WO2025257401A1

WO2025257401A1 - Novel isophthalate compounds for the treatment of a viral lung infection

Info

Publication number: WO2025257401A1
Application number: PCT/EP2025/066598
Authority: WO
Inventors: Ameni HADJ MOHAMED; Antoine GUILLON; Samir Messaoudi; Mustapha SI-TAHAR; Adeline CEZARD
Original assignee: Chru De Tours; Centre National de la Recherche Scientifique CNRS; Institut National de la Sante et de la Recherche Medicale INSERM; Universite de Tours; Universite Paris Saclay
Current assignee: Chru De Tours; Centre National de la Recherche Scientifique CNRS; Institut National de la Sante et de la Recherche Medicale INSERM; Universite de Tours; Universite Paris Saclay
Priority date: 2024-06-14
Filing date: 2025-06-13
Publication date: 2025-12-18
Anticipated expiration: 2026-12-14

Abstract

The present invention relates to a compound of formula (I) and its use in a method of treatment of a viral infection, particularly of a viral lung infection. The present invention further relates to pharmaceutical compositions comprising the compound of formula (I).

Description

NOVEL COMPOUNDS FOR THE TREATMENT OF A VIRAL INFECTION

Background

Viruses are small infectious agents that replicate only inside living cells of other organisms. They can infect all types of life forms, from animals and plants to microorganisms, including bacteria and archaea. Among them, more than 400 species of virus are known to be responsible of diseases in humans, many of them leading to serious pathologies and eventually death.

In particular, lung viral infections can be either a persistent and pervasive burden, such as influenza infections, or a sudden worldwide disruptive outbreak, such as SARS-CoV-2. This recent pandemic has shed new light on the critical importance of viruses in respiratory infections.

Influenza A virus (IAV) causes significant morbi/mortality each year, since the 1918 pandemic, and influenza pathogenesis as well as anti-influenza therapeutic strategies have been extensively investigated. The pathophysiology of influenza lung infection is the result of two phenomena: (i) the intrinsic viral pathogenicity, linked to its tropism for the airway cells of the host, and (ii) the adverse immune response of the subject, which usually comprises a hyper- inflammatory immune response. Indeed, a robust host immune response is required for the viral clearance but the massive cellular recruitment and release of cytotoxic molecules lead to lung hyper-inflammation and can be associated with lung damage, morbidity and death.

Current anti-lAV approaches, such as vaccination and antivirals, exhibit suboptimal effectiveness. The short duration of vaccine- induced immunity and the antigenic drift of influenza viruses compromise host protection. Additionally, skepticism persists regarding the efficacy of approved anti-influenza drugs like the neuraminidase inhibitor oseltamivir (Tamiflu®). Consequently, innovative strategies are crucial for improving influenza virus treatments.

The pathophysiology of influenza-related pneumonia stems from both viral pathogenicity and the host immune response. While a robust immune response is essential for viral clearance, excessive cellular recruitment and release of cytotoxic molecules lead to lung hyperinflammation, resulting in tissue damage, morbidity, and death. Given the multifaceted nature of influenza pathophysiology, involving both cytopathic viral effects and excessive inflammatory responses, there is a growing interest in exploring host-directed therapies for more effective interventions.

Recent discoveries in metabolic reprogramming of immune cells have opened new therapeutic avenues for modulating immune responses. Metabolism is fundamental to all biological functions, and the integration of metabolism with immunity, known as immunometabolism, is at the forefront of immunology research.

Building on this finding, the Applicants designed and studied a series of novel compounds for their anti-viral activity in a model of influenza viral infection. Given that influenza pathophysiology involves both cytopathic viral effects and excessive inflammatory responses, the anti-inflammatory properties of these compounds was also examined. The Applicants found that some of these compounds not only allow to inhibit influenza virus replication but also exert anti-inflammatory properties that are potent enough to disrupt the inflammatory cascades during influenza. Brief description

Accordingly, an embodiment E1 of the present disclosure relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein:

X is selected from CH and N,

Y is selected from NR₄, S, S(O), S(O)₂, S(O)(NH), Se, Se(O), Se(O)₂, Se(O)(NH), O, CO, phenyl

Z is selected from O and NH,

Ri, R₂, R₅, Re are independently selected from H, Na, K, C1-6 alkyl,

R3 is selected from C1-6 alkyl, aryl, alkyne, CO₂Rs, alkyl(CO₂Re)_n, -S-R7, -Se-R?, NR₄, halogen, sugar, aminoacid, peptide,

R₄ is selected from H, C1-6 alkyl, C1-6 alkyl-aryl,

R₇ is a C1-6 alkyl optionally substituted by m COOH moiety, m being an integer from 1 to 12, n being an integer from 1 to 12.

An embodiment E2 of the present disclosure relates to the compound according to embodiment E1 , wherein said compound is a compound of formula (la) or (lb) wherein:

Z is selected from O and NH,

R1, R₂, R₅, Re are independently selected from H, Na, K, C1-6 alkyl,

R₄ is selected from H, C1-6 alkyl, C1-6 alkyl-aryl,

An embodiment E3 of the present disclosure relates to the compound according to embodiment E1 or E2, wherein the compound is a compound of formula (Ic), (Id), (le), (If), (Ig), (Ih), (li), (Ij), (Ik), (II), (Im) or (In)

wherein:

X is selected from CH and N,

Z is selected from O and NH,

Ri, R₂, Rs, Re are independently selected from H, Na, K, C1-6 alkyl,

R3 is selected from C1-6 alkyl, aryl, alkyne, CO2R5, alkyl(CO2Re)n, -S-R7, -Se-R?, NR₄, halogen, sugar, aminoacid, peptide,

R₄ is selected from H, C1-6 alkyl, C1-6 alkyl-aryl,

An embodiment E4 of the present disclosure relates to the compound according to any one of embodiments E1 to E3, wherein the compound is selected from the group consisting of:

An embodiment E5 of the present disclosure relates to the compound according to any one of embodiments E1 to E4, for use as a medicine. An embodiment E5 of the present disclosure relates to a compound according to any one of embodiments E1 to E4, for use as a medicine.

An embodiment E6 of the present disclosure relates to a pharmaceutical composition, comprising a compound as defined in any one of embodiments E1 to E4 and a pharmaceutically acceptable support.

An embodiment E7 of the present disclosure relates to the compound according to any one of embodiments E1 to E5 or the pharmaceutical composition according to embodiment E6, for use in a method of treatment of a viral infection and/or an adverse immune response to a viral infection, the method comprising administering to a subject in need thereof an effective amount of said compound or pharmaceutical composition.

An embodiment E8 of the present disclosure relates to the compound or pharmaceutical composition for use in a method according to embodiment E7, wherein the viral infection is a lung viral infection.

An embodiment E9 of the present disclosure relates to the compound or pharmaceutical composition for use in a method according to any of embodiment E7 or E8, wherein the viral infection is caused by a respiratory virus selected from an influenza virus, a coronavirus, a respiratory syncytial virus, a parainfluenza virus, a metapneumovirus, a rhinovirus, an adenovirus, a varicella-zoster virus, a cytomegalovirus, a paramyxovirus such as Nipah virus, an enterovirus and a bocavirus.

An embodiment E10 of the present disclosure relates to the compound or pharmaceutical composition for use in a method according to any one of embodiments E7 to E9, wherein the viral infection is caused by an influenza virus.

An embodiment E11 of the present disclosure relates to the compound or pharmaceutical composition for use in a method according to any one of embodiments E7 to E10, wherein the method is applied to treat the viral infection.

An embodiment E12 of the present disclosure relates to the compound or pharmaceutical composition for use in a method according to any one of embodiments E7 to E1 1 , wherein the method is applied to treat the adverse immune response to the viral infection.

An embodiment E13 of the present disclosure relates to the compound or pharmaceutical composition for use in a method according to any one of embodiments E7 to E12, wherein the method is applied to prevent the viral infection from escalating to an adverse immune response.

An embodiment E14 of the present disclosure relates to the compound or pharmaceutical composition for use in a method according to any one of embodiments E7 to E13, wherein the adverse immune response to the viral infection is selected from a hyper-inflammatory immune response, a dyspnea, a tachypnea, a pneumonia in particular an acute pneumonia, an exacerbation of a chronic respiratory disease such as asthma or chronic obstructive pulmonary disease, a sepsis, a septic shock, a cytokine storm or an acute respiratory distress syndrome (ARDS).

An embodiment E15 of the present disclosure relates to the compound or pharmaceutical composition for use in a method according to any one of embodiments E7 to E14, wherein the compound or pharmaceutical composition is administered between 2 and 14 days post infection. Detailed description

As illustrated by examples, the inventors have demonstrated the therapeutic interest of the compound of formula (I) according to the disclosure.

Compound

An aspect of the present disclosure thus relates to a compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein:

X is selected from CH and N,

Y is selected from NR₄, S, S(O), S(O)₂, S(O)(NH), Se, Se(O), Se(O)₂, Se(O)(NH), O, CO, phenyl,

Z is selected from O and NH,

Ri, R₂, R₅, Re are independently selected from H, Na, K, C1-6 alkyl,

Rs is selected from C1-6 alkyl, aryl, alkyne, CO₂Rs, alkyl(CO₂Re)_n, -S-R7, -Se-R?, NR₄, halogen, sugar, aminoacid, peptide,

R₄ is selected from H, C1-6 alkyl, C1-6 alkyl-aryl,

In the context of the disclosure, the term “C1-6 alkyl” is to be understood as Ci alkyl, C₂ alkyl, C3 alkyl, C₄ alkyl, C5 alkyl and C& alkyl. The term “C1-6 alkyl” refers to C1-6 linear (/.e., "straightchain"), branched, or cyclic, saturated hydrocarbon chains, including for example, methyl, ethyl, propyl, isopropyl, butyl, isobutyl groups.

In the context of the disclosure, the term “aryl” refers to a group derived from a monocyclic or polycyclic aromatic hydrocarbon by removal of a hydrogen atom from a ring carbon atom, including for example phenyl, tolyl, naphtyl groups.

In the context of the disclosure, the term “alkyne” refers to an unsaturated hydrocarbon containing at least one carbon-carbon triple bond.

In the context of the disclosure, the term “halogen” refers to fluorine, chlorine, bromine, iodine.

In the context of the disclosure, the term “sugar” refers to carbohydrates which include monosaccharides, oligosaccharides and polysaccharides as well as substances derived from monosaccharides by reduction of the carbonyl group (alditols), by oxidation of one or more terminal groups to carboxylic acids, or by replacement of one or more hydroxy group(s) by a hydrogen atom, an amino group, thiol group or similar groups. It also includes derivatives of these compounds. Sugar derivatives are chemical compounds derived from sugars (carbohydrates) through structural modifications of their basic molecular framework. These modifications typically involve the addition, substitution, or removal of functional groups, while retaining the core carbon backbone characteristic of the sugar molecule. In the context of the disclosure, the term “aminoacid” refers to an organic compound containing both amino and carboxylic acids functional groups.

In the context of the disclosure, the term “peptide” refers to amides derived from two to ten amino carboxylic acid molecules (the same or different) by formation of a covalent bond from the carbonyl carbon of one to the nitrogen atom of another with formal loss of water. The term peptide applies to structures formed from a-amino acids, but it also includes those derived from any amino carboxylic acid.

In the context of the disclosure, in the terms “n being an integer from 1 to 12”, the range “1 to 12” is to be understood as covering each integer from 1 and 12 and all the ranges comprised in the range from 1 to 12, for examples 2 to 12 or 3 to 12.

