterium strains were selected from other bacterial participants using antimicrobial susceptibility test discs containing 50 μg of fosfomycin (Oxoid, Sweden). All cultured bacteria were identified by the MALDI - TOF method (Bruker Daltonik, Germany). The excellent properties of three previously isolated Corynebacterium strains (SU001 - 003) were examined, and seven additional Corynebacterium strains (SU004 - 010) were isolated and biologically tested.

[0250] Detection of ACE2, TMPRSS2, and cathepsin L expression in Caco - 2 cells after bacterial co - culture

[0251] The human colon epithelial cell line Caco-2 was previously obtained and used in this study. Caco-2 cells were cultured at 37 °C in EMEM medium (Lonza Bioscience, USA) in a humidified atmosphere containing 5% CO2, and the medium was changed every two days. Antibiotics were not added to allow bacterial co-culture. Caco-2 cells were cultured alone for four days and then an overnight culture of Corynebacterium sp. SU001-010 was added. Human Caco-2 cells and bacteria were co-cultured for an additional 24 hours.

[0252] Cells were washed and centrifuged with PBS and 0.25% trypsin. Total RNA was isolated using the innuPREP RNA Mini Kit 2.0 (Analytik Jena GmbH, Germany) according to the manufacturer's instructions. RNA concentration was determined using a NanoDrop1000 spectrophotometer (Thermo Fisher Scientific, USA). RT-PCR tests were performed using 10 - 100 ng of RNA with the PrimeScript RT Kit (Takara Bio, USA) and the resulting cDNA was amplified on a qTOWER3G (Analytik Jena GmbH, Germany) instrument in the presence of selected primers.

[0253] The primers for ACE2 were 5'-GGG ATC AGA GAT CGG AAG AAG AAA-3' forward (SEQ ID NO:1) and 5'-AGG AGG TCT GAA CAT CAT CAG TG-3' reverse (SEQ ID NO:2). The primers for TMPRSS2 were 5'-AATCGG TGT GTT CGC CTC TAC-3' forward (SEQ ID NO:3) and 5'-CGT AGT TCT CGT TCC AGT CGT-3' reverse (SEQ ID NO:4). The primers for cathepsin L were 5'-CTG GTG GTT GGC TAC GGA TT-3' CTSL forward (SEQ ID NO:5) and 5'-CTC CGG TCT TTG GCC ATC TT-3' reverse (SEQ ID NO:6). The primers for GAPDH were 5'-CTA CTG GCG CTG GCA AGG CTG T-3' forward (SEQ ID NO:7) and 5'-GCC ATG AGG TCCACC ACC CTG CTG-3' reverse (SEQ ID NO:8).

[0254] Relative mRNA expression was calculated by the change in cycle threshold (ΔΔCt) method and normalized to the geometric mean of the housekeeping gene GAPDH. The basal mRNA levels of ACE2, TMPRSS2, and cathepsin were compared to the level of the housekeeping gene GAPDH by calculating the difference between their Ct values.

[0255] Statistical analysis:

[0256] In the ACE2, TMPRSS2, and cathepsin expression studies, differences between the measured mRNA levels in different bacterial co-culture groups were calculated by two-tailed Student's t-test.

[0257] Effect of Corynebacterium on ACE2, TMPRSS2, and cathepsin expression

[0258] In the case of strains SU001 - 010, the inventors investigated how they affected the relative ACE2, TMPRSS2, and cathepsin expression in Caco-2 cells co-cultured with each strain separately.

[0259] Results

[0260] Effect of Corynebacterium on cathepsin expression

[0261] Given that the SARS-CoV-2 virus enters cells not only through ACE2 and TMPRSS2, but the cathepsin pathway is becoming increasingly important, especially in the case of the Omicron variant, we examined the effect of bacterial co-culture on protease expression in the case of previously isolated Corynebacterium strains. Corynebacterium strains significantly reduced the relative mRNA expression of cathepsin to varying degrees after 24 hours of incubation with Caco-2 cells (see Figure 2A and Figure 2B ), thus contributing to reduced ACE2 expression, reduced TMPRSS2 expression, ACE2-S protein inhibition, and the presence of lipase in addition to the previously described role of reducing the entry of the Omicron mutant into cells by the SARS-CoV-2 virus.

[0262] Corynebacterium strains also reduced the relative mRNA expression of ACE2 and TMPRSS2 (see Figure 3 and Figure 4 ). In addition, Corynebacterium strains inhibited the binding of the S1 protein and the ACE2 receptor (see Figure 5 and Figure 6 ).

