WO2025005803A1

WO2025005803A1 - Mucoadhesive formulation

Info

Publication number: WO2025005803A1
Application number: PCT/NL2024/050349
Authority: WO
Inventors: Fergus MANFORD; Yi-Han Hsieh; Maurice Mannesse
Original assignee: Leyden Laboratories B.V.
Priority date: 2023-06-28
Filing date: 2024-06-28
Publication date: 2025-01-02

Abstract

The invention relates to liquid formulations for mucosal administration comprising antibodies, bispecific antibodies or antigen-binding fragments that have broad binding activity against respiratory viruses, for example, influenza, respiratory syncytial virus and coronaviruses. The disclosure also relates to their methods of manufacture and the kits used for their administration to patients.

Description

MUCOADHESIVE FORMULATION

FIELD OF THE INVENTION

The invention relates to formulations for mucosal administration comprising antibodies, bispecific antibodies or antigen-binding fragments that have broad binding activity against respiratory viruses, for example, influenza, respiratory syncytial virus and coronaviruses. The disclosure also relates to their methods of manufacture and the kits used for their administration to patients.

BACKGROUND OF THE INVENTION

Respiratory Viruses

Influenza virus

Seasonal influenza virus epidemics cause 3 to 5 million severe cases of influenza worldwide and 650,000 deaths each year. In 2018, it was estimated that the average annual total economic burden of influenza to the healthcare system and society in the United States alone was $11.2 billion. Thus, the ongoing healthcare burden is inadequately addressed with current therapies, z.e., vaccines and antivirals. The inadequacy of these therapies has been largely due to current vaccines only being effective against a specific and very narrow spectrum of viral strains. The influenza virus also poses pandemic threats (e.g., influenza virus H5).

Respiratory syncytial virus

While Respiratory syncytial virus (RSV) very rarely causes severe disease in healthy adults, it can cause morbidity and mortality in the elderly and in those with underlying immune compromise or cardiopulmonary disease. Older adults have a similar presentation to younger adults when infected with RSV but tend to have greater symptom severity with increased risk of lower respiratory tract involvement. In particular, the elderly are more likely to experience pneumonia, respiratory distress, and death. Globally, RSV affects an estimated 64 million people and causes 160,000 deaths each year. SARS-CoV-2 virus

Coronavirus disease (COVID-19) is an infectious disease caused by the SARS-CoV-2 virus. Since the start of the CO VID-19 pandemic, there have been an estimated 767 million cases of SARS-CoV-2 virus infection and an estimated 6.9 million deaths. The continued spread of the virus is largely driven by the emergence of viral variants, which can evade the current vaccines through mutations in the spike protein.

An example of an antibody with broad binding activity against influenza is the human monoclonal antibody (mAb) CR9114 which binds to the highly conserved stem region of hemagglutinin (HA), a receptor-binding glycoprotein relevant to viral entry. CR9114 can prevent fusion of the viral envelope with the endocytic vesicular membrane, and therefore protects individuals from influenza A virus subtypes Al or A2. CR9114 may deliver in vivo protection against Influenza virus B by preventing egress of newly formed virions from infected cells. Moreover, even though CR9114 does not bind to another surface protein Neuraminidase (NA), CR9114 is capable of inhibiting NA catalytic activity which is necessary for efficient viral egress through steric hindrance.

Hence, CR9114 is eminently suited for use in a drug formulation for prophylactically and/or therapeutically treating influenza infections.

The most common reason for introducing an antibody into the nasal cavity is to provide a convenient and accessible route for rapidly and efficiently managing the localized symptoms associated with infection.

Small molecules have typically been applied to the nasal mucosa and these include antihistamines, corticosteroids, sodium cromoglicate, sympathomimetics and antiseptics/antibiotics. These molecules are administered either in liquid form (from a spray or as drops) or as creams/ointments. The nasal cavity has been used as a portal for the delivery of vaccines, particularly for vaccines against influenza infections. The presentation of an antigen to the nasal-associated lymphoid tissue can promote both cellular and humoral responses.

The nasal vestibule is the first and narrowest part of the nasal cavity, with a cross-sectional area of approximately 30 mm² on each side. The lining of the vestibule changes from skin at the entrance to a stratified squamous epithelium which extends over the anterior third of the entire nasal cavity. The nasal vestibule contains vibrissae which filter out inhaled particles with an aerodynamic droplet size greater than approximately 10 pm. Progression through the nasal cavity leads to the turbinate region. The turbinates are convoluted projections from the nasal septum which are lined with a pseudostratified columnar epithelium (80% to 90% of the total surface area of the nasal epithelium in humans) composed of mucus-secreting goblet cells, ciliated and non-ciliated cells and basal cells. The apical surfaces of the ciliated and non-ciliated cells are covered with nonmotile microvilli, which serve to increase the surface area of the epithelial cells. There are also approximately 100 motile cilia on each ciliated cell which are responsible for mucus transport. Serous and seromucous glands also contribute to nasal secretions. As air moves through the turbinate region via the meatuses, the low rate of airflow in combination with the turbulence created by the shape of the turbinates encourages the air to make contact with the highly vascularized walls, enabling it to be warmed and humidified. Particulates (5 pm to 10 pm) within the airstream, such as dust, pollen, microorganisms and pollutants, have the potential to deposit on the viscoelastic mucous gel lining the turbinate walls. The cilia, beating within the periciliary fluid, engage with the underside of the mucus and propel the gel and the deposited droplets to the nasopharynx, where they are either swallowed or expectorated. This process is termed mucociliary clearance and is able to clear mucus from the nasal turbinates at a rate of approximately 7 millimeters per minute.

Drug deposited anterior to the turbinates will remain in the nasal cavity for longer than drug deposited at the turbinates. Once drug droplets (if formulated as a suspension) or molecules (if in solution) find their way on to the mucociliary clearance mechanism, they will be cleared from the nasal cavity and therefore have a limited contact and residence. The nasal mucosa is protected from the external environment by a layer of mucus. In the nasal cavity this exists as a gel phase which is approximately 1 pm to 10 pm thick and found above a watery phase surrounding the cilia (periciliary layer) which is approximately 7 pm deep. Mucus is secreted continuously by the goblet cells and submucosal glands. Normal mucus is 97% water and 3% solids, with the latter comprising mucins (approximately 30% of the solid content), non-mucin proteins (e.g. albumin, immunoglobulins, lysozyme and lactoferrin), inorganic salts and lipids. Mucins are extremely large glycoproteins (up to 3 x 10⁶ Da per monomer) with protein regions rich in serine and threonine which are linked, by their hydroxyl side groups, to sugar chains (O-glycosylation). They are anionic (negatively charged) because most of their terminal sugars contain carboxyl or sulfate groups. These glycosylated (sugar-rich) regions are separated by regions of non-glycosylated, ‘naked’ protein, rich in cysteine residues, which are believed to form globular domains stabilized by disulfide bonds. These ‘naked’ domains are the most hydrophobic regions of mucins and probably adsorb significant amounts of lipids. They are also the most antigenic sites on mucins. Entanglement of mucin polymers leads to the formation of a mucous gel and the generation of a mesh which is stabilized by noncovalent calcium-dependent cross-linking of adjacent polymers. The sugar side chains bind large amounts of water, allowing the mucus to act as a lubricant and a reservoir for the periciliary fluid within which the cilia beat. Mucus is a viscoelastic gel with the properties of both a deformable solid (elasticity) and a viscous fluid. Cilia can transport mucus only of the appropriate viscoelasticity, and this is controlled by the level of mucus hydration. The presence of mucus at the epithelial surface of the nasal cavity provides an additional barrier to nasal residence.

A broad range of enzymes are present in the nasal cavity, including proteolytic enzymes (proteases and aminopeptidases) which provide a potential barrier to proteins.

Respiratory viruses spread mainly by drops an/or aerosols made when people with flu cough, sneeze and/or talk. These droplets can land in the mouths and/or noses of people who are nearby, and/or be inhaled into the respiratory tract or lungs. These organs are thus seen as the frontline when an individual is challenged with respiratory viruses. It is therefore desirable to trap respiratory viruses at those specific sites of the human body, so as to elicit the prophylactic and/or therapeutic treatment. The structural size and complexity of an antibody makes it susceptible to be caught up in the nasal mucociliary clearance mechanism and ultimately removed from the body. For antibodies, mucociliary clearance is likely to play a significant role in limiting the residence time required for a prophylactic and/or therapeutically delivered antibody to be effective.

There is thus a need to have an improved formulation comprising an antibody that overcomes the challenges as described herein.

Prophylactic and/or therapeutic administration of antibodies could represent a key tool in controlling transmission and infection severity of current and future strains of respiratory viruses. However, the great genetic variation within some respiratory viruses represents an obstacle to the delivery of broad protection against many if not all current and future strains.

SUMMARY OF THE INVENTION

The inventors have discovered an improved formulation comprising a prophylactic and/or therapeutic antibody, and at least one mucoadhesive compound as described herein.

In a preferred embodiment, a liquid composition comprising an antibody, a bispecific antibody or antigen binding fragment, and at least one mucoadhesive compound, wherein the concentration of the antibody, bispecific antibody or antigen-binding fragment thereof ranges from 0.001 to 75 % (w/v) and the concentration of the mucoadhesive compound ranges from 0.1 to 10 % (w/v), is disclosed.

In a preferred embodiment, the antibody is a bispecific antibody.

In a preferred embodiment, a composition is disclosed, wherein the concentration of said mucoadhesive compound ranges from 0.5 to 5 % (w/v). In a preferred embodiment, a composition is disclosed, wherein the average molecular weight of said mucoadhesive compound is at least 100 kDa.

In a preferred embodiment, a composition is disclosed, wherein said mucoadhesive compound comprises cellulose derivatives, gellan gum, guar gum, karaya gum, xanthan gum, carrageenan, alginate, pectin, dextran, chitosan, agarose, hyaluronic acid, gelatin, pectin, tragacanth, poly (vinyl pyrrolidone) such as Kollidon VA 64, poly(dimethyl siloxane), poly acrylic acid-based polymers such as poly acrylates, plyethylene glycol, sodium alginate, polycarbophil such as Noveon AA-1, polylysene, dimethylaminoethyl dextran, poly vinyl alcohol, hydroxy ethyl starch, poloxamer 407, Polycarbophil (PCP), Carbopol® 97 IP NF, Carbopol® 974P NF, Carbopol® 934 NF, Carbopol® 71GNF, amylose, sodium croscarmellose, cellulose acetate phthalate, Cellulose acetate butyrate, and/or hydroxyl ethyl cellulose (HEC).

In a preferred embodiment, a composition is disclosed, wherein the mucoadhesive compound comprises at least one selected from the group consisting of Sodium carboxyl methyl cellulose (SCMC), methyl cellulose, carboxyl methyl cellulose (CMC), hydroxyl propyl cellulose, hydroxyl propyl methylcellulose (HPMC) and ethyl cellulose.

In a preferred embodiment, a composition is disclosed, wherein the mucoadhesive compound comprises hydroxylpropyl methylcellulose (HPMC).

In a preferred embodiment, a composition is disclosed, wherein the mucoadhesive compound comprises HPMC and wherein the HPMC comprises at least one of HPMC substitution type 2910, HPMC substitution type 1828, HPMC substitution type 2208 and HPMC substitution type 2906.

In a preferred embodiment, a composition is disclosed, wherein the mucoadhesive compound comprises HPMC and wherein the HPMC comprises homogenously substituted HPMC cellulose, alternatively wherein the HPMC comprises heterogeneously substituted cellulose. In a preferred embodiment, a composition is disclosed, further comprising a preservative.

In a preferred embodiment, a composition is disclosed, wherein the preservative is selected from the group consisting of sodium acetate, benzalkonium chloride, potassium sorbate, calcium sorbate, methyl paraben, ethyl paraben, propyl paraben, phenylcarbinol, chlorolbutanol, chlorolcresol, ethylenediaminetetraacetic acid (EDTA).

In a preferred embodiment, a composition is disclosed, wherein the antibody or antigenbinding fragment thereof comprises CR9114.

In a preferred embodiment, a composition is disclosed, wherein the composition is a controlled released composition.

In a preferred embodiment, a composition is disclosed, wherein the composition is in liquid form.

In a preferred embodiment, a composition is disclosed, having a viscosity ranging from 0.001 to 10 Pa.s, as determined by a sheer stress method.

In a preferred embodiment, a composition is disclosed, wherein the viscosity of the composition ranges from 0.1 to 5 Pa.s, as determined by a sheer stress method.

In a preferred embodiment, a composition is disclosed, wherein the median droplet size (D50) of the composition ranges from 0.5 pm to 200 pm, as determined using laser diffraction assessing the distribution by volume.

In a preferred embodiment, a composition is disclosed, wherein the composition has a bimodal droplet size distribution as determined using laser diffraction assessing the distribution by volume. In a preferred embodiment, a composition is disclosed, wherein the apex of the first mode of the bimodal droplet size distribution has a droplet size ranging from 3 pm to 30 pm, as determined using laser diffraction assessing the distribution by volume.

In a preferred embodiment, a composition is disclosed, wherein the apex of the second mode has a droplet size ranging from 30 pm to 150 pm, as determined using laser diffraction assessing the distribution by volume.

In a preferred embodiment, a composition is disclosed, wherein the apexes of the bimodal composition have unequal heights.

In a preferred embodiment, a composition is disclosed, wherein the first mode has a higher apex in the bimodal composition.

In a preferred embodiment, a composition is disclosed, wherein the first mode has a lower apex in the bimodal composition.

In a preferred embodiment, a composition is disclosed, wherein the composition has a trimodal droplet size distribution as determined using laser diffraction assessing the distribution by volume.

In a preferred embodiment, a composition is disclosed, wherein the apex of the first mode of the trimodal droplet size distribution has a droplet size ranging from 3 pm to 30 pm, as determined using laser diffraction assessing the distribution by volume.

In a preferred embodiment, a composition is disclosed, wherein the apex of the second mode of the trimodal droplet size distribution has a droplet size ranging from 30 pm to 100 pm, as determined using laser diffraction assessing the distribution by volume. In a preferred embodiment, a composition is disclosed, wherein the apex of the third mode of the trimodal droplet size distribution has a droplet article size ranging from 100 pm to 150 pm, as determined using laser diffraction assessing the distribution by volume.

In a preferred embodiment, a composition is disclosed, wherein the apexes of the trimodal composition have unequal heights, as determined using laser diffraction assessing the distribution by volume.

In a preferred embodiment, a composition is disclosed, wherein the contact angles of a droplet of the composition in water, Phosphate-buffered saline (PBS), isotonic saline, and artificial saliva on the surface of the nasal mucosa are approximately 61° ± 5°, 48° ± 5°, and 57° ± 5°, respectively, as determined by atomic force microscopy (AFM).

In a preferred embodiment, a composition is disclosed, wherein the viscosity of the composition ranges from 0.001 to 10 Pa.s, as determined by a sheer stress method.

In a preferred embodiment, a kit comprising the composition is disclosed, wherein the kit further comprises at least one of the following: a pharmaceutical acceptable excipient, solvent and/or cosolvent, pH buffer, antioxidant, osmolality agent, penetration enhancer, suspending agent, and/or surfactant.

In a preferred embodiment, a spray device comprising the composition as herein described is disclosed. In a preferred embodiment, a spray device comprising the composition as herein described is disclosed, wherein the spray device is an inhaler, insufflator, or a spray pump.

In a preferred embodiment, a spray device comprising the composition as herein described is disclosed, wherein the spray is an insufflator.

In a preferred embodiment, a method for preparing the composition as herein described is disclosed.

In a preferred embodiment, a composition comprising an antibody, a bispecific antibody or an antigen binding fragment is disclosed, the concentration of the antibody, bispecific antibody or antigen binding fragment ranges from 0.00010 to 25% (w/v) and the concentration of the mucoadhesive compound ranges from 0.1 to 10 % (w/v).

In a preferred embodiment, a composition comprising an antibody, a bispecific antibody or an antigen binding fragment is disclosed, the composition further comprising at least one mucoadhesive compound, and/or at least one preservative, wherein the concentration of antibody or antigen-binding fragment thereof ranges from 0.00010 to 25% (w/v), the concentration of the mucoadhesive compound ranges from 0.1 to 10 % (w/v) and the concentration of the preservative ranges from 0.01 to 5 % (w/v).

In a preferred embodiment, a composition is disclosed, wherein the antibody or antigenbinding fragment thereof comprises CR9114 at a concentration ranging from 5 to 20% (w/v).

The inventors have discovered that a composition comprising a therapeutic antibody having the complementarity-determining regions (CDR) as disclosed herein, at least one mucoadhesive compound, at least one preservative for use in prophylactically and/or therapeutically treating diseases and/or symptoms caused by influenza A virus subtype A2, preferably influenza A subtype Al, and/or preferably influenza virus B subtypes. The inventors have discovered that a composition comprising CR9114 ranging from 0.00010 to 25 (w/v), at least one mucoadhesive compound ranging from 0.1 to 10 % (w/v), at least one preservative 0.01 to 5 % (w/v), can advantageously be used in the prophylactic and/or therapeutic treatment of diseases and/or symptoms caused by influenza A virus subtype A2, preferably influenza A subtype Al, and/or preferably influenza virus B subtypes.

In a preferred aspect of the invention, there is provided a sustained release formulation comprising CR9114 for use in preventing and/or treating a patient having influenza, the formulation comprising: (a) CR9114 or a fragment thereof in an amount of not more than 25 w/v; (b) a mucoadhesive agent ranging from 0.1 to 10 % (w/v); and (c) at least one pharmaceutically acceptable excipient, wherein the formulation is administered to the patient once daily.

DESCRIPTION OF THE DRAWINGS

Figure 1. Schematic of Experimental Protocol - Short-term exposure of the formulations of the present invention

Cilia beating frequency (CBF) assessed before the apical exposure (TO) and at 3 hours (T3h). Tissue integrity as measured by transepithelial electrical resistance (TEER) and mucociliary clearance (MCC) performed at T3h.

Figure 2. Schematic of Experimental Protocol - Repeated dose exposure of the formulations of the present invention

Schematic illustrating the long term (up to 96 hours (T96h)) plan of exposing a mixture of cells isolated from 14 different healthy nasal donors to the formulations of the present inventions by measuring transepithelial electrical resistance (TEER), LDH release, CBF, and MCC at specific time points. TEER and LDH release were assessed at T3h, T27h, T51h and T75h. Interleukin-8 (IL-8) basal medium was collected at T24h, T48h, T72h and T96h. MCC was measured at T75h. CBF was measured at TO, T3h, T24h, T27h, T48h, T5 Ih, T72h, T75h and T96h.

Figure 3. MCC assay at 3 hours (T3h) Fully differentiated human nasal epithelial cells isolated from 14 different healthy nasal donors were exposed to each of the formulations as described in Table 3a and each of the formulations were tested three times. MCC values were measures at 3 hours (T3h). Due to insufficient amount of formulation NP-018, no data was available for the cells exposed to NP-018 at T3h. In addition to exposing to the formulations as described in Table 3a, cells were exposed to hyaluronan, vehicle, Base formulation, COVITRAP™, respectively. One group of cells without being exposed to any formulations was introduced as negative control.

Figure 4. MCC assay of cells exposed for a long term to the formulations of the present invention

Fully differentiated human nasal epithelial cells isolated from 14 different healthy nasal donors were exposed to each of the formulations as described in Table 3a and each of the formulations were tested three times. MCC values were measures at 75 hours (T75h). Due to insufficient amount of formulation NP-018, the cells exposed to NP-018 were analyzed at T51h. In addition to exposing to the formulations as described in Table 3a, cells were exposed to hyaluronan, vehicle, Base formulation, COVITRAP™, respectively. One group of cells without being exposed to any formulations was introduced as negative control.

Figure 5. CBF assay at 0 (TO) and 3 hours (T3h)

Fully differentiated human nasal epithelial cells isolated from 14 different healthy nasal donors were exposed to each of the formulations as described in Table 3a and each of the formulations were tested three times. CBF values were measures at 0 (TO) and 3 hours (T3h). Due to insufficient amount of formulation NP-018, no data was available for the cells exposed to NP-018 at TO and T3h. In addition to exposing to the formulations as described in Table 3a, cells were exposed to hyaluronan, vehicle, Base formulation, COVITRAP™, respectively. One group of cells without being exposed to any formulations was introduced as negative control. Figure 6. CBF assay of cells exposed for a long term to the formulations of the present invention

Fully differentiated human nasal epithelial cells isolated from 14 different healthy nasal donors were exposed to each of the formulations as described in Table 3a and each of the formulations were tested three times. CBF was analyzed at TO, T3h, T24h, T27h, T48h, T51h, T72h, T75h, and T75h. In addition to exposing to the formulations as described in Table 3a, cells were exposed to hyaluronan, vehicle, Base formulation, COVITRAP™, respectively. One group of cells without being exposed to any formulations was introduced as negative control.

Figure 7. TEER assay at 3 hours (T3h)

Fully differentiated human nasal epithelial cells isolated from 14 different healthy nasal donors were exposed to each of the formulations as described in Table 3a and each of the formulations were tested three times. TEER values were measures at 3 hours (T3h). Due to insufficient amount of NP-018, no data was available for the cells exposed to NP-018 at T3h. In addition to exposing to the formulations as described in Table 3a, cells were exposed to hyaluronan, vehicle, Base formulation, COVITRAP™, respectively. One group of cells without being exposed to any formulations was introduced as negative control.

Figure 8. TEER assay of cells exposed for a long term to the formulations of the present invention

Fully differentiated human nasal epithelial cells isolated from 14 different healthy nasal donors were exposed to each of the formulations as described in Table 3a and each of the formulations were tested three times. TEER was analyzed at T3h, T27h, and T51h. In addition to exposing to the formulations as described in Table 3a, cells were exposed to hyaluronan, vehicle, Base formulation, COVITRAP™, respectively. One group of cells without being exposed to any formulations was introduced as negative control.

Figure 9. LDH cytotoxicity assay

Fully differentiated human nasal epithelial cells isolated from 14 different healthy nasal donors were exposed to each of the formulations as described in Table 3a and each of the formulations were tested three times. TEER was analyzed at T3h, T27h, T51h, T75h, and T96h. In addition to exposing to the formulations as described in Table 3a, cells were exposed to hyaluronan, vehicle, Base formulation, COVITRAP™, and Triton XI 00, respectively. One more group of cells without being exposed to any formulations was introduced as negative control.

Figure 10. IL-8 Basal Release assay

Fully differentiated human nasal epithelial cells isolated from 14 different healthy nasal donors were exposed to each of the formulations as described in Table 3a and each of the formulations were tested three times. TEER was analyzed at T24h, T48h, T72h, and T96h. In addition to exposing to the formulations as described in Table 3a, cells were exposed to hyaluronan, vehicle, Base formulation, and COVITRAP™, respectively. One more group of cells without being exposed to any formulations was introduced as negative control.