In some embodiments, the compound of formula (I) is a compound of formula (la) or (lb)

Z is selected from O and NH,

R1, R₂, R₅, Re are independently selected from H, Na, K, C1-6 alkyl,

R₄ is selected from H, C1-6 alkyl, C1-6 alkyl-aryl,

In some embodiments, the compound of formula (I) is a compound of formula (Ia1 ) or (Ib1 ) wherein Ri, R₂, R3 and Y are as previously defined.

In some embodiments, the compound of formula (I) is a compound of formula (Ia2) or (Ib2) wherein R1, R₂, R3 and Y are as previously defined.

In some embodiments, the compound of formula (I) is a compound of formula (Ic), (Id), (le), (If), (Ig), (Ih), (li), (Ij), (Ik), (II), (Im), or (In) wherein:

X is selected from CH and N, Z is selected from O and NH,

Ri, R₂, Rs, Re are independently selected from H, Na, K, C1-6 alkyl, R3 is selected from C1-6 alkyl, aryl, alkyne, CO2R5, alkyl(CO2Re)n, -S-R7, -Se-R?, NR₄, halogen, sugar, aminoacid, peptide,

R₄ is selected from H, C1-6 alkyl, C1-6 alkyl-aryl, R₇ is a C1-6 alkyl optionally substituted by m COOH moiety, m being an integer from 1 to 12, n being an integer from 1 to 12.

According to a particular embodiment, compounds of the invention are compounds of formula (Ida), (lea), (Ifa) or (Iga):

wherein:

X is selected from CH and N,

Z is selected from O and NH,

Ri, R2, Rs are independently selected from H, Na, K, C1-6 alkyl,

R₉ is selected from H, C1-6 alkyl optionally substituted by r COOH moiety, r being an integer from 1 to 12, or NHRu q being an integer from 0 to 12,

R11 being selected from H, acetyl, C1-6 alkyl, C1-6 alkyl-aryl.

According to a particular embodiment, compounds of the invention are of formulas (Ida), (lea), (Ifa) or (Iga), wherein q is 0, and

R₉ is C1-6 alkyl optionally substituted by r COOH moiety, r being an integer from 1 to 12.

According to a particular embodiment, compounds of the invention are of formulas (Ida), (lea), (Ifa) or (Iga), wherein q is 1 , and R₉ is H.

According to a particular embodiment, compounds of the invention are of formulas (Ida), (lea), (Ifa) or (Iga), wherein X is CH

Z is O, and q is 0.

In some embodiments, R₉ is selected from H, C1-6 alkyl optionally substituted by r COOH moiety, r being an integer from 1 to 12.

In some embodiments, compounds of the invention are of formula (Ida).

According to a particular embodiment, compounds of the invention of formula (In) have the following structure: wherein Ri, R2, R3, X and Z are as previously defined.

Formulas (le), (Ig), (Ij) and (II) designate respectively the following isomers: In some embodiments, the compound is a compound of formula (Ic1 ), ( Id 1 ), (Ie1 ), ( If 1 ), ( Ig 1 ), (Ih1), (Ii1 ), (Ij1), (Ik1), (111 ), (Im1 ) or (In1)

wherein X, Ri, R2, R3 and R4 are as previously defined.

Formulas (Ie1 ), (Ig1 ), (Ij1) and (111 ) designate respectively the following isomers:

In some embodiments, the compound is a compound of formula (Ic2), (Id2), (Ie2), (If2), (Ig2), (Ih2), (Ii2), (Ij2), (Ik2), (II2), (Im2) or (In2)

wherein X, Ri, R2, R3 and R4 are as previously defined.

Formulas (Ie2), (Ig2), (Ij2) and (112) designate respectively the following isomers: wherein Ri, R₂ and R3 are as previously defined.

In some embodiments, the compound as described herein is a compound selected from the group consisting of:

According to a particular embodiment, the compound for use according to the invention can be selected from the group consisting of:

The compounds of the present disclosure and their pharmaceutically acceptable salts may contain one or more asymmetric centers, chiral axes and chiral planes and may thus give rise to enantiomers, diastereomers, and other stereoisomeric forms and may be defined in terms of absolute stereochemistry, such as (R)- or (S)-. The present disclosure is intended to include all such possible isomers, as well as their racemic and optically pure forms. In the formulas of the compounds of the invention containing one or more asymmetric centers, the symbol * is used to designate such asymmetric center and to indicate that all isomers of such compounds are included in the compounds of the present invention.

In addition, the compounds of the present disclosure also may exist in hydrated and anhydrous forms. Hydrates of any of the formulas described herein may thus exist as a monohydrate or in the form of a polyhydrate.

As used herein, the term “pharmaceutically acceptable salt” is intended to mean base addition salts. Example of pharmaceutically acceptable salts are also described, for example, in Berge et al., “Pharmaceutical Salts”, J. Pharm. Sci. 66, 1 -19 (1977). Pharmaceutically acceptable salts may be synthesized from the parent agent that contains an acidic moiety, by conventional chemical methods. Generally, such salts and are prepared by reacting the free acid forms of these agents with a stoichiometric amount of the appropriate base in water or in an organic solvent, or in a mixture of the two. Likewise, when the parent agent contains a group such as -NH2, the pharmaceutically acceptable salts may be synthesized from the parent agent by conventional chemical methods by reacting the free -NH3+ with an anionic source in a suitable solvent.

Salts may be prepared in situ, during the final isolation or purification of the compound or by separately reacting a purified compound of the present disclosure with the desired corresponding base, and isolating the salt thus formed. For example, this approach may be implemented with the free acid form of some of the compounds of the present disclosure. The pharmaceutically acceptable salt of the compounds of the present disclosure may be selected from the group consisting of organic or inorganic salts.

For example, the pharmaceutically acceptable salt may include a sodium, potassium, calcium, magnesium, lithium, ammonium, manganese, zinc, iron, olamine, meglumine, lysine, tromethamine, or copper salt, when the compounds are amenable to be such salts. In preferred embodiments, the pharmaceutically acceptable salt of the compounds of the present disclosure may be the sodium, potassium, calcium, magnesium or lithium salt, when the compounds are amenable to be such salts. More preferably the pharmaceutically acceptable salt is sodium, when the compounds are amenable to be such salts.

For example, the pharmaceutically acceptable salt may include an acetate, benzoate, besylate, bromide, carbonate, citrate, edisylate, estolate, fumarate, gluconate, hippurate, iodide, maleate, mesylate, methylsulfate, napsylate, oxalate, pamoate, phosphate, stearate, succinate, sulfate, tartrate, tosylate, or chloride salt, when the compounds are amenable to be such salts.

All alcohol, salt and other ionic and non-ionic forms of the compounds described are included when referring to a given compound, where applicable. For example, if a compound is shown as an alcohol herein, the salt forms of the compound are also included, when the compounds are amenable to be such salts. Likewise, if a compound is shown as a salt herein, then the alcohol forms are also included. The same is also applicable to a compound having an aromatic group in one of the substituent groups, where such aromatic group on the substituent group may include a free form of a carboxylic acid. In such case, when the compound is shown as a salt herein, then the carboxylic acid free form is also included. Likewise, when the aromatic group on the substituent group is shown with a free form of a carboxylic acid, then the salt forms of the compound are also included, when the compounds are amenable to be such salts.

The compound of formula (I) according to the present disclosure can be prepared according to any chemical routes known from a skilled person, such the synthetic routes presented in the examples. It is thus understood that one skilled in the art of organic chemistry can easily synthesize the compound of formula (I) using appropriate starting materials, conventional chemicals reactions, standard and literatures procedures, and experimental conditions to synthesize the compounds of formula (I).

Compound or pharmaceutical composition for use

As illustrated by examples, the inventors have demonstrated the therapeutic interest of the compounds as described herein. Indeed, the inventors have shown that the compounds according to the present disclosure are useful in a method of treatment of a viral infection and/or an adverse immune response to a viral infection, especially of a lung viral infection such as an I AV infection.

Therefore, as it is further detailed herein the compound according to the disclosure may be useful as a drug, especially as an antiviral agent and/or an immunomodulatory agent.

Accordingly, the present disclosure relates to a compound according to the disclosure, for use as a drug or a medicine.

As further detailed herein, the present disclosure further relates to a pharmaceutical composition comprising a compound according to the disclosure and a pharmaceutically acceptable support.

Accordingly, the present disclosure also relates to a compound or pharmaceutical composition as described herein for use in a method of a viral infection and/or an adverse immune response to a viral infection, the method comprising administering to a subject in need thereof an effective amount of compound or pharmaceutical composition according to the disclosure.

Viral infection

As used in the present disclosure, the term “viral infection” relates to any condition characterized by (i) the proliferation of one or more virus in the subject, and/or (ii) one or more adverse immune response associated to or induced by said viral infection in the subject.

The viral infection is preferably a lung viral infection, i.e. a condition characterized by (i) the proliferation of one or more virus in the lungs of the subject, and/or (ii) one or more adverse immune response associated to or induced by said viral infection in the lungs of the subject. A lung is a specific organ within the respiratory system. Humans typically have two lungs. According to the present disclosure, lung viral infections may affect other organs of the respiratory system selected from the group consisting of the pharynx, the larynx or the tracheobronchial tree (composed of the trachea, the bronchi and the bronchioles).

A viral infection can cause a spectrum of symptoms, from no obvious symptoms to severe illness. The pathological changes in the lungs due to a viral infection typically include or more of cell damage (i.e. viral replication in lung cells leads to cell death and tissue damage, immune response (i.e. infiltration of immune cells such as lymphocytes, neutrophils and macrophages to combat the virus), inflammation (i.e. swelling and irritation of the lung tissues), fluid accumulation (i.e. fluid buildup in the alveoli, leading to impaired gas exchange, and mucus production (i.e. increased mucus production, which can block airways and cause breathing difficulties). These pathological changes primarily affect the respiratory system and may result in a decreased or impaired respiratory function in the subject. As it will be described below, a viral infection in particular a lung viral infection may also result in more systemic complications due to an adverse immune response associated to or induced by said viral infection in the lungs of the subject.

Virus-induced cvtopathic effects (CPE)

As used herein, the expression “virus-induced cytopathic effects” or “viral cytopathic effects” abbreviated “CPEs” refers to the visible structural changes or damage in the cells of the subject that result from the virus taking over the host cell's machinery to replicate and produce new viral particles, often disrupting normal cellular functions and ultimately leading to cell death. These changes can be observed under a microscope and are indicative of the presence and activity of a virus within the cells.

CPEs manifestations usually include one or more of cell rounding (i.e. the infected cells lose their normal shape and become rounded), cell detachment (i.e. the cells detach from the surface they are growing on), lysis (i.e. the cell membrane ruptures, leading to cell death), formation of viral inclusion bodies (i.e. formation of abnormal structures within the cell, such as aggregates of viral particles or altered host cell organelles), syncytium formation (i.e. infected cells fuse to form large multinucleated cells called syncytia), chromatin margination (i.e. chromatin in the nucleus condenses and is displaced to the edges of the nucleus), and vacuolization (i.e. formation of vacuoles or pockets within the cytoplasm).

The damage to respiratory epithelial cells, in particular, can impair the function of the respiratory tract and make it more susceptible to secondary bacterial infections. The CPEs can contribute to the pathogenesis and thus to the symptoms of the lung viral infection.