[0263] Example 2 - Combinations of Corynebacterium strains

[0264] Materials and methods

[0265] Cultivation study of Corynebacterium strains combination

[0266] The present inventors studied how each Corynebacterium strain affects the proliferation of each other. The pure bacterial cultures of each bacterial strain were suspended in saline, and their turbidity was standardized to 0.5 McFarland, and a mixture of the same proportion was constituted, which contained 4 - 4 strains of different groups of SU001 - 010 strains. 100 μL of the mixture was inoculated onto Columbia blood agar (Biolab, Hungary) and incubated at 37 °C for 48 hours in a humid atmosphere containing 5% CO2. According to the culture, it was possible to determine which Corynebacterium strains allowed other strains to grow in combination with them based on different colony morphology growth. K1 - K2 - K3, according to successful co - growth, three different combinations could be compiled. Detection of ACE2, TMPRSS2 and cathepsin L expression in Caco - 2 cells after bacterial co - culture

[0267] Previously, the human colon epithelial cell line Caco - 2 was obtained and used in this study. Caco - 2 cells were cultured in EMEM medium (Lonza Bioscience, USA) at 37 °C in a humid atmosphere containing 5% CO2, and the medium was changed every two days. No antibiotics were added to allow bacterial co - culture. Caco - 2 cells were cultured alone for four days and then an overnight culture of the K1, K2 or K3 Corynebacterium strain combination was added. The human Caco - 2 cells and bacteria were co - cultured for an additional 24 hours.

[0268] The cells were washed with PBS and 0.25% trypsin and centrifuged. Total RNA was isolated using the innuPREP RNA Mini Kit 2.0 (Analytik Jena GmbH, Germany) according to the manufacturer's instructions. The RNA concentration was determined using a NanoDrop1000 spectrophotometer (Thermo Fisher Scientific, USA). RT - PCR tests were performed using 10 - 100 ng of RNA with the PrimeScript RT Kit (Takara Bio, USA) and the resulting cDNA was amplified on a qTOWER3G (Analytik Jena GmbH, Germany) instrument in the presence of selected primers.

[0269] The primers for ACE2 are 5'-GGG ATC AGA GAT CGG AAG AAG AAA-3' forward (SEQ ID NO:1) and 5'-AGG AGG TCT GAA CAT CAT CAG TG-3' reverse (SEQ ID NO:2). The primers for TMPRSS2 are 5'-AATCGG TGT GTT CGC CTC TAC-3' forward (SEQ ID NO:3) and 5'-CGT AGT TCT CGT TCC AGT CGT-3' reverse (SEQ ID NO:4). The primers for cathepsin L are 5'-CTG GTG GTT GGC TAC GGA TT-3' CTSL forward (SEQ ID NO:5) and 5'-CTC CGG TCT TTG GCC ATC TT-3' reverse (SEQ ID NO:6). The primers for GAPDH are 5'-CTA CTG GCG CTG GCA AGG CTG T-3' forward (SEQ ID NO:7) and 5'-GCC ATG AGG TCCACC ACC CTG CTG-3' reverse (SEQ ID NO:8).

[0270] Relative mRNA expression was calculated by the cycle threshold change (ΔΔCt) method and normalized to the geometric mean of the housekeeping gene GAPDH. The basal mRNA levels of ACE2, TMPRSS2, and cathepsin were compared to the level of the housekeeping gene GAPDH by calculating the difference between their Ct values.

[0271] Statistical analysis:

[0272] In the ACE2, TMPRSS2, and cathepsin expression studies, the differences between the measured mRNA levels in different bacterial co-culture groups were calculated by two-tailed Student's t-test.

[0273] The effects of different Corynebacterium strains combinations (K1, K2, K3) on their respective biological effects: ACE2 expression, TMPRSS2 expression, cathepsin expression

[0274] In the case of the K1, K2, and K3 combinations, the inventors studied how they affected the relative ACE2, TMPRSS2, and cathepsin expression in Caco-2 cells co-cultured with each combination respectively.

[0275] Results

[0276] The effects of different Corynebacterium strains combinations (K1, K2, K3) on their respective biological effects: ACE2 expression, TMPRSS2 expression, cathepsin expression

[0277] During the research process of the present inventors, in order to detect the effect of Corynebacterium on the beneficial properties of the strains previously detected individually, they co-cultured 4 - 4 strains together and detected their combined effects.

