Neutralising antibodies against influenza viruses

Abstract

A composition comprising a monoclonal IgG antibody for use in the prophylaxis or treatment of an influenza virus infection in a mammal, wherein the composition is administered intranasally to said mammal according to a daily multidose regimen.

Description

[0001] NEUTRALISING ANTIBODIES AGAINST INFLUENZA VIRUSES

[0002] FIELD OF THE INVENTION

[0003] The present disclosure relates generally to methods for treatment or prophylaxis of infection caused by influenza virus, by administration of neutralising antibodies to the respiratory tract, including by intranasal or inhalation administration. In particular, the disclosure relates to dosing protocols for administering said neutralising antibodies.

[0004] BACKGROUND

[0005] Respiratory viruses are a group of viruses that infect the upper and / or lower respiratory tract in humans. These viruses, which are responsible for millions of illnesses and thousands of hospitalisations globally each year, include influenza virus (A and B), respiratory syncytial virus (RSV) and human metapneumovirus (hMPV).

[0006] Influenza virus causes a highly infectious respiratory illness affecting the upper and lower respiratory tracts that places a burden on healthcare systems and results in loss of life. Influenza is a leading cause of death and illness globally. Morbidity and mortality varies due to the virulence of the influenza strain and the host's exposure history, age, and immune status. In addition to seasonal epidemics, pandemic influenza strains emerge with some regularity. Due to the lack of preexisting immunity against the major viral antigens, pandemic influenza can spread quickly, often with more severe disease than seasonal influenza.

[0007] There are three types of influenza viruses: influenza A, B and C. Human influenza A and B viruses cause seasonal epidemics of disease. Influenza type C infections cause a mild respiratory illness and are not thought to cause epidemics. Influenza A viruses are divided into subtypes based on two proteins on the surface of the virus: the hemagglutinin (H) and the neuraminidase (N). There are 17 different hemagglutinin subtypes and 10 different neuraminidase subtypes. Influenza A viruses can be further broken down into different strains. Current subtypes of influenza A viruses found in people are influenza A (HINI) and influenza A (H3N2) viruses. Influenza B viruses are not divided into subtypes, but can be further broken down into two different lineages. Influenza A (H1N1), A (H3N2), and influenza B viruses are included in each year's influenza vaccine.

[0008] Current treatments for influenza are inadequate and can be ineffective. Despite widespread vaccination, susceptibility to influenza remains. The factors contributing to susceptibility include: (1) incomplete vaccination coverage; (2) vaccine formulation poorly representing the strains in circulation; (3) reduced efficacy of vaccination in the elderly; (4) the emergence of pandemic strains not represented in seasonal vaccines; and (5) emergence of drug-resistance against the currently available anti-viral therapeutics (e.g. adamantanes and Tamiflu). There is an urgent need for new influenza vaccines and therapeutics.

[0009] CR9114 is a recombinant, fully human, monoclonal antibody of the immunoglobulin G1 isotype (IgGl) produced in Chinese Hamster Ovary (CHO) cells. CR9114 has effective neutralising activity against influenza A and B viruses by binding a conserved epitope in the hemagglutinin (HA) stem of influenza virus. CR9114 was first disclosed by Dreyfus et al Science. 2012 September 14; 337(6100): 1343-1348). Since the first disclosure, CR9114 has become the focus of development for prophylactic and therapeutic approaches for combatting influenza virus infection and disease.

[0010] WO20 13 / 007770 discloses the structure, including primary amino acid sequences of the CDRs, of the CR9114 binding molecule.

[0011] Since influenza is a respiratory infection and virus particles can be transmitted though inhalation, inhaled and intranasal delivery of CR9114 has been proposed as an attractive route for administering the antibody as a pharmaceutical composition.

[0012] WO2014 / 152841 discloses neutralising anti-influenza antibodies delivered intranasally to mice. This publication concludes that direct delivery of neutralising antibody directly to the respiratory tract, by inhalation of intranasal delivery, is superior, more efficacious and affective at lower doses than systemic administration (IV or IP) of the same antibody in the same amounts. Eight different influenza virus neutralising antibodies are disclosed, including CR9114 (and a chimeric variant CA9114). WO2016 / 124768 discloses a number of studies investigating the in vivo efficacy of intranasally-administered influenza A and B multi-domain antibodies against various influenza strains (e.g. B Florida, H1N1 and H3N2). In some of the studies described in this patent publication CR9114 was used as a reference (control) antibody. Mice were given a single dose of experimental or control antibodies, at a range of single unit doses, in order to assess the efficacy of the test antibodies and determine minimum effective single doses for each of the test antibodies in the murine subjects.

[0013] There remains a need to optimise the utilisation of CR9114 as a pharmaceutical product for the prevention and treatment of influenza A and B virus infection and disease, for example, in view of no effect of a single one-off dose in the clinical trial (EUCT number: 2023-505020-57-00, Protocol Code: F21-CLD-201. Webpage: https: / / euclinicaltrials.eu / search-for-clinical- trials / ?lang=en&EUCT=2023-505020-57-00 (Date of accessing the webpage: 18th' July 2025)).

[0014] SUMMARY

[0015] According to a first aspect, the disclosure provides a composition comprising a monoclonal IgG antibody for use in the prophylaxis or treatment of an influenza virus infection in a mammal, wherein the composition is administered intranasally to said mammal according to a daily multidose regimen.

[0016] In a further aspect the disclosure provides a method for treatment or prophylaxis of an influenza virus infection in a mammal comprising administering a monoclonal IgG antibody intranasally to said mammal according to a daily multidose regimen.

[0017] DETAILED DESCRIPTION

[0018] The present disclosure provides dosage regimens for intranasally-administered compositions comprising anti-respiratory virus monoclonal IgG antibodies that are effective at neutralising respiratory viruses. The dosage regimens described herein maintain IgG concentrations in the nasal mucosa at levels that result in sustained protection against challenge from respiratory viral pathogens. By comprehending the impact of differences between mouse and non-human primate (NHP) / human nasal anatomy and the passage and clearance of IgG antibodies through and from the nasal mucosa, the inventors have devised a multidose regimen, mixed-dose (multidose followed by daily single dose) and single dose regimens that are effective in sustaining antibody concentrations in the nasal mucosa at sufficient levels to provide prolonged protection against viral challenge.

[0019] A multidose regimen comprises or consists of (i) administering a first unit dose of the composition; and then (ii) administering a second unit dose of the composition within a 24- hour period of the first dose. The multidose regimen may also be described herein as a “daily multidose” regimen.

[0020] The multidose regimen may consist of 2 unit doses a day.

[0021] The multidose regimen may consist of 3 unit doses a day.

[0022] The multidose regimen may consist of 4 unit doses a day.

[0023] DESCRIPTION OF THE FIGURES

[0024] Figure 1 shows a comparison between single (i.e. Once Daily 10 mg (O) and multiple dose (i.e. Twice Daily 2 x 10 mg (•) administration of CR9114.

[0025] Figure 2 shows viral replication (viral load (vRNA) by RT-qPCR and infectious virus by TCID50) in nasal swab (NS) of Cynomolgus macaques after IN+IT or IN-only challenge with H1N1 influenza virus strain A / Wisconsin / 67 / 2022. Figures A-C represent viral load (in copies / mL) over time (relative to time of viral challenge) across the different challenge arms. Figures D-F represent infectious virus (in TCIDso / mL) over time (relative to time of viral challenge) across the different challenge arms. Individual animals are shown with medians and interquartile ranges per timepoint. Dotted lines indicate the limit of detection (50 copies / mL and 10 TCIDso / mL for RT-qPCR and TCID50, respectively).

[0026] Figure 3 shows peak viral responses and viral AUC in the NS per challenge group. Peak viral load (vRNA) (Figure 3 A) and vRNA AUC (Figure 3B) Peak viral titer (TCID50) (Figure 3C) and TCID50 AUC (Figure 3D) are shown with group medians indicated.

[0027] Figure 4 shows viral replication (viral load (vRNA) by RT-qPCR and infectious virus by TCID50) in BAL of Cynomolgus macaques after IN+IT or IN-only challenge with H1N1 influenza virus strain A / Wisconsin / 67 / 2022. Figures 4A-4C represent viral load (in copies / mL) over time (relative to time of viral challenge) across the different challenge arms. Figures 4D- 4F represent infectious units (in TCIDso / mL) over time (relative to time viral challenge) across the different challenge arms. Individual animals are shown with the median and interquartile range per timepoint indicated. Dotted lines indicate the limit of detection (50 copies / mL and 10 TCIDso / mL for RT-qPCR and TCID50, respectively).

[0028] Figure 5 shows peak viral responses and viral AUC in the BAL per challenge group. Peak viral load (vRNA) (Figure 5A) and viral AUC (Figure 5B), Peak viral titer (TCID50) (Figure 5C) and TCID50 AUC (Figure 5D) are shown with group medians indicated..

[0029] Figure 6 shows HA-seroreactivity levels in serum following IN+IT or IN-only challenge with A / Wisconsin / 67 / 2022. Figures A-C represent anti-HA serum titers over time (relative to time of viral challenge) across the different challenge arms. Two animals M16915 (Group 1, IN+IT (open circles)) and AP986D (Group 3 IN, 0.25mL / nare (open square)) showed increased seroreactivity at baseline and reactivity increases further from day 7 onwards.

[0030] Figure 7 shows levels of recovered mAh in NS of Cynomolgus macaques after IN multi-dosing administration during the study and measured by anti-human Fc IgG ELISA. Figures 7A and 7B show individual animals per time point with the median and interquartile ranges. Figure 7C shows the IgG AUC results. Figure 7D shows the peak concentration of IgG. In Figures 7A and 7B, the x-axis represents the time in days relative to the time of viral challenge, and the y-axis represents the concentration of mAb (in pg / mL). In Figure 7C, the y-axis represents the levels of human mAb in logic pg / mL x day. In Figure 7D, the y-axis represents the concentration of mAb (in pg / mL). The dotted line represents the LoD (= 0.005 pg / mL).

[0031] Figure 8 shows levels of recovered mAb in serum of Cynomolgus macaques after IN multidosing administration during the study and measured by anti-human Fc IgG ELISA. Figures 8A and 8B show individual animals per time point with the median and interquartile ranges. Figures 8C shows the IgG AUC results. Figure 8D shows the peak concentration of IgG. In Figures 8A and 8B, the x-axis represents the time in days relative to the time of viral challenge, and the y-axis represents the concentration of mAb (in pg / mL). In Figure 8C, the y-axis represents the levels of human mAb in logic pg / mL x day. In Figure 8D, the y-axis represents the concentration of mAb (in pg / mL). The dotted line represents the LoD (= 0.005 pg / mL).

[0032] Figure 9 shows viral load (measured by RT-PCR) in NS of Cynomolgus macaques (Macaca fascicularis) after challenge with influenza A virus H1N1 A / Wisconsin / 67 / 2022 and after administration of CR9114 or sham. Figures 9A-9B represent viral load (vRNA in copies / mL) over time (relative to time of viral challenge) of the treatment and control group. Individual animals are represented per time point with the median and interquartile ranges. Figure 9C displays the viral AUC for each treatment or control group arm, along with the group medians. Figure 9D displays the peak viral load for each treatment or control group, along with the group medians. The y-axis in all panels represents the vRNA levels in copies / mL (and logic copies / mL x day in Figure 9C). The x-axis in Figures 9A-9B represents the time in days related to viral challenge (t=0). The LoD is indicated with a dotted line (= 50 copies / mL). P-values are indicated in Figures 9C and 9D.

[0033] Figure 10 shows infectious virus (measured by TCID50) in NS of Cynomolgus macaques (Macaca fascicularis) after challenge with influenza A virus H1N1 A / Wisconsin / 67 / 2022 after administration of CR9114 or sham. Figures 10A-10B represent viral titers levels (in TCIDso / mL) over time (relative to viral challenge) of the treatment and control group. Individual animals are represented per time point with the median and interquartile ranges. Figure 10C displays the viral AUC per treatment or control groups arm along with the group medians. Figure 10D shows the peak viral titer per treatment or control group, along with the group medians. The y-axis in all panels represents the TCID50 levels in TCIDso / mL (and logic TCIDso / mL x day in Figure 10C). The x-axis in Figures 10A-10B represents the time in days related to viral challenge (t=0). The LoD is indicated with a dotted line (LOD= 39 TCIDso / mL). P-values are indicated in Figures 10C and 10D.

[0034] Figure 11 shows viral replication (viral load (vRNA) by RT-qPCR and infectious virus by TCID50) in NS on Day 1 of Cynomolgus macaques (Macaca fascicularis) after challenge with influenza A virus H1N1 A / Wisconsin / 67 / 2022 and administration of CR9114 or sham. Figure 11 A represents viral load (vRNA in copies / mL) on day 1 for the treatment and control groups, along with the group medians. Figures 11B represents infectious virus (in TCIDso / mL) on day 1 for the treatment and control groups, along with the group medians. P-values are indicated.

[0035] Figure 12 shows HA-seroreactivity levels in serum of Cynomolgus macaques (Macaca fascicularis) after IN challenge with influenza A virus H1N1 A / Wisconsin / 67 / 2022 and administration of CR9114 or sham. Anti-HA serum titers are displayed for both the treatment and control groups at baseline and on day 10 after viral challenge. P-values are indicated.

[0036] Figure 13 shows prior vs posterior probability distributions of inferred parameters. Figure 13 A represents the parameter values inferred from the first-in-human PK model fitting. Figure 13B represents the parameter values inferred from the NHP PK model fitting. Figure 13C represents the parameter values inferred from the viral load fitting.

[0037] Figure 14 shows that posterior predictive check of first-in-human PK data and viral load data against model simulations show good quantitative agreement. Figure 14A shows model simulations of human CR9114 PK data versus data at all dosing regimens trialled. Grey horizontal dotted line denotes the lower limit of quantification (LLOQ = 38 ng / mL). Figure 14B shows model simulations of viral load in humans vs data taken from the Phase 1 zanamivir trials by GSK. Grey horizontal dotted line denotes the lower limit of quantification (LLOQ = 0.75 loglO TCIDso / mL).

[0038] Figure 15 shows PKPD simulation of one-off 10 mg dose of CR9114 in humans six hours prior to challenge with influenza A / H3N2 does not reduce viral load compared to untreated group. Horizontal blue dotted line denotes the challenge strain IC50. Horizontal grey dotted line denotes the lower limit of quantification of intranasal CR9114 (LLOQ = 38 ng / mL).

[0039] Figure 16 shows that posterior predictive check of NHP PK data against model simulations show good quantitative agreement. Grey horizontal dotted line denotes the lower limit of quantification (LLOQ = 32 ng / mL).