In some embodiments, the lung viral infection induces or is capable of inducing one or more viral cytopathic effects in the lungs of the subject. Therefore, in some embodiments, the compound or pharmaceutical composition as described herein is for use in a method of treatment of one or more viral cytopathic effects induced by a viral infection, in particular by a lung viral infection.

Adverse immune response

As used herein, the expression “adverse immune response”, or “host adverse immune response”, or “subject’s adverse immune response” refers to a syndrome of physiologic, pathologic, and/or biochemical abnormalities triggered by a viral infection. The expression “adverse immune response” refers also to organ dysfunction due to dysregulated host response to infection than can be defined as “sepsis”.

In some embodiments, the viral infection induces or can induce an adverse immune response in the subject. In some embodiments, the adverse immune response to the viral infection is a hyper-inflammatory immune response, a sepsis, a septic shock, or a cytokine storm.

In some embodiments, the adverse immune response to the viral infection is a multi-organ failure. Multi-organ failure may comprise heart failure, liver failure, lung failure kidney failure, or gastrointestinal (Gl) system failure.

Therefore, in some embodiments, the compound or composition according to the disclosure is for use in a method of treatment of an adverse immune response induced by a viral infection, the viral infection preferably being a lung viral infection.

Clinical manifestations of an adverse immune response to a lung viral infection usually include one or more of abnormal body temperature (typically characterized in human by a body temperature greater than 38°C or lower than 36°C), dyspnea, tachypnea (typically characterized in human by a respiratory rate greater than 20/min), bronchitis, pneumonia, sepsis-induced organ dysfunction, and acute respiratory distress syndrome (ARDS).

Biological manifestations usually include one or more of abnormal white blood cell count (typically characterized in human by a WBC count greater than 12000/mm³ or lower than 4000/mm³ or greater than 10% immature bands), excessive release of systemic cytokines such as tumor necrosis factor (TNF), interleukin-1 (IL-1 ), interleukin-6 (IL-6), interleukin-8 (IL-8), hyper-inflammatory immune response, such as a cytokine storm or cytokine release syndrome, coagulopathy (consumption of coagulation factors, fibrinogen, and platelets), hypoxemia assessed by arterial blood gas analysis (typically characterized by a PaO2/FiO2 inferior or equal to 300 mmHg). Imaging manifestation usually includes the presence of pulmonary infiltrates on chest radiograph and/or CT scan.

In some embodiments, the adverse immune response to the viral infection is a hyper- inflammatory immune response, a dyspnea, a tachypnea, a pneumonia in particular an acute pneumonia, an exacerbation of a chronic respiratory disease such as asthma or chronic obstructive pulmonary disease, a sepsis, a septic shock, a cytokine storm or an acute respiratory distress syndrome (ARDS).

In some embodiments, the adverse immune response to the lung viral infection is pulmonary hypertension or hypotension.

Immunomodulatory and antiviral properties

In some embodiments, the compound or pharmaceutical composition for use according to the disclosure has immunomodulatory properties and/or antiviral properties, preferably wherein:

- the immunomodulatory properties include the reduction or inhibition of the inflammation response to a viral infection;

- the antiviral properties include the reduction or inhibition of viral replication. Immunomodulatory properties

In some embodiments, the compound or pharmaceutical composition according to the disclosure has immunomodulatory properties. Therefore, in some embodiments, the compound or pharmaceutical composition according to the disclosure may be used as an immunomodulatory agent or an immunomodulatory medicament.

In the present disclosure, the terms “immunomodulatory agent”, “immunomodulatory medicament” or “immunomodulator” refer to a substance or composition that inhibits or reduces one or more adverse immune responses to a disease due to a virus in the subject, in particular a hyper-inflammatory immune response to a viral infection. The capacity of a substance to reduce an inflammatory response may for example be confirmed by measuring a diminution of the expression of some components of the inflammatory cascade such as IL- 6, IL-8 or tumor necrosis factor-alpha (TNF-a) and/or a diminution of the infiltration of immune cells such as lymphocytes, neutrophils and macrophages in the presence of that substance or composition.

In some embodiments, the compound reduces the concentration of interleukin-6 (IL-6) by at least 50% in an in vitro assay assessing anti-inflammatory activity, wherein human bronchial epithelial cells (e.g. BEAS-2B cells) are infected with influenza A virus (e.g. A/Scotland/20/74, H3N2 strain) and treated with the compound four hours post-infection, IL-6 levels being measured in cell culture supernatants at 20 hours post-infection, and wherein said reduction is preferably observed at a compound concentration of 5 mM or less, preferably 4.5 mM or less, more preferably 4 mM or less, even more preferably 3.5 mM or less, 3 mM or less, 2.5 mM or less, 1 .5 mM or less, 1 mM or less, or most preferably 0.5 mM or less. Such an in vitro assay for assessing anti-inflammatory activity is further detailed in the Examples. An in vitro assay for assessing anti-inflammatory activity of the compound is further described in the Examples.

In some embodiments, the immunomodulatory properties of the compound or pharmaceutical composition for use according to the disclosure include inhibiting or reducing the lung immune inflammation response triggered by a virus, in particular a respiratory virus.

Therefore, in some embodiments, the compound or pharmaceutical composition for use according to the disclosure treats the lung viral infection as an immunomodulatory medicament, more particularly by inhibiting or reducing the lung immune inflammation response triggered by the lung virus infection.

In some embodiments, the compound or pharmaceutical composition for use according to the disclosure is for treating an adverse immune response to a lung virus infection. The adverse immune response typically involves a hyper-inflammatory immune response and is preferably selected from acute exacerbations of a chronic respiratory disease such as asthma or chronic obstructive pulmonary disease, pneumonia, sepsis, septic shock, cytokine storm and acute respiratory distress syndrome. Such adverse immune response may occur in moderate to advanced cases of said lung virus infection, which may be caused by an influenza virus or a coronavirus such as SARS-CoV-2.

Antiviral properties

In some embodiments, the compound or pharmaceutical composition for use according to the disclosure has antiviral properties. Therefore, in some embodiments, the compound or pharmaceutical composition according to the disclosure may be used as an antiviral agent or an antiviral medicament.

In the present disclosure, the terms “antiviral medicament”, “antiviral agent” or “antiviral” refer to a substance or composition that inhibits or reduces virus replication in a subject. In some embodiments, the antiviral properties of the compound or pharmaceutical composition for use according to the disclosure include inhibition of viral replication.

In some embodiments, the compound or pharmaceutical composition for use according to the disclosure treats a lung viral infection as an antiviral medicament, more particularly by disrupting the life cycle of the respiratory virus.

In some embodiments, the compound reduces influenza A virus neuraminidase (NA) activity by at least 50% in an in vitro assay, wherein human bronchial epithelial BEAS-2B cells are infected with influenza A virus (e.g. A/Scotland/20/74, H3N2 strain) and treated with the compound four hours post-infection, NA activity being measured in cell culture supernatants at 20 hours post-infection, and wherein said reduction is preferably observed at a compound concentration of 5 mM or less, preferably 4.5 mM or less, more preferably 4 mM or less, even more preferably 3.5 mM or less, 3 mM or less, 2.5 mM or less, 1 .5 mM or less, 1 mM or less, or most preferably 0.5 mM or less. An in vitro assay for assessing antiviral activity of the compound is further described in the Examples.

In some preferred embodiments, the compound or pharmaceutical composition for use according to the disclosure treats the lung viral infection both as an immunomodulatory agent or medicament and as an antiviral agent or medicament.

In some embodiments, the compound for use according to the disclosure inhibits or reduces both viral replication and virus-induced inflammation. In some embodiments, the compound is an anti-inflammatory and/or antiviral compound.

Respiratory virus

In some embodiments, the viral infection is caused by a respiratory virus.

In the present disclosure, the term “respiratory virus” refers to a virus having a tropism for the airway cells and in particular for the lungs of the subject, and able to cause an adverse immune response as described herein.

In some embodiments, the respiratory virus is selected from an influenza virus, a coronavirus, a respiratory syncytial virus, a parainfluenza virus, a metapneumovirus, a rhinovirus, an adenovirus, a varicella-zoster virus, a cytomegalovirus, a paramyxovirus such as a Nipah virus, an enterovirus and a bocavirus.

Influenza virus

In some embodiments, the respiratory virus is an influenza virus.

In some embodiments, the influenza virus is an Influenza A virus, an Influenza B virus, an Influenza C virus or an Influenza D virus.

In some embodiments, the influenza A virus is of serotype H1 N1 , H1 N2, H2N2, H2N3, H3N1 , H3N2, H3N8, H5N1 , H5N2, H5N3, H5N6, H5N8, H5N9, H6N1 , H6N2, H7N1 , H7N2, H7N3, H7N4, H7N7, H7N9, H9N2, or H10N7.

In some embodiments, the influenza virus is an Influenza B virus.

In some embodiments, the Influenza B virus is of serotype Victoria or Yamagata.

In some embodiments, the influenza virus is an influenza virus usually causing human influenza, for example a H1 N1 , H2N2, H3N2, H5N1 , H7N7, H1 N2, H9N2, H7N2, H7N3, or H10N7 influenza virus.

In some embodiments, the influenza virus is an influenza virus usually causing avian influenza, for example a H5N1 or H7N9 influenza virus. In some embodiments, the influenza virus is an influenza virus usually causing swine or pig influenza such as H1 N1 , H1 N2, H2N1 , H3N1 , H3N2, H2N3, or influenza c virus.

In some embodiments, the influenza virus is an influenza virus usually causing equine influenza such as H7N7 or H3N8 influenza virus.

In some embodiments, the influenza virus is an influenza virus usually canine influenza such as H3N8 influenza virus.

In some embodiment, the compound or pharmaceutical composition for use according to the disclosure reduces or inhibits both the influenza virus replication and the immune inflammation response to an influenza virus infection.

Coronavirus

In some embodiments, the respiratory virus is a coronavirus virus of Orthocoronavirinae subfamily, as a coronavirus.

In some embodiments the coronavirus is selected from severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), severe acute respiratory syndrome coronavirus (SARS-CoV), or Middle East respiratory syndrome coronavirus (MERS-CoV) or beta-coronavirus.

In some embodiments the coronavirus is SARS-CoV-2.

In some embodiments, the compound or pharmaceutical composition for use according to the disclosure reduces or inhibits at least the immune inflammation response to the coronavirus infection, and optionally also reduces the immune inflammation response to a coronavirus infection.

Viral respiratory disease

In some embodiments, the compound or pharmaceutical composition for use according to the disclosure is for use in treating a respiratory viral disease.

As used in the present disclosure, the term “viral respiratory disease” relates to a condition or disorder that affects the respiratory system.

In some embodiments, the compound or pharmaceutical composition for use according to the disclosure is for use in treating a respiratory viral disease characterized in that it is caused by a particular respiratory virus, especially as respiratory virus as described herein.

In some embodiments, the compound or pharmaceutical composition for use according to the disclosure is for use in treating a respiratory viral disease characterized in that it is caused by a respiratory virus is selected from an influenza virus, a coronavirus, a respiratory syncytial virus, a parainfluenza virus, a metapneumovirus, a rhinovirus, an adenovirus, a varicella-zoster virus, a cytomegalovirus, a paramyxovirus such as a Nipah virus, an enterovirus and a bocavirus.

In some embodiments, the compound or pharmaceutical composition for use according to the disclosure is for use in treating a respiratory viral disease characterized in that it is caused by an influenza virus.