[0278] The following strain combinations were used: (i) combination K1 including Corynebacterium strains SU004, SU005, SU009, and SU010; (ii) combination K2 including Corynebacterium strains SU004, SU005, SU006, and SU010; and (iii) combination K3 including Corynebacterium strains SU004, SU005, SU007, and SU008.

[0279] In all three cases of these combinations, their results showed that the different Corynebacterium strains used in the combination did not affect each other's individual effects during co-culture, and surprisingly, in some cases, an effect of enhancing the beneficial results was observed.

[0280] Effect of K1 combination on mRNA expression of cathepsin, ACE2, and TMPRSS2

[0281] Combination K1 contains two Corynebacterium incertae sedis strains SU004, SU009, one Corynebacterium tuberculostearicum SU005, and one Corynebacterium propionicum SU010. When combination K1 was used, the effect of the Corynebacterium combination on the expression of cathepsin, ACE2, and TMPRSS2 was approximately the average of the effects of each bacterium individually (see Figure 7 ).

[0282] Effect of K2 combination on mRNA expression of cathepsin, ACE2, and TMPRSS2

[0283] Combination K2 contains two Corynebacterium incertae sedis strains SU004, SU006, one Corynebacterium tuberculostearicum SU005, and one Corynebacterium propionicum SU010. When combination K2 was used, the effect of the Corynebacterium combination on the expression of cathepsin and TMPRSS2 was approximately the average of the effects of each bacterium individually, and in the case of ACE2, the relative expression rate was similar to that of strain SU004 (see Figure 8 ). Effect of K3 combination on mRNA expression of cathepsin, ACE2, and TMPRSS2

[0284] Combination K3 contains two Corynebacterium incertae sedis strains SU004, SU008, one Corynebacterium tuberculostearicum SU005, and one Corynebacterium propionicum SU007. When combination K3 was used, the effect of the Corynebacterium combination on the expression of cathepsin and ACE2 was very similar to the values of the most effective strains. However, in the case of TMPRSS2 expression, combination K3 had a greater effect than each bacterium individually, showing a synergistic effect in the case of downregulation of TMPRSS2 expression (see Figure 9 ).

[0285] Example 3 - Freeze - dried preparation

[0286] Materials and methods

[0287] Study on the framework - forming properties of different solutions containing hydrocolloid polymers and carbohydrates for freeze - drying

[0288] The following excipients and excipient combinations were tested: (1) 0.05 g xanthan gum / 100 mL and vitamin A; (2) 0.1 g / 100 mL κ - carrageenan and 0.5 g / 100 mL maltodextrin and vitamin A; (3) 0.1 g / 100 mL κ - carrageenan and 0.1 g / 100 mL maltodextrin and vitamin A; (4) 0.05 g xanthan gum / 100 mL and 0.1 g / 100 mL maltodextrin and vitamin A; (5) 0.05 g xanthan gum / 100 mL and 0.5 g / 100 mL maltodextrin and vitamin A; (6) (also known as Solution A) 0.1 g / 100 mL κ - carrageenan and 0.5 g / 100 mL isomaltulose; (7) (also known as Solution B) 0.05 g xanthan gum / 100 mL and 0.5 g / 100 mL isomaltulose.

[0289] Early experiments showed that vitamin A reduced the effectiveness of the excipient combinations. Therefore, the present inventors excluded vitamin A from the excipient combinations and selected combinations (6) and (7) (also known as Solution A and Solution B respectively) for subsequent experiments.

[0290] Freeze - drying

[0291] One of the aims of the present inventors was to formulate a nasal spray probiotic and to examine the viability of the bacterial strains after freeze - drying and storage for two weeks.

[0292] The stabilization of the bacterial strains in the dry solid state was carried out using a freeze - dryer (Scanvac Coolsafe 110 - 04 freeze - dryer, LaboGene TM , Ringsted, Denmark).

[0293] The freeze - drying process generally consists of three steps, namely, freezing, primary drying, and secondary drying. During freezing, water crystallizes, and then the frozen ice is removed by sublimation. In the final secondary drying step, the remaining moisture is removed to achieve the final moisture content.

[0294] As cryoprotectants, hydrocolloid polymers and carbohydrates are used for the lyophilization of probiotics, which results in a satisfactory microbial survival rate after lyophilization. Generally, carbohydrates (e.g., trehalose, sucrose, glucose, lactose, maltodextrin) and polymers are used as cryoprotectants. [Meng XC, Stanton C, Fitzgerald GF, Daly C, Ross RP. (2008). Anhydrobiotics: The challenges of drying probiotic cultures. Food Chem., 106:1406-1416.]