[0040] Figure 17 shows that PKPD simulation of twice-daily 7.2 mg dose of CR9114 in NHPs with three days prophylactic administration prior to challenge with influenza A / H1N1 protects against infection and reduces viral load compared to untreated group. Horizontal upper grey dotted line denotes the challenge strain IC50. Horizontal lower grey dotted line denotes the lower limit of quantification of intranasal CR9114 (LLOQ = 32 ng / mL).

[0041] Figure 18 shows PKPD simulations of continuous once-daily intranasal dosing of CR9114 in humans beginning the same day as challenge with influenza A / Belgium / 4217 / 2015 H3N2. Figures 18 A, 18B and 18C show the associated efficacy at 40 mg, 60 mg, and 100 mg doses respectively. Horizontal upper grey dotted line denotes the IC50 of the influenza A / H3N2 challenge strain (7480 ng / mL). Horizontal lower grey dotted line denotes the lower limit of quantification of intranasal CR9114 (LLOQ = 38 ng / mL).

[0042] Figure 19 shows PKPD simulations of continuous once-daily intranasal dosing of CR9114 in humans beginning the same day as challenge with influenza A / Wisconsin / 67 / 2022 H1N1. Figures 19A, 19B and 19C show the associated efficacy at 5 mg, 10 mg, and 20 mg doses respectively. Horizontal upper grey dotted line denotes the IC50 of the influenza A / H1N1 challenge strain (1140 ng / mL). Horizontal lower grey dotted line denotes the lower limit of quantification of intranasal CR9114 (LLOQ = 38 ng / mL).

[0043] ANTIBODIES The disclosure can be applied to a range of neutralising monoclonal antibodies (mAbs) of the immunoglobulin G (IgG) isotype, capable of binding to a region of influenza virus in order to neutralise the virus. The virus may be influenza virus A and / or B. Preferably, the mAbs of the disclosure are fully human IgG mAbs.

[0044] PHARMACEUTICAL COMPOSITIONS

[0045] Pharmaceutical compositions of the disclosures (also referred to herein as “composition” or “compositions”) comprise the monoclonal IgG antibody formulated for administration to the nasal cavity of a subject. In an embodiment, the composition is formulated as a liquid nasal spray.

[0046] In an embodiment, the monoclonal IgG antibody is formulated with a buffer, which is used to dilute the antibody to the desired concentration. Suitable buffer solutions may contain, e.g., sodium acetate, sodium chloride, sucrose, polysorbate-20. An exemplary buffer solution is: 20mM sodium acetate; 75mM sodium chloride; 5% w / v sucrose; 0.02% w / v polysorbate-20; pH 5.5.

[0047] The disclosure is not limited to spray delivery of liquid formulations and composition of the disclosure may also be administered in other forms e. g. as a powder.

[0048] Pharmaceutical compositions of the disclosure may comprise an adjuvant or may be non- adjuvanted. In an embodiment, the composition of the disclosure does not comprise any adjuvant.

[0049] Compositions of the disclosure function prophylactically, i.e. as a vaccine, to prevent influenza viral infection prior to or during exposure to the virus.

[0050] Compositions of the disclosure may function as therapeutic agents, to reduce viral load and / or treat or reduce symptoms post-infection.

[0051] ADMINISTRATION

[0052] The pharmaceutical composition of the disclosure is administered intranasally. As used herein, “intranasal administration” means local administration to the nasopharyngeal area, via the nostrils, and preferably without being inhaled into the lungs. It is desirable to use an intranasal delivery device which delivers the composition to the nasopharyngeal area, without or substantially without it entering the lungs.

[0053] Administration to the nostrils is preferably by means of a nasal spray sprayed directly into each nostril, preferably with a pre-determined metered volume being with each spray. Administration to the nostrils can be simultaneous administration, achieved by using a device configured to administer the composition directly into each nostril at the same time, or sequential administration, achieved by achieved by using a device configured to administer the composition into one nostril, and substantially immediately thereafter using the device to administer the composition to the other nostril. In a preferred embodiment administration to each nostril is sequential.

[0054] In the context of intranasal administration, the term “unit dose” refers to total monoclonal IgG antibody dose comprised within pharmaceutical composition or formulation administered as disclosed herein which may be administered to single or both the right and left nostrils of a subject receiving the dose.

[0055] In the embodiment in the context of intranasal administration, the term “unit dose” refers to total dose of pharmaceutical composition or formulation administered to both the right and left nostrils of a subject receiving the dose. For example, if Img / lOOpL of a pharmaceutical composition is administered to each nostril then the total unit dose administered is 2mg / 200pL.

[0056] The terms “subject” and “individual” are used interchangeably herein and refer to a mammal, preferably a non-human primate (NHP) or a human. In a preferred embodiment, the subject is a human.

[0057] Human nasal anatomy is a complex system compared to non-human primates and mice. Non- human primates have a nasal structure that is somewhat similar to humans but less complex. Mice possess a relatively simple nasal anatomy with fewer and less intricate turbinates, which limits their ability to humidify and filter air as effectively as humans and non-human primates. Mucus clearance in humans is facilitated by the coordinated action of cilia and the production of mucus in the sinuses, creating an efficient system for trapping and expelling pathogens and particles. Non-human primates exhibit a similar mucus clearance mechanism to humans, but the efficiency can vary depending on species-specific anatomical differences and environmental adaptations. Mice rely heavily on ciliary action for mucus clearance, but their simpler nasal passage structure and smaller size result in a less robust system compared to the more developed mechanisms found in humans and non-human primates. We have determined that the ability to dose the nose with a liquid medicament is limited by the quantity of liquid that is retained in the nose without being swallowed or otherwise lost from the nose.

[0058] DOSAGE REGIMENS

[0059] Multi dose

[0060] An embodiment of the present disclosure is directed to multidose regimens. As used herein, the term “multidose regimen” refers to a regimen which comprises or consists of a period of administration of a pharmaceutical composition in which more than one unit dose of a pharmaceutical composition is administered to an individual in a 24-hour period. In the context of the present disclosure, a multidose regimen may include a period (or several days) during which more than one unit dose of a pharmaceutical composition is administered to an individual in a 24-hour period, followed by a further period (of days, weeks or months) during which one unit dose of a pharmaceutical composition or formulation is administered to an individual in a 24 hour period. The single daily unit dose in this further period may be the same or higher than the unit dose delivered twice a day.

[0061] In contrast, as used herein the term “single dose regimen” refers to a regimen which consists of a period of administration of a pharmaceutical composition in which one unit dose of a pharmaceutical composition or formulation is administered to an individual in a 24-hour period. In a single dose regimen there is no 24 hour period in which more than one unit dose of the pharmaceutical composition is administered.

[0062] In an embodiment of the present disclosure, the multidose regimen comprises or consists of (i) administering a first unit dose of the composition of the disclosure comprising neutralising mAb; and (ii) administering a second unit dose of said composition within a 24-hour period. Preferably, (i) and (ii) are administered 2-18 hours apart, optionally 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17 or 18 hours apart, and preferably 12 hours apart.

[0063] In another embodiment of the present disclosure, the multidose regimen comprises or consists of administering more than two, i.e. three, four, five or six unit doses of the composition of the disclosure comprising neutralising mAb to a subject in a 24hour period. Preferably, the three, four, five or six doses are administered multiple hours apart. In a preferred embodiment, three unit doses are administered 4-8 hours apart, preferably 6 hours apart.

[0064] Each dose is administered to both the right and left nostrils of the subject, and these two administered doses combine to produce a single unit dose, as described above.

[0065] In an embodiment of the present disclosure, the first and second (or all subsequent) unit doses are identical.

[0066] In an embodiment, a unit dose disclosed herein comprises 0.1 -100 mg of a monoclonal IgG antibody as disclosed herein. The dose may comprise 0.5 -100 mg, 0.5 - 80 mg, 0.5 - 60mg, 0.5 -50mg, 0.5 - 40 mg, 0.5 - 30 mg, 0.5 - 20mg, l-20mg, 2-20 mg, 3-20 mg, 4-20 mg, 5-20 mg, such as 5-15 mg of the antibody. The unit dose may comprise 0,1, 0.5, 1, 2, 3, 4, 5, 5.5 , 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, or 40 mg of the antibody.

[0067] The unit dose may be administered to each of the left and right nostrils, and therefore split between them.

[0068] The monoclonal IgG antibody disclosed herein may be provided in a unit dose volume of 10- 400pL, such as 20-300pL such as 50-200pL, such as 100, 110, 150 or 200 pL.

[0069] The total unit dose of monoclonal antibody may be 1-100 mg / 200 pl. In an embodiment, the total unit dose is 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 mg / lOOpL. In an embodiment the total unit dose is higher than 10 mg / 200 pL, e.g. is between 10-20 mg / 200 pL, e.g. 11, 12, 13, 14, 15, 16, 17,18,19 or 20 mg / 200pL.

[0070] In one embodiment the unit dose comprises less than lOOmg of the monoclonal IgG antibody disclosed herein.

[0071] In one embodiment, 5 mg of antibody / 100 pL is administered to each of the left and right nostrils, resulting in a total unit dose of 10 mg / 200 pL respectively.

[0072] In one embodiment, about 2-5 mg / 100 pL is administered to each of the left and right nostrils, resulting in a total unit dose of about 4-10 mg / 200 pL. In an embodiment, the total unit dose is 4, 5, 6, 7, 8, 9, 10 mg / pL. In a preferred embodiment, 5 mg / 100 pL administered each of the left and right nostrils, resulting in a total unit dose of 10 mg / 200 pL. In an embodiment the total unit dose is higher than 10 mg / 200 pL, e.g. 10-20 mg / 200 pL, e.g. 11, 12, 13, 14, 15, 16, 17,18,19 or 20 mg / 200pL. The total unit dose may be administered two, three, four, five or six times in a 24-hour period.

[0073] In embodiments of the disclosure where the multidose regimen comprises three or more administrations in a 24-hour period the total unit dose per administration may be lower (e.g. 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1, 1.5 mg / 200 pL, e.g. 0.1 - 2 such as 0.1 - 1 mg / 200 pL). In an embodiment, about 0.05mg / 100 pL is administered to each nostril, resulting in a total unit dose of about 0.1mg / 200 pL.

[0074] In an embodiment of the disclosure, the composition is administered according to the multidose regimen for a consecutive period of 1 to 7 days, optionally 1 to 14 days, optionally 1 to 21 days, optionally 1 to 24 weeks, optionally for 6 weeks, optionally for 8 weeks, optionally for 12 weeks, such as for 6 to 10 weeks.

[0075] The multi dose regimen may start in response to infection / symptoms of influenza infection or in advance of infection symptoms of influenza infection.

[0076] In another embodiment of the disclosure, there is provided a composition comprising a monoclonal IgG antibody for use in the prophylaxis or treatment of an influenza virus infection in a mammal, wherein the composition is administered intranasally to said mammal according to a daily multidose regimen, and wherein the use achieves or is able to achieve: at least a 10,000 fold decrease in nasal viral influenza RNA levels at days 4-7 post infection when compared with a control without the antibody; and / or a 10 fold reduction in infectious viral load in the nose within a day of infection when compared with a control without the antibody, and / or no infectious virus load detected (for example as assessed by TCID50 / ml) on day 2-6 after infection

[0077] Preferred embodiments of this specific aspect, listed as statements Group I , include:

[0078] Statements Group I:

[0079] 1 A composition comprising a monoclonal IgG antibody for use in the prophylaxis or treatment of an influenza virus infection in a mammal, wherein the composition is administered intranasally to said mammal according to a daily multidose regimen. 2 A composition for use according to statement 1, wherein the daily multidose regimen consists of administering two unit doses of the composition within a 24 hour period; or administering three unit doses of the composition within a 24 hour period; or administering four unit doses of the composition within a 24 hour period;

[0080] 3 A composition for use according to statement 2, wherein each of the first and second unit doses comprise or consist of 2-5mg / 100 pL, preferably 5mg / 100 pL, of the composition administered each of the left and right nostrils, resulting in a total unit dose of 4-10 mg / 200 pL, preferably 10 mg / 200 pL.

[0081] 4 A composition for use according to statements 2 or 3, wherein (i) and (ii) are administered 2-18 hours apart, optionally 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17 or 18 hours apart, optionally 12 hours apart.

[0082] 5 A composition for use according to any preceding statement, wherein the composition is administered according to the multidose regimen every day for a period of for 1 to 7 days, optionally for 2, 3, 4, 5, 6 or 7 days, optionally for 1 to 24 weeks, optionally for 6 weeks, optionally for 12 weeks, optionally for between 6 to 10 weeks.

[0083] 6 A composition for use according to statement 4, wherein the multidose regimen comprises administering first and second unit doses of the composition according to the regimen of claim 4 for 1 to 7 days, optionally for 2, 3, 4, 5, 6 or 7 days, followed by a single dose administration regimen, wherein a unit dose is administered once every 24 hours. 7 A composition for use according to statement 6, wherein the single dose regimen comprises administering 2-5 mg / 100 pL, preferably 5 mg / 100 pL, of the composition to each nostril, resulting in a total unit dose of 4-10 mg / 200 pL, preferably 10 mg / 200 pL.

[0084] 8 A composition for use according to any preceding statement, wherein the monoclonal IgG antibody is CR9114.

[0085] 9 A composition for use according to any preceding statement, wherein the composition is administered as a nasal spray.

[0086] 10 A composition for use according to any preceding statement, wherein the respiratory virus is influenza A or B.

[0087] 11 A composition for use according to any preceding statement, wherein the composition is administered prior to infection and / or during period of exposure to the virus and / or during a period when there is an increased risk of exposure to the virus.

[0088] 12 A composition for use according to any preceding statement, wherein the mammal is a human.

[0089] 13 A composition formulated for intranasal or inhalation administration and effective for treatment or prophylaxis of influenza viral infection in a mammal comprising monoclonal antibody CR9114 in a single unit dose of at least 10 mg / 200 pL.

[0090] 14 A composition comprising monoclonal antibody CR9114 for use in the prophylaxis or treatment of influenza virus infection in a mammal, wherein the composition is administered intranasally to said mammal as a single unit dose sufficient to achieve a threshold concentration of antibody in the nasal mucosa of at least 20 mg / L.

[0091] 15 A composition according to statement 13 or 14, wherein the mammal is a human.

[0092] 16 A method for treatment or prophylaxis of an influenza virus infection in a mammal comprising administering a monoclonal IgG antibody intranasally to said mammal according to a daily multidose regimen.