Although viral respiratory diseases can be classified according to a causative virus (e.g., influenza), they may also be classified clinically irrespective of the causal agent according to a syndrome (e.g. bronchiolitis, pneumonia, acute respiratory distress syndrome). In the present disclosure, the term “syndrome” refers to recognizable patterns of symptoms and signs that occur together and characterize a particular viral respiratory disease or condition. Thus, a viral respiratory disease may be characterized by a particular syndrome. Specific viruses commonly cause characteristic clinical manifestations (e.g., rhinovirus typically causes the common cold, respiratory syncytial virus (RSV) typically causes bronchiolitis), but can also cause the general symptoms of diseases caused by other viruses. These symptoms can include cough, sore throat, runny or stuffy nose, fever, fatigue, muscle aches, and difficulty breathing in severe cases.

"Influenza", commonly known as the flu refers to the collection of symptoms and signs associated with an influenza virus infection, primarily influenza A and B viruses. The influenza syndrome typically manifests suddenly and can range from mild to severe, sometimes leading to hospitalization and even death, particularly in high-risk groups such as young children, elderly individuals, pregnant women, and those with underlying health conditions.

"Influenza encephalitis", or “influenza-associated encephalitis”, refers to a rare yet serious neurological condition that can arise from an influenza virus infection. This complication generally follows a severe infection by specific strains of the influenza virus, particularly influenza A and B, and predominantly affects children and young adults. Clinical characteristics encompass a combination of flu symptoms and central nervous system dysfunctions, in addition to the conventional flu symptoms, there are neurological manifestations such as seizures, alterations in consciousness, diminished cognitive processing, including speech, motor paralysis or sensory loss, unusual or delirious behaviour, and shifts in mental state. The emergence of neurological complications may occur within a span of several days following the initial onset of flu symptoms.

"Influenza-like illness" (ILI) is a term used to describe a set of symptoms that are similar to those of influenza. These symptoms typically include fever, cough, sore throat, muscle aches, fatigue, and sometimes headache and nasal congestion. ILI can be caused by influenza viruses or various respiratory viruses besides influenza, including respiratory syncytial virus (RSV), adenovirus, rhinovirus, and coronaviruses.

"Bronchiolitis" refers to a respiratory condition characterized by an inflammation of the bronchioles, the smallest air passages in the lungs. It mainly occurs in infants and young children. Symptoms of bronchiolitis often start with cold-like symptoms such as a runny or stuffy nose, cough, and mild fever. As the illness progresses, symptoms can worsen to include wheezing, difficulty breathing, rapid breathing, and sometimes a decreased appetite or dehydration. It is most commonly caused by a respiratory syncytial virus (RSV) infection but may be caused by other viruses including influenza viruses, parainfluenza viruses, adenoviruses, rhinoviruses, coronaviruses and metapneumoviruses.

“Bronchitis” refers to a respiratory condition characterized by an inflammation of bronchi, which are the larger airways that branch off from the trachea and lead to the lungs. It can occur in all age groups. Common Symptoms of bronchitis are rapid breathing, wheezing, cough, nasal congestion, difficulty feeding (in infants), and sometimes fever. It is commonly caused by a respiratory syncytial virus (RSV) infection but may be caused by other viruses including influenza viruses, parainfluenza viruses, adenoviruses, rhinoviruses, coronaviruses and metapneumoviruses.

“Common cold” refers a viral infection of the upper respiratory tract, primarily caused by rhinoviruses. Other viruses, such as coronaviruses, adenoviruses, and respiratory syncytial virus (RSV), can also cause colds. It is characterized by symptoms such as runny or stuffy nose, sneezing, sore throat, cough, fatigue and fever. It is primarily caused by Rhinoviruses but may be caused by other viruses including influenza viruses, parainfluenza viruses, enteroviruses, adenoviruses, and metapneumoviruses. “Croup” refers to a respiratory condition that primarily affects infants and young children, characterized by inflammation and swelling of the upper airway, particularly the larynx (voice box) and trachea (windpipe). It typically presents with a barking cough, hoarseness, and stridor (a high-pitched sound heard during inhalation). It is primarily caused by parainfluenza viruses but may be caused by other viruses including influenza viruses, RSV or adenoviruses.

In some embodiments, the compound or pharmaceutical composition for use according to the disclosure is for use in treating a respiratory viral disease selected from influenza, influenza encephalitis, influenza-like illness, bronchiolitis, bronchitis, common cold, and croup.

Some viral respiratory diseases are characterized by syndromes arising as an adverse immune response to a viral infection. Such syndromes can include pneumonia (inflammation of the air sacs in one or both lungs), sepsis (widespread inflammation, which can lead to organ dysfunction and failure) or acute respiratory distress syndrome (lungs are unable to provide adequate oxygen to the body's tissues and/or remove carbon dioxide from the bloodstream).

“Pneumonia” refers to a respiratory condition characterized by inflammation of the air sacs (alveoli) in one or both lungs. The alveoli may fill with fluid or pus, causing symptoms such as cough, fever, chills, and difficulty breathing. Pneumonia can be caused by a variety of infectious agents, including viruses, bacteria and fungi, as well as by inhalation of certain chemicals or irritants. Viral pneumonia is commonly caused by influenza viruses, RSV, adenoviruses and coronaviruses including SARS-CoV-2 but may be caused by other viruses including parainfluenza viruses, enteroviruses, rhinoviruses, metapneumoviruses.

“Sepsis” refers to a condition characterized by widespread inflammation that may lead to tissue damage and organ failure. Sepsis can be caused by a variety of infectious agents, including viruses, bacteria and fungi. Viral sepsis is commonly caused by influenza viruses, RSV, adenoviruses and coronaviruses including SARS-CoV-2 but may be caused by other viruses including parainfluenza viruses, enteroviruses, rhinoviruses, metapneumoviruses.

“Acute respiratory distress syndrome” (ARDS) is a respiratory condition characterized by rapid onset of widespread inflammation in the lungs, leading to fluid accumulation in the air sacs (alveoli) and resulting in severe oxygenation impairment and difficulty breathing. ARDS typically occurs as a complication of another underlying illness or injury, such as pneumonia, sepsis, trauma, or inhalation injury. Viral ARDS is commonly caused by influenza viruses, RSV, adenoviruses and coronaviruses including SARS-CoV-2 but may be caused by other viruses including parainfluenza viruses, enteroviruses, rhinoviruses, metapneumoviruses.

In some embodiments, the compound or pharmaceutical composition for use according to the disclosure is for use in treating a respiratory viral disease selected from pneumonia, sepsis, or acute respiratory distress syndrome (ARDS).

Some viral respiratory syndromes and their viral cause are illustrated below:

In some embodiments, the compound or pharmaceutical composition for use according to the disclosure is for use in treating a respiratory viral disease selected from influenza, influenza encephalitis, influenza-like illness, bronchiolitis, bronchitis, common cold, croup, pneumonia, sepsis, and acute respiratory distress syndrome (ARDS).

Method of treatment

As used in this document, the term “treatment” or “therapy” refers to any action which makes it possible to reduce or suppress the symptoms associated with a pathological condition. It comprises both a curative treatment and a prophylactic treatment for a disease.

A curative treatment is defined by a treatment resulting in a cure or a treatment which relieves, improves and/or eliminates, reduces and/or stabilizes the symptoms of a disease or the suffering that it causes. The term “curative treatment" may refer to one or more of (1 ) inhibiting the disease; for example, inhibiting a disease, condition or disorder in an individual who is experiencing or displaying the pathology or symptomatology of the disease, condition or disorder (i.e., arresting further development of the pathology and/or symptomatology); and (2) ameliorating the disease; for example, ameliorating a disease, condition or disorder in an individual who is experiencing or displaying the pathology or symptomatology of the disease, condition or disorder (i.e., reversing the pathology and/or symptomatology) such as decreasing the severity of disease or reducing or alleviating one or more symptoms of the disease. In particular, with reference to the treatment of a lung viral infection, the term “curative treatment” may refer to the inhibition of the viral infection and/or of the adverse immune reaction associated to said viral infection. A prophylactic treatment comprises both a treatment resulting in the prevention of a disease and a treatment which reduces and/or delays the incidence of a disease or the risk of it occurring. The terms “improve” and “reduce” include, but do not require complete recovery or complete prevention. The term “prophylactic treatment" may refer to one or more of preventing the disease; for example, preventing a disease, condition or disorder in an individual who is at risk of experiencing or displaying the pathology or symptomatology of the disease, condition or disorder (i.e., preventing the development of the pathology and/or symptomatology); and (2) reducing and/or delaying the incidence of a disease in an individual who is experiencing or displaying the pathology or symptomatology of the disease, condition or disorder (i.e., reversing the pathology and/or symptomatology) such as decreasing the severity of disease or reducing or alleviating one or more symptoms of the disease.

In the context of the present disclosure, the term “treatment” may refer to the curative or prophylactic treatment of a viral infection and/or of an adverse immune response to a viral infection as described herein. In particular, with reference to the method of the present disclosure, the term “prophylactic treatment” may refer to the prevention of the viral infection and/or of the adverse immune reaction to said viral infection. Thus, term “treatment” may refer to one or more of:

- the curative treatment of a viral infection,

- the curative treatment of an adverse immune response to a viral infection,

- the prophylactic treatment of a viral infection, and

- the prophylactic treatment of an adverse immune response to a viral infection.

The term “treatment” may also refer to the curative or prophylactic treatment of a viral respiratory diseases as described herein.

The disclosure also provides the use of a compound according to the disclosure as described herein optionally in association with a pharmaceutically acceptable support and/or one or more active substance as described herein for the manufacture of a medicament for the treatment of a viral infection and/or of an adverse immune response to said viral infection, as described herein, wherein the viral infection is preferably a lung viral infection.

In another embodiment, the present disclosure provides a method of treatment of a viral infection and/or of an adverse immune response associated to a viral infection, the method comprising administering to a subject in need thereof an effective amount of compound or pharmaceutical composition for use according to the disclosure optionally in association with a pharmaceutically acceptable support and/or one or more active substance as described herein, wherein the viral infection is preferably a lung viral infection.

Dose

The compound or pharmaceutical composition for use according to the disclosure is administered to the patient at an effective dose. The term “effective” dose” or “therapeutically effective dose” as used herein refers to the amount required to observe a curative or prophylactic activity on the viral infection, and for example an amount required to observe an inhibition or a reduction of viral infection and/or to the adverse immune reaction to the viral infection. The amount of compound or composition to be administered and the duration of the treatment are evaluated by those skilled in the art according to criteria such as the physiological condition of the subject to be treated, the nature of the viral infection or adverse immune reaction to be treated, and the administration route used. The compound or pharmaceutical composition for use according to the disclosure can be administered in the form of a single dose or multiple doses.

In some embodiment, the compound for use according to the present disclosure is administered to said subject in a therapeutically effective dose, for example at a dose to reach a concentration of about 0.3 to about 10 mM at the site of treatment (e.g. the respiratory tract). Nevertheless, this exemplary dose can vary within wide limits and is to be suited to the individual conditions in each individual case.

Patient selection

The subject to be treated, or patient, is an animal, preferably a mammal. In other terms, the compound or pharmaceutical composition may be for human or veterinary medicine.

According to one preferred embodiment, the subject to be treated is a human, and may be an adult human or a child. In some embodiments, the subject is an aged human patient, in particular being more than 50, 60, 70, 80, 90 years old, more particularly being more than 65 years old. In some embodiments, the subject is a child, in particular a child being less than 2, 5, 7 or 10 years old.