[0295] Technologically important is the combination of these cryoprotective compounds with a composition suitable for lyophilization to ensure the survival rate of bacteria, which is characterized by a stable physical form that can be reconstituted into a sprayable liquid with an appropriate viscosity, pH value (5 - 7), and osmotic concentration (300 - 400 mosm). Preferably, the pH of the suitable composition is pH 5.5 to 6.5, and the osmotic pressure of the suitable composition is 320 to 370 mOsmol / kg.

[0296] During the formation of the lyophilized product, hydrocolloid polymers and carbohydrate derivatives are used as protectants. 2 mL of Ringer's solution containing 10 8 CFU / mL of each bacterial strain was frozen at -70 °C and stored for 24 hours. Lyophilization started with the primary drying at a temperature between -30 °C and -45 °C for a duration of no more than 16 hours. Secondary drying was carried out until the appropriate moisture content did not exceed 5%, while keeping the sample temperature not exceeding 10 °C. The entire lyophilization process took 24 - 36 hours. [Zayed, G. and Y. H. Roos. 2004. Influence of trehalose and moisture content on survival of Lactobacillus salivarius subjected to freeze-drying and storage. Process Biochem. 9:1081–1086.] See also Figure 1 。

[0297] Study on the shelf life of different solutions containing hydrocolloid polymers and carbohydrates

[0298] Two different solutions were prepared in Ringer's solution, which contained: (A) 0.1 g / 100 mL of κ-carrageenan and 0.5 g / 100 mL of isomaltulose (Solution A), or (B) 0.05 g of xanthan gum / 100 mL and 0.5 g / 100 mL of isomaltulose (Solution B).

[0299] Part of 2 mL of solution A or B was inoculated with 10 8 CFU (colony-forming units) / mL of strain SU001-010 or with a combination of strains K1, K2, and K3. After storage at room temperature for 7 and 14 days, 100 μL from the different solutions was inoculated onto Columbia blood agar (Biolab, Hungary) to determine the number of colony-forming units as the number of viable Corynebacterium strains.

[0300] Detection of the expression of ACE2, TMPRSS2, and cathepsin L in Caco-2 cells after bacterial co-culture

[0301] The human colon epithelial cell line Caco-2 was previously obtained and used in this study. Caco-2 cells were cultured in EMEM medium (Lonza Bioscience, USA) at 37 °C in a humidified atmosphere containing 5% CO2, and the medium was changed every two days. Antibiotics were not added to allow bacterial co-culture. Caco-2 cells were cultured alone for four days and then an overnight culture of Corynebacterium SU001-010 or a combination of K1, K2, or K3 in solution A or B was added. Human Caco-2 cells and bacteria were co-cultured for an additional 24 hours.

[0302] Cells were washed and centrifuged with PBS and 0.25% trypsin. Total RNA was isolated using the innuPREP RNA Mini kit 2.0 (Analytik Jena GmbH, Germany) according to the manufacturer's instructions. The RNA concentration was determined using a NanoDrop 1000 spectrophotometer (Thermo Fisher Scientific, USA). RT-PCR tests were performed using 10 - 100 ng of RNA with the PrimeScript RT kit (Takara Bio, USA) and the resulting cDNA was amplified on a qTOWER 3G (Analytik Jena GmbH, Germany) instrument in the presence of selected primers.

[0303] The primers for ACE2 are 5'-GGG ATC AGA GAT CGG AAG AAG AAA-3' forward (SEQ ID NO:1) and 5'-AGG AGG TCT GAA CAT CAT CAG TG-3' reverse (SEQ ID NO:2). The primers for TMPRSS2 are 5'-AATCGG TGT GTT CGC CTC TAC-3' forward (SEQ ID NO:3) and 5'-CGT AGT TCT CGT TCC AGT CGT-3' reverse (SEQ ID NO:4). The primers for cathepsin L are 5'-CTG GTG GTT GGC TAC GGA TT-3' CTSL forward (SEQ ID NO:5) and 5'-CTC CGG TCT TTG GCC ATC TT-3' reverse (SEQ ID NO:6). The primers for GAPDH are 5'-CTA CTG GCG CTG GCA AGG CTG T-3' forward (SEQ ID NO:7) and 5'-GCC ATG AGG TCCACC ACC CTG CTG-3' reverse (SEQ ID NO:8).