DETAILED DESCRIPTION OF THE INVENTION

For the purpose of clarity and a concise description, features may be described herein as part of the same or separate embodiments, however, it will be appreciated that the disclosure includes embodiments having combinations of all or some of the features described. References to “method for treatment”, “treatment method”, “composition for use” and “use of a composition in the manufacture of a medicament” can be used interchangeably and embodiments disclosed in relation to any one of those aspects also applies in relation to said other aspects. In other words, they all refer to medical treatments involving a composition as disclosed herein.

The present invention provides a composition comprising an antibody as described herein, and at least one mucoadhesive compound.

The present invention thereby provides the means of actively preventing and/or treating respiratory viral infections, and the means of actively performing self-prevention and/or selftreatment in any locations conveniently. The composition as described herein is capable of prophylactically and/or therapeutically treating a disease, preferably a disease and/or symptoms caused by influenza viruses.

Antibody

The term ‘antibody’, as used herein, includes reference to an intact immunoglobulin, including monoclonal antibodies, such as chimeric, humanized or human monoclonal antibodies, or to a binding molecule comprising an antigen-binding domain (such as heavy chain CDRs 1-3 of a variable domain) of an antibody as disclosed herein, or to a bispecific antibody, or an antibody that competes with an antibody as disclosed herein for specific binding to the binding partner of the immunoglobulin.

Preferably an antibody or antigen-binding fragment as described herein is capable of preventing and/or neutralizing a respiratory viral infection in an in vitro infection model, and/or in an in vivo animal infection model, and/or in a human. Preferably, the antibody or antigen-binding fragment, is human, humanized, or chimeric.

In other words, functional fragments of antibodies are also encompassed by the term ‘antibody’. Functional fragments as disclosed herein may comprise or consist of parts of or full length of the polypeptide sequence of the intact immunoglobulin, while the functional fragments are capable of binding to the immunoglobulin with sufficient affinity.

Antibodies are generally Y-shaped proteins. Within the antibody, constant domain and variable domains are generally present. The variable domain facilitates antigen binding. An antibody generally comprises two heavy chains and two light chains. Both the heavy chains and the light chains are partially constant and partially variable. Antibodies occur in a few classes: IgA, IgD, IgE, IgG and IgM. Preferably, the antibody of the invention is of the IgG, preferably IgGl, class. Some classes may be further subdivided into subclasses or isotypes. For example, the IgG class is subdivided into the subclasses IgGl, IgG2, IgG3 and IgG4. Preferably, the antibody of the invention is of the IgG, preferably IgGl, class. Antigenbinding regions, or antigen-binding fragments of an antibody, which are encompassed by the term ‘antibody’ and which are therefore part of the present invention, may include, for example, Fab, F(ab’), F(ab’)2, dAb, Fv, Fd, CDR fragments, diabodies, triabodies, tetrabodies, single-chain antibodies (scFv), scFv-Fc, bivalent single-chain antibodies such as tandem di-scFv and diabody, trivalent single-chain antibodies such as tandem tri-scFv and triabody, tandem single-domain antibodies, Fab-scFv bispecific antibodies, single-chain phage antibodies, (poly)peptides and variants thereof that contain at least a fragment of an immunoglobulin that is sufficient to confer specific antigen binding to the (poly)peptide, etc. The above fragments may be produced synthetically or by enzymatic or chemical cleavage of intact immunoglobulins or they may be genetically engineered by recombinant DNA techniques. The above fragments may be recombinantly expressed in a mammalian cell system.

Preferably, the functional fragments as disclosed herein comprise the amino acid sequences of a CDR sequence comprising SEQ ID NO: 001. Preferably wherein the first antibody (arm) comprises CDR sequences comprising any one or more of SEQ ID NO: 001, SEQ ID NO: 002, or SEQ ID NO.: 003, and the second antibody (arm) comprises CDR sequences comprising any one or more of SEQ ID NO: 004, SEQ ID NO: 005, or SEQ ID NO.: 006.

More preferably, wherein the heavy chain on the first antibody (arm) comprises CDR sequences comprising any one or more of SEQ ID NO: 001, SEQ ID NO: 002, or SEQ ID NO.: 003, the light chain on the first antibody (arm) comprises any functional humanized CDR sequences; the second antibody (arm) comprises heavy chain CDR sequences comprising any one or more of SEQ ID NO: 007, SEQ ID NO: 008, or SEQ ID NO.: 009, and the light chain on the second antibody (arm) comprises CDR sequences comprising any one or more of SEQ ID NO: 010, SEQ ID NO: 011, or SEQ ID NO.: 012.

The methods for production of antibodies and antigen-binding fragments are well-known to a person skilled in the art. The bispecific antibody or antigen-binding fragment may be conjugated or unconjugated. The bispecific antibody or antigen-binding fragment may be conjugated, linked, or otherwise physically or functionally associated with an effector moiety or tag, such as inter alia an enzyme, a liposome, a radioactive substance, a fluorophore, a toxic substance, the bispecific antibody or antigen-binding fragment may be stabilized, multimerized, humanized or otherwise manipulated.

Antibodies may be neutralizing, which includes reference to inhibition of a virus as measured by an in vitro neutralization assay, for instance in terms of viral entry and/or viral replication in the individuals as disclosed herein. Neutralization can for example be achieved by inhibiting the attachment or adhesion of the virus to the cell surface, or by inhibition of the fusion of viral and cellular membranes following attachment of the virus to the target cell or by inhibiting viral egress from cells. Neutralization does not specify the method of neutralization. Preferably, the antibody is cross-neutralizing, which includes reference to the ability of the antibodies of the invention to bind and neutralize a set of different molecules, preferably different molecules of different subtypes belonging to the Influenza family.

Preferably, the bispecific antibody or antigen-binding fragment of the invention as disclosed herein can cross-neutralize Influenza virus.

Antibodies comprise complementarity determining regions situated on the variable domains of the heavy chain and the light chain. The CDRs contribute to a large extent to the antigen binding site. Three CDRs can be distinguished, namely CDR1, CDR2 and CDR3. As each CDR can be located on either the light chain or the heavy chain, there are generally six CDRs for each antigen receptor that collectively contact the antigen: the light chain CDR1, the light chain CDR2, the light chain CDR3, the heavy chain CDR1, the heavy chain CDR2 and the heavy chain CDR3. The CDRs of type CDR3 are the most variable. The CDRs can be specific for linear epitopes, discontinuous epitopes, or conformational epitopes of proteins or protein fragments, either as present on the protein in its native conformation or, in some cases, as present on the proteins as denatured or activated. Epitopes may also consist of or comprise post-translational modifications of proteins. Antibodies of the invention that are of particular interest, are antibodies comprising CDRs that recognize influenza virus antigens, such as the hemagglutinin and neuraminidase surface antigens of an influenza virion. The bispecific antibody or antigen-binding fragment thereof as disclosed herein binds to a conserved epitope at the base of the hemagglutinin and neuraminidase surface antigens of an influenza virion. The epitope of the bispecific antibody or antigen-binding fragment thereof as disclosed herein uses light and heavy chain CDR loops.

Suitably, the bispecific antibody or antigen-binding fragment thereof of the present invention is capable of binding to two different targets, such as hemagglutinin and neuraminidase, simultaneously. Hence, the bispecific antibody or antigen-binding fragment thereof of the present invention may further reduce and/or prevent new virus variants or different virus variants from egressing and/or replicating.

The bispecific antibody or antigen-binding fragment thereof as disclosed herein can be used in isolated or non-isolated form.

Preferably, the compositions of the invention comprise a single anti-influenza virus bispecific antibody or antigen-binding fragment thereof as disclosed herein.

Furthermore, the bispecific antibody as disclosed herein can be used alone or in a mixture comprising the antibody (or variant, fragment or bispecific thereof) as disclosed herein, and/or with other antibodies that bind to an influenza virus and have an influenza virus inhibiting effect. In other words, the antibody as disclosed herein can be used in combination, e.g., as a pharmaceutical composition or co-administration of compositions comprising two or more antibodies that specifically bind influenza virus. For example, antibodies having different, but complementary activities can be combined in a single therapy to achieve a desired therapeutic or prophylactic effect, but alternatively, antibodies having identical activities can also be combined in a single therapy to achieve a desired prophylactic or therapeutic effect. Optionally, the mixture further comprises at least one other therapeutic agent.

Preferably, the bispecific antibody as disclosed herein is a human antibody.

Preferably, the compositions of the invention further comprise inhibitors of protease and aminopeptidases. Polypeptide

The term “polypeptide” as used herein, includes reference to a molecule comprising a polymer of amino acids joined together by peptide bonds which acts as an active pharmaceutical ingredient for providing the prophylactic and/or therapeutic treatment to a subject in need thereof.

Preferably, the molecule comprises an oligonucleotide, a protein and/or an antibody.

Preferably, the molecule consists of the antibody as described herein.

Preferably, the molecule comprises or consists of the monoclonal antibody known as CR9114. The composition comprising CR9114 can be used in prophylactic and/or therapeutic treatment of diseases and/or symptoms caused by at least one influenza virus as described herein.

Complementarity-determining regions (CDRs)

Preferably, Complementarity Determining Regions (CDRs) are according to Kabat et al., (1991) as described in Sequences of Proteins of Immunological Interest.

The variable binding regions comprise discrete, well-defined sub-regions known as CDRs and “framework regions” (FRs). The terms “complementarity determining region” and “CDR” t, refer to sequences of amino acids within antibody variable regions, which together confer the antigen specificity and/or binding affinity of the antibody, wherein consecutive CDRs (i.e., CDR1, CDR2 and CDR3) are separated from one another in primary structure by a framework region. There are three CDRs in each variable region (HCDR1, HCDR2, HCDR3; LCDR1, LCDR2, LCDR3; also referred to as CDRHs and CDRLs, respectively).

In a preferred embodiment, an antibody, a bispecific antibody or an antigen binding fragment is disclosed, wherein the antibody or antigen-binding fragment comprises an antigen-binding domain that binds to the stem region of hemagglutinin (HA) of the influenza virus, wherein the antigen-binding domain comprises: a heavy chain variable domain (VH) comprising a HCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 001, a HCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 002, and a HCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 003, and a light chain variable domain (VL) comprising a LCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 004, a LCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 005, and a LCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 006.

In a preferred embodiment, an antibody, a bispecific antibody or an antigen binding fragment is disclosed, wherein the antibody or antigen-binding fragment comprises an antigen-binding domain that binds to neuraminidase (NA) of the influenza virus, wherein the antigen-binding domain comprises: a heavy chain variable domain (VH) comprising a HCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 007, a HCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 008, and a HCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 009, and a light chain variable domain (VL) comprising a LCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 010, a LCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 011, and a LCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 012.

In a preferred embodiment, an antibody, a bispecific antibody or an antigen binding fragment is disclosed, wherein the antibody or antigen-binding fragment comprises an antigen-binding domain that binds to the fusion peptide of the spike protein of the SARS-CoV-2 virion., wherein the antigen-binding domain comprises: a heavy chain variable domain (VH) comprising a HCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 013, a HCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 014, and a HCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 015, and a light chain variable domain (VL) comprising a LCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 016, a LCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 017, and a LCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 018. In a preferred embodiment, an antibody, a bispecific antibody or an antigen binding fragment is disclosed, wherein the antibody or antigen-binding fragment comprises an antigen-binding domain that binds to the fusion peptide of the spike protein of the SARS-CoV-2 virion., wherein the antigen-binding domain comprises: a heavy chain variable domain (VH) comprising a HCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 019, a HCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 020, and a HCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 021, and a light chain variable domain (VL) comprising a LCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 022, a LCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 023, and a LCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 024.

In a preferred embodiment, an antibody, a bispecific antibody or an antigen binding fragment is disclosed, wherein the antibody or antigen-binding fragment comprises an antigen-binding domain that binds to the stem helix of the spike protein of the SARS-CoV-2 virion, wherein the antigen-binding domain comprises: a heavy chain variable domain (VH) comprising a HCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 025, a HCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 026, and a HCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 027, and a light chain variable domain (VL) comprising a LCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 028, a LCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 029, and a LCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 030.

In a preferred embodiment, an antibody, a bispecific antibody or an antigen binding fragment is disclosed, wherein the antibody or antigen-binding fragment comprises an antigen-binding domain that binds to the stem helix of the spike protein of the SARS-CoV-2 virion, wherein the antigen-binding domain comprises: a heavy chain variable domain (VH) comprising a HCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 031, a HCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 032, and a HCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 033, and a light chain variable domain (VL) comprising a LCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 034, a LCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 035, and a LCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 036.

In a preferred embodiment, an antibody, a bispecific antibody or an antigen binding fragment is disclosed, wherein the antibody or antigen-binding fragment comprises an antigen-binding domain that binds to the stem helix of the spike protein of the SARS-CoV-2 virion, wherein the antigen-binding domain comprises: a heavy chain variable domain (VH) comprising a HCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 037, a HCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 038, and a HCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 039, and a light chain variable domain (VL) comprising a LCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 040, a LCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 041, and a LCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 042.

Bispecific antibodies of the invention

In a preferred embodiment, we disclose an anti-influenza bispecific antibody or antigenbinding fragment thereof, wherein the bispecific antibody or antigen-binding fragment comprises: (a) a first antigen-binding domain that binds to hemagglutinin of an influenza virion, wherein the first antigen-binding domain comprises: a heavy chain variable domain (VH) comprising a HCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 001, a HCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 002, and a HCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 003, and a light chain variable domain (VL) comprising a LCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 004, a LCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 005, and a LCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 006; and (b) a second antigen-binding domain that binds to neuraminidase surface antigens of an influenza virion, wherein the second antigen-binding domain comprises: a VH comprising a HCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 007, a HCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 008, and a HCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 009, and a VL comprising a LCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 010, a LCDR 2 comprising the amino acid sequence as described in SEQ ID NO: Oil, and a LCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 012.

In a preferred embodiment, we disclose an anti-SARS-CoV-2 bispecific antibody or antigenbinding fragment thereof, wherein the bispecific antibody or antigen-binding fragment comprises: (a) a first antigen-binding domain that binds to the fusion peptide of SARS-CoV- 2, wherein the first antigen-binding domain comprises: a heavy chain variable domain (VH) comprising a HCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 013, a HCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 014, and a HCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 015, and a light chain variable domain (VL) comprising a LCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 016, a LCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 017 , and a LCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 018; and (b) a second antigen-binding domain that binds to the stem helix of SARS-CoV-2, wherein the second antigen-binding domain comprises: a VH comprising a HCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 025, a HCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 026, and a HCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 027, and a VL comprising a LCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 028, a LCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 029 , and a LCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 030.

In a preferred embodiment, we disclose an anti-SARS-CoV-2 bispecific antibody or antigenbinding fragment thereof, wherein the bispecific antibody or antigen-binding fragment comprises: (a) a first antigen-binding domain that binds to the fusion peptide of SARS-CoV- 2, wherein the first antigen-binding domain comprises: a heavy chain variable domain (VH) comprising a HCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 013, a HCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 014, and a HCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 015, and a light chain variable domain (VL) comprising a LCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 016, a LCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 017 , and a LCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 018; and (b) a second antigen-binding domain that binds to the stem helix of SARS-CoV-2, wherein the second antigen-binding domain comprises: a VH comprising a HCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 031, a HCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 032, and a HCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 033, and a VL comprising a LCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 034, a LCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 035, and a LCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 036.

In a preferred embodiment, we disclose an anti-SARS-CoV-2 bispecific antibody or antigenbinding fragment thereof, wherein the bispecific antibody or antigen-binding fragment comprises: (a) a first antigen-binding domain that binds to the fusion peptide of SARS-CoV- 2, wherein the first antigen-binding domain comprises: a heavy chain variable domain (VH) comprising a HCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 013, a HCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 014, and a HCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 015, and a light chain variable domain (VL) comprising a LCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 016, a LCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 017 , and a LCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 018; and (b) a second antigen-binding domain that binds to the stem helix of SARS-CoV-2, wherein the second antigen-binding domain comprises: a VH comprising a HCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 037, a HCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 038, and a HCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 039, and a VL comprising a LCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 040, a LCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 041, and a LCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 042. In a preferred embodiment, we disclose an anti-SARS-CoV-2 bispecific antibody or antigenbinding fragment thereof, wherein the bispecific antibody or antigen-binding fragment comprises: (a) a first antigen-binding domain that binds to the fusion peptide of SARS-CoV- 2, wherein the first antigen-binding domain comprises: a heavy chain variable domain (VH) comprising a HCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 019, a HCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 020, and a HCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 021, and a light chain variable domain (VL) comprising a LCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 022, a LCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 023, and a LCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 024; and (b) a second antigen-binding domain that binds to the stem helix of SARS-CoV-2, wherein the second antigen-binding domain comprises: a VH comprising a HCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 025, a HCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 026, and a HCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 027, and a VL comprising a LCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 028, a LCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 029 , and a LCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 030.

In a preferred embodiment, we disclose an anti-SARS-CoV-2 bispecific antibody or antigenbinding fragment thereof, wherein the bispecific antibody or antigen-binding fragment comprises: (a) a first antigen-binding domain that binds to the fusion peptide of SARS-CoV- 2, wherein the first antigen-binding domain comprises: a heavy chain variable domain (VH) comprising a HCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 019, a HCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 020, and a HCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 021, and a light chain variable domain (VL) comprising a LCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 022, a LCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 023, and a LCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 024; and (b) a second antigen-binding domain that binds to the stem helix of SARS-CoV-2, wherein the second antigen-binding domain comprises: a VH comprising a HCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 031, a HCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 032, and a HCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 033, and a VL comprising a LCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 034, a LCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 035, and a LCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 036.

In a preferred embodiment, we disclose an anti-SARS-CoV-2 bispecific antibody or antigenbinding fragment thereof, wherein the bispecific antibody or antigen-binding fragment comprises: (a) a first antigen-binding domain that binds to the fusion peptide of SARS-CoV- 2, wherein the first antigen-binding domain comprises: a heavy chain variable domain (VH) comprising a HCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 019, a HCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 020, and a HCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 021, and a light chain variable domain (VL) comprising a LCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 022, a LCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 023, and a LCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 024; and (b) a second antigen-binding domain that binds to the stem helix of SARS-CoV-2, wherein the second antigen-binding domain comprises: a VH comprising a HCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 037, a HCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 038, and a HCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 039, and a VL comprising a LCDR 1 comprising the amino acid sequence as described in SEQ ID NO: 040, a LCDR 2 comprising the amino acid sequence as described in SEQ ID NO: 041, and a LCDR 3 comprising the amino acid sequence as described in SEQ ID NO: 042.

A kit comprising the antibody of the invention

The present invention provides a kit comprising the composition as described herein and a delivery device.

For the purpose of prophylactic and/or therapeutic treatment of an infection as disclosed herein, at least one composition may be conveniently combined into a kit. Thus, the term “kit”" as disclosed herein, includes the composition as disclosed herein and at least one compound for prolonging the prophylactic and/or therapeutic effect of the antibody in a human subject, for prolonging the shelf life of the composition as described herein, for maintaining the physical and/or chemical stability of the antibody, bispecific antibody or antigen binding fragment, and/or for enhancing the administration of the composition as herein described at the site in a human subject, and means for retaining the composition and said at least one compound as herein described.

Administering and Administration

The terms ‘administering’ and ‘administration’, as used herein, include reference to the provision of one or more antibody, bispecific or antigen-binding fragment thereof and optionally one or more excipient with the aim to treat, cure, reduce, or prevent a disease or its symptoms in an individual, or to promote the individual’s well-being. Preferred methods of administration of the antibody, bispecific antibody or antigen binding fragment as disclosed herein include mucosal administration. An especially preferred method of administration is intranasal administration. An especially preferred method of administration is oral inhalation.

Individual, human subject, or subject

The term ‘individual’, “human subject” or “subject”, as used herein, includes reference to a mammal that is subject to, or a risk of suffering from viral infection. The phrase ‘an individual’, as used herein, includes reference to a mammal but not limited to a human that benefits from a specified therapy, for example, the phrase ‘an individual’ may encompass non-human primates (NHP), birds, pigs, dogs, cats, and horses.

Preferably the term ‘individual’, “human subject” or “subject”, as used herein, includes reference to a human that is subject to, or a risk of suffering from viral infection.

Infection may take place in any system, tissue or cell belonging to the host, including the host’s microbiome. Influenza virus infection and the disease influenza virus may occur in individuals of all age groups and sexes. Nonetheless, preferably, the individual is a human, in particular an elderly human such as a human that is at least 60, 65, 70, 75, 80, or at least 85 years old, or that has an increased risk of infection because of occupation or living environment. Preferably, the individual is at risk of suffering from severe illness, for example from influenza, respiratory syncytial virus and coronaviruses, once infected. In preferred embodiments, the individual has an underlying disease such as (i) a respiratory disease such as asthma, COPD, chronic bronchitis and lung emphysema, (ii) cardiovascular disease such as cardiac arrhythmia or individuals that have received cardiac surgery, (iii) diabetes, (iv) renal failure and/or (v) a disease affecting the immune system, for instance immunocompromised individuals, or higher risk of viral infection because of occupation.

An individual in need thereof

The phrase ‘an individual in need thereof, as used herein, includes reference to a mammal such as a human that benefits from a specified therapy. A treatment method of the invention may be used prophylactically, the exhibition of symptoms or indications for influenza, respiratory syncytial virus and coronavirus infection are not required. Individuals that are especially in need of the method or antibody for use of the invention, are individuals with an elevated risk of influenza, respiratory syncytial virus and coronavirus infection, individuals with an elevated risk of developing severe symptoms (illness), for example influenza or CO VID-19, and/or individuals with an elevated risk of dying from these diseases. The person skilled in the art is aware of the risk factors for an elevated risk of influenza virus or SARS- CoV-2 infection, an elevated risk of developing severe symptoms of influenza virus or SARS-CoV-2 infection, and an elevated risk of dying from influenza virus or SARS-CoV-2 infection.

Use in a method for treatment

The present invention relates inter alia to an antibody, a bispecific antibody or an antigen binding fragment, for use in a method for treatment of influenza, respiratory syncytial virus and coronavirus infection in an individual, more preferably the antibody or bispecific antibody or antigen-binding fragment thereof can be used in a method for the prophylactic and/or therapeutic treatment of influenza, respiratory syncytial virus and coronavirus infection in an individual. Influenza infection/Influenza virus infection

Preferably, the composition as herein described comprises an antibody, a bispecific antibody or an antigen binding fragment, that targets an influenza virus.