In some embodiment, the subject to be treated is a subject vulnerable to lung viral infection. As used herein “vulnerable” denotes individuals that may encounter difficulty in protecting themselves are therefore at greater risk to suffers disproportionately from a lung viral infection and its complications. Subjects vulnerable to lung viral infection include but are not limited to aged patients, in particular aged patents of 65 years old or more, children in particular children of 2 years or less, and pregnant women.

According to one embodiment, the subject to be treated is an animal other than human, preferably a domestic animal selected from the group consisting of a bird, a dog, a cat, a horse, a cow, a sheep, a pig and a non-human primate.

The domestic bird is for example a chicken, a duck, a goose or a turkey.

In some embodiments, the subject is an animal, preferably a human or a domestic bird.

The subject to be treated

Subject to be treated

The method of the present disclosure may be applied to a subject prior or after a viral infection, prior or after the appearance of an adverse immune response to said viral infection, wherein the viral infection is preferably a lung viral infection.

The method of the present disclosure can thus be applied to a subject that is infected or uninfected, as a curative and/or prophylactic treatment, at different levels of severity of the viral infection as described herein.

In some embodiments, the subject is at risk of developing an adverse immune response to a viral lung infection, for example to develop a pneumonia.

In some embodiments, the subject at risk of developing an adverse immune response to the viral infection is an aged human patient, preferably a human patient being more than 50, 60, 70, 80, 90 years old, in particular a human patient of 65 years old or more. It may also be a child, in particular child of 2 years or less, or a pregnant woman.

The subject is infected

In some embodiments, the method of the disclosure is applied to a subject infected, i.e. with a viral infection, wherein the viral infection is preferably a lung viral infection. In some embodiments, the method of the disclosure is applied to treat the viral infection. In some embodiments, the method of the disclosure is applied for the curative treatment of a viral infection.

In some embodiments, the method of the disclosure is applied to prevent the subject from developing an adverse immune response to a viral infection, preferably to a viral lung infection.

In some embodiments, the method of the disclosure is applied to prevent the subject from developing a hyper-inflammatory immune response, a dyspnea, a tachypnea, a pneumonia in particular an acute pneumonia, an exacerbation of a chronic respiratory disease such as asthma or chronic obstructive pulmonary disease, a sepsis, a septic shock, a cytokine storm or an acute respiratory distress syndrome (ARDS).

In some embodiments, the method of the disclosure is applied to prevent the viral infection from escalating to an adverse immune response.

In some embodiments, the method of the disclosure is applied for the prophylactic treatment of an adverse immune response to a viral infection.

In some embodiments, the method of the disclosure is applied for the curative treatment of a viral infection and for the prophylactic treatment of an adverse immune response to the viral infection.

The subject is infected and presents an adverse immune reaction to the infection

In some embodiments, the method of the disclosure is applied to treat an adverse immune response caused by a viral infection, wherein the viral infection is preferably a lung viral infection.

In some embodiments, the method of the disclosure is applied for the curative treatment of an adverse immune response to a viral infection.

In some embodiments, the method of the disclosure is applied for the curative treatment of a viral infection and for the curative treatment of an adverse immune response to the viral infection.

The subject is uninfected.

In some embodiments, the method of the disclosure is applied to a subject uninfected with a viral infection.

In some embodiments, the method of the disclosure is applied for the prophylactic treatment ofa lung viral infection and/or of an adverse immune response to the viral lung infection.

Severity/stage of the viral infection

The method can be applied at various stages of the viral infection.

The inventors have shown for some of the compounds according to the disclosure, that, surprisingly, in a murine model of IAV infection, these compounds were effective even when administered at a distance from the infection (2 or even 4-5 days post-infection). This result strongly suggests that these compounds can treat an advanced stage lung infection. As used herein, “advanced stage lung infection”, refers to patient with lung infection that needs oxygen therapy. This is particularly advantageous because current antiviral treatments, such as Tamiflu®, are ineffective alone on advanced IAV infections.

In some embodiments, the treatment according to the disclosure is for treating a subject at an advanced stage of a viral infection, preferably at an advanced stage of a viral lung infection. In these embodiments, the compound or pharmaceutical composition for use according to the disclosure is preferably administered to the subject at between to 2 to 4 days, preferably at between 4 to 14 days post infection, preferably at between 5 to 11 days post infection, more preferably at 7 to 1 1 days post infection. In some embodiments, the compound or pharmaceutical composition for use according to the disclosure is preferably administered to the subject at between 4 to 1 1 days post infection, more preferably 4 to 7 days post infection.

In some other embodiments, the treatment according to the disclosure is for treating a subject at an early stage of the viral infection.

In these embodiments, the compound or pharmaceutical composition for use according to the disclosure is preferably administered to the subject immediately, one hour, 6 hours, 12 hours, 1 day, 2 days, 3 days post infection, between 8 to 24 hours post infection, more preferably between 10 to 20 hours post infection, even more preferably at 12 to 16 hours post infection.

In some embodiments, the method according to the disclosure is applied for preventing a lung viral infection from escalating to an adverse immune response associated to a viral lung infection in a subject that is suffering from a lung viral infection.

Without wanting to be bound by any theory, the inventors believe that the compound or pharmaceutical composition for use according to the disclosure is probably most effective when given as soon as possible. Yet, as mentioned above the inventors have found that quite surprisingly, in the context of a IAV infection it is still effective 4 days after infection, which, from a practical and/or clinical point of view, is a significant advantage over the existing antiviral treatments such as Tamiflu® (oseltamivir).

The method of the present disclosure can be applied to subjects at various stages of the viral infection, i.e. to subjects presenting a variety of severity of viral lung infection and/or adverse immune response to lung viral infection.

The severity of the subject state may be assessed by a score according to the following ordinal scale where increasing numbers denote increased severity: _ _ _ _ In some embodiment, the subject has a severity assessed as uninfected/ambulatory.

In some embodiment, the subject has a severity assessed as hospitalized but without ventilatory support.

In some embodiment, the subject has a severity assessed as hospitalized with ventilatory support.

Combination therapy

The compound or pharmaceutical composition for use according to the disclosure according to the disclosure can be used as a sole active ingredient or in combination with one or more active substances. The compound or pharmaceutical composition for use according to the disclosure and said active substance(s) can be administered simultaneously or sequentially.

In the present disclosure, the term “administering” means administration of a sole therapeutic agent or in combination with another therapeutic agent.

According to one embodiment, the compound or pharmaceutical composition for use according to the disclosure is used in combination with one or more active substance selected from the group consisting of antivirals, antibiotics, and/or antalgics.

In some embodiments, the compound or pharmaceutical composition for use according to the disclosure is used in combination with one or more antivirals, in particular antivirals of standard therapy of respiratory system viral infections, such as a neuramidase inhibitor (e.g. oseltamivir (Tamiflu®), zanamivir (Relenza®), peramivir (Rapivab®), favipiravir, remdesivir, ribavirin, interferon alpha 2a or 2b, molnupiravir, sotrovimab, casirivimab/imdevimab, baloxavir marboxil (Xofluza®)).

In some embodiments, the compound or pharmaceutical composition for use according to the disclosure is used in combination with one or more antibiotics used in the treatment of bacterial co-infection, in particular antibiotics of standard antibiotherapy such as penicillins, cephalosporins, fluoroquinolones, aminoglycosides, glycopeptides, carbapenems, and macrolides.

In some embodiments, the compound or pharmaceutical composition for use according to the disclosure is used in combination with one or more antalgics such as acetaminophen (paracetamol), nefopam, tramadol, and opioids.

Routes of Administration

The compound or pharmaceutical composition for use according to the disclosure may be administered via any known administration route, including intrapulmonary, systemically (parenterally, intravenously, etc.), orally, rectally, topically or subcutaneously.

In some preferred embodiments, the compound or pharmaceutical composition for use according to the disclosure is administered intrapulmonary, nasally, orally, enterally, intravenously, intramuscularly and subcutaneously.

In some preferred embodiments, the compound or pharmaceutical composition for use according to the disclosure is administered intrapulmonary. As used herein, “intrapulmonary”, refers to an administration route allowing to deliver the compound or pharmaceutical composition for use according to the disclosure to the lungs and/or bronchi, where it particularly concentrates at the alveolar and/or bronchial epithelium.

In some embodiments, the compound or pharmaceutical composition for use according to the disclosure may be administered as an aerosol of a powder or aqueous solution or aqueous suspension, in particular using a nebulizer or a dry powder inhaler.

In some embodiments, the compound or pharmaceutical composition for use according to the disclosure is administered as an aerosol of a powder or aqueous solution or aqueous suspension in subject spontaneously breathing or receiving supplemental oxygen (including high oxygen devices) or being assisted by mechanical ventilation (non-invasive or invasive).

The term “aerosol”, as used in the present disclosure, refers to a dispersion of solid particles or liquid droplets in a gas adapted for targeting the lower airway passages, and preferably the lungs. A nebulizer is defined as a device capable of aerosolizing a liquid material (solution or dispersion) in the form of inhalable liquid droplets. The nebulizer allows the administration of said composition by means of a mask or a tip disposed on the mouth and/or the nose of the subject.

Administration regimen

In some embodiment, the compound or pharmaceutical composition for use according to the disclosure and optionally the one or more additional active substance(s) as described herein is or are administered as a single dose, or in a fractionated dose regimen, simultaneously, separately, or sequentially.

In some embodiment, the compound or pharmaceutical composition for use according to the disclosure and optionally the one or more additional active substance(s) as described herein is or are administered to the subject in a fractionated dose regimen.

In some embodiments, the fractionated dose regimen as described herein comprises 2 to 10 fractionated doses.

In a preferred embodiment, the fractionated dose regimen as described herein is administered once daily or once every two days.

In an embodiment, the fractionated doses as described herein are administered with a time lapse between two fractionated doses comprised between 4h and 48h, preferably between 4h and 12h, more preferably between 4h and 10h, for example with a time lapse of 6 hours.

Pharmaceutical composition

When employed as pharmaceutical, the compound according to the disclosure can be administered in the form of pharmaceutical composition. As described herein, the pharmaceutical composition according to the disclosure may be for human or veterinary use.

The pharmaceutical composition according to the disclosure comprises a compound according to the disclosure and a pharmaceutically acceptable support.

In the context of the disclosure, the term “pharmaceutically acceptable support denotes substances such as excipients, carriers, adjuvants, buffers or the like which are conventionally used in combination with the active ingredient(s), for the preparation of a medicament. The choice of such supports depends essentially on the route of administration envisaged. Pharmaceutically acceptable supports include diluents (fillers, bulking agents, e.g. lactose, microcrystalline cellulose), disintegrants (e.g. sodium starch glycolate, croscarmellose sodium), binders (e.g. PVP, HPMC), lubricants (e.g. magnesium stearate), glidants (e.g. colloidal SIO₂), solvents/co-solvents (e.g. aqueous vehicle, Propylene glycol, glycerol), buffering agents (e.g. citrate, gluconates, lactates), preservatives (e.g. Na benzoate, parabens (Me, Pr and Bu), BKC), anti -oxidants (e.g. BHT, BHA, Ascorbic acid), wetting agents (e.g. polysorbates, sorbitan esters), thickening agents (e.g. methylcellulose or hydroxyethylcellulose), sweetening agents (e.g. sorbitol, saccharin, aspartame, acesulfame), humectants (e.g. propylene, glycol, glycerol, sorbitol). Other suitable pharmaceutically acceptable supports are inter alia described in Remington’s Pharmaceutical Sciences, 15^th Ed., Mack Publishing Co., New Jersey (1991 ) and Bauer et ak, Pharmazeutische Technologic, 5^th Ed., Govi-Verlag Frankfurt (1997). The person skilled in the art knows will readily be able to choose suitable pharmaceutically acceptable supports, depending, e.g., on the formulation and administration route of the pharmaceutical composition.