[0093] 17 A method according to statement 16, wherein the daily multidose regimen consists of administering two unit doses of the composition within a 24 hour period; or administering three unit doses of the composition within a 24 hour period; or administering four unit doses of the composition within a 24 hour period;

[0094] 18 A method according to statement 17, wherein each of the first and second unit doses comprise or consist of 2-5 mg / 100 pL, preferably 5 mg / 100 pL, of the composition administered each of the left and right nostrils, resulting in a total unit dose of 4-10 mg / 200 pL, preferably 10 mg / 200 pL.

[0095] 19 A method according to statement 17 or 18, wherein (i) and (ii) are administered 2-18 hours apart, optionally 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17 or 18 hours apart, optionally 12 hours apart.

[0096] 20 A method according to any of statements 17-19, wherein the composition is administered according to the multidose regimen for a period of 1 to 24 weeks, optionally for 6 weeks, optionally for 12 weeks, optionally for between 6 to 10 weeks. 21 A method according to statement 19, wherein the multidose regimen comprises administering first and second unit doses of the composition according to the regimen of statement 19 for 1 to 7 days, optionally for 2, 3, 4, 5, 6 or 7 days, followed by a single dose administration regimen, wherein a unit dose is administered once every 24 hours.

[0097] 22 A method according to statement 21, wherein the single dose regimen comprises administering 2-5 mg / 100 pL, preferably 5 mg / 100 pL, of the composition to each nostril, resulting in a total unit dose of 4-10 mg / 200 pL, preferably 10 mg / 200 pL.

[0098] 23 A method according to any of statements 16-22, wherein the monoclonal IgG antibody is CR9114.

[0099] 24 A method according to any of statements 16-23, wherein the composition is administered in as a nasal spray.

[0100] 25 A method according to any of statements 16-24, wherein the respiratory virus is influenza A or B.

[0101] 26 A method according to any of statements 16-25, wherein the composition is administered prior to infection and / or during period of exposure to the virus and / or during a period when there is an increased risk of exposure to the virus.

[0102] 27 A method according to any of statements 16-26, wherein the mammal is a human.

[0103] 28 A nasal delivery device for delivering a composition to the human nasal cavity, said device comprising a composition comprising monoclonal antibody CR9114, wherein the device is metered such that a single unit dose is administered as two 5 mg / 100 pL doses applied sequentially to each nostril as a nasal spray. Single dose

[0104] The present disclosure also contemplates a single dose regimen.

[0105] According to this aspect, a dose of the antibody composition of the disclosure is administered intranasally once per 24-hour period. At least 2 doses are delivered, once on each of 2 consecutive days. This regimen may also be described as continuous once daily dosing or continuous single daily dosing herein.

[0106] In an embodiment, a unit dose for delivery once per 24 hour period comprises 0.1-100 mg of a monoclonal IgG antibody as disclosed herein. The dose may comprise 0.5 -lOOmg, 0.5 - 80 mg, 0.5 - 60mg, 0.5 -50mg, 0.5 - 40 mg, 0.5 - 30 mg, 0.5 - 20mg, l-20mg, 2-20 mg, 3-20 mg, 4-20 mg, 5-20 mg, such as 5-15 mg of the antibody. The unit dose may be or comprise 0.1, 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19 or 20 mg of the antibody as disclosed herein. The unit dose may be or comprise 25mg, 30 mg, 35 mg, 40mg, 45mg or 50mg, 60mg, 70mg, 80mg, 90mg or lOOmg of antibody as disclosed herein.

[0107] The total dose may be provided in a volume of 20- 400uL, such as such as 50-300uL, such as 100 - 200uL. Suitable volumes for a single daily dose may be 100, 150 or 200 pL.

[0108] The unit dose may be administered to each of the left and right nostrils, and therefore split between them.

[0109] The monoclonal IgG antibody disclosed herein may be provided in a unit dose volume of 10- 400pL, such as 20-300 pL such as 50-200pL, such as 100, 150 or 200 pL.

[0110] The total unit dose may be 1-100 mg / 200 pL. In an embodiment, the total unit dose is 0.5, 1, 2, 3, 4, 5, 6, 7, 8, 9 orlO mg / lOOpL. In an embodiment the total unit dose is higher than 10 mg / 200 pL, e.g. is between 10-20 mg / 200 pL, e.g. 11, 12, 13, 14, 15, 16, 17,18,19 or 20 mg / 200pL.

[0111] In one embodiment the unit dose comprises less than lOOmg of the monoclonal IgG antibody disclosed herein. In a preferred embodiment, 5 mg / 100 pL is administered to each of the left and right nostrils, resulting in a total unit dose of 10 mg / 200 pL respectively

[0112] According to this aspect, in one embodiment, a total unit dose of up to 10mg / 200 pL of the antibody composition of the disclosure is administered as a 5mg / 100pL dose into each nostril once per 24-hour period.

[0113] The total daily unit dose for a single dose regimen may be 5, 6, 7, 8, 9 or 10 mg / 200 pL.

[0114] In an embodiment the total unit dose is higher than 10 mg / 200 pL, e.g. 10-20 mg / 200 pL, e.g. 11, 12, 13, 14, 15, 16 17,18,19 or 20 mg / 200 pL.

[0115] In an embodiment of the disclosure, the composition is administered according to the single dose regimen for more than one day, such as for a consecutive period of 1 to 3 days, 1 to 4 days, 1-5 days, 1 to 6 days, a consecutive period of 1 to 7 days, a consecutive period of 1 to 8 days, a consecutive period of 1 to 9 days, optionally 1 to 14 days, optionally 1 to 21 days, optionally 1 to 24 weeks, optionally for 6 weeks, optionally for 8 weeks, optionally for 12 weeks, such as for 6 to 10 weeks.

[0116] A dose for a single dose regimen may be higher than a dose used in multiple daily dosing. For example, a single daily dose regimen may be at least lOmg per dose of monoclonal antibody.

[0117] It will be appreciated that dosing frequency and unit dose are interrelated variables, and multiple daily dosing may use lower dosages of antibody per dose when compared with consecutive daily dosing unit doses, to deliver the same overall daily antibody dose.

[0118] The single dose regimen may start in response to infection / symptoms of influenza infection or in advance of infection symptoms of influenza infection.

[0119] Preferred embodiments of this specific aspect, listed as statements Group II, include:

[0120] Statements

[0121] 1 A composition comprising a monoclonal IgG antibody for use in the prophylaxis or treatment of an influenza virus infection in a mammal, wherein the composition is administered intranasally to said mammal according to once a day regimen for at least 2 consecutive days.

[0122] 2 A composition for use according to statement 1, wherein the unit dose comprises or consist of 0.5-100 mg of monoclonal IgG antibody, and the composition is administered to each of the left and right nostrils. . 3 A composition for use according to statement 1 or 2 wherein the composition is administered according to the once a day regimen every day for a period of for 2 days, for 3 days, for 4 days, for 5 days, for 6 days, for 7 days, optionally for 1 to 24 weeks, such as 1-4 weeks, 1-6 weeks, 1-8 weeks, 1-10 weeks, 1-12 weeks, 1-14 weeks, such as optionally for 6 weeks, optionally for 12 weeks, optionally for between 6 to 10 weeks.

[0123] 4 A composition for use according to any preceding statement , wherein the monoclonal IgG antibody is CR9114.

[0124] 5 A composition for use according to any preceding statement, wherein the composition is administered as a nasal spray.

[0125] 6 A composition for use according to any preceding statement , wherein the influenza virus is influenza A or B.

[0126] 7 A composition for use according to any preceding statement, wherein the composition is administered prior to infection and / or during period of exposure to the virus and / or during a period when there is an increased risk of exposure to the virus.

[0127] 8 A composition for use according to any preceding statement , wherein the mammal is a human.

[0128] 9 A composition formulated for intranasal or inhalation administration and effective for treatment or prophylaxis of influenza viral infection in a mammal comprising monoclonal antibody CR9114 in a single unit dose of 0.5-100mg such as 5mg or at least 5mg, lOmg or at least lOmg, 15 mg or at least 15mg or 20mg or at least 20 mg,

[0129] 10 A composition formulated for intranasal or inhalation administration and effective for treatment or prophylaxis of influenza viral infection in a mammal comprising monoclonal antibody CR9114 in a single unit dose of 5 mg / 200 pL or at least 5 mg / 200 pL, 10 mg / 200 pL or at least 10 mg / 200 pL, 15 mg / 200 pL or at least 15 mg / 200 pL or 20mg / 200 pL or at least 20mg / 200 pL.

[0130] 11 A composition comprising monoclonal antibody CR9114 for use in the prophylaxis or treatment of influenza virus infection in a mammal, wherein the composition is administered intranasally to said mammal as a single unit dose sufficient to achieve a threshold concentration of antibody in the nasal mucosa of at least 20 mg / L. 12 A composition according to statements according to anyone of 9-11, wherein the mammal is a human.

[0131] 13 A method for treatment or prophylaxis of an influenza virus infection in a mammal comprising administering a monoclonal IgG antibody intranasally to said mammal according to a once a day regimen for at least 2 consecutive days.

[0132] 14 A method according to statement 13, wherein dose comprises or consists of 0.5-100mg of monoclonal IgG antibody, administered to each of the left and right nostrils. .

[0133] 15 A method according to any of statements 13-14, wherein the composition is administered according to the once a day regimen for a period of for 2 days, for 3 days, for , for 5 days, for 6 days, for 7 days, optionally for 1 to 24 weeks, such as 1-4 weeks, 1-6 weeks, 1-8 weeks, 1-10 weeks, 1-12 weeks, 1-14 weeks, optionally for 6 weeks, optionally for 12 weeks, optionally for between 6 to 10 weeks.

[0134] 16 A method according to any of statements 13-15, wherein the monoclonal IgG antibody is CR9114.

[0135] 17 A method according to any of statements 13-16, wherein the composition is administered as a nasal spray.

[0136] 18 A method according to any of statements 13-17, wherein the influenza virus i s influenza A or B.

[0137] 19 A method according to any of statements 13-18, wherein the composition is administered prior to infection and / or during period of exposure to the virus and / or during a period when there is an increased risk of exposure to the virus.

[0138] 20 A method according to any of statements 13-19, wherein the mammal is a human.

[0139] 21 A nasal delivery device for delivering a composition to the human nasal cavity, said device comprising a composition comprising monoclonal antibody CR9114, wherein the device is metered such that a single unit dose is administered as two 5 mg / 100 pL doses applied sequentially to each nostril as a nasal spray.

[0140] Mixed dose regimen An embodiment of the disclosure provides a mixed dosage regimen, comprising administering first, second (and optionally further additional) unit doses of the composition according to a multidose regimen described above, such as for 1 to 7 days, optionally for 2, 3, 4, 5, 6 or 7 days, followed by a single dose administration regimen, wherein a first unit dose is administered to each nostril once every 24 hours. The single dose may be administered for a consecutive period of 1 to 7 days, optionally 1 to 14 days, optionally 1 to 21 days, optionally 1 to 24 weeks, optionally for 6 weeks, optionally for 8 weeks, optionally for 12 weeks, such as for 6 to 10 weeks.

[0141] For multidose, mixed dose and single dose regimens, an objective of the disclosure is to maintain a threshold antibody concentration within the nasal mucosa of a human subject of at least 20 mg / L for the period of exposure to the virus. This is achieved by the dosage regimens of the disclosure. In an embodiment, this is achieved by the dosage regimens of the disclosure when the composition comprises the anti-influenza virus IgG mAb CR9114. Suitably the dosing regimen keeps the concentration of CR9114 or other anti-influenza antibody in the nasal mucosa above the IC50 or ICso value for the target or each target of the antibody. Antibody titres of the nasal mucosa may be determined by, for example, sampling through nasal swabs. Samples may be analyzed by ELISA or similar methods and in-vitro neutralization assay (e.g. Pseudovirus neutralization).

[0142] Prophylaxis

[0143] For the purposes of prophylaxis the antibody (composition comprising the antibody) of any aspect disclosed herein may be delivered in advance of potential exposure to influenza. For example, if a person is travelling to a region with an influenza outbreak, antibody doses may be taken in advance of travel.

[0144] Embodiments of the invention include the composition of the invention for use in a treatment regimen of the invention delivered at least one day before travel to a region with influenza virus, or before potential exposure to an influenza virus, such as at least 2, at least 3, at least 4, at least 5, at least 6 or at least 7 days before travel to a region with influenza virus or before potential exposure to an influenza virus.

[0145] PHARMACEUTICAL KITS The disclosure further provides a pharmaceutical kit comprising an intranasal administration device as described herein containing a vaccine formulation according to the invention. In an embodiment the device is a spray device that delivers the composition the form of a liquid spray into each nostril. In an embodiment, the device is configured such that each spray delivers a pre-defined metered volume of the composition into the nostrils.

[0146] Preferred devices for intranasal administration of the compositions of the disclosure are spray devices. Suitable commercially-available nasal spray devices include Accuspray™ (Becton Dickinson) and nebulisers.

[0147] Preferred spray devices for intranasal use are devices for which the performance of the device is not dependent upon the pressure applied by the user. These devices are known as pressure threshold devices. Liquid is released from the nozzle only when a threshold pressure is applied. These devices make it easier to achieve a spray with a regular droplet size. Pressure threshold devices suitable for use with the present invention are known in the art and are described for example in WO 91 / 13281 and EP 311 863 B and EP 516 636. Such devices are commercially available from Pfeiffer GmbH and are also described in Bommer, R. Pharmaceutical Technology Europe, Sept 1999.

[0148] Preferred intranasal devices produce droplets (measured using water as the liquid) in the range 1 to 200pm, preferably 10 to 120pm. Below 10pm there is a risk of inhalation, therefore it is desirable to have no more than about 5% of droplets below 10pm. Droplets above 120pm do not spread as well as smaller droplets, so it is desirable to have no more than about 5% of droplets exceeding 120pm.

[0149] INFLUENZA STRAINS

[0150] The Monoclonal CR9114 has been shown to fully protect mice against challenges with lethal doses of influenza A H1N1, H3N2 , H3N1, H5N1, and H9N2 in line with its ability to neutralize the same subtypes in vitro (B). CR9114 grants in vivo protection in mice challenged with lethal doses of influenza virus B (Florida / 4 / 2006 and Malaysia / 2506 / 2004 and Phuket / 3037 / 2013) and H2 (A / Ann Arbor / 6 / 1960 and A / swine / MO / 4296424 / 06) , despite not showing any in vitro neutralizing activity against influenza B and H2. See Figure 1 of Beukenhorst et al The influenza hemagglutinin stem antibody CR9114: Evidence for a narrow evolutionary path towards universal protection - Frontiers in Biology, Mini Review PUBLISHED 02 November 2022 DOI 10.3389 / fviro.2022.1049134. CR9114 therefore is a broadly neutralizing human monoclonal antibody capable of neutralisation of a wide range of influenza strains.