The terms ‘influenza virus infection’, ‘flu virus infection’, and ‘flu’ as used herein, includes reference to the pathological or non-pathological, preferably pathological, entrance and residence of an influenza virus of any type in a human host. The infecting virus may replicate within the host, its cells or the cells of its microbiome. The infecting virus may or may not cause a disease, for example influenza. The infection may or may not be able to be detected by methods for virus infection detection known in the art. The infected individual may or may not be aware of the infection. Typical, but non-exclusive locations of the human body where, for example Influenza virus, may be located in an infected individual, are the respiratory system and/or cells thereof and/or the cardiovascular system and/or cells thereof. The term ‘influenza virus infection’, as used herein, further includes reference to the entrance and residence of a part of an influenza virus of any type that is able to cause viral replication in a human host. The term ‘influenza virus infection’ encompasses symptoms or disease following the infection, e.g. influenza.

The term ‘influenza virus’, as used herein, includes reference to a negative-sense singlestranded RNA virus belonging to the family of Influenza viruses.

Preferably, an antibody, bispecific antibody or antigen binding fragment as disclosed herein is capable of specifically binding to the hemagglutinin and neuraminidase of an influenza virus, in particular influenza A virus subtype A2, influenza A virus subtype Al and/or influenza virus B in both the Yamagata and Victoria lineages.

Preferably, an antibody, bispecific antibody or antigen binding fragment as disclosed herein is capable of neutralizing an influenza virus, in particular, in particular influenza A virus subtype A2, influenza A virus subtype Al and/or influenza virus B in both Yamagata and Victoria lineages. Preferably, an antibody, bispecific antibody or antigen binding fragment as disclosed herein is capable of neutralizing at least one or more, preferably two or more, preferably three or more, preferably four or more, even more preferably five or more influenza virus subtypes.

The composition is capable of providing prophylactic and/or therapeutic effects on symptoms caused by influenza viruses A subtypes Al, A2 and/or influenza B subtypes. The composition as described herein is capable of being conveniently administered through various administration routes as described herein. Moreover, the composition as described herein is comprised in a medical device.

Coronaviruses

Preferably, the composition as herein described comprises an antibody, bispecific antibody or antigen binding fragment, that targets a coronavirus.

Preferably, the composition as herein described comprises an antibody, bispecific antibody or antigen binding fragment, that targets any one or more of the preferred viruses selected from the alpha coronavirus 229E, the alpha coronavirus NL63, the beta coronavirus OC43, the beta coronavirus HKU1, the beta coronavirus MERS-CoV (that causes Middle East Respiratory Syndrome, or MERS), the beta coronavirus SARS-CoV (that causes severe acute respiratory syndrome, or SARS), or the novel coronavirus SARS-CoV-2 (that causes coronavirus disease 2019, or COVID-19).

The terms ‘COVID-19 infection’, ‘SARS-CoV-2 infection’, and ‘CO VID’ as used herein, includes reference to the pathological or non-pathological, preferably pathological, entrance and residence, in a host, of a SARS-CoV-2 virus or any variant of interest (VOIs) and variants of concern (VOCs) as identified by the World Health Organization (WHO). The infecting virus may replicate within the host, its cells or the cells of its microbiome. The infecting virus may or may not cause a disease, for example COVID-19. The infection may or may not be able to be detected by methods for virus infection detection known in the art. The infected individual may or may not be aware of the infection. Typical, but non-exclusive locations of the human body where, for example SARS-CoV-2 virus, may be located in an infected individual, are the respiratory system and/or cells thereof and/or the cardiovascular system and/or cells thereof. The term ‘COVID-19 infection’, as used herein, further includes reference to the entrance and residence of a part of a SARS-CoV-2 virus of any type that is able to cause viral replication in a human host. The term ‘SARS-CoV-2 virus infection’ encompasses symptoms or disease following the infection, e.g. COVID-19.

Respiratory Syncytial Virus infection

Preferably, the composition as herein described comprises an antibody, bispecific antibody or antigen binding fragment, that targets an orthopneumovirus.

Preferably the composition as herein described comprises an antibody, bispecific antibody or antigen binding fragment, that targets any one or more of the preferred viruses selected from bovine respiratory syncytial virus (BRSV), respiratory syncytial virus (RSV) in particular human respiratory syncytial virus A2 (HRSV-A2) and human respiratory syncytial virus Bl (HRSV-B1).

Human respiratory syncytial virus (HRSV) is the most common and researched orthopneumovirus in view of the importance to humans. RSV is the leading viral agent among pneumoviruses in pediatric upper respiratory diseases globally. Novel pneumoviruses continue to be discovered.

Prophylactic treatment

The term ‘prophylactic treatment’, as used herein, includes reference to a treatment for preventing infection of an individual with a respiratory viral infection virus, or preventing symptoms after infection with a respiratory virus, or preventing severe symptoms after known infection with a respiratory virus with or without symptoms, or preventing hospitalization and death after infection with a respiratory virus.

Prevention of an infection is preferably performed by administration of an antibody, bispecific antibody or antigen binding fragment prior to exposure with a respiratory virus i.e. pre-exposure prophylaxis. ‘Prophylactically’ therefore preferably means prior to virus exposure. Nonetheless, it may also involve administration after infection, for example for reducing the replication or spread, or increasing clearance of the virus i.e. post-exposure prophylaxis.

Infected individuals may present with no symptoms. Alternatively, infected individuals may present with symptoms.

Preferably, post-exposure prophylaxis involves administering an antibody, bispecific antibody or antigen binding fragment after respiratory viral exposure to prevent symptomatic disease.

Preferably, post-exposure prophylaxis involves administering an antibody, bispecific antibody or antigen binding fragment after respiratory viral exposure to prevent severe disease, in particular hospitalization.

In a treatment method of the invention, prophylactic treatment involves administration of an antibody, bispecific antibody or antigen binding fragment against a respiratory virus at a point in time when the individual is not infected with the respiratory virus. In some embodiments, an individual in need thereof is not (yet) infected with a respiratory virus.

Therapeutic treatment

The term ‘therapeutic treatment’, as used herein, includes reference to treatment of a viral infection (including influenza virus disease, COVID-19 and RSV) after viral infection has taken place. A viral infection involves the entry of the body by the virus, and/or the replication of the virus in the body and/or the spreading of the virus to cells, tissues or locations in the body that were previously uninfected. A viral infection may cause one or more disease, but may also be latent, in other words may reside in the body without causing symptoms or disease. Mucosal administration

The mucosa comprises membranes that line cavities in the human body, covering internal organs, and consist of one or more layers of epithelial cells and loose connective tissue, which may produce mucus. Mucosal epithelial cells can secrete mucus, a protective fluid which can prevent and/or reduce pathogens and/or dirt from entering the body. At the same time, the mucus may keep the body tissues of a subject hydrated. The term ‘mucosally’, as used herein, may also be referred to as ‘mucosal administration’, and includes reference to a route of administration in which the composition as described herein is prophylactically and/or therapeutically provided to the mucosa, for example, mucosa found in the sites including but not limited to the nose, mouth, respiratory track, lungs, vagina, rectum or stomach. The mucous membrane lubricates and protects these organs and cavities from abrasive particles and bodily fluids, as well as invasive pathogens.

Preferably, the composition as described herein is administered mucosally. A preferred route of administration includes mucosal administration.

Preferably, the composition as described herein is a mucosal composition.

Intranasal

The term ‘intranasally’, as used herein, may also be referred to as ‘nasal administration’, and includes reference to a route of administration in which an antibody, bispecific antibody or antigen binding fragment as described herein or the composition as described herein is provided into the upper respiratory tract and/or lower respiratory tract, preferably through the nostrils, as part of a prophylactic and/or therapeutic treatment as disclosed herein. Nasal delivery is similar to pulmonary administration as it provides a non-invasive route of delivery to the mucosal surfaces. Nasal delivery avoids needles and allows repeat administration from a single device. Nasal delivery involves inspiration via the nose but where the composition as disclosed herein is primarily collected in the nasal cavity and turbinates. Preferably, the administration provides for an antibody, a bispecific antibody or an antigen binding fragment in the nasal cavity. Nasal administration can either be a form of topical administration or systemic administration, as the antibody, bispecific antibody or antigen binding fragment as disclosed herein are locally delivered and may go on to have either local or systemic effects. Preferably, nasal administration of the composition as disclosed herein, is in a form suitable for topical administration. Preferably, the composition as disclosed herein is delivered to the mucous membrane lining the nasal cavity. Preferred routes of administration include mucosal and especially intranasal administration.

Preferably, intranasal administration as disclosed herein may be performed using a liquid composition as described herein. Preferably the intranasal administration is performed using a nasal spray. Preferably the intranasal administration is performed using nose drops.

In some embodiments, the composition is delivered to the nasal cavity via the oral route. For example, RetroNose uses a breath-actuated pressurised metered-dose inhaler (pMDI) to administer drugs through the buccal cavity during the nasal expiratory phase. Such methods allow the droplets comprising an antibody, a bispecific antibody or antigen binding fragment to enter the nasal cavities through the pharynx. Preferably, the composition of the invention is administered intranasally.

Mucociliary clearance

Particulates within the inhaled airstream, such as dust, pollen, microorganisms and pollutants, have the potential to deposit on the viscoelastic mucous gel lining the turbinate walls. The cilia, beating within the periciliary fluid, engage with the underside of the mucus and propel the mucus and the deposited droplets to the nasopharynx, where they are either swallowed or expectorated. This process is termed mucociliary clearance. Under normal physiological conditions, the mucociliary clearance mechanism is able to clear mucus from the nasal turbinates at a rate of approximately 7 millimeters per minute. An efficient mucociliary clearance process reduces the residence time of any antibody, bispecific antibody or antigen binding fragment. A reduced nasal residence time reduces the ability of the antibody, bispecific antibody or antigen binding fragment to sequester any respiratory viruses before they infect a subject. In a preferred embodiment, a composition is disclosed comprising an antibody, bispecific antibody or antigen binding fragment, and at least one mucoadhesive compound, wherein the composition reduces the rate of mucociliary clearance to a rate of less than 20 millimeters per minute, preferably less than 19 millimeters per minute, preferably less than 18 millimeters per minute, preferably less than 17 millimeters per minute, preferably less than 16 millimeters per minute, preferably less than 15 millimeters per minute, preferably less than 14 millimeter per minute, preferably less than 13 millimeters per minute, preferably less than 12 millimeters per minute, preferably less than 11 millimeters per minute, preferably less than 10 millimeters per minute, preferably less than 9 millimeters per minute, preferably less than 8 millimeter per minute, preferably less than 7 millimeters per minute, preferably less than 6 millimeters per minute, preferably less than 5 millimeters per minute, preferably less than 4 millimeters per minute, preferably less than 3 millimeters per minute, preferably less than 2 millimeter per minute, preferably less than 1 millimeter per minute.

In a preferred embodiment, a composition is disclosed comprising an antibody, bispecific antibody or antigen binding fragment, and at least one mucoadhesive compound, wherein the composition reduces the rate of mucociliary clearance preferably by up to 5%, preferably up to 10%, preferably up to 15%, preferably up to 20%, preferably up to 25%, or preferably up to 30%, preferably up to 35%, preferably up to 40%, preferably up to 45%, preferably up to 50%, preferably up to 55%, or preferably up to 60%, preferably up to 65%, preferably up to 70%, preferably up to 75%, preferably up to 80%, preferably up to 85%, or preferably up to 90%, preferably by up to 95% of the original physiological mucociliary clearance speed.

Mucosal desiccant

Reduced nasal residence time reduces the ability of the antibody, bispecific antibody or antigen binding fragment to sequester any respiratory viruses before they infect a subject. Extending the nasal residence time of any administered antibody, bispecific antibody or antigen binding fragment increases the chances of viral sequestration and thus enhances the efficacy of these molecules. The inventors have surprisingly discovered that a formulation strategy involving some, but not complete, mucosal desiccation enhances the efficacy of the administered antibodies, bispecific antibodies or antigen binding fragments. In a preferred embodiment, a composition is disclosed comprising an antibody, bispecific antibody or antigen binding fragment and at least one mucosal desiccant.

In a preferred embodiment, a composition is disclosed comprising an antibody, bispecific antibody or antigen binding fragment as described herein and at least one mucosal desiccant.

In a preferred embodiment, the mucosal desiccating agent possesses hygroscopic properties.

Preferred mucosal desiccating agents may include, but are not limited to, sodium carboxymethylcellulose, calcium carboxymethylcellulose, colloidal silica dioxide, and combinations thereof.

A preferred mucosal desiccating agent comprises magnesium sulphate.

A preferred mucosal desiccating agent comprises sodium zeolite.

An especially preferred mucosal desiccating agent comprises hydroxypropyl methylcellulose (HPMC).

In a preferred embodiment, the mucosal desiccating agent may comprise at least one of hydroxypropyl methyl cellulose substitution type 2910, hydroxypropyl methylcellulose substitution type 1828, hydroxypropyl methylcellulose substitution type 2208 and hydroxypropyl methylcellulose substitution type 2906.

In a preferred embodiment, the mucosal desiccating agent may comprise any of the following preferred combinations, the hydroxypropyl methylcellulose substitution type 1828 and hydroxypropyl methylcellulose substitution type 2208, hydroxypropyl methylcellulose substitution type 1828 and hydroxypropyl methylcellulose substitution type 2906, hydroxypropyl methylcellulose substitution type 1828 and hydroxypropyl methylcellulose substitution type 2910, hydroxypropyl methylcellulose substitution type 2208 and hydroxypropyl methylcellulose substitution type 2906, hydroxypropyl methylcellulose substitution type 2208 and hydroxypropyl methylcellulose substitution type 2910, or hydroxypropyl methylcellulose substitution type 2906 and hydroxypropyl methylcellulose substitution type 2910.

Preferably the mucosal desiccant reduces the water content of the mucus to less than 95%, preferably to less than 90%, preferably to less than 85%, preferably to less than 80%, preferably to less than 75%, preferably to less than 70%, preferably to less than 65%, preferably to less than 60%, preferably to less than 55%, preferably to less than 50%, preferably to less than 45%, preferably to less than 40%, preferably to less than 35%, preferably to less than 30%, preferably to less than 25%, preferably to less than 20%, preferably to less than 15%, preferably to less than 10%, preferably to less than 5% of the original water content.

Preferably the mucosal desiccant reduces the water content of the mucus to the desired range within less than 3 hrs, preferably to less than 2 hrs, preferably less than 1 hr, preferably less than 50 minutes, preferably less than 45 minutes, preferably less than 40 minutes, preferably less than 35 minutes, preferably less than 30 minutes, preferably less than 25 minutes, preferably less than 20 minutes, preferably less than 15 minutes, preferably less than 10 minutes, preferably less than 5 minutes, preferably less than 4 minutes, preferably less than 3 minutes, preferably less than 2 minutes, preferably less than 1 minute.

In a preferred embodiment, the composition may comprise a demulcent. Preferred demulcents include pectin, glycerin (glycerol/glycerine) or honey.

Oropharyngeal administration

The term ‘oropharyngeal administration’, as used herein, may also be referred to as delivery to the part of the pharynx that lies between the soft palate and the hyoid bone and includes reference to a route of administration in which a drug is provided via either the mouth or nasal passages as part of a prophylactic and/or therapeutic treatment. Oropharyngeal administration may, for example, be used for drugs in the form of liquid droplets or aerosols. Preferably, the antibody, bispecific antibody or antigen binding fragment as described herein are administered by oropharyngeal administration.

Oral inhalation

The term ‘oral inhalation’, as used herein, may also be referred to as ‘mouth inhalation’, and includes reference to a route of administration in which the antibody, bispecific antibody or antigen binding fragment as described herein is provided through the mouth to the upper and/or lower respiratory tract such as lungs, as part of a prophylactic and/or therapeutic treatment of the invention. Oral inhalation may for example be applied for the antibody, bispecific antibody or antigen binding fragment in the form of liquid droplets or aerosols.

Preferably, the antibody, bispecific antibody or antigen binding fragment is administered by oral inhalation.

Oral inhalation may for example be applied for an antibody, bispecific antibody or antigen binding fragment in the form of liquid droplets or aerosols.

Oral inhalation may include the use of an inhaler. The inhaler may be involved in the achievement of the dose that was determined. The drug that is administered by oral inhalation may reach the lung but may also partially be cleared out by exhalation.

Administration by oral inhalation as disclosed herein may be performed using a medicament comprising aerosols in liquid form. Liquid droplets or aerosols comprising droplets smaller than 3 pm in diameter will primarily reach the respiratory region of the lung and will therefore be absorbed better than larger droplets. The medicament may comprise adjuvants. These adjuvants may for example be salts, oils, cytokines, emulsifiers, buffering agents, carbohydrates and combinations thereof. Dosage forms

Preferred dosage forms for mucosal administration of an administered antibody, bispecific antibody or antigen binding fragment of this invention may include ointments, pastes, creams, lotions, gels, foams, powders, solutions, or sprays.

Preferably, liquid nasal formulations may comprise aqueous solutions. Preferably, liquid nasal formulations may also comprise suspensions, emulsions, liposomes, and microspheres.

Other preferred liquid formulations may comprise liposomes, microspheres, mixed aqueous- organic formulations, aqueous formulations, and retentive formulations (commonly referred to as a gel).

A preferred dosage form for mucosal administration of an administered antibody, bispecific antibody or antigen binding fragment of this invention comprises a retentive formulation.

Dosages

The term “Nominal Dose” of a formulation as used herein is the total mass of antibody, bispecific antibody or antigen binding fragment present in the metered form presented by the device in question. For example, the Nominal Dose might be the mass of antibody, bispecific antibody or antigen binding fragment present in the metering chamber of a premetered spray device. The Nominal Dose is also referred to as the Metered dose.

Such “nominal dosages” are also referred to as “flat dosages” or in contrast to dosages based on the weight of the patient. Nominal dosages have the advantage that the medicament can be packaged in a single-unit dose. A single dose of an antibody, bispecific antibody or antigen binding fragment according to the present invention can provide protection from respiratory virus infection for several days and may be provided “on demand” or “as needed”. For example, an individual may administer antibody before leaving the house or before coming into contact with other individuals. In a preferred embodiment, intranasal nominal dosages range from 0.01 mg to 90 mg of an antibody, bispecific antibody or antigen binding fragment (i.e. 0.01 to 90% w/v when administered as a 100 pL aqueous formulation). In a preferred embodiment, intranasal dosages may range from 0.01 mg to 40 mg of an antibody, bispecific antibody or antigen binding fragment (i.e. 0.01 to 40% w/v when administered as a 100 pL aqueous formulation). Preferably such doses are administered as a 100 pL aqueous formulation. Preferably such doses are administered as a 100 pL aqueous formulation per nostril.

In a preferred embodiment, intranasal nominal dosages range from 0.01 mg to 5 mg, from 0.01 mg to 6 mg, from 0.01 mg to 7 mg, from 0.01 mg to 8 mg, from 0.01 mg to 9 mg, from 0.01 mg to 10.0 mg, from 0.01 mg to 11 mg, from 0.01 mg to 12 mg, from 0.01 mg to 13 mg, from 0.01 mg to 14 mg, from 0.01 mg to 15 mg, from 0.01 mg to 16 mg, from 0.01 mg to 17 mg, from 0.01 mg to 18 mg, from 0.01 mg to 19 mg, from 0.01 mg to 20 mg, from 0.01 mg to 21 mg, from 0.01 mg to 22 mg, from 0.01 mg to 23 mg, from 0.01 mg to 24 mg, or from 0.01 mg to 25 mg, from 0.01 mg to 26 mg, from 0.01 mg to 27 mg, from 0.01 mg to 28 mg, from 0.01 mg to 29 mg, from 0.01 mg to 30 mg, from 0.01 mg to 31 mg, from 0.01 mg to 32 mg, from 0.01 mg to 33 mg, from 0.01 mg to 34 mg, or from 0.01 mg to 35 mg of an antibody, bispecific antibody or antigen binding fragment as a 100 pL aqueous formulation.

In a preferred embodiment, orally inhaled nominal dosages also range from 0.01 mg to 90 mg of an antibody, bispecific antibody or antigen binding fragment. In a preferred embodiment, oral inhalation dosages also range from 0.01 mg to 40 mg of an antibody, bispecific antibody or antigen binding fragment. Preferably such aqueous formulation doses are administered in volumes ranging from 25 pL to 500 pL. Preferably such aqueous formulation doses are administered in volumes ranging from 100 pL to 250 pL.

In a preferred embodiment, orally inhaled nominal dosages range from 0.01 mg to 5 mg, from 0.01 mg to 6 mg, from 0.01 mg to 7 mg, from 0.01 mg to 8 mg, from 0.01 mg to 9 mg, from 0.01 mg to 10.0 mg, from 0.01 mg to 11 mg, from 0.01 mg to 12 mg, from 0.01 mg to 13 mg, from 0.01 mg to 14 mg, from 0.01 mg to 15 mg, from 0.01 mg to 16 mg, from 0.01 mg to 17 mg, from 0.01 mg to 18 mg, from 0.01 mg to 19 mg, from 0.01 mg to 20 mg, from 0.01 mg to 21 mg, from 0.01 mg to 22 mg, from 0.01 mg to 23 mg, from 0.01 mg to 24 mg, or from 0.01 mg to 25 mg, from 0.01 mg to 26 mg, from 0.01 mg to 27 mg, from 0.01 mg to 28 mg, from 0.01 mg to 29 mg, from 0.01 mg to 30 mg, from 0.01 mg to 31 mg, from 0.01 mg to 32 mg, from 0.01 mg to 33 mg, from 0.01 mg to 34 mg, or from 0.01 mg to 35 mg of an antibody, bispecific antibody or antigen binding fragment thereof as a 100 pL aqueous formulation.

Timing and intervals

The term “dosage” as used herein, refers to the amount of antibody, bispecific antibody or antigen binding fragment thereof to be given at a particular time (e.g., over the course of a 24-hour, 12-hour, 30-minute period, etc.). A dose refers to a single dosing episode, whether the dose is a unit dosage form or multiple unit dosage forms taken together (e.g., administration of two or more nasal administrations). A dosage includes reference to a pharmaceutical dosage form wherein the medicament is packaged for administration as, e.g., a single-unit dose or multiple-unit dose. A dosage may also be administered as, e.g., one or more drops of an antibody-comprising composition (e.g., nasal drops) or one or more sprays of an antibody-comprising composition (e.g., nasal sprays).