[0304] Relative mRNA expression was calculated by the cycle threshold change (ΔΔCt) method and normalized to the geometric mean of the housekeeping gene GAPDH. The basal mRNA levels of ACE2, TMPRSS2, and cathepsin were compared to the level of the housekeeping gene GAPDH by calculating the difference between their Ct values.

[0305] Statistical analysis:

[0306] In the ACE2, TMPRSS2, and cathepsin expression studies, the differences between the measured mRNA levels in different bacterial co-culture groups were calculated by two-tailed Student's t-test.

[0307] The effects of different solutions containing hydrocolloid polymers and carbohydrates on the individual biological effects of the SU001-010 strain or the K1-K3 composition on ACE2 expression, TMPRSS2 expression, and cathepsin expression

[0308] In the case of the SU001-SU010 strains, or the K1, K2, and K3 combinations in solution A or B, the inventors studied how they affect the relative ACE2, TMPRSS2, and cathepsin expression in Caco-2 cells cultured separately with each strain or combination.

[0309] Results

[0310] Effect of different excipients (xanthan gum and κ-carrageenan) on the combination of Corynebacterium strains K1, K2, K3

[0311] Various combinations K1, K2 and K3 found to be effective were studied with different excipients to determine the effect of the excipients on the bacterial combinations. The present inventors further studied the effects of two excipients, xanthan gum, an anionic polysaccharide, and κ-carrageenan on the biological activity of Corynebacterium strains.

[0312] Study on the effect of freeze-drying on the growth of Corynebacterium

[0313] The present inventors further examined the effect of lyophilization on different Corynebacterium strains as well as on combinations K1, K2 and K3. After lyophilization, the present inventors determined the colony counts of each bacterium and of combinations K1, K2, K3, respectively. Their research results showed that all bacteria and their combinations maintained their original bacterial numbers.

[0314] Table 1 Effect of lyophilization

[0315]

[0316] They further analyzed the survival rate of Corynebacterium strains, and the Corynebacterium strains could survive for 14 days in all combinations K1, K2 and K3.

[0317] Effect of xanthan gum on mRNA expression of cathepsin, ACE2, and TMPRSS2

[0318] The present inventors further analyzed the effect of xanthan gum alone and in mixtures with combinations K1, K2 and K3 on the relative expression of ACE2, TMPRSS2 and cathepsin. However, xanthan gum itself reduced the expression of ACE2, TMPRSS2 and cathepsin to a lesser extent compared to the combinations of K1, K2 and K3.

[0319] Interestingly, the use of K1 and K3 in combination with xanthan gum did not affect their original effects. However, surprisingly, in the case of combination K2, a synergistic effect with xanthan gum was observed, since the combination of K2 and xanthan gum downregulated the expression of ACE2, TMPRSS2 and cathepsin to a greater extent than when alone (see Figure 10 ).

[0320] Effect of κ-carrageenan on mRNA expression of cathepsin, ACE2, and TMPRSS2

[0321] They further analyzed the effect of κ-carrageenan alone and in mixtures with combinations K1, K2 and K3 on the relative expression of ACE2, TMPRSS2 and cathepsin. κ-carrageenan alone also significantly reduced the expression of ACE2, TMPRSS2 and cathepsin.

[0322] However, interestingly, no additional effect was produced in the case of the combination of K1 with κ-carrageenan. Surprisingly, a synergistic effect was observed in the case of K2 and κ-carrageenan, and when K2 and κ-carrageenan were used together, the expression of all ACE2, TMPRSS2, and cathepsin was downregulated to a greater extent. In the case of K3, κ-carrageenan did not affect the downregulation effect of the combination of K3 on the expression of ACE2, TMPRSS2, and cathepsin (see Figure 11 ).

[0323] Industrial Applicability

[0324] The Corynebacterium culture according to the present invention, or a combination thereof, or a freeze-dried preparation thereof is used for treating or preventing respiratory diseases of the upper respiratory tract. The Corynebacterium culture according to the present invention, or a combination thereof, or a freeze-dried preparation thereof is also used for treating or preventing respiratory tract infections, respiratory tract colonization, or respiratory tract diseases. Preferably, the respiratory tract infections, respiratory tract colonization, or respiratory tract diseases are of the upper respiratory tract. The Corynebacterium culture according to the present invention, or a combination thereof, or a freeze-dried preparation thereof is used for preventing diseases caused by infectious agents in the respiratory tract, preferably the upper respiratory tract, of a host.