[0151] SEQUENCES

[0152] Heavy chain CR9114 CDR1 region

[0153] SEQ ID NO.: 001

[0154] NYAIS

[0155] Heavy chain CR9114 CDR2 region

[0156] SEQ ID NO.: 002

[0157] GISPIFGSTAYAQKFQG

[0158] Heavy chain CR9114 CDR3 region

[0159] SEQ ID NO.: 003

[0160] HGNYYYYSGMDV

[0161] Light chain CR9114 CDR1 region

[0162] SEQ ID NO.: 004

[0163] SGSDSNIGRRSVN

[0164] Light chain CR9114 CDR2 region

[0165] SEQ ID NO.: 005

[0166] SNDQRPS Light chain CR9114 CDR3 region

[0167] SEQ ID NO.: 006

[0168] AAWDDSLKGAV

[0169] Heavy chain CR9114 variable domain

[0170] SEQ ID NO.: 007

[0171] QVQLVQSGAEVKKPGSSVKVSCKSSGGTSNNYAISWVRQAPGQGLDWMGGISPIFGS

[0172] TAYAQKFQGRVTISADIFSNTAYMELNSLTSEDTAVYFCARHGNYYYYSGMDVWGQ

[0173] GTTVTVSS

[0174] Light chain CR9114 variable domain

[0175] SEQ ID NO.: 008

[0176] QSALTQPPAVSGTPGQRVTISCSGSDSNIGRRSVNWYQQFPGTAPKLLIYSNDQRPSVV PDRFSGSKSGTSASLAISGLQSEDEAEYYCAAWDDSLKGAVFGGGTQLTVL

[0177] CR9114 Heavy Chain Sequence

[0178] SEQ ID NO.: 009

[0179] QVQLVQSGAEVKKPGSSVKVSCKSSGGTSNNYAISWVRQAPGQGLDWMGGISPIFGS

[0180] TAYAQKFQGRVTISADIFSNTAYMELNSLTSEDTAVYFCARHGNYYYYSGMDVWGQ

[0181] GTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGV

[0182] HTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKRVEPKSCDKTHT

[0183] CPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVE

[0184] VHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKA

[0185] KGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPP

[0186] VLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0187] CR9114 Light chain Sequence SEQ ID NO.: 010

[0188] QSALTQPPAVSGTPGQRVTISCSGSDSNIGRRSVNWYQQFPGTAPKLLIYSNDQRPSVV PDRFSGSKSGTSASLAISGLQSEDEAEYYCAAWDDSLKGAVFGGGTQLTVLGQPKAA PSVTLFPPSSEELQANKATLVCLISDFYPGAVTVAWKADSSPVKAGVETTTPSKQSNN KYAASSYLSLTPEQWKSHRSYSCQVTHEGSTVEKTVAPTECS

[0189] EXAMPLES

[0190] Example 1 : Phase 1 Clinical Study (Single Dose)

[0191] A Phase 1 Randomized, Double-Blind, Placebo-Controlled, Single-Center, First-in-Human, Single Ascending Dose Study to Evaluate the Safety, Tolerability, and Pharmacokinetics of Intranasally Administered CR9114 in Healthy Volunteers (Part 1) with double-blind dose expansion to evaluate multiple doses of CR9114 (Part 2) and extended duration of exposure (Part 3). Trial Parts 1 and 2 also contained open-label PK cohorts to assess the intranasal pharmacokinetics of CR9114.

[0192] The cohorts consisted of 5 (Cohort IS) or 4 (Cohorts 2S and 3S) healthy participants who received a single IN administration of CR9114 at 1 of 3 dose levels or placebo. The planned single doses of CR9114 for dose escalation in Part 1 are provided in the table below. The approval to proceed to the next dose level was provided by the protocol safety review team (PSRT; consisting of the principal investigator and the Sponsor’s medical monitor) to the site. A sentinel dosing approach was applied to the first cohort (Cohort IS). Dosing was performed in a staggered approach.

[0193] Part 1

[0194] Part 1 evaluated the safety, tolerability, and PK of single ascending doses (SAD) of CR9114 in up to 3 dose levels. The safety cohorts were randomized, double-blinded and placebo- controlled. The PK cohorts were open-labeled.

[0195] Safety Cohorts

[0196] The cohorts consisted of 5 (Cohort IS) or 4 (Cohorts 2S and 3S) healthy participants who received a single IN administration of CR9114 at 1 of 3 dose levels or placebo. The planned single doses of CR9114 for dose escalation in Part 1 are provided in the table below. The approval to proceed to the next dose level was provided by the protocol safety review team (PSRT; consisting of the principal investigator and the Sponsor’s medical monitor) to the site. A sentinel dosing approach was applied to the first cohort (Cohort IS). Dosing was performed in a staggered approach.

[0197] PK Cohorts Each cohort consisted of 8 healthy participants who received a single IN administration of CR9114 at 1 of 3 dose levels. The planned single doses of CR9114 for dose escalation in Part 1 are provided below:

[0198] Table 1: Planned Single Doses of CR9114 for Part 1 aTwo sentinel participants were randomized at a 1 : 1 ratio (CR9114:placebo) and the remaining participants at a 2:1 ratio.

[0199] Part 2

[0200] Part 2 evaluated the safety, tolerability and PK of multiple doses of CR9114 for 5 days, up to 2 doses per day. The safety cohorts were randomized, double-blinded and placebo-controlled. The PK cohorts were open-labeled.

[0201] Safety Cohorts

[0202] Cohorts 4S (once daily [QD]) and 5S (twice daily [BID]) consisted of 4 healthy participants, each, who received repeated IN administration of CR9114 or placebo for 5 days at the MAD as determined in Cohorts IS, 2S and 3S. The approval to move to cohort (4S) was provided by the PSRT based on safety results from Cohorts IS, 2S and 3S. The approval to move to BID dosing (5S) was provided by the PSRT based on safety data from the QD (4S) dosing cohort.

[0203] A dose level of 10 mg / 200 pL (i.e., 5 mg / 100 pL per nostril) for 5 days was decided by the PSRT for Cohort 4S (QD) and 5S (BID).

[0204] PK Cohorts

[0205] Cohorts 4P (QD) and 5P (BID) consisted of 8 healthy participants, each, who received repeated IN administration of CR9114 for 5 days at the MAD as determined in Cohorts IS, 2S and 3S.

[0206] A dose level of 10 mg / 200 pL (ie., 5 mg / 100 pL per nostril) for 5 days was decided by the PSRT for Cohort 4P (QD) and 5P (BID).

[0207] Table 2: Planned Doses of CR9114 for Part 2 recommendations regarding the opening of subsequent cohorts at the preplanned or at a modified (lower) dose and dose regimen.

[0208] Part 3

[0209] Safety Cohorts

[0210] Cohort 6S evaluated the safety and tolerability of multiple doses of CR9114 QD or BID for 14 days. The dosing regimen was selected based on the analysis of the observed safety profile and IN PK of CR9114 in the SAD cohorts. This cohort was randomized, double-blinded and placebo controlled.

[0211] The approval to move to the 14-day dosing cohort (6S) was provided by the PSRT based on safety result from 4S and 5S Cohorts.

[0212] Cohort 6S consisted of 12 healthy participants who received repeated IN administration of CR9114 or placebo for 14 days at the MAD as determined in Cohorts IS, 2S and 3S.

[0213] A dose level of 10 mg / 200 pL (ie., 5 mg / 100 pL per nostril) for 14 days BID was decided by the PSRT for Cohort 6S.

[0214] Table 3: Planned Doses of CR9114 for Part 3 aThe PSRT reviewed the accumulated blinded data at prespecified times and made recommendations regarding the opening of subsequent cohorts at the preplanned or at a modified (lower) dose and dose regimen. Endpoints

[0215] The Primary Endpoint

[0216] The Primary endpoint of this clinical study was to evaluate the safety and tolerability of intranasal (IN) administration of CR9114 as single or repeat dose in healthy adults.

[0217] The Secondary Endpoint

[0218] The Secondary endpoints were to evaluate the pharmacokinetic (PK) profile of IN administration of CR9114 as single or repeat dose in healthy adults.

[0219] Study Design

[0220] Type: Randomized, double-blind, placebo-controlled; Center: Single-center; Participants: Healthy adults; Dosing: Single ascending doses of CR9114 via intranasal administration.

[0221] Inclusion Criteria

[0222] Healthy volunteers aged 18-55, Body Mass Index (BMI) between 19 and 30 kg / m2, No clinically significant medical history or laboratory abnormalities. A total of 73 participants were randomized and treated.

[0223] Test Product, Dose, Mode of Administration, Batch Number(s)

[0224] CR9114 is a recombinant, fully human monoclonal antibody (mAb) of the IgGl isotype produced in Chinese Hamster Ovary cells that binds to the hemagglutinin (HA) stem of influenza virus. The concentration of the drug product is 50 mg / mL and is to be diluted with the formulation buffer (20mM Sodium Acetate; 75mM Sodium Chloride; 5% w / v Sucrose; 0.02% w / v Polysorbate-20, pH 5.5).

[0225] Placebo, Dose, Mode of Administration

[0226] Formulation buffer (20mM Sodium Acetate; 75mM Sodium Chloride; 5% w / v Sucrose; 0.02%w / v Polysorbate-20, pH 5.5).

[0227] Trial Duration Trial duration for Parts 1 and 2 was up to 13 weeks per participant, screening up to 28 days before first IN administration of investigational product (IP) on Day 1 and 8 weeks of followup after last administration. Trial duration for Part 3 was up to 14 weeks per participant, screening up to 28 days before first IN administration of IP on Day 1 and 8 weeks of followup after last administration.

[0228] Trial Evaluations

[0229] Pharmacokinetics

[0230] Blood samples were collected for analysis of CR9114 in serum. Nasal mucosal lining fluid samples were collected for analysis of CR9114 in the nasal mucosal lining fluid. The following PK parameters were determined and analyzed for both serum and nasal mucosal lining fluid (PK cohorts only) per cohort by non-compartmental analysis (NCA) and NCA based on composite profiles (per cohort): Cmax, tmax, Ctrough, Cmin, AUCco, AUClast, z, tl / 2, Cmax, dose norm, AUCoo , dose norm, and AUCiast, dose norm- The parameters calculated for nasal mucosal lining fluid could be adjusted based on the detectable levels of CR9114 and additional PK parameters could be calculated as appropriate.

[0231] Immunogenicity

[0232] The immune responses against CR9114 were evaluated by assessing the presence of anti-drug antibodies (AD As) in serum and assessing the titer in ADA confirmed samples as indicated in the time and event schedule.

[0233] Exploratory Immunology and Immunogenicity

[0234] Nasal mucosal lining fluid and saliva samples were collected for exploratory immunology and immunogenicity analysis of CR9114, if appropriate, according to the time points in the time and event schedule. Exploratory immunogenicity endpoints could be evaluated by quantifying anti-CR9114 IgG in nasal mucosal lining fluid and saliva samples.

[0235] Exploratory immunology endpoints could be evaluated by assessing influenza virus-specific Ab function in nasal mucosal lining fluid through ADCC, ADCP, and ADCVI assays.

[0236] Safety The trial evaluated safety and tolerability through assessment of AEs, clinical laboratory tests, vital signs, and physical examinations (including ear, nose, throat, [ENT] examination and nasopharyngeal examination) at time points as outlined in the Time and Events Schedule.

[0237] Results Subject Disposition

[0238] A total of 73 participants were randomized in 3 trial parts, 37 participants in Part 1, 24 participants in Part 2, and 12 Participants in Part 3. All participants completed the trial as planned.

[0239] Table 4: Part 1 (Safety Analysis Set) N = number of participants; n=number of participants with that observation Table 5: Part 2 (Safety Analysis Set)

[0240] Table 6: Part 3 (Safety Analysis Set) Pharmacokinetic Results

[0241] After a single IN dose of CR9114 at doses of 0.2 to 10 mg / 200 pL, CR9114 was quantifiable in nasal mucosal lining fluid for all participants at the first sampling time point after dosing, with, on average, the highest CR9114 concentrations observed at 6 hours postdose. A higher dose resulted in higher CR9114 nasal mucosal lining fluid exposure, with a dose-proportional increase for mean urea-normalized nasal mucosal lining fluid Cmax and AUCs over the 0.2 to 10 mg / 200 pL dose range. In serum, few participants showed sporadic low (<15 ng / mL) concentrations of CR9114. The mean CR9114 serum concentrations were below the lower limit of quantification (BLQ) for all dose groups. After multiple IN doses of CR9114 10 mg / 200 pL for 5 days, CR9114 nasal mucosal lining fluid trough concentrations were rather stable between Day 2 and Day 6 after BID dosing; more variability was observed after QD dosing. The CR9114 nasal mucosal lining fluid concentrations were much higher after BID dosing compared with QD dosing. Mean CR9114 serum concentrations were BLQ after QD dosing (Cohort 4P). After BID dosing (Cohort 5P), accumulation was observed, with the CR9114 trough serum concentrations increasing up to Day 5. After multiple IN BID doses of CR9114 10 mg / 200 pL for 14 days, low levels of serum CR9114 were measured from Day 3 onwards, with the highest mean level on Day 15.

[0242] Table 7: Urea-normalized Nasal Mucosal Lining Fluid Ctrough Concentrations (ng / mL) of

[0243] CR9114 (Part 2) (Pharmacokinetic Analysis Set)

[0244] BLQ: Below limit of quantification (<32 ng / mL); NC: Not calculable Table 8: Urea-normalized Nasal Mucosal Lining Fluid Ctrough Concentrations (ng / mL) of

[0245] CR9114 (Part 2) (Pharmacokinetic Analysis Set)

[0246] BLQ: Below limit of quantification (<32 ng / mL); NC: Not calculable

[0247] The difference in median, minimum and maximum values appearing in Table 7 (CR9114 Once Daily 10 mg) and Table 8 (CR9114 Twice Daily 2 x 10 mg) are clearly evident in Figure 1.

[0248] Immunogenicity Results

[0249] No AEs with a clinically suspected relationship to ADA have been observed throughout the trial across all dose levels tested, including the highest cumulative dosed Cohort 6S. No treatment-induced ADA have been found. Pre-existent ADA were observed in one participant (S197- MAD5D-QD group) out of 64. A slight increase but within the Minimum Significant Ratio (MSR) of the ADA assay was observed post dose, which is therefore not considered to be a treatment boosted increase. The PK profile (both serum and nasal mucosal lining fluid) of this participant is not different from the PK profile of other participants exposed to the same dose level. Two AEs have been reported for this participant (sore throat and clavicular luxation due to injury), which both have been assessed as not related to the study drug and also have no reasonable clinical relationship with ADA.