In order to provide long-lasting protection, the antibody, bispecific antibody or antigen binding fragment may be administered on a regular basis. For example, the antibody, bispecific antibody or antigen binding fragment is administered once, or at least twice per month. In a preferred embodiment, the antibody, bispecific antibody or antigen binding fragment thereof is administered once, or at least once per week, e.g., twice weekly. In a preferred embodiment, the antibody, bispecific antibody or antigen binding fragment is administered once, or at least once per day. As is clear to a skilled person, less antibody, bispecific antibody or antigen binding fragment may be administered when the antibody, bispecific antibody or antigen binding fragment is administered more frequently (e.g., daily). In a preferred embodiment, between 0.01 mg to 35 mg of bispecific antibody or antigenbinding fragment thereof is administered per week (e.g., once or twice weekly or daily). Prior to

The term ‘prior to’, as used herein, includes reference to the administration of an antibody, bispecific antibody or antigen binding fragment before an individual has been exposed to, or is infected with, a respiratory virus.

Preferably, the antibody, bispecific antibody or antigen binding fragment as disclosed is administered to an individual up to 24 hours prior to viral exposure, for example between zero and 24 hours before the individual has been exposed to said respiratory virus.

Preferably, the antibody, bispecific antibody or antigen binding fragment as disclosed is administered to an individual up to 48 hours prior to viral exposure, for example between zero and 48 hours before the individual has been exposed to said respiratory virus.

In a preferred embodiment, the antibody, bispecific antibody or antigen binding fragment is administered 2 days or more prior to viral exposure. In a preferred embodiment, the bispecific antibody or antigen-binding fragment thereof is administered 3 days or more prior to viral exposure. In a preferred embodiment, the bispecific antibody or antigen-binding fragment thereof is administered 4 days, or more prior to viral exposure. In a preferred embodiment, the bispecific antibody or antigen-binding fragment thereof is administered 5 days or more prior to viral exposure. In a preferred embodiment, the bispecific antibody or antigen-binding fragment thereof is administered 6 days or more prior to viral exposure. In a preferred embodiment, the bispecific antibody or antigen-binding fragment thereof is administered 7 days or more prior to viral exposure.

Delivery devices

The term “device,” as used herein, refers to an apparatus capable of delivering a drug to patient in need thereof.

Thus, the ability to efficiently formulate, process, package, and deliver the liquid composition with a minimal loss of drug is critical. With liquid drug delivery, both the delivered dose efficiency, i.e. the percentage of drug from a unit dose receptacle which is aerosolized and delivered from a delivery device, and the median droplet size distribution, i.e. the deviation from the median size, are critical to the successful delivery of liquid aerosol to a patient's lungs.

Aparticularly promising approach for the pulmonary delivery of liquid aerosol drugs utilizes a hand-held device with a hand pump for providing a source of pressurized gas. The pressurized gas is abruptly released through a liquid aerosol dispersion device, such as a venturi nozzle, and the dispersed liquid aerosol made available for patient inhalation.

Examples of standard nasal spray devices include those offered by Aptar Pharma’s CPS multi-dose spray pump platform for precise, reliable and targeted preservative-free nasal drug delivery. Such devices comprise a reservoir which holds multiple doses of the nasal spray formulation ( e.g., 50, 100, 150, 200, 60, or 120 doses), a closure (e.g., screw, crimp, or snap-on), and an actuator which delivers anywhere from 45 to 1000 pL (e.g. 50, 100, 140, 150, or 200 pL) of fluid per actuation to comprise a single dose. The actuator may be configured to count doses, deliver gel formulations, deliver in an upside-down configuration, etc. In traditional spray pump systems, antimicrobial preservatives are typically required to maintain microbiological stability in liquid formulations. Metered spray pumps have dominated the nasal drug delivery market since they were introduced. The pumps typically deliver 100 pL (25-250 pL) per spray, and they offer high reproducibility of the emitted dose and plume geometry in in vitro tests.

Sprays and plume geometry

The droplet size and plume geometry can vary within certain limits and depend on the properties of the pump, the formulation, the orifice of the actuator, and the force applied. The droplet size distribution of a nasal spray is a critical parameter, since it significantly influences the in vivo deposition of the drug in the nasal cavity. The droplet size is influenced by the actuation parameters of the device and the formulation. The prevalent median droplet size should be between about 30 and about 100 pm . If the droplets are too large (> about 120 pm), deposition takes place mainly in the anterior parts of the nose, and if the droplets are too small (< about 10 pm ), they can possibly be inhaled and reach the lungs and oral cavity, which should be avoided because of safety reasons. In its capacity as a surfactant, benzalkonium chloride and alkylmaltosides (e.g., a tetradecyl maltoside (TDM), a dodecyl maltoside (DDM), etc.) can affect the surface tension of droplets from a delivered nasal spray plume, producing spherical or substantially spherical droplets having a narrow droplet size distribution (DSD), as well as the viscosity of a liquid formulation.

Plume geometry, droplet size and DSD of the delivered plume subsequent to spraying may be measured under specified experimental and instrumental conditions by appropriate and validated and/or calibrated analytical procedures known in the art. These include photography, laser diffraction, and impaction systems (cascade impaction, NGI). Plume geometry, droplet size and DSD can affect pharmacokinetic outcomes such as Cmax, Tmax, and dose proportionality.

Optimal droplet sizes for a plume can be those that ensure the maximum amount of antibody, bispecific antibody or antigen binding fragment thereof is applied to the nasal epithelium. Minimizing the amount of very small droplets in the plume can reduce the amount of the plume that enters into the esophagus or lungs. This can reduce or eliminates side effects and ensure maximal delivery to the intended site of action. Minimizing the amount of larger drops can prevent loss of the active ingredient due to dripping out of the nose. Larger drops can also result in the formulation dripping into the back of the throat, which can cause irritation and delivery of the antibody, bispecific antibody or antigen binding fragment thereof to undesired regions.

Provided herein are multi-dose nasal spray devices that deliver a dosage unit of one or more antibody, bispecific antibody or antigen binding fragment thereof in a plume when actuated.

The plume can be characterized according to one or more parameters such as total volume, droplet size distribution, spray pattern, and plume geometry. The droplet size distribution can be characterized according to the percentage of droplets having a size of less than 10 pm. When characterized using parameters such as Dio, D50, D90, or a combination thereof, these indicate the size below which 10%, 50% or 90% of all droplets are found using, for example, Malvern Panalytical's Spraytec laser diffraction system which allows measurement of spray droplets and spray droplets size distributions in real-time.

The Span can be calculated from these numbers according to the following formula:

Span = (D9O^— D )/D₅₀.

Spray pattern measures the ovality of the spray, which can be calculated from the ratio of maximum to minimum cross sections diameter of the plume at a distance from the spray device. Plume geometry measures the plume angle at the origin of the plume. Plume geometry can be measured at two distances from the origin of the plume, for example, at two side views 90° relative to each other. Plume geometry can also be calculated from the spray pattern. The volume of the plume, the droplet size distribution, the spray pattern and plume geometry can all effect the treatment efficacy of active ingredients delivered by nasal spray. The multi-dose nasal spray devices and plumes disclosed herein can optimize treatment efficacy by controlling the delivery of the active ingredient to the correct target site.

As a preferred embodiment, disclosed herein is a multi-dose nasal spray device for delivery of an antibody, bispecific antibody or antigen binding fragment to a human's nasal epithelium that delivers a dosage unit in a plume upon actuation, wherein the dosage unit comprises from 0.01 mg to 90 mg of an antibody, bispecific antibody or antigen binding fragment in a pharmaceutically acceptable carrier comprising one or more excipients; and wherein the plume is characterized by one or more of the following preferred features:

(i) a total plume volume of from 25 pL to 250 pL,

(ii) a droplet size distribution characterized by less than 5% of the droplets in the plume having a size of less than 5 pm,

(iii) a droplet size distribution characterized by a Dw of greater than 10 pm, wherein 10% of the droplets in the plume have a size less than the Dw, (iv) a droplet size distribution characterized by a D50 of from 30 to 70 pm, wherein 50% of the droplets in the plume have a size less than the D50,

(v) a droplet size distribution characterized by a D90 of less than 200 pm, wherein 90% of the droplets in the plume have a size less than the D90, and

(vi) a plume span of from 1 to 6.

The multi-dose nasal spray devices disclosed herein can deliver a dosage unit in a plume that has a total volume of from 25 to 200 pL. “Total plume volume,” when used in connection with a plume, is the total liquid volume of all the droplets in the plume. The total volume can be determined by weighing the multi-dose nasal spray device before delivery of the dosage unit to the patient, actuating the device to deliver a dose, and then weighing the multidose nasal spray device after the actuation.

In a preferred embodiment, the plume is characterized by the total plume volume that is from 25 pL to 250 pL. In a preferred embodiment, the total plume volume is from 75 pL to 125 pL. In a preferred embodiment, the total plume volume is from 90 pL to 110 pL.

In a preferred embodiment, the plume is characterized by the total plume volume that is from 25 pL to 75 pL. In a preferred embodiment, the total plume volume is from 40 pL to 60 pL. In a preferred embodiment, the total plume volume is from 45 pL to 55 pL.

In a preferred embodiment, the liquid nasal composition is an aqueous solution. In a preferred embodiment, the liquid nasal composition is a suspension. In a preferred embodiment, the liquid nasal composition is an emulsion. In a preferred embodiment, the liquid nasal composition comprises liposomes. In a preferred embodiment, the liquid nasal composition comprises microspheres. In a preferred embodiment, the liquid nasal composition comprises mixed aqueous-organic formulations.

In a preferred embodiment, the liquid nasal composition comprises dispersible retentive formulations (i.e. dispersible gels). Preferably, the plume is characterized by less than 4% of the droplets in the plume having a size of less than 10 pm as determined using laser diffraction assessing the plume distribution by volume. Preferably, the plume is characterized by less than 3% of the droplets in the plume having a size of less than 10 pm as determined using laser diffraction assessing the plume distribution by volume. Preferably, the plume is characterized by less than 2% of the droplets in the plume having a size of less than 10 pm as determined using laser diffraction assessing the plume distribution by volume. The laser diffraction system referred to above, may for example, include the use of Malvern Panalytical's Spraytec laser diffraction system.

In a preferred embodiment, the plume is characterized by the Dio that is greater than 15 pm, preferably the plume is characterized by the Dw that is greater than 17.5 pm, preferably the plume is characterized by the Dw that is from 12.5 pm to 30 pm, preferably the plume is characterized by the D that is from 15 pm to 25 pm as determined by laser diffraction, for example, by using Malvern Panalytical's Spraytec laser diffraction system.

In a preferred embodiment, the plume is characterized by the D50 that is from 40 pm to 60 pm, preferably the plume is characterized by the D50 that is from 30 pm to 60 pm, preferably the plume is characterized by the D50 that is from 30 pm to 50 pm, preferably the plume is characterized by the D50 that is from 30 pm to 40 pm, preferably the plume is characterized by the D50 that is about 30 pm, 32.5 pm, 35 pm, 37.5 lam, 40 pm, 42.5 pm, 45 pm, 50 pm, 55 pm, 60 pm, 65 pm, or 70 pm as determined by laser diffraction, for example, by using Malvern Panalytical's Spraytec laser diffraction system.

In a preferred embodiment, the plume is characterized by the D90 that is less than 175 pm, preferably the plume is characterized by the D90 that is less than 150 pm, preferably the plume is characterized by the D90 that is less than 125 pm, preferably the plumeis characterized by the D90 that is less than 100 pm, preferably the plume is characterized by the D90 that is less than 90 pm, preferably the plume is characterized by the D90 that is from 75 pm to 199 pm, preferably the plume is characterized by the D90 that is from 75 pm to 175 pm, preferably the plume is characterized by the D90 that is from 75 pm to 150 pm, preferably the plume is characterized by the D90 that is from 75 pm to 125 pm, preferably the plume is characterized by the D90 that is from 75 pm to 100 pm, preferably the plume is characterized by the D90 that is about: 75 m, 80 pm, 85 pm, 90 pm, 100 pm, 110 pm, 120 pm, 130 pm, 140 pm, 150 pm, 160 pm, 170 pm, 180 pm, or 190 pm as determined by laser diffraction, for example, by using Malvern Panalytical's Spraytec laser diffraction system.

In a preferred embodiment, the plume is characterized by the span that is from 1 to 5 In a preferred embodiment, the plume is characterized by the span that is from 1 to 4. In a preferred embodiment, the plume is characterized by the span that is from 1 to 3. In a preferred embodiment, the plume is characterized by the span that is from 1 to 2. In some embodiments, the plume is characterized by the span that is about: 1, 1.1, 1.2, 1.3, 1.4, 1.5, 1.6, 1.7, 1.8, 1.9, 2, 2.1, 2.2, 2.3, 2.4, 2.5, 2.6, 2.7, 2.7, 2.9, 3, 3.25, 3.5, 3.75, 4, 4.5, 5, 5.5, or 6.

The spray pattern and geometry of the plume can affect the efficacy of treatment. For example, an irregular shape can result in uneven coating of the nasal epithelium and a resulting reduction in therapeutic efficacy. In another example, too narrow of a plume can reduce the area of the nasal epithelium that is coated by the plume. Conversely, too wide of a plume can direct the plume towards unintended targets such as the back of the throat.

In certain embodiments, the formulation when dispensed by actuation from the device will produce a uniform circular plume with an ovality ratio close to 1. Ovality ratio is calculated as the quotient of the maximum diameter (D_max) and the minimum diameter (Dmin) of a spray pattern taken orthogonal to the direction of spray flow (e.g., from the “top”). In certain preferred embodiments, the ovality ratio is less than ± 2.0. In certain preferred embodiments, the ovality ratio is less than ± 1.5. In certain preferred embodiments, the ovality ratio is less than ± 1.3. In certain preferred embodiments, the ovality ratio is less than ± 1.2. In certain preferred embodiments, the ovality ratio is less than ± 1.1.

In a preferred embodiment, the plume is further characterized by having an ovality of from 0.7 to 1. In a preferred embodiment, the plume is further characterized by having an ovality of from 0.8 to 1. In a preferred embodiment, the plume is further characterized by having an ovality of from 0.9 to 1. In a preferred embodiment, the plume is further characterized by having an ovality of about 1.

In a preferred embodiment, the plume is further characterized by having a geometry of from 30° to 90°. In a preferred embodiment, the plume is further characterized by having a geometry of from 45° to 75°. In a preferred embodiment, the plume is further characterized by having a geometry of about: 30 °, 35°, 40°, 45°, 50°, 55°, 60°, 65°, 70°, 75°, 80°, 85°, or 90°.

Liquid compositions

Preferably, the composition of the invention is a water-based composition such as an aqueous liquid.

Preferably, the liquid composition of the invention further comprises one or more salts, for example sodium chloride is a preferred salt.

Preferably, the liquid composition of the invention may further comprise one or more buffering agents, for example sodium acetate is a preferred buffering agent.

Preferably, the liquid composition of the invention may further comprise one or more carbohydrates, such as sucrose, emulsifiers, or combinations thereof. pH buffering/adjusting agents

The pH of the composition of the invention is preferably between 4 and 8, more preferably the pH is between 5.5 and 7.5. Preferably the pH of the composition of the invention is between 6.8 and 7.5.

Preferably, the liquid composition of the invention has a pH of about 4.0, 4.1, 4.2, 4.3, 4.4, 4.5, 4.6, 4.7, 4.8, 4.9, 5.0, 5.1, 5.2, 5.3, 5.4, 5.5, 5.6, 5.7, 5.8, 5.9, 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 7.7, 7.8, 7.9, 8.0. 8.1, 8.2, 8.3, 8.4, 8.5, 8.6,

8.7, 8.8, 8.9, or 9.0.

Preferably, the liquid composition of the invention has a pH of about 6.5 or 7.0.

Preferably, a pH buffering agent is selected from any one or more of ascorbic acid, citric acid, dimethyl succinate, disodium hydrogen phosphate, hydrochloric acid, magnesium carbonate, magnesium silicate, magnesium sulphate, magnesium trisilicate, sodium acetate, sodium bicarbonate, sodium carbonate, sodium carboxymethylcellulose, sodium chloride, phosphate buffer, sodium pyrophosphate, tetrasodium pyrophosphate, triethanolamine, trisodium citrate, trisodium phosphate and zinc citrate.

An especially preferred pH buffering agent comprises tetrasodium pyrophosphate.

An especially preferred pH buffering agent comprises sodium acetate.

Preferably, a pH buffering agent is present in the liquid composition in an amount ranging from about 0.01% to about 10% w/v.

Preferably, the liquid composition comprises pH adjusting agents and can be selected from any one or more of acids described herein (e.g. hydrochloric acid, citric acid), buffers (e.g. phosphate, acetate, and citrate buffers), and bases (e.g. sodium hydroxide, sodium citrate, sodium bicarbonate, sodium carbonate).

A preferred pH buffering agent comprises histidine. A preferred pH buffering agent comprises glycine. A preferred pH buffering agent comprises sodium hydroxide. A preferred pH buffering agent comprises hydrochloric acid. A preferred pH buffering agent comprises sodium acetate anhydrous. A preferred pH buffering agent comprises acetic acid, in particular glacial acetic acid, which is capable of adjusting and/or stabilizing the pH value of the composition as described herein. Preferably, pH buffering agent comprises acetic acid, in particular glacial acetic acid and sodium acetate anhydrous.

Preferably, the pH buffering agent comprises citric acid, succinic acid or acetic acid, or an amino acid such as glycine or histidine. Preferably, the pH buffering agent comprises sodium hydroxide or potassium hydroxide or an organic base such as tris (hydroxymethyl) aminomethane.

Isotonicity adjusting agents

Isotonicity adjusting agents may be included to achieve isotonicity with body fluids e.g. fluids of the nasal cavity, thereby resulting in reduced levels of irritancy.

Preferred isotonicity adjusting agents comprise any one or more of calcium chloride, sodium chloride, potassium chloride, glycerin, sucrose, dextrose, xylitol and mannitol.

Preferably, isotonicity agent may be included in the composition in an amount of between about 0.1 and 10% (w/v), such as about 4.5% by weight based on the total weight of the composition.

Preferred isotonicity adjusting agents comprise any one or more of mannitol, dextrose, sucrose, for preparation of isotonic or hypertonic intranasal solution.

A preferred concentration of the tonicity agent is preferably within the range from 1.0 to 20% w/v.

Stabilizing agents

In a preferred embodiment, the composition as described herein comprises at least one stabilizer (also called a stabilizing agent). Preferably, said at least one stabilizer comprised in the composition is capable of enhancing the physical and/or chemical stability of the composition as described herein, stabilizing the conformation of the polypeptide as described herein, stabilizing the composition as described herein during the storage, transportation, production process such as lyophilization, freeing and/or thawing, and/or in body.

Preferable stabilizing agents include any one or more of the following, 2- methyl-2,4- pentanediol (MPD), alanine, arginine, diethyl glycol, dimethyl sulfoxide (DMSO), ethylene glycol, leucine, polyvinyl alcohol (PVA), proline, propylene glycol, sodium chloride, sodium stearate, and magnesium stearate.

Preferable stabilizing agents include sugar. Preferred stabilizing sugars include any one or more of the following, dextrose, lactose, mannitol, raffinose, sorbitol, sucrose, and trehalose (also called as threalose).

An especially preferred stabilizing sugars comprises or consists of trehalose. Trehalose is capable of stabilizing protein conformation, for example antibody conformation, due to its effect on the structure and properties of solvent water better than other sugars.

Preferably, at least two stabilizers are comprised in the composition as described herein. Said at least two stabilizers are selected from the group consisting of sucrose, trehalose (also called as threalose), leucine, raffinose, alanine, arginine, sodium stearate, sodium chloride and mannitol. Suitably, the two stabilizers comprised in the composition as described herein are in a ratio ranging from 60: 1 to 1 :60, more suitably from 2: 1 to 1 :2.

In one preferred embodiment, the stabilizer comprises sucrose and/or sodium chloride. The stabilizers are capable of adjusting and/or stabilizing the osmolality of the composition as described herein. Also, the stabilizers are capable of maintaining and/or improving the stability of the antibody, bi specific antibody or antigen binding fragment as described herein. Surfactants

In a preferred embodiment, the composition as described herein comprises at least one surfactant.

Preferable surfactants include any one or more of the following, alkoxylated alcohols, alkoxylated tributylphenols, alkoxylated tri styrylphenols, alkylaminethoxylates, alkynaphtalenesulphonate ethoxylated and/or propoxylated alcohol phosphate ester, calcium dodecylbenzene sulfonate, calcium salt of alkylaryl sulfonates, castor oil ethoxylate, cetylbetaine, cocamidopropyl-betaine, commercial lung surfactants (e.g. ALEC ™), copolymers of ethylene, dipalmitoyl phosphatidylcholine (DPPC), dipalmitoyl phosphatidylethanolamine (DPPE), dipalmitoyl phosphatidylinositol (DPPI), disodium methyl ofeyl-taurate, especially alkylphenols, ethoxylated alcohols, ethoxylated and/or propoxylated alkylaryl phosphate ester, ethoxylated and/or propoxylated di- or tri styrylphenol phosphate, ethoxylated and/or propoxylated di- or tri -styryl phenol sulfate, ethoxylated fatty acids, ethoxylated poly adducts of ethylene oxide and propylene oxide, ethoxylated propoxylated alcohols, ethoxylated propoxylated polyaryl phenol, ethoxylated ricinoleic acid triglycerides, isostearamidopropyl dimethylamine, isostearamidopropyl- betaine (e.g. lauroamidopropyl), lauroamidopropyl-betaine, lauryl-sarcosine, laurylsulfobetaine, linoleamidopropyl-betaine, linoleyl-betaine, linoleyl-sarcosine, linoleyl- sulfobetaine, Monaquat™ ISIES, myristamidopropyl-betaine, myristarnidopropyl, myristylbetaine, myristyl-sarcosine, myristyl-sulfobetaine, palmidopropyl, palmidopropyl-betaine, phenyl sulfonate, phosphatidylglycerol (PG), phospholipids, phosphoric esters of polyethoxylated phenols or alcohols or polycarboxylate, pluronics, Pluronic™ F-68, pol oxamer 188, Pol oxamers, polyethyl glycol, polycondensates of ethylene oxide with fatty alcohols or with fatty acids or with fatty amines, polypropyl glycol, Polysorbate, Polysorbate 20, polysorbate-40, polysorbate-60, polysorbate-65, polysorbate-80, polysorbate-85, propylene glycol, salts of lignosulphonic acid, salts of phenylsulphonic, salts of polyacrylic acids, sodium dodecyl sulfate (SDS), sodium laurel sulfate (SLS), sodium methyl cocoyl, sodium octyl glycoside, sorbitan esters and their ethoxylates, sorbitol esters, stearyl- sarcosine, stearyl-sulfobetaine, substituted phenols, sulphosuccinic ester salts, suphosuccinate, tristyryphenol ethoxylate, and Triton™. A preferred surfactant is a polysorbate (e.g., polysorbate-20, polysorbate- 40, polysorbate- 60, polysorbate-65, polysorbate-80, polysorbate-85, or a combination thereof). The Polysorbate may be added into the composition as described herein for reducing, preventing and/or stopping the potential for agitation and/or for freeze/thaw induced aggregation. In some embodiments, the Polysorbate added in an amount of about 0.01 to 10% (v/v).