[0325] References

[0326] Kulkarni&Shaw(2012).Formulation and characterization of nasalsprays.Inhalation Magazine,June 2012Lappan&Peacock,2019.Corynebacterium andDolosigranulum:future probiotic candidates for upper respiratory tractinfections.Microbiology Australia,40(4),172–177.

[0327] De Maio F,Posteraro B,Ponziani FR,Cattani P et al.(2020).Nasopharyngeal Microbiota Profiling of SARS-CoV-2 InfectedPatients.Biological Procedures Online 22,18.doi:10.1186 / s12575-020-00131-7.

[0328] Man WH, de Steenhuijsen Piters WA, Bogaert D (2017). The microbiota of the respiratory tract: Gatekeeper to respiratory health. Nature Reviews Microbiology, 15(5), 259–270. doi:10.1038 / nrmicro.2017.14.

[0329] Meng XC, Stanton C, Fitzgerald GF, Daly C, Ross RP. (2020). Anhydrobiotics: The challenges of drying probiotic cultures. Food Chem., 106:1406-1416.

[0330] Mostafa HH, Fissel JA, Fanelli B, Bergman Y et al. (2020). Metagenomic next-generation sequencing of nasopharyngeal specimens collected from confirmed and suspect COVID-19 patients. MBio, 11(6), 1–13. doi:10.1128 / mBio.01969-20.

[0331] Nardelli C, Gentile I, Setaro M, Di Domenico C, et al. (2020). Nasopharyngeal Microbiome Signature in COVID-19 Positive Patients: Can We Definitively Get a Role to Fusobacterium periodonticum? Frontiers in Cellular and Infection Microbiology, 11(February), 1–7. doi:10.1038 / nrmicro.2017.14.

[0332] Rosas-Salazar C, Kimura KS, Shilts MH, Strickland BA et al. (2020). SARS-CoV-2 infection and viral load are associated with the upper respiratory tract microbiome. J Allergy Clin Immunol. 2021;147(4):1226-1233.e2. doi:10.1038 / nrmicro.2017.14.

[0333] Tchoupou Saha et al., 2022. Profile of the Nasopharyngeal Microbiota Affecting the Clinical Course in COVID-19 Patients. Front. Microbiol. 13:871627. doi:10.3389 / fmicb.2022.871627

[0334] Zayed, G. and Y. H. Roos. 2004. Influence of trehalose and moisture content on survival of Lactobacillus salivarius subjected to freeze-drying and storage. Process Biochem. 9:1081–1086.

Claims

1. A Corynebacterium culture, the Corynebacterium culture being used for preventing the infection of a coronavirus in a subject, wherein, The Corynebacterium culture is capable of reducing the expression of cathepsin.

2. The Corynebacterium culture used according to claim 1, wherein, The Corynebacterium culture capable of reducing the expression of cathepsin is a Corynebacterium culture that reduces the relative expression of cathepsin mRNA after incubation with Caco-2 cells for 24 hours.

3. A Corynebacterium culture, the Corynebacterium culture being used for preventing the infection of a coronavirus in a subject, wherein, The coronavirus uses the cathepsin pathway, preferably mainly uses the cathepsin pathway, to enter the cells of the subject, and wherein the Corynebacterium culture is capable of reducing the expression of cathepsin. Preferably, the Corynebacterium culture capable of reducing the expression of cathepsin is a Corynebacterium culture that reduces the relative expression of cathepsin mRNA after incubation with Caco-2 cells for 24 hours.

4. The Corynebacterium culture used according to any one of the preceding claims, wherein, The coronavirus is SARS-CoV-2.

5. The Corynebacterium culture used in any of the preceding claims, wherein, The coronavirus is the SARS-CoV-2 Omicron variant.

6. The Corynebacterium culture used in any one of the preceding claims, wherein, The Corynebacterium culture comprises at least one or at least two, preferably at least three, more preferably four Corynebacterium strains selected from the group consisting of Corynebacterium accolens, Corynebacterium propionicum, and Corynebacterium tuberculostearicum.