[0250] Adverse Events Summary The results of the trial show that intranasal administration CR9114 up to doses of 10 mg / 200 pL in dosing regimens up to 14 days BID was generally safe and well tolerated in healthy participants.

[0251] Single dose IN dosing of 0.2, 2 and 10 mg / 200 pL showed CR9114 concentrations in the nose up to 2 to 15 days after dosing, depending on dose level. A high variability in concentrations was found between participants. CR9114 was rapidly cleared from the nasal mucosal lining fluid. A dose-proportional increase in CR9114 in the nasal mucosal lining fluid was found over the 0.2 to 10 mg / 200 pL CR9114 dose range.

[0252] After a single IN dose of CR9114 up to 10 mg / 200 pL, in general, CR9114 was not quantifiable in serum, indicating very limited systemic exposure. After BID dosing of 10 mg / 200 pL CR9114, serum levels of CR9114 could be detected after 2 days. Levels slightly increased after IN BID dosing of 10 mg / 200 pL CR9114 for 14 days though overall CR9114 serum levels remained very low. Steady-state of CR9114 serum concentrations were not yet reached after BID doses of CR9114 10 mg / 200 pL for 14 days.

[0253] Exclusion Criteria

[0254] Participants meeting any of the following criteria were excluded from participation in this trial. Evidence of any active or chronic disease or condition that could have interfered with, or for which the treatment of might have interfered with the conduct of the trial, or that would have posed an unacceptable risk to the participant in the opinion of the investigator (following a detailed medical history, physical examination, vital signs [systolic and diastolic blood pressure, pulse rate, body temperature] and 12-lead ECG).

[0255] Excluded active or chronic diseases or conditions (history or current) included Immunocompromised (known or expected immune deficiency, disease, or use of medication that may affect the immune system) or evidence of autoimmune disorder (deemed clinically relevant by the investigator).

[0256] Bleeding disorder that was diagnosed by a physician (e.g., factor deficiency, coagulopathy or platelet disorder that requires special precautions). Note: A participant who stated that he or she had easy bruising or bleeding but did not have a formal diagnosis and had intramuscular injections and blood draws without any adverse experience, was eligible. History of a splenectomy. Seizure disorder: a participant who had had a seizure in the last 3 years prior to screening was excluded. (Not excluded: a participant with a history of seizures who had neither required medications nor had a seizure for 3 years prior to screening.) Chronic respiratory or cardiac diseases, e.g., sarcoidosis, chronic obstructive pulmonary disease, uncontrolled hypertension or arrythmia. Active infections requiring parenteral antibiotic, antiviral, or antifungal therapy within 30 days prior to screening. Conduction disturbance (complete left or complete right bundle branch block or nonspecific intraventricular conduction disturbance with QRS >120 msec, PR interval >220 ms, any second or third degree AV block, or corrected QT interval (QTc) prolongation [>450 ms]). Significant repolarization (ST segment or T wave) abnormality. Significant atrial or ventricular arrhythmia; frequent atrial or ventricular ectopy (e.g., frequent premature atrial contractions, 2 premature ventricular contractions in a row). ST elevation consistent with ischemia; or evidence of past or evolving myocardial infarction.

[0257] Trial Intervention

[0258] Intervention Administered

[0259] Manufacturing, packaging, and labeling of CR9114 and placebo was done under the responsibility of the Sponsor.

[0260] CR9114 and placebo were prepared in accordance with Good Manufacturing Practice (GMP) as required by the ICH-Good Clinical Practice (GCP) guideline (ICH-E6[R2]). A copy of the certificate of analysis of CR9114 and placebo accompanied CR9114 and placebo to the clinical site. Investigational Products (IP) are described in Table 9. Any deviation from the intervention regimen defined in the protocol had to be documented in the eSource system.

[0261] The concentration of the drug product was 50 mg / mL and was to be diluted with the formulation buffer (20mM Sodium Acetate; 75mM Sodium Chloride; 5% w / v Sucrose; 0.02%w / v Polysorbate-20, pH 5.5). The formulation buffer was used as placebo. The IP preparation process was detailed in the Pharmacy Manual.

[0262] The exact date and time of administration of the IP had to be recorded in the eSource system. Table 9: Description of Investigational Products aBID dosing regimen consisting of a morning dose and an evening dose.

[0263] Pharmacokinetic Measurements

[0264] Sample collection and handling (Blood, Nasal Mucosal Lining Fluid, and Saliva Samples)

[0265] Throughout the trial, nasal mucosal lining fluid samples, venous blood samples, and saliva samples were collected for analysis of CR9114 in nasal mucosal lining fluid, serum, and optionally saliva according to the time points defined in the time and event schedule. No water, meal, gum, or tobacco intake was allowed 30 min prior to saliva sampling. The exact date and time of sampling and of administration of the IP had to be recorded in the eSource system.

[0266] Blood samples were collected by venipuncture or via indwelling cannula in the forearm into vacuum tubes and was immediately chilled (ice bath).

[0267] Details on processes for collection, processing, storage, shipment, and destruction of these samples could be found in the laboratory manual that was provided to the site.

[0268] Bioanalysis

[0269] Nasal mucosal lining fluid and serum samples for determination of the concentration of CR9114 were analyzed by Resolian UK under the responsibility of the Sponsor, using a validated analytical method.

[0270] Samples that remained after protocol-specific assessments had been performed could be used for further exploratory work on PK, metabolites, serum protein binding, protein analysis, immunogenicity, and biochemistry.

[0271] Immunogenicity Measurements

[0272] Throughout the trial, venous blood samples were collected for analysis of immunogenicity against CR9114, according to the time points defined in the time and event schedule. Exploratory Immunology and Immunogenicity

[0273] Throughout the trial, nasal mucosal lining fluid and saliva samples were collected for exploratory immunology and immunogenicity analysis of CR9114, if appropriate, according to the time points defined in the time and event schedule.

[0274] Safety Measurements and Evaluations

[0275] The safety assessment in this trial were based on AEs, clinical laboratory tests, vital signs, 12- lead ECG, and physical examination as described in the following sections. Adverse events were monitored continuously from informed consent until the last trial-related activity. At regular intervals during the trial, participants were asked non-leading questions to determine the occurrence of any AEs. All AEs reported spontaneously during the course of the trial were recorded as well.

[0276] Discussion and overall conclusions

[0277] This was a randomized, double-blind, placebo-controlled, single-center, first-in-human, single ascending dose safety and tolerability trial (Part 1) with double-blind dose expansion to evaluate multiple doses of CR9114 (Part 2) and extended duration of exposure (Part 3). Trial Parts 1 and 2 also contained open-label PK cohorts to assess the IN PK of CR9114.

[0278] Part 1 evaluated the safety, tolerability and PK of SAD of CR9114 in up to 3 dose levels, ie, 0.2 mg / 200 pL, 2 mg / 200 pL, and 10 mg / 200 pL. Part 2 evaluated the safety, tolerability and PK of multiple doses of CR9114 QD or BID for 5 days.

[0279] Cohort 6S evaluated the safety and tolerability of multiple doses of CR9114 BID for 14 days

[0280] Safety

[0281] In Part 1, 17 (45.9%) participants reported TEAEs. No fatal AEs, SAEs, or AEs leading to discontinuation were reported. No Grade 3 or 4 TEAEs were reported. The most frequently reported TEAEs were headache (in 9 [27.3%] participants in the all CR9114 group), nausea (in 2 [6.1%] participants in the all CR9114 group), and back pain (in 2 [6.1%] participants in the all CR9114 group). One (9.1%) participant in the CR9114 0.2 mg / 200 pL group was reported with a TEAE headache (Grade 1) 1 day after dosing, which was considered possibly related to the trial treatment by the investigator. This TEAE was considered resolved after a duration of 2 days. No concomitant medication was administered for this AE.

[0282] In Part 2, 12 (50.0%) participants reported TEAEs. No fatal AEs, SAEs, or AEs leading to discontinuation were reported. No Grade 3 or 4 TEAEs were reported. The most frequently reported TEAEs were headache (in 4 [18.2%] participants in the all CR9114group) and oropharyngeal pain (in 3 [13.6%] participants in the all CR9114 group). Four (16.7%) participants were reported with a TEAE that was considered related to the trial treatment by the investigator. Grade 1 epistaxis which was considered possibly related to the trial treatment was reported in the CR9114 5-day QD group. This TEAE was reported as resolved after a duration of less than 1 day. Grade 1 nasal congestion which was considered possibly related to the trial treatment was reported in the CR9114 5-day QD group. This TEAE was reported as resolved after a duration of less than 1 day. Grade 1 nasal discomfort which was considered possibly related to the trial treatment was reported in the CR9114 5-day BID group. This TEAE was reported as resolved after a duration of less than 1 day. Grade 1 nasal dryness which was considered possibly related to the trial treatment was reported in the CR9114 5-day BID group. This TEAE was reported as resolved after a duration of 6 days.

[0283] In Part 3, 6 (50.0%) participants reported TEAEs. No fatal AEs, SAEs, or AEs leading to discontinuation were reported. No Grade 3 or 4 TEAEs were reported. The most frequently reported TEAEs were headache (in 2 [16.7%] participants in the CR9114 group), diarrhoea (in 2 [16.7%] participants in the CR9114 group), and oropharyngeal pain (in 2 [16.7%] participants in the CR9114 group). None of the TEAEs were considered treatment-related.

[0284] None of the treatment-emergent laboratory abnormalities and vital signs abnormalities were considered clinically relevant and none were reported as TEAEs.

[0285] Pharmacokinetics

[0286] After a single IN dose of CR9114 at doses of 0.2 to 10 mg / 200 pL, CR9114 was quantifiable in nasal mucosal lining fluid for all participants at the first sampling time point after dosing, with, on average, the highest CR9114 concentrations observed at 6 hours postdose. A higher dose resulted in higher CR9114 nasal mucosal lining fluid exposure, with a dose proportional increase for mean urea-normalized nasal mucosal lining fluid Cmax and AUCs over the 0.2 to 10 mg / 200 pL dose range. In serum, few participants showed sporadic low (<15 ng / mL) concentrations of CR9114. The mean CR9114 serum concentrations were BLQ for all dose groups.

[0287] After multiple IN doses of CR9114 10 mg / 200 pL for 5 days, CR9114 nasal mucosal lining fluid concentrations were much higher after BID dosing compared with QD dosing. Mean CR9114 serum concentrations were BLQ after QD dosing (Cohort 4P).

[0288] After BID dosing (Cohort 5P), accumulation was observed, with the CR9114 trough serum concentrations increasing up to Day 5 though remaining very low and close to the detection limit.

[0289] After multiple IN BID doses of CR9114 10 mg / 200 pLfor 14 days, low levels of serum CR9114 were measured from Day 3 onwards, with the highest mean level on Day 15.

[0290] Overall Conclusions

[0291] The results of the trial show that intranasal administration CR9114 up to doses of 10 mg / 200 pL in dosing regimens up to 14 days BID was generally safe and well tolerated in healthy participants.

[0292] Single dose IN dosing of 0.2, 2 and 10 mg / 200 pL showed CR9114 concentrations in the nose up to 2 to 15 days after dosing, depending on dose level. A high variability in concentrations was found between participants.

[0293] CR9114 was rapidly cleared from the nasal mucosal lining fluid. A dose proportional increase in CR9114 in the nasal mucosal line fluid was found over the 0.2 to 10 mg / 200 pL CR9114 dose range.

[0294] After a single IN dose of CR9114 up to 10 mg / 200 pL, in general, CR9114 was not quantifiable in serum indicating very limited systemic exposure. After BID dosing of 10 mg / 200 pL CR9114, serum levels of CR9114 could be detected after 2 days. Levels slightly increased after IN BID dosing of 10 mg / 200 pL CR9114 for 14 days though overall CR9114 serum levels remained very low. Steady-state of CR9114 serum concentrations were not yet reached after BID doses of CR9114 10 mg / 200 pL for 14 days. Example 2: Antibody according to the invention (NHP - PK of CR9114 administered Intranasally (IN) in a multi-dosing regimen)

[0295] The aim of this study is to assess the bioavailability and half-life of a monoclonal antibody according to the invention, having a heavy chain CDR1 as SEQ ID NO: 001, heavy chain CDR2 as SEQ ID NO: 002, heavy chain CDR3 as SEQ ID NO: 003, a light chain CDR1 as SEQ ID NO: 004, a light chain CDR2 as SEQ ID NO: 005, and a light chain CDR3 as SEQ ID NO: 006, following intranasal administration in aNon-Human Primate (NHP) model and naive animals for treatment with human IgGs

[0296] Animals are treated with an intranasal dose range of the test antibody described above 0.01 to 5 mg (“Low Dose”) or more than 5, for example, 6 to 10 mg or 10 mg to 14 mg (“High Dose”) nominal dose at study day 0, in a dose regimen that is once daily, (QD), twice daily (BID) or three times daily (TID), for consecutive days up to 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, or 14 days

[0297] Materials and Methods

[0298] The monoclonal antibody according to the invention is provided in a sodium acetate buffer (e.g. 20 mM Sodium Acetate, 75 mM Sodium Chloride, 5% sucrose, 0.02% PS20, pH 5.5) and is diluted to the final concentration for administration ranging from 0,01 to 5 mg (“Low Dose”) or more than 5, for example, 6 to 10 mg or 10 to 14 mg (“High Dose”) nominal dose, administered as 50 to 100 pL or 100 to 500 pL per nare.

[0299] Samples are taken throughout the study duration, sample types include nasal sampling (for example nasal swabs, wash or scrapes), nasopharyngeal samples, serum and / or bronchoalveolar lavage (BAL) samples. Sampling period includes study duration, in a time frame between day -1 to the end of the study (7-21 days follow-up period). Also, study presamples are taken to measure baseline levels.

[0300] Samples are analyzed for antibody titers and / or neutralization titers by ELISA or similar methods and in-vitro neutralization assay (e.g. Pseudovirus neutralization).

[0301] Animals The NHP subjects are used at a weight of c. 2-22 kg (e.g. 5 kg per animal) on commencement of the study. Between 3-12 animals are allocated to each treatment group.

[0302] Study design

[0303] The dose of the monoclonal antibody applied in the current example is based on the maximal dosing.

[0304] Table (10): Intranasal experimental detail (administration at day 0)

[0305] The NHP subjects are treated via the intranasal route of administration with the antibody at a dose between 0.01 - 5 mg or 6 - 14 mg nominal dose. After antibody administration on day 0, samples are taken once daily throughout the study duration before mAb administration and animals are monitored for clinical observations (e.g. weight loss and temperature) until the end of the study (e.g. Day 10-21).