A preferred source of surfactants for use in the composition, includes lung surfactants which may be obtained by using ALEC ™ which comprise phospholipids, for example, mixtures of DPPC (dipalmitoyl phosphatidylcholine) and PG (phosphatidyl glycerol).

The surfactant may be present in an amount ranging from about 0.01% to about 20 % w/v of the liquid composition.

Emulsifying or solubilizing agents

In a preferred embodiment, the composition as described herein comprises at least one emulsion stabilizer. A preferred emulsion stabilizer comprises or consists of cellulose microcrystalline.

Spreading agents

In a preferred embodiment, the composition as described herein comprises at least one spreading agent.

A preferred spreading agent includes any one or more of the following polyoxyethylene alkyl ether, trisiloxane ethoxylate, polysorbates, ethoxylated tri styrylphenol phosphate, sodium lauryl sulphate, sodium methyl oleoyl taurate, tridecyl alcohol ethoxylate or mixtures thereof.

The spreading agents may be present in an amount ranging from about 0.01% to about 10 % w/v of the liquid composition. Dispersing agents

In a preferred embodiment, the composition as described herein comprises at least one dispersing agent.

A preferred dispersing agent includes lignin derivatives, in particular lignin-based hydrogels which may be used for the controlled release of both hydrophobic and hydrophilic compounds due to lignin naturally containing both hydrophobic and hydrophilic groups.

The dispersing agent may be present in an amount ranging from about 0.01% to about 15 % w/v of the liquid composition.

Antifoaming agents

In a preferred embodiment, the composition as described herein comprises at least one antifoaming agent.

A preferred anti-foaming agent includes any one or more of the following selected from the group comprising perfluroalkylphosphonic acids, polydimethyl siloxane or mixtures thereof.

The anti-foaming agent may be present in an amount ranging from about 0.01% to about 5 % w/v of the liquid composition.

Water-soluble nonionic triblock copolymers

In a preferred embodiment, the composition as described herein comprises at least one water- soluble nonionic triblock copolymer.

A preferred water-soluble nonionic triblock copolymer includes any one or more of the following selected from the group comprising polyvinylpyrrolidone (PVP) in particular Povidone K30, and Poloxamers, in particular Poloxamer 188. A preferred Poloxamer, in particular poloxamer 188. The water-soluble nonionic triblock copolymer may be present in an amount ranging from about 0.01% to about 5 % w/v of the liquid composition.

Preservatives

In a preferred embodiment, the composition as described herein comprises at least one preservative.

The term “preservative” as used herein, includes reference to that a compound is capable of essentially reducing and/or stopping bacterial action in the composition and/or during the process of preparing the composition, thus facilitating the preserving the quality, further prolonging the shelf life, and/or enhancing the physical and/or chemical stability of the composition as described herein

A preferred preservative includes any one or more of the following selected from the group comprising sodium acetate, benzyl alcohol, benzalkonium chloride, benzethonium chloride, potassium sorbate, calcium sorbate, methyl paraben, phenylethyl alcohol, ethyl paraben, propyl paraben, phenylcarbinol, chlorolbutanol, chlorolcresol, ethylenediaminetetraacetic acid (EDTA), octadecyldimethylbenzyl ammonium chloride, and hexamethonium chloride.

A preferred preservatives includes benzalkonium chloride.

The preservative may be present in an amount ranging from about 0.01% to about 5 % w/v of the liquid composition.

Taste masking agents

In a preferred embodiment, the composition as described herein comprises at least one taste masking agent. A preferred taste masking agent is menthol. A preferred taste masking agent may also comprise a sweetening agent and may include any one or more of the following: glucose dextrose, glycerol, saccharin and/or sorbitol.

The taste masking agent may be present in an amount ranging from about 0.01% to about 5 % w/v of the liquid composition.

Osmotic agents

In a preferred embodiment, the composition as described herein comprises at least one osmotic agent.

Osmotic agents are osmotically active ions or molecules that are poorly absorbed by the mucosal epithelium and thereby obligate water retention within, for example, the nasal cavity, to maintain isotonicity with plasma.

Preferred viscosity enhancing agents include any one or more of the following selected from the group comprising magnesium salts, phosphate and sulfate salts, poorly absorbed disaccharides, such as lactulose, sugar alcohols, such as mannitol and sorbitol, and polyethylene glycol.

The one osmotic agent may be present in an amount ranging from about 0.01% to about 5 % w/v of the liquid composition.

Viscosity enhancers

In a preferred embodiment, the composition as described herein comprises at least one viscosity enhancing agents.

A preferred viscosity enhancing agents includes any one or more of the following selected from the group comprising hydroxypropyl cellulose (HPC), hydroxyethyl cellulose (HEC), Pectin, Polyvinyl pyrrolidine (PVP) and mixtures thereof and hydroxypropyl methylcellulose (HPMC). Especially preferred viscosity enhancing agents include any one or more of the following selected from the group comprising HPMC 1828, HPMC 2208, HPMC 2906 and HPMC 2910.

The viscosity enhancing agent may be present in an amount ranging from about 0.01% to about 75 % w/v of the liquid composition. Preferably, the viscosity enhancing agent may be present in an amount ranging from about 0.01% to about 20 % w/v of the liquid composition, preferably from about 0.01% to about 10 % w/v of the liquid composition.

Antioxidants

In a preferred embodiment, the composition as described herein comprises at least one antioxidant.

A preferred antioxidant includes any one or more of the following selected from the group comprising ascorbic acid, N-acetyl cysteine (NAC), glutathione, co-enzyme Q-10, superoxide dismutase (SOD), and a combination thereof.

The antioxidant may be present in an amount ranging from about 0.01% to about 20 % w/v of the liquid composition, preferably from about 0.01% to about 10 % w/v of the liquid composition.

Suspension stabilisers

In a preferred embodiment, the composition as described herein comprises at least one suspension stabiliser.

A preferred suspension stabiliser includes any one or more of the following selected from the group comprising: 1,2- dipalmitoylphosphatidycholine (DPPC), benzalkonium chloride, block copolymers of oxyethylene and oxypropylene, cetyl alcohol, cetyl pyridinium chloride, Diethylene glycol dioleate, ethyl oleate, glyceryl monolaurate, glyceryl monooleate, glyceryl monoricinoleate, glyceryl monostearate, isopropyl myristate, lauryl polyoxyethylene ether, natural lecithin, oleyl polyoxyethylene ether, phosphatidylcholine (PC), PLURONICS, poly(vinyl alcohol), polyethylene glycol, polyoxyethylene their derivatives, polyoxylene sorbitan fatty acid esters (TWEENS), polyoxyethylene sorbitan monolaurate (e.g. TWEEN 808), polyoxyethylene sorbitan monooleate (e.g. TWEEN 208), polyoxyethylene sorbitan monopalmitatc (e.g. TWEEN 408), polyoxyethylene sorbitan monostearate (e.g. TWEEN 608), polyoxyethylene sterates, polyoxylene alkyl ethers, polyvinyl alcohol (PVA), sorbitan monolaurate, sorbitan mono-oleate, sorbitan trioleate, stearyl alcohol, stearyl polyoxyethylene ether, synthetic lecithin, tetrahydrofurfuryl oleate, and Vitamin-PEG and their derivatives.

The formulations of the present invention may be stabilised by appropriate selection of pH as discussed previously.

A preferred suspension stabilizer is HPMC which may be formulated into granules and forms colloids when dissolved in water.

A preferred suspension stabilizer employes repulsive inter-droplet forces. For example, the plurality of droplets in suspension may be electrically or electrostatically charged. When two charged particles encounter each other, they repel each other, thus preventing collision and aggregation.

In preferred embodiments, the stabilizer may act as a surface- altering agent. The surfacealtering agents described herein that may act as stabilizers may be, for example, polymers or surfactants. Examples of polymers include those suitable for use in the coating of the droplets of the invention, such as poly(vinyl alcohol), metal stearates and PLURONICS. An especially preferred surface-altering agent is magnesium stearate.

In preferred embodiments, the stabilizing agent may comprise or consist of PVA enabling encapsulation of a wide variety of compounds by emulsification techniques. In a preferred embodiment, the suspension stabilizer may be present in an amount ranging from about 0.01% to about 20 % w/v of the liquid composition, preferably from about 0.01% to about 10 % w/v of the liquid composition.

Carriers

In a preferred embodiment, the composition as described herein comprises at least one acceptable carrier.

When formulated as a liquid composition, the composition may further comprise a pharmaceutically acceptable carrier. In some preferred embodiments, the pharmaceutically acceptable liquid carrier comprises water. In a preferred embodiment, the pharmaceutically acceptable carrier may comprise saline. In an especially preferred embodiment, the pharmaceutically acceptable liquid carrier comprises water, a buffering agent, a flavoring agent, a humectant, a penetration enhancer, a pH adjusting agent, a preservative, a solvent or co-solvent, a surfactant, a tonicity adjusting agent, a viscosity adjusting agent, or a combination thereof.

Preferred vehicles or carriers for administration to a subject include any one of lipid or lipidderived delivery vehicles, such as liposomes, solid lipid nanoparticles, oily suspensions, submicron lipid emulsions, lipid microbubbles, inverse lipid micelles, cochlear liposomes, lipid microtubules, lipid microcylinders, or lipid nanoparticles (LNP) or a nanoscale platform.

Force Control Agents

In a preferred embodiment, the composition as described herein comprises at least one force control agent.

Other preferred force control agents include sodium benzoate, hydrogenated oils which are solid at room temperature, talc, titanium dioxide, aluminium dioxide, silicon dioxide and starch. Also useful as force control agents are film-forming agents, fatty acids and their derivatives, as well as lipids and lipid-like materials. Humectants

In a preferred embodiment, the composition as described herein comprises at least one humectant.

When formulated the composition includes any one or more of the following preferred humectants selected from the group comprising: propylene glycol, hexylene glycol, butylene glycol, aloe vera gel, alpha hydroxy acids such as lactic acid, glyceryl triacetate, lithium chloride, polymeric polyols such as polydextrose, sodium hexametaphosphate E452i, sugar alcohols (sugar polyols) such as glycerol, sorbitol, xylitol and maltitol.

A preferred humectant comprises glycerol for improving smoothness and providing lubrication to the composition.

In a preferred embodiment, the humectant may be present in an amount ranging from about 0.01% to about 20 % w/v of the liquid composition, preferably from about 0.01% to about 10 % w/v of the liquid composition.

Droplet size

The term “droplet size” as used herein, can be used interchangeably with the term “droplet diameter”.

Preferably, the droplet size of each of the components comprised in the composition as described herein is reduced to a desired size by the methods known by the skilled in the art.

Preferably, the liquid as described herein comprises or consists of a homogeneous mixture. Preferably, the median droplet of the liquid ranges from 0.5 pm to 500 pm, as determined by laser diffraction. The mass median droplet size for nasal delivery is preferably in excess of 5 pm and more preferably greater than 10 pm as this reduces the material passing through the nasal cavity and maximises its deposition in the nasal turbinates.

Preferably, the median droplet size of the liquid composition as described herein ranges from 0.5 pm to 400 pm, suitably from 0.5 pm to 300 pm, suitably from 0.5 pm to 200 pm, suitably from 1 pm to 200 pm, suitably from 10 pm to 200 pm, suitably from 20 pm to 200 pm, suitably from 30 pm to 200 pm, as determined by laser diffraction.

Preferably, the liquid composition as described herein has a bimodal droplet size distribution.

Preferably, the liquid composition as described herein has a trimodal droplet size distribution.

Wetting agents

In a preferred embodiment, the composition as described herein comprises at least one wetting agent.

A preferred wetting agents includes any one or more of the following selected from the group comprising: alkyl phenol ethoxylate, fatty oil ethoxylate, phenyl naphthalene sulphonates, alkyl naphthalene sulfonates, sodium alkyl naphthalene sulfonate, sodium salt of sulfonated alkyl carboxylate, polyoxyalkylated ethyl phenols, polyoxy ethoxylated fatty alcohols, polyoxyethoxylated fatty amines, lignin derivatives, alkane sulfonates, alpha olefin sulfonates, alkylbenzene sulfonates, salts of polycarboxylic acids, salts of esters of sulfosuccinic acid, octyl phenol ether sulphate, anionic phosphate esters, disodium laureth sulfosuccinate, diisodecyl sodium sulfosuccinate, alkyl naphthalene sulfonates, alkylbenzenesulfonates, alkylpolyglycol ether sulfonates, alkyl ether phosphates, alkyl ether sulfates and alkyl sulfosuccinic monoesters, dioctyl sulfosuccinate sodium salt, Cl 2- 15 ethoxylated alcohols, C12-15 alcohol ethoxylate (Atlox 4894), disodium hexadecyl sulphate or mixtures thereof. Preferable wetting agents may include any one or more of cellulose derivatives, dextran 70, gelatine, liquid polyols, polyvinyl alcohol (PVA) and Povidone, also known as polyvinylpyrrolidone (PVP).

Preferably, the wetting agent may be present in an amount ranging from about 0.01% to about 15 % v/v of the composition. Preferably, the wetting agent may be present in an amount ranging from about 0.01% to about 10 % v/v of the liquid composition, preferably from about 0.01% to about 5 % v/v of the liquid composition.

Mucoadhesive compounds

In an especially preferred embodiment, the composition as described herein comprises at least one mucoadhesive compound.

The mucoadhesive properties as described herein include but not limited to for example, adhesion of a polymer with the mucus layer where the mucoadhesive compound forms a physical barrier on the mucosal surface. Retarding or slowing the transit of a composition comprising an antibody, bispecific antibody or antigen binding fragment as described herein is a key aspect of the invention as disclosed herein. The retarding or slowing of the antibody, bispecific antibody or antigen binding fragment’s transit afford these therapeutic molecules greater opportunity for viral sequestration and thus enhances the therapeutic and/or prophylactic efficacy of the antibody, bispecific antibody or antigen binding fragment as described herein.

A preferred mucoadhesive compound includes any one or more of the following selected from the group comprising cellulose derivatives, gellan gum, guar gum, karaya gum, xanthan gum, carrageenan, alginate, pectin, dextran, chitosan, agarose, hyaluronic acid, gelatin, tragacanth, vinyl polymers, vinyl copolymers, vinyl alcohols, alkoxy polymers, polyethylene oxide polymers, poly (vinyl pyrrolidone) such as Kollidon VA 64, poly(dimethyl siloxane), poly acrylic acid-based polymers such as poly acrylates, plyethylene glycol, polyethers, sodium alginate, acrylic acid copolymers, polycarbophil such as Noveon AA-1, polylysene, dimethylaminoethyl dextran, poly vinyl alcohol, hydroxy ethyl starch, amylose, sodium croscarmellose, cellulose acetate phthalate, Cellulose acetate butyrate, poloxamer 407, polycarbophil (PCP), Carbopol® 97 IP NF, Carbopol® 974P NF, Carbopol® 934 NF, Carbopol® 934P NF and/or Carbopol® 71GNF.

Preferably, the composition as described herein comprises or consists of a cellulose derivative selected from the group consisting of sodium carboxyl methyl cellulose (SCMC), methyl cellulose, carboxyl methyl cellulose (CMC), hydroxyl propyl cellulose, hydroxyl propyl methyl cellulose (HPMC), ethyl cellulose, hydroxyl ethyl cellulose (HEC), Pharmacoat 603, Pharmacoat 606, Pharmacoat 615, Methocel E5, Methocel E8, Methocel E10, Methocel E15, Methocel E50, and Methocel 60SH50, and the like.

The term “derivative” and “the like” as described herein, include reference to cellulose compounds which possess substantially the same properties and/or utilities as one of the cellulose compounds as described herein.

Preferably, the cellulose derivatives as described herein are soluble in water, isotonic saline, and/or artificial saliva.

Preferably, the concentration of the mucoadhesive compound in the composition as described herein ranges from 0.1 to 20 % v/v. Preferably, the concentration of the mucoadhesive compound in the composition as described herein ranges from 0.25 to 10% v/v.

An especially preferred mucoadhesive compound in the composition as described herein is HPMC. Preferably, the concentration of HPMC in the composition as described herein ranges from 0.1 to 20 % v/v. Preferably, the concentration of the HPMC in the composition as described herein ranges from 0.25 to 10% v/v, preferably from 0.5 to 5% v/v.

Suitably, said at least one mucoadhesive compound in the composition as described herein comprises or consists of polyethylene oxide-based polymers. Preferably, the composition as described herein comprises at least two mucoadhesive compounds as described herein.

Preferably, the composition as described herein comprises or consists of HPMC and polyethylene oxide based polymers. Preferably, HPMC and polyethylene oxide-based polymers are in the ratio ranging from 10: 1 to 1 : 10.

In one preferred embodiment, the average molecular weight of said at least one mucoadhesive compound is in the range of 1 x 10³ - 5 x 10⁵ KDa.

Preferably, the mucoadhesive comprises a cellulose derivative.

Preferably the cellulose derivative is selected from the group comprising hydroxypropyl methylcellulose (HPMC), methylcellulose (MC), hydroxypropyl cellulose (HPC), hydroxyethyl cellulose (HEC), microcrystalline cellulose and carboxymethyl cellulose (CMC).

Preferably the cellulose derivative comprises at least one of hydroxypropyl methylcellulose (HPMC), methylcellulose (MC), hydroxypropyl cellulose (HPC), hydroxyethyl cellulose (HEC) and carboxymethyl cellulose (CMC).

Preferably the cellulose derivative comprises a homogenously substituted cellulose. Preferably the cellulose derivative comprises a heterogeneously substituted cellulose.

Chitosan

In a preferred embodiment, the composition as described herein comprises chitosan and/or chitosan derivatives, as a further example of a mucoadhesive compound.

The term “chitosan” as used herein, refers to a family of amino polysaccharides composed of N-acetyl-D-glucosamine and D-glucosamine and its units linked by 1-4-P-glycosidic bonds. This family of amino polysaccharides can be differentiated according to their degree of deacetylation (ranging from 2% to 60%) and molecular weight (ranging from 50 to 2000 kDa).

The -OH and NH2 groups of chitosan lead to the capability of forming hydrogen and covalent bonding. The cationic nature provides strong electrostatic interaction with negatively charged components of mucus, such as sialic acid and/or epithelial surfaces. Hydrogen bonding and hydrophobic interaction also play a role in the mucoadhesion of chitosan.

Preferably, chitosan derivatives includes any one or more of the following selected from the group comprising trimethyl chitosan, trimethyl chitosan derivatives, triolated chitosan, and/or triolated chitosan derivatives.

Preferably, chitosan derivatives comprise trimethyl chitosan (TMC), PEGlyted TMC, PEG- bearing thiolated TMC, thiolated chitosan, methylated N-aryl chitosan derivatives, methylated N-(4-N,N-dimethylaminocinnamyl) chitosan chloride, methylated N-(4- pyridylmethyl) chitosan chloride, carboxymethyl chitosan, chitosan-cysteine, chitosan-N- acetyl-cysteine, chitosan-thioglycolic acid (Chitosan-TGA), chitosan-4-thiobutylamidine (Chitosan-TBA), chitosan-thioethylamidine, chitosan-glutathione, pre-activated (S- protected) thiolated chitosans, thiolated methylated dimethylaminobenzyl chitosan, acrylated chitosan, half-acetylated chitosan wherein the degree of acetylation = 50 ± 5 mol%, glycol chitosan, palmitoyl glycol chitosan, hexanoyl glycol chitosan, chitosan-enzyme inhibitors, chitosan-EDTA, chitosan-enzyme inhibitors-EDTA, chitosan-catechol conjugate, methyl pyrrolidinone chitosan, and itosancyclodextrin,

The term “chitosan-enzyme inhibitors” as described herein, means the covalent attachment of enzyme inhibitors to mucoadesive compounds such as chitosan and/or chitosan derivatives. Suitable enzyme inhibitors include inhibitors for reducing and/or stopping the enzymatic cleavage of chitosan. Examples of enzyme inhibitors include but not limited to antipain, chymostatin, elastatinal, and/or Bowman-Birk inhibitor. Enzyme inhibitors are capable of reducing the enzymatic degradation and thereby further prolonging the retention time of the antibody, bispecific antibody or antigen binding fragment on the mucus.

Preferably, chitosan derivatives have substantially similar and/or improved mucoadhesive characters to chitosan. The mucoadhesive character of chitosan can be evaluated by a range of techniques including but not limited to mucin-particle interaction, tensile strength, and/or flow-through technique coupled with fluorescence microscopy.

Hyaluronic acid

In a preferred embodiment, the composition as described herein comprises hyaluronic acid (HA) and/or hyaluronic acid derivatives, as a further example of a mucoadhesive compound.

Hyaluronic acid is a naturally occurring and water soluble polysaccharide comprising disaccharide units of D-glucuronic acid (GlcUA) and N-acetyl-D-glucosamine (GlcNAc), which are alternately linked, forming a linear polymer. High molecular weight hyaluronic acid may comprise 100 to 10,000 disaccharide units.

As hyaluronic acid exhibits a relatively high molecular weight and strong interactions with water, hyaluronic acid is viscous in aqueous solution even low concentrations. Hence, hyaluronic acid is suitable for being used as a mucoadhesive in drug formulations.

Preferably, the hyaluronic acid derivatives can be naturally occurring or synthesized.

Hyaluronic acid often occurs naturally. Preferred hyaluronic acid derivatives include any one or more of the following selected from the group comprising sodium hyaluronate, potassium hyaluronate, magnesium hyaluronate, and/or calcium hyaluronate.

Preferably, synthesized hyaluronic acids comprise chemically-modified hyaluronic acids, such as cross-linked hyaluronic acid. The mucoadhesive properties of a synthesized hyaluronic acid are essentially the same and/or improved compared to a naturally-occurring hyaluronic acid. The properties include but not limited to mucoadhesive properties, stability, improved bioavailability, and/or enhanced resistance to degradation.

A preferred synthetic hyaluronic acid is thiolated hyaluronic acid which is obtained through modifying the carboxylic and/or hydroxyl groups on hyaluronic acid and attaching a sulfhydryl ligand onto the backbone of hyaluronic acid. The mucoadhesive property of thiolated hyaluronic acid mainly results from the disulfide bonds between the sulfhydryl moieties of the backbone and the cysteine-rich residues of the mucus.

Pullulan

In a preferred embodiment, the composition as described herein comprises pullulan and/or pullulan derivatives, as a further example of a mucoadhesive compound..