In some embodiments, the compound according to the disclosure may be in encapsulated form, by being, for example, introduced into microspheres or microcapsules which are reservoirs consisting of a core of active ingredient surrounded by a membrane of coating material. The polymers forming the coating material may be of natural origin (gelatin, chitosan, etc.), semisynthetic origin (cellulose derivatives, etc.) or synthetic origin, such as the lactic and glycolic acid copolymers commonly used. The compounds of the disclosure may also be encapsulated in polymers such as those mentioned above in the form of a film. The compounds of the disclosure may also be encapsulated in nanoparticles, which are colloidal systems of which the size is between 10 and 1000 nm, based on biodegradable polymers, or on lipids capable of retaining one or more active molecules by sequestration and/or adsorption.

The pharmaceutical composition according to the disclosure preferably comprises an amount of compound according to the disclosure of between 5 pg and 1000 mg, preferably between 1 and 500 mg, preferably between 5 and 100 mg.

The ratio between the amounts by weight of compound according to the disclosure and of pharmaceutically acceptable support may be between 5/95 and 95/5, preferably between 20/80 and 80/20.

The pharmaceutical composition according to the disclosure may for example be formulated as a tablet, capsule, granule, powder, sachet, reconstitutable powder, dry powder inhaler and/or chewable. Such solid formulations may comprise excipients and other ingredients in suitable amounts. Such solid formulations may contain e.g. cellulose, cellulose microcrystalline, polyvidone, magnesium stearate and the like.

In some preferred embodiments, the pharmaceutical composition is for inhalation. In this case, the dosage can preferably be reduced because of the application of the drug directly to the site of action, i.e. the lungs.

The present disclosure also relates to a method for treating a lung infection, comprising the administration, to a subject, of an effective amount of compound or pharmaceutical composition for use according to the disclosure and/or of a pharmaceutical composition containing the same.

A subject of the present disclosure is also the use of compound according to the disclosure in the context of the preparation of a pharmaceutical composition, especially of a pharmaceutical composition intended for the treatment of viral infection, preferably of a viral lung infection, as it is described herein.

Other aspects and advantages of the present disclosure will emerge upon reading the examples which follow, which should be considered to be non-limiting illustrations. Legends of the figures

Figure 1. Epithelial tolerance of the AHM compounds.

(Figure 1a and Figure 1 b) Human bronchial epithelial BEAS-2B cells were treated with the indicated concentrations (mM) of the compounds for 20 hours, and epithelial tolerance was assessed using the MTS assay.

Figure 2. Inhibition of viral replication and inflammation by AHM compounds

Human bronchial epithelial BEAS-2B cells were infected with influenza A virus (IAV; A/Scotland/20/74, H3N2 strain) at an MOI of 1 for 4 hours and then treated with the indicated concentrations (mM) of AHM compounds or the control agent (CA) for 16 hours. (Figure 2a and Figure 2b) Viral particle production was assessed using a neuraminidase activity (NA) assay. (Figure 2c and Figure 2d) hlL-6 production in the cell supernatant was measured by ELISA. All data are presented as the mean ± SEM from three independent experiments.

Figure 3. AHM92 and AHM90 Prevent Mortality in a Preclinical Animal Model

(Figure 3a) 7-week-old female C57BI/6 mice were infected intranasally with 100 PFLI of A/Scotland/20/74 (H3N2) virus (IAV) and treated with 75 mM CA, 75 mM AHM90, or 37.5 mM AHM92 starting on day 2 post-infection. Survival rates were monitored daily. (Figure 3b) In a separate experiment, mice were infected and treatment was initiated on day 4 and 5 postinfection using the same concentrations of CA, AHM90, and AHM92. Survival data are presented as mean ± SEM and are cumulative from two independent experiments (n = number of individual animals). Statistical analysis was performed using the Log-rank (Mantel-Cox) test. Significance is indicated as follows: *p < 0.05, **p < 0.005, ***p < 0.0005, ****p < 0.0001 .

EXAMPLES

Example 1 : New chemical structures for the treatment of influenza infection Abstract

Influenza poses a significant global health threat, annually claiming approximately 500,000 lives worldwide and imposing a considerable socioeconomic burden. Current anti-influenza strategies often have suboptimal efficacy. Given the multifaceted nature of influenza pathophysiology, involving both cytopathic viral effects and excessive inflammatory responses, there is a growing interest in exploring host-directed therapies for more effective interventions. In this context, cis-aconitate (CA) has recently emerged as a potent immuno-regulatory and anti-viral agent (unpublished patent application PCTEP2023/085945). Accordingly, new compounds were designed (AHM compounds). The synthesized derivatives and their activities against IAV were evaluated both in vitro and in vivo, using appropriate cellular and animal models. It was revealed that compounds efficiently inhibit virus replication in human airway epithelial cells. Additionally, these compounds exhibit potent anti-inflammatory properties, disrupting the inflammatory cascades associated with IAV infection. Notably, AHM90 and AHM92 show the most potent anti-lAV effects, with half-maximal effective concentration (EC50) values of 3.4mM and 0.95mM, respectively. Furthermore, in a mouse model of lethal influenza-induced pneumonia, it was demonstrated that AHM90 and AHM92 significantly enhance animal survival rates within a clinically relevant timeframe. In conclusion, our findings underscore promising avenues for the development of new compounds for the treatment of influenza.

Introduction

Influenza A virus has consistently caused significant morbidity and mortality since the 1918 pandemic [1 ], triggering extensive investigations into therapeutic strategies. Current anti-lAV approaches, i.e. vaccination and antivirals, exhibit suboptimal effectiveness. Indeed, the short duration of vaccine- induced immunity, coupled with the intrinsic antigenic drift of influenza viruses, compromises host protection [2, 3]. Skepticism also persists regarding the real efficacy of approved anti-influenza drugs such as the neuraminidase inhibitor oseltamivir/Tamiflu® [4, 5]. Accordingly, innovative strategies are crucial for improved treatment of influenza virus infections.

The pathophysiology of influenza-related pneumonia stems from the intrinsic viral pathogenicity and the immune response. While a robust host immune response is necessary for viral clearance, excessive cellular recruitment and release of cytotoxic molecules lead to lung hyper-inflammation, associated with tissue damage, morbidity, and death [6, 7],

Interestingly, the recent discovery of metabolic reprogramming of immune cells has opened new therapeutic avenues for modulating immune responses [8-12], The inventors and others have demonstrated that hosts develop metabolic counter-measures in response to infection. Several molecules conventionally linked to metabolic processes have emerged as potent antimicrobial and immunoregulatory agents [8, 13-18]. For instance, using an integrated approach combining metabolomics, in vitro, and in vivo infection assays, the inventors recently unveiled the inhibitory effect of TCA-derived cis-aconitic acid (CA) on influenza virus infection (patent application PCTEP2023/085945). In an effort to explore a new chemical space for identifying more active compounds against influenza infection, the inventors designed and synthesized a series of novel derivatives. The inventors further examined the anti-influenza activity of the synthesized compounds using appropriate in vitro and in vivo screening tests.

1.1/Material and Methods

Chemistry:

Preparation of AHM03:

Propylamine was dissolved in a 1 ml of DCM, then DIPEA (0.22 mmol) was added. A solution of benzene-1 ,3,5-tricarbonyl trichloride (300 mg, 1.13 mmol) in DCM was then added to the reaction mixture at 0°C. The reaction was stirred two hours at room temperature. After reaction completion, the solvent was evaporated and the crude was purified by flash column chromatography (DCM/MeOH, 96/4). The desired product was recovered as a white solid in 80% yield (298 mg). ¹H NMR (400 MHz, DMSO-cfe) 5 8.64 (t, J = 5.6 Hz, 3H), 8.36 (s, 3H), 3.25 (dt, J = 7.3, 6.1 Hz, 6H), 1.55 (p, J = 7.3 Hz, 6H), 0.91 (t, J = 7.4 Hz, 9H). ¹³C NMR (101 MHz, DMSO-cfe) 5 165.35, 134.92, 128.38, 41 .08, 22.27, 20.37, 11 .48, 10.86.

Preparation of AHM44: In a 50 ml round bottom were solubilized trimesic acid (150 mg, 0.713) and NaHCOs (3 eq) in DMF (5ml). Then iodopropanfluore (7eq, 5 mmol) was added and the mixture was stirred at room temperature overnight. After reaction completion, the sovent was removed under vacuum and the crude was washed with water and extracted with DCM then concentrated and purified by flash column chromatography (Cyhex/EtOAc : 50/50). The desired product was recovered as a white solid in 84% yield (132 mg).

Preparation of AHM37:

A round bottom with a stirrer was charged with dimethyl 5-bromoisophthalate (50 mg, 0.18 mmol), 4-fluorophenylboronic acid (2eq, 51 ,2 mg, 0.336 mmol),

Tetrakis(triphenylphosphine)palladium (3 mol%, 5mg), K2CO3 (1 mg) and 1 mL of dioxane. The reaction was refluxed overnight and after the reaction completion, the mixture was cooled at room temperature and concentrated under vacuum. The crude was washed with water and extracted with DCM and concentrated. The desired compound was recovered as yellow solid after purification by column chromatography (DCM/EtOAc : 90/10) in 98% yield. ¹H NMR (400 MHz, CDCI3) 5 ¹H NMR (300 MHz, CDCI3) 5 8.67 (t, J = 1 .6 Hz, 1 H), 8.43 (d, J = 1 .6 Hz, 2H), 7.74 - 7.50 (m, 2H), 7.19 (t, J = 8.6 Hz, 1 H), 4.00 (s, 6H). ¹³C NMR (101 MHz, CDCI3) 5 171.91 , 165.86, 137.61 , 134.12, 131.50, 128.53, 52.68, 52.10, 52.03, 34.04, 34.01 , 33.25, 28.71.

Preparation of AHM39:

A round bottom with a stirrer was charged with dimethyl 5-aminoisophthalate (250 mg, 1 ,18 mmol), 1 -(bromomethyl)-4-methylbenzene (2eq, 436.7 mg, 2.26 mmol), (3 mol%, 5mg), Na2CC>3 (1 mg) and Nal (mg, mmol,) 1 mL of DMF. The reaction was stirred at 90°C overnight and after the reaction completion, the mixture was cooled at room temperature and concentrated under vacuum. The crude was washed with water and extracted with DCM and concentrated. The desired compound was recovered as a yellowish solid after purification by column chromatography (DCM/EtOAc : 80/20) in 87% yield (457 mg). ¹H NMR (400 MHz, CDCI3) 5 7.98 (t, J = 1.4 Hz, 1 H), 7.62 (d, J = 1.4 Hz, 2H), 7.16 - 7.02 (m, 8H), 4.64 (s, 4H), 3.87 (s, 6H), 2.33 (s, 6H). ¹³C NMR (101 MHz, CDCI3) 5 167.03, 149.48, 136.97, 134.56, 131 .51 , 129.59, 126.91 , 1 18.78, 1 17.44, 53.83, 52.35, 21 .22.