[0306] Antibody administration

[0307] The test antibody is administered to each nare (e.g. pipette or spray or other atomizer device) according to the treatment schedule (Table 10), between 50 to 500 pL per nare.

[0308] Data analysis and statistical methods Bioavailability of the IN administered test antibody is determined by ELISA in NS, Serum and BAL and is summarized by group and timepoint using standard statistical tools as appropriate to the statistical distribution of the measurements.

[0309] The half-life of the mAb concentration in NS and the time required for reaching steady state and mAb concentration at steady state is determined

[0310] Results - mAb bioavailability

[0311] Following the IN administration of the antibody according to the invention in a multi-dosing of the test antibody it can be detected in the nose. The steady state concentration in NS is reached between 1-3 days post-administration in the three treated groups. The concentration of the test antibody at steady state is expected to be higher for the group treated with a higher dose and the antibody fluctuations around steady state smaller in the BID treated group.

[0312] Example 3. Antibody according to the invention (NHP - PK of CR9114 administered aerosolized (nebulization) in a multi-dosing regimen)

[0313] The aim of this study is to assess the bioavailability and half-life of a monoclonal antibody according to the invention, having a heavy chain CDR1 as SEQ ID NO: 001, heavy chain CDR2 as SEQ ID NO: 002, heavy chain CDR3 as SEQ ID NO: 003, a light chain CDR1 as SEQ ID NO: 004, a light chain CDR2 as SEQ ID NO: 005, and a light chain CDR3 as SEQ ID NO: 006, following intranasal administration in aNon-Human Primate (NHP) model and naive animals for treatment with human IgGs.

[0314] Animals are treated with a nebulized dose range of the test antibody described above 1-20 mg, 20-40 mg, 40-60 mg, 60-80 mg, 80-100 mg, 100-125 mg, 125-150mg, 150-175mg, 175-200 mg, 200-225 mg nominal dose at study day 0, in a dose regimen that is once daily, (QD), twice daily (BID) or three times daily (TID), for consecutive days up to 14 days.

[0315] Materials and Methods The monoclonal antibody according to the invention is provided in a sodium acetate buffer (e.g. 20 mM Sodium Acetate, 75 mM Sodium Chloride, 5% sucrose, 0.02% PS20, pH 5.5) and is diluted to the final concentration for administration ranging from 1-20 mg, 20-40 mg, 40-60 mg, 60-80 mg, 80-100 mg, 100-125 mg, 125-150mg, 150-175mg, 175-200 mg, 200-225 mg nominal dose, administered with a nebulization device and mask or similar device.

[0316] Samples are taken throughout the study duration, sample types include nasal sampling (for example nasal swabs, wash or scrapes), nasopharyngeal samples, serum and / or bronchoalveolar lavage (BAL) samples. Sampling period includes study duration, in a time frame between day -1 to the end of the study (7-21 days follow-up period). Also, study presamples are taken to measure baseline levels.

[0317] Samples are analyzed for antibody titers and / or neutralization titers by ELISA or similar methods and in-vitro neutralization assay (e.g. Pseudovirus neutralization).

[0318] Animals

[0319] The NHP subjects are used at a weight of c. 2-22 kg (e.g. 5 kg per animal) on commencement of the study. Between 3-12 animals are allocated to each treatment group.

[0320] Study design

[0321] The dose level of the monoclonal antibody applied in the current example is based on the possible maximal dosing.

[0322] Table (11): Intranasal experimental detail (administration at day 0)

[0323] *0f note, the realized concentration in the lung is likely ~6% of the administered dose.

[0324] The NHP subjects are treated via the nebulized route of administration with the antibody at a dose between 1 - 225 mg nominal dose. After antibody administration on day 0, samples are taken once daily throughout the study duration before mAb administration and animals are monitored for clinical observations (e.g. weight loss and temperature) until the end of the study (e.g. Day 10-21).

[0325] Antibody administration

[0326] The test antibody is administered by nebulization (e.g. nebulizer device and mask or similar device) according to the treatment schedule (Table 11).

[0327] Data analysis and statistical methods

[0328] Bioavailability following nebulization of test antibody will be determined by ELISA measurements in NS, nasopharyngeal, BAL and Serum, and will be summarized by group and timepoint using standard statistical tools as appropriate to the statistical distribution of the measurements. Determine if steady state is reached following the different mAb administration regimens.

[0329] Results - mAb bioavailability

[0330] Following the nebulization of the test antibody in a multi-dosing regimen, deposition of the antibody is detectable in the BAL and from nasal swabs. Conclusion

[0331] In this NHP model, the PK of the test antibody according to the invention administered nebulized in a multidosing regimen as 1-225 mg nominal dose QD and / or BID is expected to show that antibody deposition is detectable in the BAL.

[0332] Example 4: Antibody according to the invention (NHP - Efficacy study of CR9114 administered IN in a multi-dosing regimen against a flu challenge)

[0333] The aim of this study is to assess the pre- and post-exposure efficacy of a monoclonal antibody according to the invention, having a heavy chain CDR1 as SEQ ID NO: 001, heavy chain CDR2 as SEQ ID NO: 002, heavy chain CDR3 as SEQ ID NO: 003, a light chain CDR1 as SEQ ID NO: 004, a light chain CDR2 as SEQ ID NO: 005, and a light chain CDR3 as SEQ ID NO: 006, following intranasal administration in aNon-Human Primates (NHP) model in which each subject is pre-screened for the absence of serum antibodies against the flu challenge virus.

[0334] Animals are treated with an intranasal dose range of the test antibody described above 0,01 to 5 mg (“Low Dose”) or more than 5mg, for example, 6-10mg or 10- less than 15 mg (“High Dose”) nominal dose or control at study in a dose regimen that is once daily, (QD), twice daily (BID) or three times daily (TID), for consecutive days up to 14 days. Animals are challenged with influenza virus between 1-3 days post first mAb administration which will be considered t=0 of the study.

[0335] Materials and Methods

[0336] The monoclonal antibody according to the invention is provided in a sodium acetate buffer (e.g. 20 mM Sodium Acetate, 75 mM Sodium Chloride, 5% sucrose, 0.02% PS20, pH 5.5) and is diluted to the final concentration for administration ranging from 0.01 to 5 mg or more than 5 mg, for example 6 to 10 mg, or 10 to 14 mg nominal dose, administered as 50-100 pL or 100 to 500 pL per nare.

[0337] The NHP subjects allocated to the Control Group (Sodium acetate buffer) are administered with vehicle (e.g. 20 mM Sodium Acetate, 75 mM Sodium Chloride, 5% sucrose, 0.02% PS20, pH 5.5), administered as 50-100 pL or 100 to 500 pL per nare. All NHPs receive intranasal challenge with Influenza virus (e.g H1N1) at day 0.

[0338] Samples are taken throughout the study duration, sample types include nasal sampling (for example nasal swabs, wash or scrapes), nasopharyngeal samples, serum and / or bronchoalveolar lavage (BAL) samples. Sampling period includes study duration, in a time frame between one day before the first antibody administration to the end of the study (10-21 days follow-up period). Also, pre-samples are taken to measure baseline levels.

[0339] Samples are analyzed for quantification of viral titers (e.g. RT-PCR) and / or functionality titers of replicating virus (e.g. by TCID50, plaque assays or similar assays), as well as antibody titers and neutralization titers by ELISA or similar methods and in-vitro neutralization assay (e.g. Pseudovirus neutralization).

[0340] Animals

[0341] The NHP subjects are used at a weight of c. 2-12 kg (e.g. 5kg per animal) on commencement of the study. Between 4-12 animals are allocated to each treatment group.

[0342] Study design

[0343] The dose level of the monoclonal antibody applied in the current example is based on the maximal dosing.

[0344] Table (12): Intranasal experimental detail (Animals are challenged with influenza between 1-3 days post first mAb administration which will be considered t=0).

[0345] The NHP subjects are treated via the intranasal route of administration with the antibody at a dose between 0.01 - 5 mg or 6 - 14 mg nominal dose. On Day 0, all NHP subjects are challenged with a dose of Influenza (e.g. H1N1, H5N1) and assessed for virology measurements and clinical observations (e.g. weight loss and temperature) until the end of the study (e.g. Day 10-21).

[0346] Antibody administration

[0347] The test antibody is administered to each nare (e.g. pipette or spray or other atomizer device) according to the treatment schedule (Table 12), between 50 to 500 pL per nare.

[0348] Virus administration

[0349] The virus material is defrosted prior to administration. Once defrosted, the material is diluted and each animal receives between 50 to 500 pL of virus per nare (e.g. pipette or syringe or spray or other atomizer device).

[0350] Data analysis and statistical methods

[0351] Virological measurements will be used for comparisons between treatment and control groups.

[0352] Descriptive statistics will be performed for viral loads (RT-PCR and / or functional titers or replicating virus) and antibody levels (ELISA), where applicable and summarized by group and timepoint using standard statistical tools as appropriate to the statistical distribution of the measurements.

[0353] The primary outcome parameter is anticipated to be a reduction in viral load, summarized as Area Under the Curve (vAUC) in the nose compared to the control. Example 5. Use of monoclonal antibody CR9114 according to the invention (NHP - Efficacy study of CR9114 administered nebulized in a multi -dosing regimen against a flu challenge)

[0354] The aim of this study is to assess the pre- and post-exposure efficacy of a monoclonal antibody according to the invention, having a heavy chain CDR1 as SEQ ID NO: 001, heavy chain CDR2 as SEQ ID NO: 002, heavy chain CDR3 as SEQ ID NO: 003, a light chain CDR1 as SEQ ID NO: 004, a light chain CDR2 as SEQ ID NO: 005, and a light chain CDR3 as SEQ ID NO: 006, following nebulized administration in aNon-Human Primates (NHP) model in which each subject is pre-screened for the absence of serum antibodies against the flu challenge virus.

[0355] Animals are treated with a nebulized dose range of the test antibody described above 1-225 mg nominal dose at study day 0, in a dose regimen that is once daily, (QD), twice daily (BID) or three times daily (TID), for consecutive days up to 14 days.

[0356] Materials and Methods

[0357] The monoclonal antibody according to the invention is provided in a sodium acetate buffer (e.g. 20 mM Sodium Acetate, 75 mM Sodium Chloride, 5% sucrose, 0.02% PS20, pH 5.5) and is diluted to the final concentration for administration ranging from 1 - 225 mg nominal dose, administered with a nebulization device and mask or similar device.

[0358] The NHP subjects allocated to the Control Group (Sodium acetate buffer) are administered with vehicle (e.g. 20 mM Sodium Acetate, 75 mM Sodium Chloride, 5% sucrose, 0.02% PS20, pH 5.5).

[0359] All NHPs receive preferably an Intratracheal + Intranasal challenge or Intranasal with Influenza virus (e.g. H1N1) at day 0.

[0360] Samples are taken throughout the study duration, sample types include nasal sampling (for example nasal swabs, wash or scrapes), nasopharyngeal samples, serum and / or bronchoalveolar lavage (BAL) samples. Sampling period includes study duration, in a time frame between one day before the first antibody administration to the end of the study (10-21 days follow-up period). Also, pre-samples are taken to measure baseline levels.

[0361] Samples are analyzed for quantification of viral titers (e.g. RT-PCR) and / or functionality titers of replicating virus (e.g by TCID50, plaque assays or similar assays), as well as antibody titers and neutralization titers by ELISA or similar methods and in-vitro neutralization assay (e.g. Pseudovirus neutralization).

[0362] Animals The NHP subjects are used at a weight of c. 2-12 kg (e.g. 5kg per animal) on commencement of the study. Between 5-12 animals are allocated to each treatment group.

[0363] Study design

[0364] The dose level of the monoclonal antibody applied in the current example is based on the maximal dosing.

[0365] Table (13): Nebulized experimental detail (Animals are challenged with influenza between 1-3 days post first mAb administration which will be considered t=0)

[0366] The NHP subjects are treated via the nebulized route of administration with the antibody at a dose between 1-225 mg nominal dose. On Day 0, all NHP subjects are challenged with a dose of Influenza virus (e.g. H1N1, H5N1) and assessed for virology measurements and clinical observations (e.g. weight loss and temperature) until the end of the study (e.g. Day 10-21).

[0367] Antibody administration The test antibody is administered by nebulization (e.g. nebulizer device and mask or similar device) according to the treatment schedule (Table 13).

[0368] Virus administration

[0369] The virus material is defrosted prior to administration. Once defrosted, the material is diluted and each animal received between 0-5000 pL Intratracheally, and 50 to 1000 pL of virus per nare and / or 50- 1000 pL of virus per nare only.

[0370] Data analysis and statistical methods

[0371] Virological measurements will be used for comparisons between treatment and control groups. Descriptive statistics will be performed for viral loads (RT-PCR and / or functional titers) and mAb levels (ELISA), where applicable and summarized by group and timepoint using standard statistical tools as appropriate to the statistical distribution of the measurements.

[0372] The primary outcome parameter is expected to be a reduction in viral load, summarized as Area Under the Curve (vAUC) in the nose compared to the control.

[0373] Results - virological measurements

[0374] Treatment with the different nebulization regimens of test antibody is expected to provide a reduction in virological measurements in the upper and lower airways, compared to the control group.

[0375] Conclusion

[0376] In this Influenza NHP model, the prophylactic nebulized administration of 1-225 mg nominal dose QD and / or BID in a multi-dosing regimen of test antibody according to the invention is expected to provide a reduction in virological measurements in the lung as compared with control group NHPs.

[0377] Example 6.: H1N1 NHP Intranasal model set up

[0378] The aim of this study was to establish an intranasal-only (IN) influenza challenge model using A / Wisconsin / 67 / 2022 (H1N1) virus in Cynomolgus macaques (Macaca fascicular is). The study aimed to demonstrate viral replication in the upper airways / nasal cavity by determining viral titers in NS and BAL following IN challenge.

[0379] Materials and Methods

[0380] Influenza A virus H1N1 A / Wisconsin / 67 / 2022, propagated and titrated in the animal handling facility, was used for the study. Animals

[0381] The NHP subj ects, Cynomolgus macaques Macaca fascicularis) were used at a weight ranging between 3 to 9 kg (e.g. 3.8 kg per animal) on commencement of the study. Serum antibodies against the H1N1 flu challenge virus were measured at baseline before viral challenge. All challenge groups consisted of 6 animals each (in total, 18 animals).

[0382] Study design

[0383] Three groups were established for challenge via IT+IN route and IN only route with two different challenge volumes. All groups received the same amount of total viral particles (106TCID50), see Table 14. Thus, animals were challenged with A / Wisconsin / 67 / 2022 virus via the IN+IT route (group 1) (0.5 mL / nare, plus ImL intratracheal), or via the IN route only using either 0.5 mL / nare (group 2), or 0.25 mL / nare (group 3). The challenge virus was administered as a liquid using a syringe. After the challenge, the animals were monitored for 14 days.