Pullulan is a natural, non-toxic, n on-immunogenic, non-carcinogenic and non-mutagenic polymer. Pullulan has a linear and unbranched structure with molecular formula (CeHioOsjn and molecular weight ranging from 4.5 x 10⁴ to 6 x 10⁵ Da. The chemical structure of Pullulan comprises a linkage pattern with two a-(l- ) and one a-(l— >6) glycosidic bonds in maltotriose repeating units (G3). There are nine hydroxyl groups on glucopyranose rings of G3 units.

Owning to its monomer configuration, pullulan has favorable properties such as renewable origin, biocompatibility, and/or hydrophilic nature, making it useful in pharmaceutical applications, such as carrying molecules to a target site in a subject. However, pullulan has a relatively short degradation time, which can be improved by chemical modification.

The hydroxyl groups on pullulan provide reactive sites to induce new functionalities through the formation of bonds with different groups of atoms by means of sulfation, oxidation, etherification, phosphatation, and/or esterification reactions. Suitably, pullulan can be modified by replacing at least one of the nine hydroxyl groups on its glucopyranose rings with a functional group including but not limited to hydrophobic cholesterol, aldehyde, carboxylic group, carb oxym ethyl, alkyl group, N-phenylur ethane groups, N-hexylur ethane groups, sulfite, polyethylene glycol, and polyamines. Modified pullulans can be referred to as pullulan derivatives within the present invention.

Preferably, pullulan derivatives comprises includes any one or more of the following selected from the group comprising cholesterol bearing pullulan, pullulan acetate, carboxymethyl pullulan, aminated carboxymethyl pullulan, succinylated pullulan, pullulan-b- polyetheramine, pullulan folate, maleilated pullulan, diethylene triamine penta acetic acid pullulan, all trans retinoic acid bearing pullulan, poly(DL-Lactide-co-glycolide)-grafted pullulan, pullulan-g-poly(L-lactide), pullulan/deoxycholic acid, diethylaminoethylamine pullulan, polyethylenimine pullulan, pullulan-g-poly(l-lysine), vitamin H modified pullulan acetate, O-Urocanyl pullulan, biotin modified cholesterol-pullulan, N-urocanyl pullulan, oligo (methacryloyl sulfadimethoxine) grafted pullulan acetate, dibutylaminopropyl carbamate-pullulan octanoate, polyaminated pullulans such as pullulan-tris(2- aminoethyl)amine (Pul-TAEA) and pullulan-polyethyleneimine (Pul-PEI), thiolated pullulans, and/or preactivated thiolated pullulan.

Methylcellulose

In a preferred embodiment, the composition as described herein comprises methylcellulose (MC), as a further example of a mucoadhesive compound.

Methylcellulose is characterized by the average degree of substitution (DS) which is defined as the average number of hydroxyl groups substituted per anhydroglucose unit.

A preferred methylcellulose has a heterogeneous substituent distribution along its backbone. Preferably, methylcellulose comprises 20% to 40% methoxy groups.

A preferred methylcellulose has a viscosity ranging (0.005 to 75 N s/m²) at 2% concentration, corresponding to average molecular weight range of 10 to 220 kDa. Sodium Carboxymethylcellulose

In a preferred embodiment, the composition as described herein comprises sodium carboxymethylcellulose (NaCMC), as a further example of a mucoadhesive compound.

Sodium carboxymethylcellulose is the sodium salt of carboxymethylcellulose and/or cellulose gum, an anionic derivative.

A preferred molecular weight range for sodium carboxymethylcellulose is from 21 kDa to 500 kDa, preferably from 50 kDa to 400 kDa, preferably from 100 kDa to 300 kDa, preferably from 150 kDa to 250 kDa.

Preferably, the viscosity of the sodium carboxymethylcellulose ranges from 0.005 to 2 N s/m², preferably from 0.010 to 1.75 N s/m², from 0.025 to 1.5 N s/m², preferably from 0.05 to 1 N s/m²

Preferably, the sodium carboxymethylcellulose may be present in an amount ranging from about 0.01% to about 15 % v/v of the composition. Preferably, the sodium carboxymethylcellulose may be present in an amount ranging from about 0.01% to about 10 % v/v of the liquid composition, preferably from about 0.01% to about 5 % v/v of the liquid composition.

Hydroxyethyl cellulose

In a preferred embodiment, the composition as described herein comprises hydroxyethyl cellulose (HEC), as a further example of a mucoadhesive compound.

Hydroxyethyl cellulose is a preferred viscosity enhancing agent.

Preferably, the hydroxyethyl cellulose may be present in an amount ranging from about 0.01% to about 15 % v/v of the composition. Preferably, the hydroxyethyl cellulose may be present in an amount ranging from about 0.01% to about 10 % v/v of the liquid composition, preferably from about 0.01% to about 5 % v/v of the liquid composition.

Hydroxypropyl methylcellulose

In an especially preferred embodiment, the composition as described herein comprises hydroxypropyl methylcellulose (HPMC), otherwise known as hypromellose, as a further example of a mucoadhesive compound.

HPMC is a solid powder and forms colloids when dissolved in water. An HPMC solution acts to swell and absorb water, thereby expanding the thickness of any resulting HPMC layer. Swelling is enhanced for higher molecular weight HPMC grades. The molecular weights and molecular weight distributions of HPMC samples can be measured using gel permeation chromatography.

In a preferred embodiment, the use of hydroxypropyl methylcellulose as a mucosal desiccating agent, is disclosed.

In a preferred embodiment, the use of hydroxypropyl methylcellulose as a mucosal barrier, is disclosed.

In a preferred embodiment, the use of hydroxypropyl methylcellulose as a mucosal adhesive, is disclosed.

In a preferred embodiment, the use of hydroxypropyl methylcellulose as a protease and aminopeptidase inhibitor, is disclosed.

In a preferred embodiment, hydroxypropyl methylcellulose may conjugate with a protease inhibitor. When formulating, preferably, HPMC is separately mixed with glycerin as this significantly aids in the dissolution of HPMC. Once dissolved, the mixture of HPMC and glycerin is then added to any subsequent formulation mixture.

Hydroxypropyl methylcellulose - degree of substitution

HPMC is a partly O-methylated and O-2-hydroxypropylated cellulose ether.

In addition to the various molecular weights, HPMC is provided in a variety of substituted forms that vary with respect to the methyl and hydroxypropyl content. The United States Pharmacopoeia (USP) distinguishes four different types of HPMC, classified according to their relative -OCH₃ and -OCH₂CH(OH)CH₃ content: HPMC 1828, HPMC 2208, HPMC 2906 and HPMC 2910. The first two numbers indicate the percentage of methoxy-groups, the last two numbers the percentage of hydroxypropoxy-groups. This degree of substitution is the average level of methoxy substitution on the cellulose chain. Since there are maximum of three possible substitution sites with each cellulose molecule, this average value is a real number ranging from 0 to 3. However, the extent of the substitution is often expressed in percentages. In addition to various substitution ratios, an HPMC polymer is available in as well as in a wide range of molecular weights and viscosity grades. The critical congealing temperature is inversely related to both the solution concentration of HPMC and the concentration of the methoxy group within the HPMC molecule, which in turn depends on both the degree of substitution of the methoxy group and the molar substitution. In other words, the higher the concentration of the methoxy group, the lower the critical temperature. The inflexibility/viscosity of the resulting HPMC-containing composition, however, is directly related to the concentration of the methoxy group, i.e. the higher the methoxy group concentration, the more viscous or less flexible the resulting composition.

In a preferred embodiment, the composition as described herein comprises HPMC in form that has high methoxy and hydroxypropyl content.

In a preferred embodiment, the composition as described herein comprises HPMC in form that has high methyl and medium hydroxypropyl content. In a preferred embodiment, the composition as described herein comprises HPMC in form that has medium methyl and hydroxypropyl content.

Hydroxypropyl methylcellulose - substitution type - HPMC 1828

In a preferred embodiment, the composition as described herein comprises HPMC substitution type 1828 (“HPMC 1828”). HPMC substitution type 1828 has a high viscosity and thus eminently suited for mucosal administration.

In a preferred embodiment, the composition as described herein comprises HPMC with 16.5% to 20% methoxy groups by weight, and 23% to 32% hydroxypropyl groups by weight.

Preferably, the HPMC 1828 may be present in an amount ranging from about 0.01% to about 15 % w/v of the composition. Preferably, the HPMC 1828 may be present in an amount ranging from about 0.01% to about 10 % w/v of the liquid composition, preferably from about 0.01% to about 5 % w/v of the liquid composition.

64. Hydroxypropyl methylcellulose - substitution type - HPMC 2208

In a preferred embodiment, the composition as described herein comprises HPMC substitution type 2208 (“HPMC 2208”). HPMC substitution type 2208 has a relatively low viscosity and thus eminently suited for mucosal administration.

In a preferred embodiment, the composition as described herein comprises HPMC with 19.0% to 24.0% methoxy groups by weight, and 4.0% to 12.0% hydroxypropyl groups by weight.

Preferably, the HPMC 2208 may be present in an amount ranging from about 0.01% to about 15 % w/v of the composition. Preferably, the HPMC 2208 may be present in an amount ranging from about 0.01% to about 10 % w/v of the liquid composition, preferably from about 0.01% to about 5 % w/v of the liquid composition. Hydroxypropyl methylcellulose - substitution type - HPMC 2906

In a preferred embodiment, the composition as described herein comprises HPMC substitution type 2906 (“HPMC 2906”). HPMC substitution type 2906 has a relatively high viscosity and thus eminently suited for mucosal administration.

In a preferred embodiment, the composition as described herein comprises HPMC with 27.0% to 30.0% methoxy groups by weight, and 4.0% to 7.5% hydroxypropyl groups by weight.

Preferably, the HPMC 2906 may be present in an amount ranging from about 0.01% to about 15 % w/v of the composition. Preferably, the HPMC 2906 may be present in an amount ranging from about 0.01% to about 10 % w/v of the liquid composition, preferably from about 0.01% to about 5 % w/v of the liquid composition.

Hydroxypropyl methylcellulose - substitution type - HPMC 2910

In a preferred embodiment, the composition as described herein comprises HPMC substitution type 2910 (“HPMC 2910”). HPMC substitution type 2910has a relatively high viscosity and thus eminently suited for mucosal administration.

In a preferred embodiment, the composition as described herein comprises HPMC with 28% to 30.0% methoxy groups by weight, and 4.0% to 12% hydroxypropyl groups by weight.

Preferably, the HPMC 2910may be present in an amount ranging from about 0.01% to about 15 % w/v of the composition. Preferably, the HPMC 2910 may be present in an amount ranging from about 0.01% to about 10 % w/v of the liquid composition, preferably from about 0.01% to about 5 % w/v of the liquid composition. Hydroxypropyl methylcellulose HPMC1828, HPMC2208, HPMC2906 and

HPMC2910

In a preferred embodiment, the composition as described herein comprises HPMC 1828 and HPMC 2208. Preferably, the HPMC 1828 and HPMC 2208 may be present in a combined amount ranging from about 0.01% to about 15 % w/v of the composition. Preferably, the HPMC 1828 and HPMC 2208 may be present in a combined amount ranging from about 0.01% to about 10 % w/v of the liquid composition, preferably from about 0.01% to about 5 % w/v of the liquid composition, also preferably from about 0.05% to about 3%w/v of the liquid composition.

In a preferred embodiment, the composition as described herein comprises HPMC 1828 and HPMC 2906. Preferably, the HPMC 1828 and HPMC 2906 may be present in a combined amount ranging from about 0.01% to about 15 % w/v of the composition. Preferably, the HPMC 1828 and HPMC 2906 may be present in a combined amount ranging from about 0.01% to about 10 % w/v of the liquid composition, preferably from about 0.01% to about 5 % w/v of the liquid composition, also preferably from about 0.05% to about 3%w/v of the liquid composition.

In a preferred embodiment, the composition as described herein comprises HPMC 1828 and HPMC 2910. Preferably, the HPMC 1828 and HPMC 2910 may be present in a combined amount ranging from about 0.01% to about 15 % w/v of the composition. Preferably, the HPMC 1828 and HPMC 2910 may be present in a combined amount ranging from about 0.01% to about 10 % w/v of the liquid composition, preferably from about 0.01% to about 5 % w/v of the liquid composition, also preferably from about 0.05% to about 3%w/v of the liquid composition.

In a preferred embodiment, the composition as described herein comprises HPMC 2208 and HPMC 2906. Preferably, the HPMC 2208 and HPMC 2906 may be present in a combined amount ranging from about 0.01% to about 15 % w/v of the composition. Preferably, the HPMC 2208 and HPMC 2906 may be present in a combined amount ranging from about 0.01% to about 10 % w/v of the liquid composition, preferably from about 0.01% to about 5 % w/v of the liquid composition, also preferably from about 0.05% to about 3%w/v of the liquid composition.

In a preferred embodiment, the composition as described herein comprises HPMC 2208 and HPMC 2910. Preferably, the HPMC 2208 and HPMC 2910 may be present in a combined amount ranging from about 0.01% to about 15 % w/v of the composition. Preferably, the HPMC 2208 and HPMC 2910 may be present in a combined amount ranging from about 0.01% to about 10 % w/v of the liquid composition, preferably from about 0.01% to about 5 % w/v of the liquid composition, also preferably from about 0.05% to about 3%w/v of the liquid composition.

In a preferred embodiment, the composition as described herein comprises HPMC 2906 and HPMC 2910. Preferably, the HPMC 2906 and HPMC 2910 may be present in a combined amount ranging from about 0.01% to about 15 % w/v of the composition. Preferably, the HPMC 2906 and HPMC 2910 may be present in a combined amount ranging from about 0.01% to about 10 % w/v of the liquid composition, preferably from about 0.01% to about 5 % w/v of the liquid composition, also preferably from about 0.05% to about 3%w/v of the liquid composition.

In a preferred embodiment, the composition as described herein comprises at least two selected from the group consisting of HPMC 1828, HPMC 2208, and HPMC 2906. Preferably, the at least two HPMCs may be present in a combined amount ranging from about 0.01 to about 15 % w/v of the composition. Preferably, the at least two HPMCs may be present in a combined amount ranging from about 0.01% to about 10 % w/v of the liquid composition, preferably from about 0.01% to about 5 % w/v of the liquid composition, also preferably from about 0.05% to about 3%w/v of the liquid composition.

Hydroxypropyl methylcellulose - heterogeneously substituted

In a preferred embodiment, the composition as described herein comprises HPMC that is a heterogeneously substituted HPMC. A heterogeneously substituted HPMC comprises clustered and adjoining methoxy and/or hydroxypropyl substituted molecules. Hydroxypropyl methylcellulose - homogenously substituted

In a preferred embodiment, the composition as described herein comprises HPMC that is a homogenously substituted HPMC. A homogenously substituted HPMC comprises evenly dispersed and few, if any, adjoining methoxy and/or hydroxypropyl substituted molecules.

Hydroxypropyl methylcellulose - Number Average Molecular Weight (Mn)

In a preferred embodiment, the composition as described herein comprises HPMC that has a Number Average Molecular Weight (Mn) ranging from 0.3 X 10⁴ to 10.5 X 10⁴ Daltons. Preferably, the Number Average Molecular Weight (Mn) of the HPMC disclosed herein ranges from approximately 1 X 10⁴ to 9 X 10⁴ Daltons. Preferably Number Average Molecular Weight (Mn) of the HPMC as herein disclosed ranges from approximately 2.5 X 10⁴ to 8.6 X 10⁴, or 86,000 Daltons.

Hydroxypropyl methylcellulose - Weight Average Molecular Weight (Mw)

In a preferred embodiment, the composition as described herein comprises HPMC that has a Weight Average Molecular Weight (Mw) ranging from 0.5 X 10⁶ to 75 X 10⁶ Daltons.

Preferably, the Weight Average Molecular Weight (Mw) of the HPMC disclosed herein ranges from approximately 1 X 10⁶ to 6 X 10⁶ Daltons. Preferably Weight Average Molecular Weight (Mw) of the HPMC as herein disclosed ranges from approximately 2.5 X 10⁶ to 5 X 10⁶, or 5,00,000 Daltons.

Preferably, the Dispersity (Mw/Mn) of the HPMC as herein disclosed ranges from 9.0 to 350.0

Hydroxypropyl methylcellulose - Peak Molecular Weight

In a preferred embodiment, the composition as described herein comprises HPMC that has a Peak Molecular Weight ranging from 5 X 10⁴ to 16 X 10⁴ Daltons. Preferably, the Peak Molecular Weight of the HPMC disclosed herein ranges from approximately 6 X 10⁴ to 14 X 10⁴ Daltons. Preferably Peak Molecular Weight of the HPMC as herein disclosed ranges from approximately 2.5 X 10⁴ to 5 X 10⁴, or 50,000 Daltons.

Hydroxypropyl methylcellulose - Viscosity

In a preferred embodiment, the composition as described herein comprises HPMC that has viscosity between 2.0 (N s/m²) i.e. 2000 (mPa s) to 120.0 (N s/m²) i.e. 120,000 (mPa s), 2 % in H2O at 20 °C. Preferably the viscosity of the HPMC as herein disclosed is between 4.0 (N s/m²) i.e. 4000 (mPa s) to 100.0 (N s/m²) i.e. 100,000 (mPa s), 2 % in H2O at 20 °C. Preferably the viscosity of the HPMC as herein disclosed is between 10.0 (N s/m²) i.e. 10,000 (mPa s) to 80.0 (N s/m²) i.e. 80,000 (mPa s), 2 % in H2O at 20 °C. Preferably the viscosity of the HPMC as herein disclosed is between 25.0 (N s/m²) i.e. 25,000 (mPa s) to 50.0 (N s/m²) i.e. 50,000 (mPa s), 2 % in H2O at 20 °C. The viscosity of a 2% w/v solution of each sample can be measured using the standard tests, for example as those set out in the United States Pharmacopeia.

In a preferred embodiment, the composition as described herein comprises combination of HPMCs, the composition having a viscosity between 0.001 (N s/m²) i.e. 1 (mPa s) to 0.050 (N s/m²) i.e. 50 (mPa s) at 20 °C.

In a preferred embodiment, the composition as described herein comprises a combination of HPMCs, the composition having a viscosity between 0.002 (N s/m²) i.e. 2 (mPa s) to 0.045 (N s/m²) i.e. 45 (mPa s) at 20 °C.

In a preferred embodiment, the composition as described herein comprises a combination of HPMCs, the composition having a viscosity between 0.003 (N s/m²) i.e. 3 (mPa s) to 0.040 (N s/m²) i.e. 40 (mPa s) at 20 °C.

In a preferred embodiment, the composition as described herein comprises a combination of HPMCs, the composition having a viscosity between 0.004 (N s/m²) i.e. 4 (mPa s) to 0.030 (N s/m²) i.e. 30 (mPa s) at 20 °C. In a preferred embodiment, the composition as described herein comprises a combination of HPMCs, the composition having a viscosity between 0.005 (N s/m²) i.e. 5 (mPa s) to 0.025 (N s/m²) i.e. 25 (mPa s) at 20 °C.

Methods for preparing the composition | The Composition

In a preferred embodiment, the invention is directed to a process for the production of a formulation comprising an antibody, bispecific antibody or antigen binding fragment, comprising:

(a) dissolving an antibody, bispecific antibody or antigen binding fragment in a suitable solvent to form a solution;

(b) adding a first quantity of a first anti-solvent to said solution;

(c) dispersing a first quantity of HPMC in said solution;

(d) adding a second quantity of said first anti-solvent to said solution;

(e) dispersing a second quantity of HPMC in said solution; and

(f) adding a second anti-solvent to said solution so as to combine said antibody, bispecific antibody or antigen binding fragment with said HPMC and thereby form said formulation, wherein said second anti-solvent is a combination consisting essentially of organic solvent.

In a preferred embodiment, a method of making a composition is disclosed, wherein an antibody, bispecific antibody or antigen binding fragment as herein described is preformulated with a hydrophobic phase such that the antibody, bispecific antibody or antigen binding fragment is associated with and/or encapsulated within the hydrophobic phase. The formulated hydrophobic phase with the antibody, bispecific antibody or antigen binding fragment is then further formulated with a composition comprising a mucoadhesive compound.

In a preferred embodiment, a method of making a composition is disclosed, wherein an antibody, bispecific antibody or antigen binding fragment as herein described is preformulated with anyone of lipid particles, lipid microparticles, and/or lipid nanoparticles such that the antibody, bispecific antibody or antigen binding fragment is associated with and/or encapsulated within anyone of the lipid particles, lipid microparticles and/or lipid nanoparticles. The formulated lipid particles, lipid microparticles, or lipid nanoparticles with the antibody, bispecific antibody or antigen binding fragment is then further formulated with a composition comprising a mucoadhesive compound.

In a preferred embodiment, we disclose a method of preparing a composition comprising anyone of lipid particles, lipid microparticles and/or lipid nanoparticles and an antibody, bispecific antibody or antigen binding fragment, wherein the antibody, bispecific antibody or antigen binding fragment is associated with and/or encapsulated within anyone of the lipid particles, lipid microparticles and /or lipid nanoparticles, comprising the steps of:

(a) providing a first liquid composition, wherein the first liquid composition comprises an organic solution of one or more lipids;

(b) providing a second liquid composition, wherein the second liquid composition comprises an aqueous solution of antibody, bispecific antibody or antigen binding fragment;

(c) mixing the first liquid and the second liquid such as to form a third liquid composition, wherein the third liquid composition comprises anyone of newly formed lipid particles, lipid microparticles and/or lipid nanoparticles comprising the one or more lipids and anyone of the antibody, bispecific antibody or antigen binding fragment;

(d) further formulating the third liquid composition with a composition comprising a mucoadhesive compound such as to form a fourth liquid and obtain a physically stable formulation wherein the antibody, bispecific antibody or antigen binding fragment remains associated with and/or encapsulated within anyone of the lipid particles, lipid microparticles and/or lipid nanoparticles. Preferably, the mixing of the first and second liquids in stl(c) is performed using mixing devices selected from static mixers, microfluidic mixing platforms, jet impingement reactors, T-piece mixers, Y-shaped mixers, or multi-inlet vortex mixers. In a preferred embodiment, the mixing of the first liquid and the second liquid inlep (c) is performed using a T-piece, a Y-piece, a static mixer, or a microfluidic mixer.

Preferably, the mixing of the third liquid with the composition comprising a mucoadhesive compound in step (d) is performed using mixing devices selected from static mixers, microfluidic mixing platforms, jet impingement reactors, T-piece mixers, Y-shaped mixers, or multi-inlet vortex mixers. In a preferred embodiment, the mixing of the third liquid and the composition comprising a mucoadhesive compound in step (d) is performed using a T- piece, a Y-piece, a static mixer, or a microfluidic mixer.

Without wishing to be bound by theory, the physically stable formulation comprises lipid particles encapsulating some or all the antibody, bispecific antibody or antigen binding fragment, suspended within the composition comprising the mucoadhesive compound.