General protocol of the Buchwald-Hartwig-Migita coupling of thiols with iodide: round bottom with a stirrer was charged with the ester iodide compound (1 eq) , the thiol derivative (1 .5 eq), PdG3-Xanthphos (5 mol%). And dioxane. Upon stirring the reaction mixture at room temperature, EtsN was added to the medium. After reaction completion (1 -2 hours), the mixture was concentrated under vacuum. The crude was washed with water, extracted with EtOAC, concentrated and purified by column chromatography (DCM/EtOAc) to give the desired product.

Preparation of AHM240:

Following the general procedure, AHM 240 was recovered as a white solid in 87% yield (325 mg). ¹H NMR (300 MHz, MeOD) 5 8.31 (s, 2H), 4.49 (dd, J = 8.7, 6.3 Hz, 1 H), 3.98 (s, 6H), 3.11 - 2.71 (m, 2H). ¹³C NMR (75 MHz, MeOD) 5 173.37, 165.98, 152.46, 149.39, 126.94, 53.46, 44.33, 36.88.

Preparation of AHM86:

Following the general procedure, AHM AHM86 was recovered as a white solid in 79% yield (145 mg). ¹H NMR (300 MHz, CDCI3) 5 8.62 (t, J= 1 .6 Hz, 1 H), 8.42 - 8.26 (m, 2H), 5.26 (t, J = 9.3 Hz, 1 H), 5.05 (m, 2H), 4.80 (d, J= 10.0 Hz, 1 H), 4.33 - 4.10 (m, 2H), 3.79 (ddd, J= 10.1 , 5.0, 2.4 Hz, 1 H), 2.12 (s, 3H), 2.08 (s, 3H), 2.04 (s, 3H), 2.01 (s, 3H). ¹³C NMR (75 MHz, CDCI3) 5 170.81 , 170.22, 169.46, 169.34, 165.61 , 137.49, 133.73, 131.58, 130.36, 85.45, 76.21 , 74.00, 70.01 , 68.24, 62.16, 52.70, 20.69.

Preparation of AHM90:

Following the general procedure, AHM90 was recovered as a white solid in 79 yield (169 mg). ¹H NMR (300 MHz, MeOD) 5 8.56 (t, J = 1.6 Hz, 1 H), 8.38 (d, J = 1.6 Hz, 2H), 4.09 (dd, J = 8.9, 6.1 Hz, 1 H), 3.97 (s, 6H), 3.00 - 2.70 (m, 2H). ^13C NMR (75 MHz, MeOD) 5 174.14, 173.41 , 166.85, 138.92, 136.28, 132.79, 130.92, 53.08, 47.27, 37.52.

Preparation of AHM230:

Following the general procedure, AHM230 was recovered as an orange oil in 84% (167 mg). ¹H NMR (300 MHz, CDCI3) 5 8.41 (t, J = 1.6 Hz, 1 H), 8.10 (d, J = 1.6 Hz, 2H), 3.88 (s, 6H), 3.63 (s, 3H), 3.20 (t, J = 7.3 Hz, 2H), 2.59 (t, J = 7.3 Hz, 2H). ¹³C NMR (101 MHz, CDCI3) 5 171.91 , 165.86, 137.61 , 134.12, 131.50, 128.53, 52.68, 52.10, 52.03, 34.04, 34.01 , 33.25, 28.71.

Preparation of AHM231 :

Following the general procedure, AHM231 was recovered as a brown solid in 73% yield (162 mg). ¹H NMR (300 MHz, CDCI3) 5 8.42 (t, J= 1.5 Hz, 1 H), 8.14 (d, J= 1.6 Hz, 2H), 6.21 (d, J = 7.3 Hz, 1 H), 4.84 (dt, J= 7.3, 4.6 Hz, 1 H), 3.88 (s, 6H), 3.55 (s, 3H), 3.48 (d, J = 4.7 Hz, 2H), 1.88 (s, 3H). ¹³C NMR (75 MHz, MeOD) 5 166.95, 138.80, 135.54, 132.83, 129.36, 53.55, 53.08, 52.97, 36.11 , 22.23.

General protocol of saponification:

The ester compounds were solubilized in the appropriate solvent, then an aqueous solution of NaOH was added. After reaction completion, the reaction medium was acidified using an acid resin (Dowex 50WX8) or HCI until pH =1 -2. The reaction mixture was then filtered and concentrated. The crude was purified by preparatuve HPLC.

Preparation of AHM84:

Following the general procedure, the compound AHM84 was obtained from the saponification of compound AHM39 as a white solid in 74% yield (138%). ¹H NMR (300 MHz, MeOD) 5 7.90 (t, J = 1 .4 Hz, 1 H), 7.54 (d, J = 1 .4 Hz, 2H), 7.27 - 7.01 (m, 8H), 4.60 (s, 4H), 2.28 (s, 6H). ¹³C NMR (101 MHz, MeOD) 5 176.33, 150.19, 139.69, 137.31 , 137.11 , 130.04, 128.26, 1 16.99, 54.66, 21.10.

Preparation of AHM232:

Following the general procedure, the compound AHM232 was obtained from the saponification of compound AHM230 as a white solid in 90% yield (77 mg). ¹H NMR (300 MHz, MeOD) 5 8.44 (t, J = 1.5 Hz, 1 H), 8.17 (d, J = 1.5 Hz, 2H), 3.26 (d, J = 7.1 Hz, 1 H), 2.65 (t, J = 7.1 Hz, 2H). ¹³C NMR (75 MHz, MeOD) 5 175.00, 168.27, 139.24, 134.71 , 133.36, 129.39, 34.80, 29.50.

Preparation of AHM301-F2:

Following the general procedure, the compound AHM301 -F2 was obtained from the saponification of compound AHM230 as a yellowish solid in 34% yield (31 mg). ¹H NMR (400 MHz, MeOD) 5 8.45 (t, J = 1.5 Hz, 1 H), 8.16 (d, J = 1.5 Hz, 2H), 3.67 (s, 3H), 3.29 (t, J = 7.0 Hz, 2H), 2.69 (t, J = 7.0 Hz, 2H). ¹³C NMR (101 MHz, MeOD) 5 173.60, 168.38, 138.89, 134.84, 133.48, 129.51 , 52.32, 34.83, 29.48. Preparation of AHM92:

Following the general procedure, the compound AHM92 was obtained from the saponification of compound AHM90 as a white solid in 90% yield (80 mg). ¹H NMR (300 MHz, MeOD) 5 8.59 (t, J= 1 .6 Hz, 1 H), 8.37 (d, J= 1 .6 Hz, 2H), 4.08 (dd, J= 8.9, 6.0 Hz, 1 H), 3.00 - 2.69 (m, 2H). ¹³C NMR (75 MHz, MeOD) 5 173.81 , 168.05, 139.27, 135.59, 133.49, 131 .58, 47.07, 37.40

Preparation ofAHM243:

Following the general procedure, the compound AHM243 was obtained from the saponification of compound AHM240 as an orange solid in 82% yield (70 mg). ¹H NMR (300 MHz, MeOD) 5 7.97 (s, 2H), 4.51 (dd, J= 8.3, 6.0 Hz, 1 H), 3.07 (m, 2H). ¹³C NMR (75 MHz, MeOD) 5 173.52, 167.38, 153.45, 149.46, 129.90, 129.20, 126.25, 44.61 , 37.05.

Preparation of AHM118:

Following the general procedure, the compound AHM1 18 was obtained from the saponification of compound AHM86 as a white solid in 73% yield (45 mg). ¹H NMR (400 MHz, D2O) 5 8.12 (t, J= 1 .6 Hz, 1 H), 8.02 (d, J= 1 .6 Hz, 2H), 3.80 (d, J= 2.2 Hz, 1 H), 3.65 (dd, J= 12.5, 5.5 Hz, 1 H), 3.50 - 3.40 (m, 2H), 3.37 - 3.21 (m, 2H). ¹³C NMR (75 MHz, D2O) 5 174.24, 137.57, 134.54, 131.97, 128.62, 87.14, 79.98, 77.33, 71.78, 69.38, 66.64, 60.85.

Preparation of AHM 108:

Following the general procedure, AHM 108 was obtained from the saponification of AHM37 as a white solid in 66% yield (28 mg). ¹H NMR (300 MHz, MeOD) 5 8.62 (s, 1 H), 8.37 (d, J = 1 .5 Hz, 4H), 7.76 - 7.66 (m, 4H), 7.22 (t, J = 8.8 Hz, 3H).

Viruses. Mouse adapted-influenza A/Scotland/20/74 (H3N2) was generously given by Pr. Sylvie van der Werf’s team (Pasteur Institute, Paris, France).

Cell culture. In vitro experiments were performed using human bronchial epithelial BEAS-2B cells, cultured in F-12K Medium supplemented with 10% FBS and 100 LI/mL penicillin, 100 pg/mL streptomycin. All cells were mycoplasma-free. BEAS-2B cells were infected in medium without FBS for 4 hours with IAV Scotland at MOI=1 . Four hours after the challenge, cells were washed with PBS and incubated for 4 h or 16 h with different concentrations of metabolites.

Neuraminidase (NA) assay. The assay measures the release of a 4-methylumbelliferone fluorescent product from the 2'-(4-Methylumbelliferyl)-a-D-N-acetylneuraminic acid sodium salt hydrate (MU-NANA) substrate. 67 pL of cell supernatant was incubated with 33 pL of MU- NANA (50 pM) in black 96-well micro-plates. Fluorescence was immediately measured in a kinetic assay over 1 h at Ex = 355 nm and Em = 460 nm.

ELISA. Cells supernatants were centrifuged 5 min at 500 g and supernatants were stored at - 80°C. DuoSet ELISA (Human IL-6) was performed according to the manufacturer’s (R&D Systems) instructions.

Animal infection and treatment. 7-week-old female C57BI/6 mice were challenged intranasally with 100 pfu of A/Scotland/20/74 (H3N2) IAV, and treated or not with compounds (75mM of CA or 75 mM AHM90 or 37.5mM of AHM92). Survival was monitored daily.

Study approval. All animal experimentations were performed according to the national governmental guidelines and were approved by our local and national ethics committee (CEEA.19, #201604071220401 -4885). Statistical analysis. Statistical analyses were performed using GraphPad Prism. All statistical analysis were calculated on raw data. Data are reported as mean ± SEM. Statistical values, including the number of replicates (n) and the statistical test used, can be found in the figure legends. *p < 0.05, **p < 0.005, ***p < 0.0005, ****p < 0.0001 . For in vitro experiments, n = the number of separate experiments. For in vivo work, n = the number of individual animals.

1 .2/ Results

Defining new chemical space to discover novel drug candidates for anti-influenza therapy.

To identify new derivatives with a higher antiviral activity against influenza A virus than the cisaconitate reference, we designed series of cyclic compounds (AHM).

In vitro assessment of epithelial tolerance of the candidate compounds.

Influenza viruses predominantly infect the epithelial cells lining the respiratory tract and replicate to produce new viral particles. Given that the antiviral properties of the newly designed AHM molecules would be tested on epithelial cells, the tolerance of 16 molecules was first assessed using in vitro human bronchial epithelial BEAS-2B cells. For each compound, the highest non-cytotoxic dosage was determined, confirmed through MTS viability assays.

The results are presented on Figure 1a and Figure 1 b.

In vitro assessment of the candidate compounds to inhibit viral replication and prevent hyper-inflammation.

The anti-influenza properties of the AHM compounds was then assessed in vitro. Human bronchial epithelial BEAS-2B cells were infected with influenza A virus (IAV; A/Scotland/20/74, H3N2 strain) and treated with the compounds 4 hours post-infection (p.i.) in two sets of experiments (respectively Figure 2a, Figure 2c and Figure 2b, Figure 2d)

Assessment of the anti-viral properties. To evaluate the release of new virions, the neuraminidase (NA) activity was measured in cell supernatants at 20 hours p.i.