7. A Corynebacterium culture for preventing infection by the SARS-CoV-2 virus in a subject, wherein, The Corynebacterium culture comprises at least one or at least two, preferably at least three, more preferably four Corynebacterium strains selected from the following: Corynebacterium accolens strain SU004 or its derivatives, variants, or mutants with accession number NCAIM P(B)001500 deposited at NCAIM (National Collection of Agricultural and Industrial Microorganisms, Institute of Food Science and Technology, Szent István University, H-1118 Budapest, Somlóyi út 14-16, Hungary) on January 17, 2022, wherein its derivatives, variants, or mutants are capable of reducing cathepsin expression; Corynebacterium tuberculostearicum strain SU005 or its derivatives, variants, or mutants with accession number NCAIM P(B)001501 deposited at NCAIM on April 27, 2022, wherein its derivatives, variants, or mutants are capable of reducing cathepsin expression; Corynebacterium accolens strain SU006 or its derivatives, variants, or mutants with accession number NCAIM P(B)001502 deposited at NCAIM on April 27, 2022, wherein its derivatives, variants, or mutants are capable of reducing cathepsin expression; Corynebacterium propionicum strain SU007 or its derivatives, variants, or mutants with accession number NCAIM P(B)001504 deposited at NCAIM on April 27, 2022, wherein its derivatives, variants, or mutants are capable of reducing cathepsin expression; Corynebacterium accolens strain SU008 or its derivatives, variants, or mutants with accession number NCAIM P(B)001505 deposited at NCAIM on April 27, 2022, wherein its derivatives, variants, or mutants are capable of reducing cathepsin expression; The Corynebacterium accolens strain SU009 or its derivatives, variants or mutants with the accession number NCAIM P(B)001506 deposited at NCAIM on May 25, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression; The Corynebacterium propionicum strain SU010 or its derivatives, variants or mutants with the accession number NCAIM P(B)001507 deposited at NCAIM on May 25, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression; Preferably, any derivatives, variants or mutants capable of reducing cathepsin expression are derivatives, variants or mutants that reduce the relative expression of cathepsin mRNA after incubation with Caco-2 cells for 24 hours, respectively.

8. The Corynebacterium culture used according to claim 3, wherein, Preferably, the coronavirus is the SARS-CoV-2 virus, more preferably the SARS-CoV-2 Omicron variant, and the Corynebacterium culture comprises at least one of the following The Corynebacterium accolens strain SU001 or its derivatives, variants or mutants with the accession number NCAIM P(B)001495 deposited at NCAIM (National Collection of Agricultural and Industrial Microorganisms, Institute of Food Science and Technology, Szent István University, H-1118 Budapest, Somlóu ut 14-16, Hungary) on June 22, 2021, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression; The Corynebacterium propionicum strain SU002 or its derivatives, variants or mutants with the accession number NCAIM P(B)001496 deposited at NCAIM on June 22, 2021, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression; The Corynebacterium propionicum strain SU003 or its derivatives, variants or mutants with the accession number NCAIM P(B)001497 deposited at NCAIM on June 22, 2021, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression; The Corynebacterium accolens strain SU004 or its derivatives, variants or mutants with the accession number NCAIM P(B)001500 deposited at NCAIM on January 17, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression; The Corynebacterium tuberculostearicum strain SU005 or its derivatives, variants or mutants with the accession number NCAIM P(B)001501 deposited at NCAIM on April 27, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression; The Corynebacterium accolens strain SU006 or its derivatives, variants or mutants with the accession number NCAIM P(B)001502 deposited at NCAIM on April 27, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression; Corynebacterium propionicum strain SU007 or its derivatives, variants or mutants with accession number NCAIM P(B)001504 deposited at NCAIM on April 27, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression; Corynebacterium accolens strain SU008 or its derivatives, variants or mutants with accession number NCAIM P(B)001505 deposited at NCAIM on April 27, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression; Corynebacterium accolens strain SU009 or its derivatives, variants or mutants with accession number NCAIM P(B)001506 deposited at NCAIM on May 25, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression; Corynebacterium propionicum strain SU010 or its derivatives, variants or mutants with accession number NCAIM P(B)001507 deposited at NCAIM on May 25, 2022, wherein the derivatives, variants or mutants thereof are capable of reducing cathepsin expression; Preferably, any derivatives, variants or mutants capable of reducing cathepsin expression are derivatives, variants or mutants that reduce the relative expression of cathepsin mRNA after incubation with Caco-2 cells for 24 hours, respectively.

9. A Corynebacterium accolens strain SU004, which was deposited at NCAIM on January 17, 2022, with accession number NCAIM P(B)001500.