In a preferred embodiment, we disclose a formulation comprising a composition comprising the mucoadhesive compound, the formulation further comprising lipid particles encapsulating or associated with anyone one of an antibody, bispecific antibody or antigen binding fragment, wherein the lipid particles are suspended within a composition comprising the mucoadhesive compound.

In a preferred embodiment, the mucoadhesive compound in step (d) comprises or consists of hydroxypropyl methyl cellulose.

Sustained release

Preferably, the composition is administered in the form of a sustained release preparation. Other expressions like “extended release”, “controlled release”, “modified release” or “delayed release” “preparation” or “formulation” are understood herein to have the same meaning as “sustained release preparation”. Such preparations can in principle be in any form conceivable to the skilled person and include pharmaceutical forms, as long as a sustained release is ensured.

Preferably sustained release preparations encompass all pharmaceutical forms that create a steady antibody, bispecific antibody or antigen binding fragment release profile making the antibody, bispecific antibody or antigen binding fragment available over an extended period of time following application to the patient. Such an extended period of time may be between preferably 10, 20, 30, 40, 50 or 60 minutes and about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or preferably 24 hours. Extended release may also be defined functionally as the release of over preferably 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 or preferably 99 percent (%) of the antibody, bispecific antibody or antigen binding fragment after about 10, 20, 30, 40, 50 or 60 minutes and about 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23 or preferably 24 hours. Extended release as used herein may also be defined as making the antibody, bispecific antibody or antigen binding fragment available to the patient regardless of uptake, as some antibody, bi specific antibody or antigen binding fragment may never be absorbed by the patient.

Composition or formulation

The term “composition” or “formulation” as used herein, refers to any preparations which are in such a form as to permit the biological activity of the antibody, bispecific antibody or antigen binding fragment to be effect, and thereby are capable of being administered to a subject for prophylactic and/or therapeutic use, for example, prevention and/or treatment of respiratory viral infections, and/or symptoms caused by respiratory viruses as described herein.

The composition as described herein is formulated so that antibody, bispecific antibody or antigen binding fragment is capable of being delivered to a specific site in the human body, such as the mucosal surfaces, in particular the nasal cavity, oropharyngeal region, upper respiratory tract, respiratory tract, and/or lungs, and remain in place at the intended site for an appreciable time. Hence, with the composition as described herein, the bioavailability of the antibody is improved and thereby the composition of the present invention can provide prophylactic and/or therapeutic treatment of a disease. That is, the composition of the present invention is capable of contributing to an improved prophylactic and/or therapeutic treatment efficacy.

In a preferred embodiment, the present invention provides a composition comprising at least one antibody, bispecific antibody or antigen binding fragment, and at least one mucoadhesive compound. Preferably, the composition further comprises at least one preservative.

In a preferred embodiment, described herein is an aqueous formulation suitable for intranasal administration comprising an antibody, bispecific antibody or antigen binding fragment, water and one or more ingredients selected from mucoadhesive compound, antioxidants, stabilizing agents, surfactants, isotonicity agents and pH adjusting agents.

In a preferred embodiment, described herein is an aqueous formulation suitable for intranasal administration comprising an antibody, bispecific antibody or antigen binding fragment, water and one or more ingredients selected from alkylglycosides, chitosan, alkylcyclodextrins, benzalkonium chloride, sodium chloride and EDTA.

In a preferred embodiment, described herein is an aqueous formulation suitable for intranasal administration comprising an antibody, bispecific antibody or antigen binding fragment, water and one or more ingredients selected from dodecyl maltoside (DDM), tetradecyl maltoside (TDM), benzalkonium chloride, sodium chloride, hydrochloric acid and EDTA.

In a preferred embodiment, described herein is an aqueous formulation suitable for intranasal administration comprising an antibody, bispecific antibody or antigen binding fragment, water and one or more ingredients selected from dodecyl maltoside (DDM), benzalkonium chloride, sodium chloride and EDTA.

In a preferred embodiment, described herein is an aqueous formulation suitable for intranasal administration comprising an antibody, bispecific antibody or antigen binding fragment, water, dodecyl maltoside (DDM) and one or more ingredients selected from benzalkonium chloride, sodium chloride, pH adjusting agents and EDTA.

In a preferred embodiment, described herein is an aqueous formulation suitable for intranasal administration comprising an antibody, bispecific antibody or antigen binding fragment, water, benzalkonium chloride and one or more ingredients selected from dodecyl maltoside (DDM), sodium chloride, pH adjusting agents, and EDTA.

In some embodiments, the pharmaceutically acceptable liquid carrier comprises water and one or more excipient(s) selected from the group comprising a mucoadhesive compound, a buffering agent, a flavoring agent, a humectant, a penetration enhancer, a pH adjusting agent, a preservative, a solvent or co-solvent, a surfactant, a tonicity adjusting agent, a viscosity adjusting agent, or a combination thereof.

In a preferred aspect, the present invention provides a formulation comprising an antibody, bispecific antibody or antigen binding fragment and one or more excipient(s) selected from the group comprising a mucoadhesive compound, an emulsifier, pH stabilizer, a dispersing agent, stabilizing agent, wetting agent, spreading agent, anti-foaming agent, rheology modifier, solvent, capsule forming agent, quenching agent, super spreader, an antifreezing agent, a biocide, an anti-caking agent, an inert carrier or combination thereof.

The composition as described herein is formulated so that during storage under the temperature ranging from -80 °C to 30 °C or to room temperature, during transportation, and/or in human body, the quality and/or the structure of an antibody including but not limited to the antibody as described herein is stable. That is, the degradation and/or deactivation of the antibody is reduced, stopped and/or prevented, the physical and/or chemical stability is retained, biological activity of the antibody is maintained, and/or the shelf life of the composition is prolonged.

Preferably, the composition as described herein is capable of delivering molecules with a high molecular weight. Such molecules include but not limited to oligonucleotides and/or polypeptides such as proteins and/or antibodies or antigen-binding fragment thereof. In a preferred embodiment, said at least polypeptide comprises or consists of at least one antibody or antigen-binding fragment thereof as disclosed herein. More preferably, said at least polypeptide comprises or consists of CR9114. Thereby, the composition of the present invention is capable of prophylactically and/or therapeutically treating a disease or symptoms caused by at least one influenza viruses.

More preferably, the composition as described herein is capable of delivering at least one antibody.

More preferably, the composition as described herein is capable of delivering CR9114.

In a preferred embodiment, a composition comprising an antibody, bispecific antibody or antigen binding fragment, at least one mucoadhesive compound, and at least one preservative, wherein the concentration of the antibody, bispecific antibody or antigen binding fragment ranges from 0.001 to 250 % (w/v) and the concentration of the mucoadhesive compound ranges from 0.1 to 10 % (w/v).

The concentration of an antibody, bispecific antibody or antigen binding fragment comprised in the liquid composition as described herein is provided at a concentration ranging from 0.001, 0.03, 0.05, 0.1, 5, 10, 15, 20, 25, 30, 35, 40, or 45 % (w/v), or at most 250, 230, 200, 180, 160, 140, 120, 100, or 90, 85, 80, 75, 70, 65, 60, 55 or 50 % (w/v). Preferably, the concentration of the antibody, bispecific antibody or antigen binding fragment comprised in the liquid composition as described herein is provided at a concentration ranging from 0.1 % (w/v) to 20 % (w/v).

Preferably, the concentration of the mucoadhesive compound in the liquid composition as described herein is provided at a concentration ranging from 0.1, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 4.5, or 5% (w/v), or at most 10, 9.5, 9, 8.5, 8, 7.5, 7, 6.5, 6, or 5.5 % (w/v). In one preferred embodiment, the composition is a liquid formulation for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment (0.025 % w/v), HPMC (3% w/v), sodium acetate 20 mM, sodium chloride 75 mM, sucrose 5% (w/v), polysorbate 20 (0.02% w/v), pH 5.5.

In one preferred embodiment, the composition is a liquid formulation for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment (0.025 % w/v), HPMC (4% w/v), sodium acetate 20 mM, sodium chloride 75 mM, sucrose 5% (w/v), polysorbate 20 (0.02% w/v), pH 5.5.

In one preferred embodiment, the composition is a liquid formulation for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment (0.025 % w/v), HPMC (5% w/v), sodium acetate 20 mM, sodium chloride 75 mM, sucrose (5% w/v), polysorbate 20 (0.02% w/v), pH 5.5.

In one preferred embodiment, the composition is a liquid for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment (0.025 % w/v), HPMC (3% w/v), sodium acetate 20 mM, sodium chloride 75 mM, sucrose (5% w/v), polysorbate 40 (0.02% w/v), pH 5.5.

In one preferred embodiment, the composition is a liquid for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment of (0.025 % w/v), HPMC (3% w/v), sodium acetate 20 mM, sodium chloride 75 mM, sucrose (5% w/v), polysorbate 60 (0.02% w/v), pH 5.5.

In one preferred embodiment, the composition is a liquid for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment of (0.025 % w/v), HPMC (3% w/v), sodium acetate 20 mM, sodium chloride 75 mM, sucrose (5% w/v), 0.02% (w/v) of mixtures of DPPC (dipalmitoyl phosphatidylcholine) and PG (phosphatidyl glycerol), pH 5.5. In one preferred embodiment, the composition is a liquid for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment of (0.025 % w/v), HPMC (3% w/v), sodium acetate 20 mM, sodium chloride 75 mM, trehalose (5% w/v), polysorbate 20 (0.02% w/v), pH 5.5.

In one preferred embodiment, the composition is a liquid for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment of (0.025 % w/v), HPMC (3% w/v), sodium acetate 20 mM, sodium chloride 75 mM, trehalose (5% w/v), polysorbate 40 (0.02% w/v), pH 5.5.

In one preferred embodiment, the composition is a liquid for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment of (0.025 % w/v), HPMC (3% w/v), sodium acetate 20 mM, sodium chloride 75 mM, trehalose (5% w/v), polysorbate 60 (0.02% w/v), pH 5.5.

In one preferred embodiment, the composition is a liquid for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment of (0.025 % w/v), HPMC (3% w/v), sodium acetate 20 mM, sodium chloride 75 mM, trehalose (5% w/v), 0.02% (w/v) of mixtures of DPPC (dipalmitoyl phosphatidylcholine) and PG (phosphatidyl glycerol), pH 5.5.

In one preferred embodiment, the composition is a liquid for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment of (0.025 % w/v), HPMC (3% w/v), sodium acetate 20 mM, sucrose 75 mM, trehalose (5% w/v), polysorbate 20 (0.02% w/v), pH 5.5.

In one preferred embodiment, the composition is a liquid for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment of (0.025 % w/v), HPMC (3% w/v), sodium acetate 20 mM, sucrose 75 mM, trehalose (5% w/v), polysorbate 40 (0.02% w/v), pH 5.5. In one preferred embodiment, the composition is a liquid for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment of (0.025 % w/v), HPMC (3% w/v), sodium acetate 20 mM, sucrose 75 mM, trehalose (5% w/v), polysorbate 60 (0.02% w/v), pH 5.5.

In one preferred embodiment, the composition is a liquid for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment of (0.025 % w/v), HPMC (3% w/v), sodium acetate 20 mM, sucrose 75 mM, trehalose (5% w/v), 0.02% (w/v) of mixtures of DPPC (dipalmitoyl phosphatidylcholine) and PG (phosphatidyl glycerol), pH 5.5.

In one preferred embodiment, the composition is a liquid for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment of (0.025 % w/v), HPMC (3% w/v), benzalkonium chloride 20 mM, sodium chloride 75 mM, sucrose (5% w/v), polysorbate 20 (0.02% w/v), pH 5.5.

In one preferred embodiment, the composition is a liquid for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment of (0.025 % w/v), HPMC (3% w/v), benzalkonium chloride 20 mM, sodium chloride 75 mM, sucrose (5% w/v), polysorbate 40 (0.02% w/v), pH 5.5.

In one preferred embodiment, the composition is a liquid for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment of (0.025 % w/v), HPMC (3% w/v), benzalkonium chloride 20 mM, sodium chloride 75 mM, sucrose (5% w/v), polysorbate 60 (0.02% w/v), pH 5.5.

In one preferred embodiment, the composition is a liquid for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment of (0.025 % w/v), HPMC (3% w/v), benzalkonium chloride 20 mM, sodium chloride 75 mM, sucrose (5% w/v), 0.02% (w/v) of mixtures of DPPC (dipalmitoyl phosphatidylcholine) and PG (phosphatidyl glycerol), pH 5.5. In one preferred embodiment, the composition is a liquid for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment of (0.025 % w/v), HPMC (3% w/v), benzalkonium chloride 20 mM, sodium chloride 75 mM, trehalose (5% w/v), polysorbate 20 (0.02% w/v), pH 5.5.

In one preferred embodiment, the composition is a liquid for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment of (0.025 % w/v), HPMC (3% w/v), benzalkonium chloride 20 mM, sodium chloride 75 mM, trehalose (5% w/v), polysorbate 40 (0.02% w/v), pH 5.5.

In one preferred embodiment, the composition is a liquid for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment of (0.025 % w/v), HPMC (3% w/v), benzalkonium chloride 20 mM, sodium chloride 75 mM, trehalose (5% w/v), polysorbate 60 (0.02% w/v), pH 5.5.

In one preferred embodiment, the composition is a liquid for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment of (0.025 % w/v), HPMC (3% w/v), benzalkonium chloride 20 mM, sodium chloride 75 mM, trehalose (5% w/v), 0.02% (w/v) of mixtures of DPPC (dipalmitoyl phosphatidylcholine) and PG (phosphatidyl glycerol), pH 5.5.

In one preferred embodiment, the composition is a liquid for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment of (0.025 % w/v), HPMC (3% w/v), benzalkonium chloride 20 mM, sucrose 75 mM, trehalose (5% w/v), polysorbate 20 (0.02% w/v), pH 5.5.

In one preferred embodiment, the composition is a liquid for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment of (0.025 % w/v), HPMC (3% w/v), benzalkonium chloride 20 mM, sucrose 75 mM, trehalose (5% w/v), polysorbate 40 (0.02% w/v), pH 5.5. In one preferred embodiment, the composition is a liquid for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment of (0.025 % w/v), HPMC (3% w/v), benzalkonium chloride 20 mM, sucrose 75 mM, trehalose (5% w/v), polysorbate 60 (0.02% w/v), pH 5.5.

In one preferred embodiment, the composition is a liquid for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment of (0.025 % w/v), HPMC (3% w/v), benzalkonium chloride 20 mM, sucrose 75 mM, trehalose (5% w/v), 0.02% (w/v) of mixtures of DPPC (dipalmitoyl phosphatidylcholine) and PG (phosphatidyl glycerol), pH 5.5.

In one preferred embodiment, the composition is a liquid for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment of (0.025 % w/v), HPMC (3% w/v), EDTA 20 mM, 75 mM sodium chloride 75 mM, sucrose (5% w/v), polysorbate 20 (0.02% w/v), pH 5.5.

In one preferred embodiment, the composition is a liquid for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment of (0.025 % w/v), HPMC (3% w/v), EDTA 20 mM, 75 mM sodium chloride 75 mM, sucrose (5% w/v), polysorbate 40 (0.02% w/v), pH 5.5.

In one preferred embodiment, the composition is a liquid for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment of (0.025 % w/v), HPMC (3% w/v), EDTA 20 mM, 75 mM sodium chloride 75 mM, sucrose (5% w/v), polysorbate 60 (0.02% w/v), pH 5.5.

In one preferred embodiment, the composition is a liquid for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment of (0.025 % w/v), HPMC (3% w/v), EDTA20 mM, sodium chloride 75 mM, sucrose (5% w/v), 0.02% (w/v) of mixtures of DPPC (dipalmitoyl phosphatidylcholine) and PG (phosphatidyl glycerol), pH 5.5.

In one preferred embodiment, the composition is a liquid for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment of (0.025 % w/v), HPMC (3% w/v), EDTA 20 mM, sodium chloride 75 mM, trehalose (5% w/v), polysorbate 20 (0.02% w/v), pH 5.5.

In one preferred embodiment, the composition is a liquid for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment of (0.025 % w/v), HPMC (3% w/v), EDTA 20 mM, sodium chloride 75 mM, trehalose (5% w/v), polysorbate 40 (0.02% w/v), pH 5.5.

In one preferred embodiment, the composition is a liquid for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment of (0.025 % w/v), HPMC (3% w/v), EDTA 20 mM, sodium chloride 75 mM, trehalose (5% w/v), polysorbate 60 (0.02% w/v), pH 5.5.

In one preferred embodiment, the composition is a liquid for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment of (0.025 % w/v), HPMC (3% w/v), EDTA 20 mM, sodium chloride 75 mM, trehalose (5% w/v), 0.02% (w/v) of mixtures of DPPC (dipalmitoyl phosphatidylcholine) and PG (phosphatidyl glycerol), pH 5.5.

In one preferred embodiment, the composition is a liquid for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment of (0.025 % w/v), HPMC (3% w/v), EDTA 20 mM, sucrose 75 mM, trehalose (5% w/v), polysorbate 20 (0.02% w/v), pH 5.5.

In one preferred embodiment, the composition is a liquid for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment of (0.025 % w/v), HPMC (3% w/v), EDTA 20 mM, sucrose 75 mM, trehalose (5% w/v), polysorbate 40 (0.02% w/v), pH 5.5.

In one preferred embodiment, the composition is a liquid for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment of (0.025 % w/v), HPMC (3% w/v), EDTA 20 mM, sucrose 75 mM, trehalose (5% w/v), polysorbate 60 (0.02% w/v), pH 5.5.

In one preferred embodiment, the composition is a liquid for nasal administration. The composition comprises an antibody, bispecific antibody or antigen binding fragment of (0.025 % w/v), HPMC (3% w/v), EDTA 20 mM, sucrose 75 mM, trehalose (5% w/v), 0.02% (w/v) of mixtures of DPPC (dipalmitoyl phosphatidylcholine) and PG (phosphatidyl glycerol), pH 5.5

Viscosity

Viscosity is a measure of resistance to deformation and may be determined by employing a sheer stress method as known to a person skilled in the art. The viscosity of a dosage unit can affect the residence time at the site of administration, for example, in the nasal cavity.

As a preferred embodiment, when formulated as a liquid composition the composition comprises a viscosity modifier, otherwise known as a “viscosity regulating agent”, commonly understood to be a thickener or gelling agent.

Preferably, when formulated as a liquid composition, the composition has a higher kinematic viscosity than water at the same temperature.

Preferred viscosity modifiers may include any one or more of polysaccharides, carbomers, acrylic polymers, such as Carbopol ®, polyvinyl alcohol and other vinylic polymers, povidone, Co-Polyvidone (Kollidon VA64), cetyl alcohols, bentonite, diatomaceous earth, montmorillonite, attapulgite, cellulose and cellulose derivatives thereof, such as, hydroxymethyl cellulose, hydroxymethyl cellulose, hydroxyethyl cellulose and hydroxypropyl methylcellulose, gellan gum, guar gum, karaya gum, xanthan gum, polyvinyl alcohol, silica, colloidal silicon dioxide, such as Aerosil® 200 or Cab-O-Sil®, such as Cab- O-Sil® M-5P, lipophilic silicon dioxide, such as Aerosil® R972, hydrated clay minerals, magnesium aluminum silicate, polysaccharide gel, hydrophobic fumed silica, organic derivative of hectorite clay, stearic acid, glyceryl behenate, wax, beeswax, petrolatum, triglycerides, lanolin and suitable mixtures thereof.

Preferred viscosity modifiers may include any one or more of colloidal silicon dioxide such as Aerosil® 200, SiCh and polyvinyl alcohol.

Preferred viscosity modifiers may include any one or more of methylcellulose, carboxymethyl cellulose (CMC), Me-OH-Pr cellulose, microcrystalline cellulose (MCC), sodium carboxymethyl cellulose (Na CMC).

Preferred viscosity modifiers that lower the viscosity may include polyethylene glycol 400 (PEG 400)

Especially preferred viscosity modifiers may include any one or more of methylcellulose A4M

Preferred viscosity modifiers may include any one or more of methylcellulose, hydroxymethyl cellulose, hydroxyethyl cellulose and hydroxypropyl methylcellulose.

An especially preferred viscosity modifier is hydroxypropyl methylcellulose.

As a preferred embodiment, when formulated as a liquid composition the composition comprises viscosity modifiers in an amount ranging from about 0.01 % to about 10% w/v of the composition. In some embodiments, the pharmaceutically acceptable liquid carrier comprises or further comprises from about 0.1% to about 10% w/v carboxymethyl cellulose.

In some embodiments, the pharmaceutically acceptable liquid carrier comprises or further comprises from about 0.1% to about 15% w/v Me-OH-Pr cellulose.

In some embodiments, the pharmaceutically acceptable liquid carrier comprises or further comprises from about 0.1% to about 2% w/v microcrystalline cellulose. In some embodiments, the pharmaceutically acceptable liquid carrier comprises or further comprises from about 0.1% to about 5% w/v sodium carboxymethyl cellulose (Na CMC).

Preferably, when formulated as a liquid composition, the composition has a kinematic viscosity of from 0.5 to 2 (m²/s) at 20° C.

Preferably, when formulated as a liquid composition, the composition has a kinematic viscosity of from 0.5 to 2 m²/s, from 0.5 to 1.5 m²/s, 0.5 to 1.25 m²/s, 0.5 to 1.1 m²/s, 0.5 to 1 m²/s, 0.5 to 0.9 m²/s, 0.5 to 0.75 m²/s, 0.75 to 2 m²/s, 0.75 to 1.5 m²/s, 0.75 to 1.25 m²/s, 0.75 to 1.1 (m²/s), 0.75 to 1 (m²/s), 0.75 to 0.9 (m²/s), 0.9 to 2 (m²/s), 0.9 to 1.5 (m²/s), 0.9 to 1.25 (m²/s), 0.9 to 1.1 (m²/s), 0.9 to 1 (m²/s), 1 to 2 (m²/s), 1 to 1.5 (m²/s), 1 to 1.25 (m²/s), 1 to 1.1 (m²/s), 1.1 to 2 (m²/s), 1.1 to 1.5 (m²/s), 1.1 to 1.25 (m²/s), 1.25 to 2 (m²/s), 1.25 to 1.5 (m²/s), or 1.5 to 2 (m²/s) at 20° C.

In some embodiments, the dosage unit has a kinematic viscosity of 0.9 to 1.25 (m²/s) at 20' C.