The results are presented in Figure 2a and Figure 2b.

Most of the compounds assayed show some antiviral activity against IAV. Two compounds particularly outperformed CA: (i) AHM90 reduced NA activity by 93%, and (ii) AHM92 achieved similar NA activity reduction to CA but at a much lower concentration (3.4mM vs. 0.95mM).

Assessment of the anti-inflammatory properties. To evaluate the anti-inflammatory properties, the hlL-6 production was measured in cell supernatants at 20 hours p.i.

The results are presented in Figure 2c and Figure 2d.

Interestingly, several compounds, including AHM90 and AHM92, could modulate like CA the immune response triggered by viral infection, reducing IL-6 production in influenza-infected epithelial cells down to 20% (Figure 2c). In conclusion, several compounds including AHM90 and AHM92 not only inhibit influenza viral replication but also control the associated hyperinflammation.

In Figure 2b and Figure 2d, it can be observed a dose effect of reduced neuraminidase activity and hlL-6 secretion for each compound at concentrations lower than or equal to those used for CA (3.4 mM). Several compounds appear to possess very promising antiviral and antiinflammatory properties, such as AHM231 , which significantly inhibits neuraminidase activity and hlL-6 production at a dose of just 1 mM. AHM92 and AHM90 effectively protect mice from IAV pneumonia.

The above in vitro studies demonstrated that among other compounds, both AHM92 and AHM90 possess particularly strong antiviral and anti-inflammatory properties. This dual action is particularly advantageous in combating influenza virus infections, as severe cases are attributed to both viral replication-induced cellular damage and an exaggerated inflammatory response [6, 7],

To validate those protective properties in vivo, we infected mice with a lethal dose of IAV and evaluated the efficacy of AHM92 and AHM90 in preventing mortality. Notably, it is known that some previous drug candidates failed to translate to clinical success because they were tested simultaneously with viral inoculation, unlike the delayed treatment approach more representative of human scenarios. Therefore, we postponed the dosing of AHM92 and AHM90 by at least two days to mimic the typical delay between symptom onset and treatment in humans.

The results are presented on Figure 3.

Dosina at day 2 post infection. Remarkably, AHM92 and AHM90 increased survival rates by 80% and 40%, respectively (Figure 3a). These findings indicate that AHM92 and AHM90 provide protection against influenza infection in a manner comparable to CA. Additionally, the effective dose of AHM92 was 50% lower than that required for CA and AHM90.

Dosina at days 4 and 5 post-infection. Considering the average symptom-to-hospitalization time for severe influenza in humans is approximately 4 days [21], we further delayed treatment until day 4 post-infection, administering a second dose on day 5 post-infection to better simulate clinical practice. AHM92 demonstrated significant efficacy under these conditions, increasing the survival rate from 0% to approximately 25% (Figure 3b). Although AHM90 did not increase survival at this later administration time, it statistically delayed death by three days.

The results show that the compounds of the invention demonstrate remarkable antiviral and anti-inflammatory properties. The in vivo experiments confirmed their efficacy in shielding the host from severe influenza pneumonia, even when administered at a late stage of infection. At this step, conventional treatment with oseltamivir proved completely ineffective.

References

1. Centers for Disease Control and Prevention, National Center for Immunization and Respiratory Diseases (NCIRD) (2021 ) Influenza (Flu)

2. World Health Ornagization (2023) Influenza (Seasonal)

3. WHO | WHO guidelines for pharmacological management of pandemic (H1 N1 ) 2009 influenza and other influenza viruses.

4. Aliberti S, Dela Cruz CS, Amati F, et al (2021 ) Community-acquired pneumonia. Lancet 398:906-919. https://doi.Org/10.1016/S0140-6736(21 )00630-9

5. Jefferson T, Jones MA, Doshi P, et al (2014) Neuraminidase inhibitors for preventing and treating influenza in adults and children. Cochrane Database of Systematic Reviews 2018:. https://doi.Org/10.1002/14651858.CD008965.pub4

6. Herold S, Becker C, Ridge KM, Budinger GRS (2015) Influenza virus-induced lung injury: pathogenesis and implications for treatment. Eur Respir J 45:1463-1478. https://doi.Org/10.1183/09031936.00186214

7. Tavares LP, Teixeira MM, Garcia CC (2017) The inflammatory response triggered by Influenza virus: a two edged sword. Inflamm Res 66:283-302. https://doi.org/10.1007/s0001 1 - 016-0996-0

8. O’Neill LAJ, Kishton RJ, Rathmell J (2016) A guide to immunometabolism for immunologists. Nat Rev Immunol 16:553-565. https://doi.org/10.1038/nri.2016.70

9. Pearce EJ, Pearce EL (2018) Immunometabolism in 2017: Driving immunity: all roads lead to metabolism. Nat Rev Immunol 18:81-82. https://doi.org/10.1038/nri.2017.139

10. Rambold AS, Pearce EL (2018) Mitochondrial Dynamics at the Interface of Immune Cell Metabolism and Function. Trends Immunol 39:6-18. https://doi.org/10.1016/j-it.2017.08.006

11. Cezard A, Monard S, Brea-Diakite D, et al (2021 ) [Metabokines reviewed: Essential mediators of anti-infectious immunity]. Med Sci (Paris) 37:342-348. https://doi.org/10.1051/medsci/2021031

12. Rao M, Dodoo E, Zumla A, Maeurer M (2019) Immunometabolism and Pulmonary Infections: Implications for Protective Immune Responses and Host-Directed Therapies. Front Microbiol 10:962. https://doi.org/10.3389/fmicb.2019.00962

13. Guillon A, Arafa El, Barker KA, et al (2020) Pneumonia recovery reprograms the alveolar macrophage pool. JCI Insight 5:e133042, 133042. https://doi.org/10.1 172/jci.insight.133042

14. Palsson-McDermott EM, O’Neill LAJ (2020) Targeting immunometabolism as an antiinflammatory strategy. Cell Res 30:300-314. https://doi.org/10.1038/s41422-020-0291 -z

15. Guillon A, Brea-Diakite D, Cezard A, et al (2022) Host succinate inhibits influenza virus infection through succinylation and nuclear retention of the viral nucleoprotein. The EMBO Journal 41 :. https://doi.Org/10.15252/embj.2021108306

16. Martinez-Reyes I, Chandel NS (2020) Mitochondrial TCA cycle metabolites control physiology and disease. Nat Common 1 1 :102. https://doi.org/10.1038/s41467-019-13668-3 17. Soto-Heredero G, Gomez de Las Heras MM, Gabande-Rodnguez E, et al (2020) Glycolysis - a key player in the inflammatory response. FEBS J 287:3350-3369. https://doi.org/10-11 11/febs.15327

18. Cezard A, Guillon A, Si-Tahar M (2021 ) [Identification of a metabolic immune regulator in the host that protects against influenzal pneumonia]. Rev Mai Respir 38:567-571. https://doi.Org/10.1016/j.rmr.2O21 .05.003

19. Bruneau A, Roche M, Hamze A, et al (2015) Stereoretentive Palladium-Catalyzed Arylation, Alkenylation, and Alkynylation of 1 -Thiosugars and Thiols Using Aminobiphenyl Palladacycle Precatalyst at Room Temperature. Chemistry A European J 21 :8375-8379. https://doi.Org/10.1002/chem.201501050

20. Montoir D, Amoura M, Ababsa ZEA, et al (2018) Synthesis of aryl-thioglycopeptides through chemoselective Pd-mediated conjugation. Chem Sci 9:8753-8759. https://doi.org/10.1039/C8SC02370K

21 . Lhommet C, Garot D, Grammatico-Guillon L, et al (2020) Predicting the microbial cause of community-acquired pneumonia: can physicians or a data-driven method differentiate viral from bacterial pneumonia at patient presentation? BMC Pulm Med 20:62. https://doi.Org/10.1186/S12890-020-1089-y

Claims

1. A compound of formula (I) or a pharmaceutically acceptable salt thereof, wherein:

X is selected from CH and N,

Y is selected from NR₄, S, S(O), S(O)₂, S(O)(NH), Se, Se(O), Se(O)₂, Se(O)(NH), O,

CO, phenyl

Z is selected from O and NH,

Ri, R₂, R₅, Re are independently selected from H, Na, K, C1-6 alkyl,

R₄ is selected from H, C1-6 alkyl, C1-6 alkyl-aryl,

2. The compound according to claim 1 , wherein said compound is a compound of formula (la) or (lb) wherein:

CO, phenyl

Z is selected from O and NH,

R1, R₂, R₅, Re are independently selected from H, Na, K, C1-6 alkyl,

R₄ is selected from H, C1-6 alkyl, C1-6 alkyl-aryl,

3. The compound according to claim 1 or 2, wherein the compound is a compound of formula (Ic), (Id), (le), (If), (Ig), (Ih), (li), (li), (Ik), (II), (Im) or (In)

wherein:

X is selected from CH and N,

Z is selected from O and NH,

Ri, R₂, Rs, Re are independently selected from H, Na, K, C1-6 alkyl,

R₄ is selected from H, C1-6 alkyl, C1-6 alkyl-aryl,

4. The compound according to any one of claims 1 to 3, wherein the compound is selected from the group consisting of:

5. The compound according to any one of claims 1 to 4, wherein the compound is selected from the group consisting of:

6. A compound according to any one of claims 1 to 5, for use as a medicine.

7. A pharmaceutical composition, comprising a compound as defined in any one of claims 1 to 5 and a pharmaceutically acceptable support.

8. The compound according to any one of claims 1 to 6 or the pharmaceutical composition according to claim 7, for use in a method of treatment of a viral infection and/or an adverse immune response to a viral infection, the method comprising administering to a subject in need thereof an effective amount of said compound or pharmaceutical composition.

9. The compound or pharmaceutical composition for use in a method according to claim 7, wherein the viral infection is a lung viral infection preferably caused by a respiratory virus selected from an influenza virus, a coronavirus, a respiratory syncytial virus, a parainfluenza virus, a metapneumovirus, a rhinovirus, an adenovirus, a varicella-zoster virus, a cytomegalovirus, a paramyxovirus such as Nipah virus, an enterovirus and a bocavirus.

10. The compound or pharmaceutical composition for use in a method according to any one of claims 7 to 9, wherein the viral infection is caused by an influenza virus.

11 . The compound or pharmaceutical composition for use in a method according to any one of claims 7 to 10, wherein the method is applied to treat the viral infection.

12. The compound or pharmaceutical composition for use in a method according to any of claims 7 to 1 1 , wherein the method is applied to treat the adverse immune response to the viral infection.

13. The compound or pharmaceutical composition for use in a method according to any one of claims 7 to 12, wherein the method is applied to prevent the viral infection from escalating to an adverse immune response.

14. The compound or pharmaceutical composition for use in a method according to any one of claims 7 to 13, wherein the adverse immune response to the viral infection is selected from a hyper-inflammatory immune response, a dyspnea, a tachypnea, a pneumonia in particular an acute pneumonia, an exacerbation of a chronic respiratory disease such as asthma or chronic obstructive pulmonary disease, a sepsis, a septic shock, a cytokine storm or an acute respiratory distress syndrome (ARDS).

15. The compound or pharmaceutical composition for use in a method according to any one of claims 7 to 14, wherein the compound or pharmaceutical composition is administered between 2 and 14 days post infection.