10. A Corynebacterium tuberculostearicum strain SU005, which was deposited at NCAIM on April 27, 2022, with accession number NCAIM P(B)001501.

11. A Corynebacterium accolens strain SU006, which was deposited at NCAIM on April 27, 2022, with accession number NCAIM P(B)001502.

12. A Corynebacterium propionicum strain SU007, which was deposited at NCAIM on April 27, 2022, with accession number NCAIM P(B)001504.

13. A Corynebacterium accolens strain SU008, which was deposited at NCAIM on April 27, 2022, with accession number NCAIM P(B)001505.

14. A Corynebacterium accolens strain SU009, which was deposited at NCAIM on May 25, 2022, with accession number NCAIM P(B)001506.

15. A Corynebacterium propionicum strain SU010, which was deposited at NCAIM on May 25, 2022, with accession number NCAIM P(B)001507.

16. A Corynebacterium strain or any of its derivatives, variants or mutants capable of reducing cathepsin expression according to any one of claims 9 to 15, for use in preventing infection by SARS-CoV-2 virus in a subject.

17. A Corynebacterium culture according to any one of claims 1 to 7, wherein, The Corynebacterium culture comprises four Corynebacterium strains selected from: (i) Corynebacterium accolens SU004, Corynebacterium tuberculostearicum SU005, Corynebacterium accolens SU009 and Corynebacterium propionicum SU010, or any of its derivatives, variants or mutants capable of reducing cathepsin expression; (ii) Corynebacterium accolens SU004, Corynebacterium tuberculostearicum SU005, Corynebacterium accolens SU006 and Corynebacterium propionicum SU010, or any of its derivatives, variants or mutants capable of reducing cathepsin expression; and (iii) Corynebacterium accolens SU004, Corynebacterium tuberculostearicum SU005, Corynebacterium propionicum SU007 and Corynebacterium accolens SU008, or any of its derivatives, variants or mutants capable of reducing cathepsin expression.

18. A Corynebacterium culture for preventing infection with the SARS-CoV-2 virus in a subject, wherein, The Corynebacterium culture comprises four Corynebacterium strains, which are Corynebacterium accolens SU004, Corynebacterium tuberculostearicum SU005, Corynebacterium propionicum SU007 and Corynebacterium accolens SU008.

19. A freeze-dried preparation for preventing the infection of coronavirus in a subject, wherein, The lyophilized preparation comprises a Corynebacterium culture or strain as defined in any of the preceding claims, and a cryoprotectant.

20. The lyophilized preparation according to claim 19, (i) wherein the coronavirus is SARS-CoV-2, and (ii) Among them, the lyophilized preparation comprises a Corynebacterium culture or strain as defined in any of claims 7 to 16, and (iii) wherein the cryoprotectant comprises a hydrocolloid polymer, and a carbohydrate or carbohydrate derivative, preferably, wherein the carbohydrate or carbohydrate derivative is selected from trehalose, sucrose, glucose, lactose, mannitol, sorbitol, inulin, maltodextrin and isomaltulose, preferably selected from maltodextrin and isomaltulose, and preferably, wherein the hydrocolloid polymer is selected from hydroxypropyl methylcellulose, hydroxyethyl cellulose, carboxymethyl cellulose, guar gum, carrageenan and xanthan gum, preferably selected from κ-carrageenan and xanthan gum.

21. The lyophilized preparation used according to claim 20, wherein, The carbohydrate or carbohydrate derivative is isomaltulose at a concentration of 0.5 g / 100 mL to 2.5 g / 100 mL, and the hydrocolloid polymer is κ-carrageenan at a concentration of 0.1 g / 100 mL to 0.5 g / 100 mL or xanthan gum at a concentration of 0.05 g / 100 mL to 0.25 g / 100 mL, and / or wherein, The Corynebacterium culture comprises from 10 3 CFU / mL to 10 11 CFU / mL of Corynebacterium strains, preferably from 10 3 CFU / mL to 10 11 CFU / mL of each Corynebacterium strain.

22. The lyophilized preparation according to any of claims 19 to 21, wherein: The freeze-dried preparation comprises a Corynebacterium culture, the Corynebacterium culture comprising four Corynebacterium strains, namely Corynebacterium accolens SU004, Corynebacterium tuberculostearicum SU005, Corynebacterium accolens SU006 and Corynebacterium propionicum SU010; and The cryoprotectant comprises κ-carrageenan or xanthan gum.

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