[0384] Table (14): Experimental detail (administration at day 0)

[0385] After the challenge, the animals were monitored throughout the study for 14 days. Samples were taken at days 1, 2, 4, 7, 10 and 14 post challenge, from the nose (via nasal swabs, NS), blood (serum), and lungs (via Bronchoalveolar lavage, BAL) to determine viral load via RT- qPCR (copies / mL), infectious virus via TCID50 (TCIDso / mL), and seroreactivity (serum titer). Additionally, baseline samples were taken before the challenge (pre-study samples). Individual nasal samples were taken from both nostrils (i.e. right and left) at indicated time points and before BAL samples were taken.

[0386] Animal Sedation or Anesthesia

[0387] For restraint purposes and to reduce stress during all procedures, the animals were anesthetized with an intramuscular (IM) injection of ketamine hydrochloride (10-25 mg / kg) or other suitably approved anesthetics, as per the animal facility protocols.

[0388] Viral administration

[0389] All challenge groups received the same total amount of virus of 1 x 106TCID50, from the animal facility stock A / Wisconsin / 67 / 2022 (H1N1, Lot no. 11152023-TO, stock titer of 8.3 x 106TCIDso / mL determined in MDCK-SIAT1 cells). See Table (14) for details on administration volumes.

[0390] For IN challenge, virus was administered using a 1 mL syringe with either 0.5 or 0.25 mL for each nare. For IT challenge, 1 mL of the viral challenge dose was administered intratracheally using a 3 mL syringe.

[0391] Clinical monitoring

[0392] Standard cage-side or individual observations were performed throughout the study period. Animals were monitored for body weight, temperature, responsiveness, nasal discharge, appetite, respiratory rate, respiratory effort, cough, and fecal consistency.

[0393] Virology analysis

[0394] RT-qPCR and TCIDso’s were performed to determine viral load and quantify infectious virus respectively, in different body compartments (e.g. nose, lungs, serum).

[0395] Seroconversion: Antibody concentration Anti-HA ELISA analyzed seroreactivity in the serum.

[0396] Data analysis and statistical methods

[0397] Viral load and infectious titers in NS and BAL, and seroreactivity in serum were summarized by group and time point using standard statistical tools as appropriate to the statistical distribution of the measurements.

[0398] Peak was defined as the highest response per animal across all sample timepoints. The Area Under the Curve (AUC) was calculated using the loglO-transformed response (adjusted for the limit of quantitation) and the time post-challenge (expressed in days), using the trapezoidal method. Baseline values are measured before viral challenge. T=0 is set as the time when the viral challenge was performed. Differences between the experimental groups and the reference group were addressed qualitatively.

[0399] Results

[0400] Viral replication in NS

[0401] Administration of the virus by IN-only resulted in increased viral load (RT-qPCR), infectious virus (TCID50) levels (see Figure 2), peak responses (peak viral load and peak titer) and AUC (see Figure 3) in the NS compared to IN+IT challenge.

[0402] The IN-only groups exhibited high viral loads that peaked between days 1 - 7 in all animals (median peak on day 4); the increase in viral load, which continued until the peak was reached, suggested active replication of the inoculated virus in the nasal compartment. The differences in challenge volume of groups challenged by the IN-only route did not affect viral load or infectious virus levels (see Figure 3).

[0403] Viral replication in BAL

[0404] The viral load (RT-qPCR) in the bronchoalveolar lavage (BAL) following an IN-only challenge was decreased compared to an IN+IT challenge (see Figure 4A-C and Figure 5A). In IN-only challenge groups, virus replication in the BAL is observed from day 1 onwards, suggesting flow-through of part of the inoculum into the lung. Infectious virus (peak titer) in BAL from IN-only group 3 (0.25 mL / nare) seems decreased compared to IN+IT group 1 (Figure 5C), and no differences in AUC were observed across groups (Figure 5D).

[0405] Seroreactivity

[0406] Anti -HA serum titers show that at day 10 and 14, seroreactivity of all serum samples increased compared to baseline (Figure 6). Two animals showed increased seroreactivity at baseline compared to the rest of the macaques in the study; however, these high baseline titers did not have a consistent effect on viral replication.

[0407] Clinical observations

[0408] No deviations were reported for the clinical parameters monitored (score 0 during the experiment). No intervention by the veterinarian was required.

[0409] Conclusion

[0410] IN-only challenge with A / Wisconsin / 67 / 2022 (H1N1) resulted in productive viral infection in upper respiratory tract, with increased viral loads and infectious virus in the nasal compartment for the IN-only challenge when compared to IN+IT challenge. Delivering the inoculum directly to the nose leads to higher nasal viral loads compared to dividing the inoculum between the nose and trachea.

[0411] During IN-only challenge, the peak viral load and AUC in the lower airways (in the BAL) was lower compared to the IN+IT group. Following the IN-only challenge, the viral load detected in the lower respiratory tract was lower than in the upper respiratory tract. No clear difference in viral loads or viral titer were observed in the NS or BAL with respect to the IN-only challenge volume.

[0412] Signs of seroreactivity were detected as of day 10 in all groups, and serum titers continued to increase until the last time point tested (day 14), indicating that animals mounted an adaptive immune response to the viral challenge, irrespective of administration route or volume. In conclusion, the IN-only H1N1 challenge method results in viral replication that is more, but not solely, localized to the upper airways compared to IN+IT.

[0413] Example 7.: CR9114 (NHP - Efficacy study of CR9114 administered IN in a multi-dosing regimen against a flu challenge)

[0414] The aim of this study was to assess the pre- and post-exposure efficacy of a monoclonal antibody according to the invention, having a heavy chain CDR1 as SEQ ID NO: 001, heavy chain CDR2 as SEQ ID NO: 002, heavy chain CDR3 as SEQ ID NO: 003, a light chain CDR1 as SEQ ID NO: 004, a light chain CDR2 as SEQ ID NO: 005, and a light chain CDR3 as SEQ ID NO: 006, following intranasal administration in a Non-Human Primate (NHP) model (Cynomolgus macaques, Macaca fascicu laris). For each animal, serum antibodies against the flu challenge virus are measured as baseline in samples collected before the study start (before mAb administration and virus challenge).

[0415] Animals were treated with an intranasal dose of 3.6 mg / nare (equivalent to 7.2 mg nominal administration with the nasal stray per animal / timepoint) of the antibody according to the invention using the multi-dosing scheme, as indicated in Table 15. The mAb was administered both before and after viral challenge (day 0). The dosing occurred twice daily (BID) on days - 3, -2, -1, one dose on day 0 (challenge day) in the afternoon, followed by BID administration from day 1 until day 9. Three days after the first mAb administration, animals were challenged via IN route with IxlO6TCID50 H1N1 A / Wisconsin / 67 / 2022 in the morning of DO, which is considered t=0 of the study.

[0416] For this study, protection was defined as a significant reduction in the vAUC of the viral loads determined by RT-PCR (vRNA) (i.e., in the nasal samples (NS) of the treatment group compared to the control group.

[0417] Table (15). Experimental schedule of antibody and virus challenge administration

[0418] Note: Time frame between am (morning) administration and pm (afternoon) administration is ~8 hours. All time points are relative to virus challenge.

[0419] Materials and Methods

[0420] The monoclonal antibody according to the invention was provided in a sodium acetate buffer (e.g. 20 mM Sodium Acetate, 75 mM Sodium Chloride, 5% sucrose, 0.02% PS20, pH 5.5). It was administered at 3.6 mg in 70 pl per nare (equivalent to 7.2 mg in 140 pl in the nose) using a nasal spray device. The NHP subjects allocated to the Control (sham) group were administered with vehicle (phosphate Saline buffer (PBS) formulation), administered as 70 pL per nare (140 pL in the nose), using a nasal spray device.

[0421] All NHPs received an intranasal challenge (0.5 mL / nare, 1 x 106TCID50 in total) with influenza virus A / Wisconsin / 67 / 2022 (H1N1) at day 0 in the morning. For the IN challenge, the virus was administered using a 1 mL syringe with 0.5 mL for each nare (equivalent to 1 mL per nose).

[0422] Samples were taken during the study at baseline and days 1, 2, 4, 7, and 10 post-challenge. Sample types include nasal sampling (nasal swabs) and serum. Additionally, pre-samples were taken to measure baseline levels before the study begins (i.e. before first antibody administration and viral challenge).

[0423] Samples were analyzed via: RT-PCRto assess viral loads, TCID50 to determine infectious virus, and ELISA to measure the concentration of the antibody according to the invention, and seroconversion.

[0424] Animals

[0425] The NHP subjects, Cynomolgus macaques Macaca fascicularis), had an average weight ranging between 3.1 and 5.2 kg at the commencement of the study. Eight animals were allocated to the treatment and control group (total animal number n=16).

[0426] Study design

[0427] Table (16): Scheme for antibody administration and virus challenge per treatment group.

[0428] Clinical monitoring was conducted throughout the study. The NHP subjects in Group 1 were treated via the intranasal route of administration with the antibody according to the invention at a dose of 3.6 mg per nare (equivalent to 7.2 mg nominal dose per administration with the nasal spray per animal / timepoint, following the dosing scheme specified in Table 15). Group

[0429] 2 served as the control receiving IN sham treatment.

[0430] On Day 0, in the morning, all NHP subjects were challenged intranasally with IxlO6TCID50 of Influenza (H1N1). Nasal samples (swabs) and serum samples were taken after the challenge at days 1, 2, 4, 7, and 10. Nasal swabs were used for virology measurements (RT-PCR and TCID50) as well as human IgG quantification. Serum samples were used for the quantification of human IgG and for detection of seroconversion.

[0431] Animal Sedation or Anesthesia For restraint purposes, and to reduce stress for all procedures, the animals were anesthetized with intramuscular (IM) injection of Ketamine HC1 (10-25 mg / kg) or other suitably approved anesthetics according to the animal facility protocols.

[0432] Antibody administration

[0433] For this, the antibody was administered intranasally using a nasal spray device. The test antibody was administered to each nare according to the treatment schedule (Table 15), 70 pL per nare (140 pL per nose).

[0434] Virus administration

[0435] The viral stock titer of A / Wisconsin / 67 / 2022 (H1N1, Lot no. 11152023-TO, 8.3 x 106TCIDso / mL) was determined in MDCK-SIAT1 cells. The virus material was defrosted before administration. Once defrosted, the material was diluted to achieve the appropriate challenge dose, and each animal received 500 pL of virus per nostril (e.g., using a syringe for each nostril) for a total challenge dose of 1 x 106TCID50.

[0436] Clinical monitoring

[0437] Standard cage-side or individual observations were performed throughout the study period. Animals were monitored for body weight, temperature, responsiveness, nasal discharge, appetite, respiratory rate, respiratory effort, cough, and fecal consistency.

[0438] Virology analysis

[0439] RT-qPCR and TCIDso’s were performed to determine total viral load and quantify infectious virus in different body compartments (e.g. nose, lungs, serum).

[0440] Antibody concentration and Seroconversion

[0441] Anti-human Fc IgG ELISA was performed to determine the IgG levels in different samples. Seroreactivity was analyzed by anti -HA ELISA in the serum. Data analysis and statistical methods

[0442] Virological measurements (viral titer determined by TCID50 and viral load determined by qPCR) were used for comparisons between treatment and control groups.

[0443] Descriptive statistics were performed for viral loads (RT-PCR), viral titers (TCID50) and antibody levels (ELISA), where applicable and summarized by group using standard statistical tools as appropriate to the statistical distribution of the measurements.

[0444] The primary objective, a reduction in viral AUC compared to the control group, was determined by measuring total viral load in the nose using RT-PCR.

[0445] Additional exploratory analyses compared peak responses (viral load and viral titer) between treatment group and control group as well as AUC for viral titer in NS samples. Peak values are defined as the maximum value per animal across all time points. The Area Under the Curve (AUC) was calculated using the loglO-transformed response (adjusted for the limit of quantitation) and the time post-challenge (expressed in days), using the trapezoidal method. Baseline values are considered before mAb administration or viral challenge. T=0 is defined as the moment when the viral challenge was performed. AUC’s and peak values of the treatment group were compared to the Sham control group using Welch's t-test or Exact Wilcoxon-Mann-Whitney Test, in case of negatives.

[0446] Results

[0447] Human IgG antibody PK in NS & Serum

[0448] NS and serum samples were analyzed by anti-human Fc IgG ELISA to assess the concentration of the IN-administered mAb from DI to DIO (Figure 7).

[0449] A steady IgG concentration was detected in the nose in the group treated with the antibody according to the intervention (median concentration from day 1 to day 10 was >100 pg / mL, Figure 7A). With the multidosing scheme, the steady state of mAb in the nose of the treatment group was generally maintained until the end of the study, day 10. No human IgG was detected in nasal samples from the control group at any time point tested (Figure 7B). The IgG AUC (Figure 7C) and peak IgG concentration (Figure 7D) in the NS for the treatment group was statistically significantly higher compared to the control (p<0.001 for both datasets). In the serum (Figure 8), human IgG was detected in the serum samples NHPs treated IN with the antibody according to the invention, but the concentration was lower than in the nasal compartment, did not reach a steady state, and declined from D4 onwards. The median concentration in serum from day 1 to day 10 was < 1 pg / mL in NHPs treated IN with the antibody according to the invention (Figure 8A). No human IgG was detected in the sera from the control group at any of the tested time points (Figure 8B). The IgG AUC (Figure 8C) and peak IgG concentration (Figure 8D) in the sera for the treatment group were statistically significantly higher compared to the control group (p=0.026 for both datasets).

[0450] Viral replication in NS

[0451] IN challenge with A / Wisconsin / 67 / 2022 (H1N1) resulted in productive viral infection in the nasal compartment of the control group (Figure 9B and Figure 10B). The viral load (RT-qPCR) in the NS following IN challenge was reduced in the treatment group as compared to the control group (Figure 9A-B). Both AUC (Figure 9C) and peak viral loads (Figure 9D) were statistically significantly reduced in the antibody -treated group compared to the control group (p<0.001 and p=0.003, respectively). Importantly, a statistical significant reduction in viral load and infectious titer in the NS was detected in the treatment group compared to the control group one day after viral infection (p=0.005 and p=0.046, respectively) (Figure 11).

[0452] In addition, viral titer in the nasal compartment was determined as an exploratory endpoint (Figure 10). Thus, exploratory analyses were performed to compare peak titers and AUC, in NS samples from the treatment group with those from the control group. The viral titers in the NS following an IN challenge were decreased in the treatment group as compared to the control group (Figure 10A-B). Both AUC (Figure 10C) and peak titer (Figure 10D) were significantly reduced in the antibody-treated group compared to the control group (p=0.047 for both datasets).