SEQUENCES

Heavy chain CDR1 region

SEQ ID NO.: 001 NYAIS Heavy chain CDR2 region

SEQ ID NO. : 002 GISPIFGSTAYAQKFQG

Heavy chain CDR3 region SEQ ID NO : 003 HGNYYYYSGMDV

Light chain CDR1 region

SEQ ID NO. : 004 SGSDSNIGRRSVN Light chain CDR2 region

SEQ ID NO. : 005 SNDQRPS

Light chain CDR3 region

SEQ ID NO. : 006 AAWDDSLKGAV

Heavy chain CDR1 region

SEQ ID NO. : 007 DYAMS

Heavy chain CDR2 region SEQ ID NO : 008 GLNWNGDITAYTDSVKG

Heavy chain CDR3 region

SEQ ID NO. : 009 TSSWGDYTRGPEPKITWYFDL Light chain CDR1 region

SEQ ID NO : 010 RASQGIDGYLA

Light chain CDR2 region SEQ ID NO : Oil AASLLQS

Light chain CDR3 region

SEQ ID NO.: 012 QHLDSYPLFT

Heavy chain CDR1 region

SEQ ID NO : 013 DYRIH

Heavy chain CDR2 region

SEQ ID NO. : 014 RMNPKSGDTNFAQKFQG

Heavy chain CDR3 region

SEQ ID NO. : 015 LLIVGGFDPLDDFEV

Light chain CDR1 region

SEQ ID NO.: 016 SGTSSDVGGYNFVS

Light chain CDR2 region

SEQ ID NO : 017 EVTKRPS

Light chain CDR3 region

SEQ ID NO.: 018 SSYGGTNNLL

Heavy chain CDR1 region

SEQ ID NO : 019 GYAMH

Heavy chain CDR2 region

SEQ ID NO. : 020 VISRDARNKYYADSVKG Heavy chain CDR3 region

SEQ ID NO. : 021 LIIPGITEPGSPDALDI Light chain CDR1 region

SEQ ID NO. : 022 RASQDISKWLA

Light chain CDR2 region

SEQ ID NO. : 023 AASSLQS

Light chain CDR3 region

SEQ ID NO. : 024 QQASSFPWSIT

Heavy chain CDR1 region SEQ ID NO : 025 YFYLH

Heavy chain CDR2 region

SEQ ID NO. : 026 IINPRGDGTRYAQKFQG Heavy chain CDR3 region

SEQ ID NO. : 027 GADHGAFDI

Light chain CDR1 region

SEQ ID NO. : 028 RASQSVRRNYFA

Light chain CDR2 region

SEQ ID NO. : 029 DASTRAT Light chain CDR3 region

SEQ ID NO. : 030 QQYDSSPPMYI

Heavy chain CDR1 region SEQ ID NO. : 031 SHYMH

Heavy chain CDR2 region

SEQ ID NO. : 032 IINPSGSGTAYGQKFQG Heavy chain CDR3 region

SEQ ID NO. : 033 GSGGLFAY

Light chain CDR1 region

SEQ ID NO. : 034 RASQIVRSNYLA

Light chain CDR2 region

SEQ ID NO.: 035 GASSRAT

Light chain CDR3 region SEQ ID NO : 036 LQYDSSPPTYI

Heavy chain CDR1 region

SEQ ID NO. : 037 SYYMH Heavy chain CDR2 region

SEQ ID NO. : 038 LITPSGDDTYYAQRFQG Heavy chain CDR3 region

SEQ ID NO. : 039 MSRAGGFDV

Light chain CDR1 region

SEQ ID NO. : 040 RASQSITGRYLA

Light chain CDR2 region

SEQ ID NO.: 041 GESSRVT

Light chain CDR3 region

SEQ ID NO. : 042 QHFASSPPTYT

EXAMPLES

Monoclonal antibodies (mAbs) are powerful therapeutics that address unmet prophylactic and/or therapeutic treatment needs. Local delivery of these mAbs, used in treatment as described herein via the inhalation route, is capable of reducing the amount of dose and/or the frequency of dosing, limiting systemic exposure of healthy tissues, and lessening adverse events, while improving patient compliance and lowering cost of treatment. Additional beneficial effects of inhalation route administration include the large surface area for being in contact with the mAbs.

As described above, pulmonary delivery also offers the advantage of delivering the composition as described herein at high concentrations in the respiratory tissues and/or organs. Also, the distribution profile of the aerosolised composition of the present invention is desirable for prophylactically and/or therapeutically treatment of respiratory diseases.

Components in the compositions as tested in the Examples of the present invention are readily available from, for example, Dow Chemicals (U.S.A) or International Flavors & Fragrances (IFF, U.S. A.). The exemplary mucoadhesive compounds used in the Examples are HPMC 1828 (METHOCEL™ J12MS, IFF), HPMC 2208 (METHOCEL™ K3, IFF), HPMC 2906 (METHOCEL™ F4M, IFF), and HPMC2910 (METHOCEL™ E15LV, IFF).

Example 1. Preparation of liquid compositions

The liquid compositions according to Table 1 are prepared to enable evaluation of the formulation properties. Table 1. Preparation of liquid compositions

*Additional excipients such as amino acids, preservatives, and/or stabilizers can be added into the compositions.

The formulations are filled in single-dosing and multiple-dosing devices, for example Aptar VP7, Aptar UDSp Device, Aptar UDSp Blister Device (Aptar, the United Kingdom), respectively. Example 2. Preparation of different HPMC concentrations in liquid compositions

Liquid compositions (approximatively 5 g) are prepared according to Table 2a to 2d for evaluating their properties.

5 Table. 2a. Liquid compositions comprising different concentrations of HPMC2208

Table. 2b. Liquid compositions comprising different concentrations of HPMC1828

Table. 2c. Liquid compositions comprising different concentrations of HPMC2906

10

Table. 2d. Liquid compositions comprising different concentrations of HPMC2910

Example 3. Preparation of liquid compositions comprising two HPMCs

Liquid compositions (approximatively 5 g) are prepared according to Table 3 or Table 3 a for evaluating their properties.

Table. 3. Two different HPMCs comprised in the liquid compositions

Table. 3a. Two different HPMC comprised in the liquid compositions

Base formulation, as described in Table 3a, comprises Sodium chloride : 0.22% w/v; sodium phosphate dibasic: 0.12% w/v; sodium phosphate monobasic: 1.14% w/v; glycerin: 2.5% w/v; benzalkonium chloride: 0.02% w/v; and has a pH value of 6.5.

Vehicle, as described in Table 3a, comprises saline solution (NaCl 0.9% w/v).

Example 4. Mucociliary clearance testing

The aim of this assay is to perform a short-term (for example, from 1 to 4 days) evaluation of the effect of the compositions as prepared according to Tables 1-3 on mucociliary clearance, cilia beating frequency (CBF) and mucus secretion on a fully differentiated human airway epithelium (MucilAir™, Epithelix, Swtizerland).

Cilia Beating Frequency

CBF is measured by an experimental system consisting of three parts: a camera (for example, Sony XCD V60 Firewire), a peripheral component interconnect (PCI) card and a specific package of software. Images are captured at high frequency rate (125 fps) at room temperature and the cilia beating frequency is then calculated using Epithelix software. CBF values may be subject to fluctuations due to parameters such as temperature, mucus viscosity or liquid (such as a buffered saline solution) applied on the apical surface of the MucilAir™ 3D epithelial model.

Mucociliary Clearance and mucus secretion

The purpose of these tests is to analyse the residence time of the compositions as prepared according to Tables 1-3 on the mucosa of the nasal and/or oral cavity. The mucolytic activity is evaluated through mucociliary clearance analysis on Day 1 and Day 4, and mucin secretion by mucin quantification on Day 1 and Day 4. The mucociliary clearance (“MCC”) can be monitored by techniques known by the skilled in the art, such as by using a Sony XCD-U100CR camera connected to an Olympus BX51 microscope with a 5* objective. Polystyrene microbeads of 30 pm diameter (Sigma, 84135) are added on the apical surface of MucilAir™. Microbead movements are video tracked at 2 frames per second for 30 images at room temperature. Three movies were taken per insert. Average beads movement velocity (Q m/sec) is calculated with ImageProPlus 6.0 software.

Example 5. Assessment of the stability of the liquid composition

This assessment is to evaluate the performance of the compositions in various storage conditions (25°C/60% RH, 40°C/75% RH), temperature (5°C, -20°C) and time-points (1 month, 2 months, 3 months). In addition, the compositions as prepared according to Tables 1-3 are subject to 5 freeze-thaw cycles. Freezing is carried out at -20°C and thaw at not controlled ambient conditions.

Example 6. Nasal case deposition assessment of the liquid formulation

This test is to evaluate the distribution of the compositions on the mucosal surfaces. Abroad distribution of the drug on the mucosal surfaces can be desirable for drugs intended for local action or systemic absorption and for vaccines.

A nasal cast male Caucasian model produced by Aptar (the United States) is used to determine the regional deposition of the formulations. This nasal cast is an accurate model made after a male scanner with sections of 0.5 mm, composed by flexible nostrils (devices nozzle can be turned and placed like in human nostril) and validated by tomography. The measurement method is reliable and accurate with no leakage between the blocks of the model, and has been validated with in vivo results.

The nasal cast is designed to simulate a male human model and thereby comprises various parts with substantially similar functions to the nose, frontal sinus and nasal valve, maxillary sinus, frontal sinus, floor of nasal cavity, turbinates, ethmoids, maxillary sinus, sphenoids, and floor of nasal cavity of a natural person. The compositions are sprayed into the nasal cast model employing Aptar CPS pump, respectively. The CR9114 deposited in each region of interest is collected and quantified by SEC and/or HPLC.

Example 7. Rheological Profiling of the compositions

The viscosity of the compositions as prepared according to Tables 1-3 can be measured by any known or otherwise effective technique employed to determine viscosity, such as those described in ASTM DI 824-87, ASTM DI 084-88, and/or ASTM D2196-86. Typical viscometers employed to measure viscosity include the Brookfield Syncho-Lectric Viscometer (Brookfield Ametek, the U.S.A.) and the Haake Viscometer (Thermofisher, the U.S.A.).

The viscosity of the compositions is determined by shear rates. Casson method of data analysis is applied to derive the plastic viscosity value for the composition. The formulations prepared according to Tables 1-3 are tested in triplicate employing Kinexus (NETZSCH- Geratebau GmbH, DE), respectively, and the mean value is obtained. The compositions are ideally tested at 5°C and 25°C so that the flow properties across the temperature range of interest.

Example 8. Spray Pattern (SP) and Plume Geometry (PG)

The plume of the compositions as prepared according to Tables 1-3 is characterized (CPS lOOpL, Aptar, UK). A pulsed laser technique (Oxford Laser, UK) is also employed.

For Spray Pattern (SP) the laser plane is positioned at 3 and 6 cm from the pump nozzle and cut horizontally while a high-speed image is recorded. This allows to analyse the following parameters:

-Minimum dimeter (Dmin);

-Maximum diameter (Dmax);

-Ovality ration (Dmin/Dmax);

-Area (Equation: n x Dmin x Dmax). Plume width, length and angle are evaluated for Plume Geometry (PG) at a single distance which allows to capture the whole plume emitted. Analysis is performed 9 times for each formulation, 3 times per distance employing automated actuation (Vereo® actuator NSx, Proveris Scientific, US). The parameters for the automated actuation are: pump (CPS lOOpL), velocity (70 mm/s), acceleration (3000 mm/s²), hold time (300 ms), and symmetric profile.

SP and PG tests are performed at T=0 for low and high concentration formulations at 3 and 6 cm (SP) and at 6 cm (PG).

Example 9. Mucociliary Clearance (MCC) assay, Cilia Beating Frequency (CBF) assay, Transendothelial Electrical Resistance (TEER) assay, and Lactate Dehydrogenase (LDH) analysis

The aim of this assay was to evaluate the local tolerance and mucolytic effects of the formulations as described in Table 3 a by using fully differentiated human nasal epithelial cells cultured at the air-liquid interface. Epithelia (MucilAir™-Pool) were reconstituted with a mixture of cells isolated from 14 different healthy nasal donors. The formulations were applied apically to mimic exposure on the nasal mucosa via inhalation and removed after 3 hours to mimic mucociliary clearance.

Two series were carried out in parallel.

Series 1 was performed for acute dose exposure for assessing CBT before apical exposure (TO) and at 3 hours (T3h), and for assessing TEER and MCC at 3 hours (Figure 1).

Series 2 was performed for repeated dose exposure for assessing TEER and LDH cytotoxicity analysis at 3 hours (T3h), 27 hours (T27h), 51 hours (T5 Ih) and 75 hours (T75h). Basal medium for IL-8 release was collected and analysed at T24h, T48h, T72h and T96h. CBF was measured at TO, T3h, T24h, T27h, T48h, T51h, T72h, T75h, and T96h. MCC was measured at T75h (Figure 2). In each of Series 1 and 2, eleven groups of fully differentiated human nasal epithelial cells were tested. One group of cells was tested with Formulation NP-015 which is a commercially available product called COVITRAP™ (Hibiocy Co., Thailand) comprising 3% w/v of HPMC 2910, the base formulation, and the antibody against SARS-CoV-2.

Seven groups of cells were tested with Formulations NP-016 - NP-022 comprises the mucoadhesive compound(s) as described in Table 3a and the base formulation, respectively.

One group of cells was tested with the base formulation as described in Table 3a. One group of cells was tested with vehicle as described in Table 3a which comprises saline solution (NaCl 0.9% w/v). One group of cells was untreated with any formulation.

10 pl of each of the formulations, as described herein, was applied on the apical side of Mucil Air™-Pool .

Each of the formulations, as described in Table 3 a, was repeated three times.

Example 9a. Mucociliary Clearance (MCC) assay

The mucociliary clearance was monitored using a Sony XCD-U100CR camera connected to an Axiovert 200M microscope (Zeiss) with a 5x objective. Polystyrene microbeads (30pm diameter - Sigma, 84135) were added on the apical surface of Mucil Air™-Pool. Microbeads movements were video tracked at 2 frames per second for 30 images at room temperature. Three movies are taken per insert. Average bead movement velocity (pm/sec) are calculated with the ImageProPlus 6.0 software.

The MCC values are typically comprised between 40 to 60 pm/s for healthy human donors. MCC values are linked to several factors such as (but not limited too): CBF values and mucus properties (i.e. rheological behavior). The results of MCC at 3 hours and 75 hours are shown in Figures 3 and 4, respectively. At 3 hours, the cells treated with, for example, NP-016, NP-017 or NP-019, showed lower MCC values than NP-020 or NP-021. That demonstrated that NP-016, NP-017 or NP-019 is capable of delaying the clearance of formulations from the nasal mucosa, and hence prolonging the retention of formulations on the nasal mucosa.

Due to lack of material, the group of cells treated with NP-018 was not measured at 3 hours.

Table 4: MCC Assay at 3 hrs

Table 5: MCC Assay at 75 hrs

Example 9b. Cilia beating frequency (CBF) assay

The measurement of cilia beating frequency comprised three parts: a camera connected to a microscope, a PCI card and a specific package of software based on Fourier spectral analysis. The Cilia beating frequency is expressed as Hz. 256 images movies were captured at high frequency rate (125 frames per second). Subsequently, cilia beating frequency was calculated using an Epithelix software (Cilia-X). The CBF values are typically comprised between 4 and 8 Hz. CBF value is temperature dependent, so measurements were done in similar temperature conditions. The results of CBF at 0 hour, 3 hours, 24 hours, 27 hours, 48 hours, 51 hours, 72 hours, 75 hours and 96 hours are shown in Figures 5 and 6.

Table 6: CBF Assay at 0 and 3 hrs

Table 7: CBF Assay at 0 hour, 3 hours, 24 hours, 27 hours, 48 hours, 51 hours, 72 hours, 75 hours and 96 hours

Example 9c. Transendothelial Electrical Resistance (TEER) assay

The change of TEER reflects the integrity/state of epithelia. For example, if holes were present or if cellular junction were broken, the TEER values would be generally below 100 Q.cm². In contrast, when epithelia are not damaged, the TEER values are typically comprised between 200 to 600 Q.cm². A notable decrease of the TEER value (but > 100 Q.cm2) could be observed in certain cases. This change generally reflects an activation of the ion channels. A drastic increase of the TEER value reflects a blockage of the ion channel activity or a destruction of the ciliated cells.

After addition of 200 pl of liquid solution (generally NaCl 0.9% or culture media) to the apical compartment of MucilAir™ cultures, resistance was measured with an EVOMX voltohm-meter (World Precision Instruments UK, Stevenage) for each condition. Resistance values (Q) were converted to TEER (Q.cm2) using the following formula: TEER (Q.cm2) = (resistance value (Q) -100(Q)) x 0.33 (cm2), where 100 Q is the resistance of the membrane and 0.33 cm2 is the total surface of the epithelium.

The results of TEER assay are shown in Figures 7 and 8. The cells treated with any one of the formulations as listed in Table 3a, were within the range of 200 to 600 Q.cm². Hence, no destruction of the cells was observed.

Table 8: TEER 3 hrs

Table 9: TEER at 3, 27, 51, 75 and 96 hrs

Example 9d. Lactate Dehydrogenase (LDH) analysis

Quantification by dosing the LDH released in the medium by dead cells. Cytotoxicity LDH Assay Kit-WST (Dojindo, Japan) was used for this quantification.

LDH is a stable cytoplasmic enzyme that is rapidly released into the culture medium upon rupture of the plasma membrane. 100 pl basolateral medium collected at each time-point was incubated with the reaction mixture of the LDH Assay Kit-WST. The amount of the released LDH was then quantified by measuring the absorbance of each sample at 490 nm with a microplate reader. To determine the percentage of cytotoxicity, the following equation was used (A= absorbance values): Cytotoxicity (%) = (A (exp value)-A (low control)/A (high control)-A (low control))* 100.

The high control value was obtained by 10 % Triton X-100 apical treatment (24 hours). Triton X-100 causes a massive LDH release and corresponds to 100 % cytotoxicity.

The negative controls (non-treated and vehicle) show a low daily basal LDH release, <5 %, which is due to a physiological cell turnover in MucilAir™.

The result of LDH assay is shown in Figure 9. Triton X-100 was used in the assay as negative control. It was observed that apical exposure to Triton X-100 lead to a massive LDH release, which represented 100% cytotoxicity. The cells exposed to any one of the formulations as described in Table 3a displayed normal LDH value of < 5%. Hence, the formulation as described in Table 3 a represented no cytotoxicity and considered safe to be applied on human cells.

The result is shown in Figure 9. No cytotoxicity was observed in all the cells exposed to the formulations as described in Table 3a for up to 96 hours. That means that the formulations of the present invention did not pose long-term toxicity to human cells.

Example 9e. Interleukin-8 (IL-8) Basal Release assay

The release of IL-8 is measured by ELIS A technique. Lyophilised standard is reconstituted, aliquoted and stored at -80 °C. Each ELISA plate contained a standard curve. Samples are diluted with the appropriate assay diluent at the appropriate dilution rate, washing steps were performed with an automatic microplate washer and absorbance was measured at 450 nm.

The IL-8 values are typically comprised between 20 to 50 ng/ml for healthy donors in unstimulated conditions.

The result is shown in Figure 10. It is observed that all the cells exposed to the formulations as described in Table 3a remained in a stable and healthy conditions for up to 72 hours. In particular, the cells exposed to NP-017 remained stable and healthy from T24h to T96h, compared to the cells exposed to the Base formulation as described in Table 3a.

Example 10. Viscosity

The viscosity of the formulations as described in Table 3a were measured. Each of the formulations were tested three times. The result is shown in Table 11.

Table 11. Viscosity of the formulations of the present invention

Claims

1. A liquid composition comprising an antibody, a bispecific antibody or an antigenbinding fragment thereof, and at least one mucoadhesive compound, wherein the concentration of the antibody, bispecific antibody or antigen-binding fragment thereof ranges from 0.001 to 75 % (w/v) and the concentration of the mucoadhesive compound ranges from 0.1 to 10 % (w/v).

2. A composition according to Claim 1, wherein the concentration of said mucoadhesive compound ranges from 0.5 to 5 % (w/v).

3. A composition according to any one of Claims 1 or 2, wherein the average molecular weight of said mucoadhesive compound is at least 100 kDa.

4. A composition according to any one of Claims 1 to 3, wherein said mucoadhesive compound comprises at least one selected from the group consisting of cellulose derivatives, amylose, gellan gum, guar gum, karaya gum, xanthan gum, carrageenan, alginate, pectin, dextran, chitosan, agarose, hyaluronic acid, gelatin, pectin, tragacanth, poly (vinyl pyrrolidone) such as Kollidon VA 64, poly(dimethyl siloxane), poly acrylic acid-based polymers such as poly acrylates, plyethylene glycol, sodium alginate, polycarbophil such as Noveon AA-1, polylysene, dimethylaminoethyl dextran, poly vinyl alcohol, hydroxy ethyl starch, Carbopol 97 IP NF, Carbopol 974P NF, Carbopol 71G NF, and/or hydroxyl ethyl cellulose (HEC).

5. A composition according to Claims 1 to 4, wherein the mucoadhesive compound comprises at least one selected from the group consisting of Sodium carboxyl methyl cellulose (SCMC), methylcellulose, carboxylmethylcellulose (CMC), hydroxylpropyl cellulose, hydroxylpropyl methylcellulose (HPMC) and ethyl cellulose.

6. A composition according to any one of Claims 1 to 5, wherein the mucoadhesive compound comprises hydroxylpropyl methylcellulose (HPMC).

7. A composition according to any one of Claims 1 to 6, wherein the mucoadhesive compound comprises HPMC and wherein the HPMC comprises at least one of HPMC substitution type 2910, HPMC substitution type 1828, HPMC substitution type 2208 and HPMC substitution type 2906.

8. A composition according to any one of Claims 1 to 7, wherein the mucoadhesive compound comprises HPMC and wherein the HPMC comprises homogenously substituted HPMC cellulose, alternatively wherein the HPMC comprises heterogeneously substituted cellulose.

9. A composition according to any one of Claims 1 to 8, further comprising a preservative.

10. A composition according to Claim 9, wherein the preservative is selected from the group consisting of sodium acetate, benzalkonium chloride, potassium sorbate, calcium sorbate, methyl paraben, ethyl paraben, propyl paraben, phenylcarbinol, chlorolbutanol, chlorolcresol, Ethylenediaminetetraacetic acid (EDTA).

11. A composition according to any one of Claims 1 to 10, wherein the antibody, bispecific antibody or antigen-binding fragment thereof comprises CR9114.

12. A composition according to any one of Claims 1 to 11, wherein the composition is a controlled released composition.

13. A composition according to Claims 1 to 12, having a viscosity ranging from 2 to 120 (N s/m²) as determined by a sheer stress method.

14. A composition according to Claims 1 to 13, wherein the viscosity of the composition ranges from 25 to 50 (N s/m²), as determined by a sheer stress method.

15. An inhaler comprising the composition according to Claims 1 to 14.