[0453] Seroreactivity

[0454] Anti -HA serum titers show that at day 10 after challenge, seroreactivity of serum from sham group increased compared to baseline (p=0.004) (Figure 12). In contrast, animals treated with the antibody according to the intervention did not show a significant increase in serum anti-HA (Hl) titers from baseline to post-challenge (p=0.146).

[0455] Clinical observations

[0456] No deviations were reported for responsiveness, discharge, respiratory rate, respiratory effort, cough, fecal consistency (score 0 during the experiment). Following viral challenge, the appetite of animals from both groups was reduced, with a trend that CR9114 had increased appetite compared to sham. No intervention by the veterinarian was required.

[0457] Conclusion

[0458] The intranasal administration twice-daily of the antibody according to the invention showed that the antibody concentration in the nose reached steady state at day 1 or earlier, and it was sustained throughout the study until day 10. Low mAb serum concentrations were detected without reaching steady state. A significant reduction in viral load and infectious titer in the NS was detected on day 1 after intranasal administration twice-daily of the antibody according to the invention. Additionally, twice-daily administration of the antibody according to the invention significantly reduced viral AUC of RNAin the nasal samples determined by RT-PCR (i.e. viral loads of influenza A virus H1N1 A / Wisconsin / 67 / 2022). Therefore, treatment with the twice-daily dosing intranasal regimen of antibody protected against H1N1 challenge.

[0459] As exploratory endpoints, twice-daily multi-dosing of the antibody according to the invention significantly reduced the peak viral load in nasal samples and decreased infectious virus quantified by viral AUC and peak titer.

[0460] Example 8,: Predicting in vivo efficacy of intranasal CR9114 in humans and non-human primates

[0461] Materials and Methods

[0462] Mathematical model Intranasal pharmacokinetics of CR9114 are described by a one-compartment linear elimination model. For a sequence of N instantaneous doses administered at times dvd2, ... , dN, the predicted intranasal CR9114 concentration C at time t is given by where Dtis the dose amount at time d V is the volume of distribution, keiis the antibody elimination rate constant, and lt>dis the indicator function which is equal to 1 if t > d and 0 otherwise.

[0463] Intranasal viral dynamics are modelled by an adapted target-cell limited model which is solved subject to initial conditions

[0464] T = To, V = V0at t = 0, (3) where T is the number of target cells available to infect, and V denotes the infectious viral load. In Equation (2), / 3 denotes the infection rate, r is the rate of virus production by infected cells, y is the viral decay rate, and £ is the efficacy of the drug in reducing virus infection rate. We assume that drug efficacy varies over time proportionally to the instantaneous antibody concentration, such that where 1C5Ois the half-maximal inhibitory concentration of CR9114 required to neutralize a specific influenza challenge strain in vitro.

[0465] Data availability

[0466] To parameterize Equation 1 to describe intranasal PK of CR9114 in humans, intranasal PK data from the Phase 1 PK study of CR9114 was used. To parameterize Equation (1) to describe intranasal PK of CR9114 in NHPs, intranasal PK data from the Lovelace Biomedical study of 6 cynomolgus macaques was used.

[0467] To parameterize Equations 2-3 to describe intranasal viral load dynamics in humans, infectious viral load (TCIDso / mL) data of influenza A / Texas / 91 H1N1 was used from thirteen individuals belonging to placebo groups from four zanamivir (Relenza) Phase 1 trials (NAIA1001 - NAIA 1004) conducted by GSK. It is assumed that the inferred viral kinetics from this data will be representative of other influenza A subtypes.

[0468] To parameterize Equation 4 for in vivo efficacy, the IC50 for A / Wisconsin / 67 / 2022 H1N1 was taken to be 1.14 pg / mL, and for A / Belgium / 4217 / 2015 H3N2 to be 7.48 pg / mL, as per internal neutralization data.

[0469] Model parameter inference

[0470] Bayesian inference was used to fit each of the three datasets to the corresponding model. In each instance, a pooled fitting approach was used to determine the aggregated cohort behavior. Bayesian inference approaches assume that parameters take a probabilistic distribution, accounting for the marked variation between participants over time. A log-normal error model was selected for each observable output based on the large variability in concentration measurements that vary over several orders of magnitude logyi(t)~JV'(log (t), cr2), (5) denotes the observable concentration measurement, C£denotes the model-predicted concentration measurement, and <J is the standard deviation associated with the log-normal error. For each model fit, posterior convergence diagnostics including Effective Sample Size (ESS), Markov Chain Standard Error (MCSE), and Gelman-Rubin diagnostic (R-hat), were computed using the ArviZ python package to assess the independence of the samples and ensure accurate convergence (Table 17). Additionally, autocorrelation and pairwise parameter plots were visually inspected to confirm that the posterior parameter estimates were independent. A comparison between prior and posterior distributions was performed (Figure 13) to verify that the data sufficiently informed the posterior distributions. Finally, posterior predictive checks ensured that posterior parameter distributions accurately describe the observed data (Figures 14,16).

[0471] Table (17).Summary of model priors and posterior fitting metrics demonstrating goodness of fit with each dataset.

[0472] Data includes Markov Chain Standard Error (MCSE), Effective Sample Size (ESS), and Gelman-Rubin diagnostic (R-hat). Table (17A). Priors and fitting metrics for the human PK data

[0473] Table (17B). Priors and fitting metrics for the NHP PK data

[0474] Table (17C). Priors and fitting metrics for the viral load data

[0475]

[0476] Pharmacokinetics (PK)

[0477] First-in-human PK

[0478] All human PK data from the first-in-human PK study was fitted simultaneously to Equation (1) using Bayesian inference methods implemented in the open-source python package, chi, assuming a log-normal error model. Uniform prior probability distributions were prescribed as per Table 17A.

[0479] Posterior parameter distributions were sampled using the Haario-Bardenet Adaptive Covariance Markov Chain Monte Carlo (MCMC) sampler implemented in PINTS with 20,000 iterations across three parallel chains. Chains were visually inspected to ensure sufficient mixing, and the first 5,000 warm-up iterations were discarded. For posterior predictive plots (Figure 14A), we drew 10,000 random samples from the posterior parameter distribution to simulate Equation (1). For efficacy simulations (Figures 15,18), we drew 1000 random samples from the posterior parameter distribution to simulate Equation (1).

[0480] NHP PK

[0481] All NHP PK data was simultaneously fitted to Equation (1) using Bayesian inference methods implemented in the open-source python package, CmdStanPy, assuming a log-normal error model. Concentration measurements at or below the lower limit of quantification (32 ng / mL) were censored. Prior probability distributions were prescribed as per Table 17B.

[0482] Posterior parameter distributions were sampled using the No-U-Turn (NUTS) MCMC sampler with 10,100 iterations across four parallel chains. Chains were visually inspected to ensure sufficient mixing, and the first 1000 warm-up iterations were discarded. For posterior predictive plots (Figure 16), we drew 1000 randoms samples from the posterior parameter distribution to simulate Equation (1). For efficacy simulations (Figure 17), we drew 1000 random samples from the posterior parameter distribution to simulate Equation (1). Middle panel in Figure 17 demonstrates 100% efficacy.

[0483] Viral load dynamics

[0484] Viral load data was simultaneously fitted to Equation (2) solved subject to Equation (3) using Bayesian inference methods implemented in the open-source python package, CmdStanPy, assuming a log-normal error model. For inference, the initial number of target cells Toin humans was fixed at 4* 108as per literature estimates, and the efficacy £ was fixed to 0. Viral load measurements at or below the lower limit of concentration (0.75 logic TCIDso / mL) were censored. Prior probability distributions were prescribed as per Table 17C.

[0485] Posterior parameter distributions were sampled using the NUTS MCMC sampler with 2,000 iterations across four parallel chains. Chains were visually inspected to ensure sufficient mixing, and the first 1000 warm-up iterations were discarded. For posterior predictive plots (Figure 14B), we drew 1000 random samples from the posterior parameter distribution to simulate Equations (2)-(3). For efficacy simulations (Figures 15,17,18), we drew 1000 random samples from the posterior parameter distribution to simulate Equations (2-3).

[0486] The viral load fitting results from human viral load data were adapted to simulate viral load kinetics in NHPs. The initial target cell concentration Towas reduced proportionally to the ratio of the surface area between humans and NHPs, which was taken to be 4.5. Similarly, the inferred initial virus concentration Voin humans was multiplied by 4.5 for NHPs. Infection rate / 3 and viral decay rate y were scaled by 6 and 1.2 respectively to show good qualitative agreement with viral load kinetics observed in NHPs in the absence of CR9114.

[0487] Efficacy simulations

[0488] The efficacy of CR9114 to reduce viral load in both NHPs and humans was simulated by solving Equations (1) - (4) parameterized by inferred and literature-derived parameter estimates summarized above. To simulate new dosing regimens not tested experimentally (Figure 18), the dose amount and frequency of dosing was prescribed as a model input. In simulations representing untreated groups, viral load simulations are performed taking the efficacy parameter E=0. For treated groups, efficacy was related to CR9114 concentration and ICso values as per Equation (4). Log-normal error is simulated on model outputs using corresponding standard deviations inferred from model fitting.

[0489] Results

[0490] Validation of CR9114 PK modelling in humans

[0491] Model outputs show good quantitative agreement with human PK data. At least 95% of the datapoints lie within the 95% model confidence interval across all dose amounts and dosing regimens tested (Figure 14A), indicating that our PK model is predictive of intranasal CR9114 pharmacokinetics in humans. Similarly, model outputs show good quantitative agreement with viral load measurements (Figure 14B), indicating that our pharmacodynamic (PD) model is predictive of intranasal viral load dynamics.

[0492] With these PK and PD model parameterizations, we validate our efficacy model by comparing our predicted viral load with that of the Phase 2A trial in humans in the same trial setup. That is, we simulate the impact of a single one-off dose of 10 mg of CR9114 six hours prior to challenge with influenza A / Belgium / 4217 / 2015 H3N2 in humans. Model outputs show significant overlap between the viral load in treated and untreated groups, meaning there was no effect in reducing viral load (Figure 15). This is because the intranasal concentration of CR9114 rapidly drops below the IC50 of CR9114 against the H3N2 challenge strain. This accurately recreates the outcome of the Phase 2A trial in humans with the same trial setup, which also shows no effect of a single one-off 10 mg dose in reducing viral load if administered six hours before challenge. As such, we conclude that our model accurately predicts the intranasal efficacy of CR9114 in humans against H3N2.

[0493] Validation of CR9114 PK modelling in NHPs

[0494] Model outputs showed good quantitative agreement with NHP PK data. At least 95% of the datapoints lie within the 95% model confidence interval (Figure 16), indicating that our PK model is predictive of intranasal CR9114 pharmacokinetics in NHPs.

[0495] We then validated our efficacy model by comparing our predicted viral load in the presence of CR9114 to that of the NHP efficacy trial in the same trial setup. That is, we simulated the impact of twice-daily dosing of 7.2 mg of CR9114 with three days prophylactic administration prior to challenge with influenza A / Wisconsin / 67 / 2022 H1N1 in NHPs. Model output showed that 100% of PK concentrations remain above the neutralizing IC50 of the challenge strain at all timepoints, and therefore achieveed near-100% efficacy (Figure 17). Consequently, the viral load in the treated group decreases immediately following challenge, and remains below quantifiable limits (Figure 17). This accurately recreates the outcome of the NHP efficacy trial with the same trial setup, which showed unquantifiable infectious viral load at all measured timepoints across all monkeys treated with CR9114. As such, we concluded that our model accurately predicts the intranasal efficacy of CR9114 in NHPs against H1N 1.

[0496] Simulations of continuous once-daily CR9114 dosing in humans

[0497] Using our PKPD model parameterization, we simulated the efficacy of continuous once-daily dosing of CR9114 in reducing viral load, with administration starting on the day of challenge. Simulations showed that once-daily dosing >40 mg is successful against the influenza A / H3N2 challenge agent in decreasing the overall viral load compared to the untreated group, with median viral load curves showing a clear monotonic decrease to below quantifiable levels (Figure 18). All doses lead to >50% of simulated CR9114 concentrations remaining above the IC50 for the H3N2 challenge strain, and are successful in reducing the area under the viral load curve and peak viral load compared to untreated groups across all simulated timepoints. With once-daily 40 mg dosing, median efficacy drops to -40% prior to re-dosing. Conversely, 100 mg dosing leads to a -60% median efficacy prior to re-dosing. Therefore, we showed that a higher dose leads to an increase in efficacy and a decrease in viral load.

[0498] Additionally, we showed that consecutive once-daily dosing >5 mg is successful against influenza A / H1N1 in reducing viral load (Figure 19). Again, all doses lead to >50% of simulated CR9114 concentrations remaining above the IC50 for the H1N 1 challenge strain, and are successful in reducing the area under the viral load curve and peak viral load compared to untreated groups across all simulated timepoints.

[0499] Conclusion

[0500] Our PKPD modelling has been shown to accurately predict the intranasal pharmacokinetics of CR9114, and the associated viral load reduction, in NHPs and humans. Simulations predict that continuous once-daily dosing beginning on the same day as challenge is sufficient to successfully reduce viral load in humans challenged with influenza A / H3N2 and A / H1N 1.

Claims

CLAIMS1 A composition comprising a monoclonal IgG antibody for use in the prophylaxis or treatment of an influenza virus infection in a mammal, wherein the composition is administered intranasally to said mammal according to a daily multidose regimen.2 A composition for use according to claim 1, wherein the daily multidose regimen consists of administering two unit doses of the composition within a 24 hour period; or administering three unit doses of the composition within a 24 hour period; or administering four unit doses of the composition within a 24 hour period;3 A composition for use according to claim 2, wherein each of the first and second unit doses comprise or consist of 0.5 - 100 mg of the monoclonal IgG antibody, such as 2-5mg / 100 pL of the composition, preferably 5mg / 100 pL, of the composition administered to each of the left and right nostrils, resulting in a total unit dose of 4-10 mg / 200 pL, preferably 10 mg / 200 pL.4 A composition for use according to claims 2 or 3, wherein two unit doses are administered within a 24 hour period and the first and second unit doses are administered 2- 18 hours apart, optionally 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17 or 18 hours apart, optionally 12 hours apart.5 A composition for use according to any preceding claim, wherein the composition is administered according to the multidose regimen every day for a period of for 1 to 7 days, optionally for 2, 3, 4, 5, 6 or 7 days, optionally for 1 to 24 weeks, optionally for 6 weeks, optionally for 12 weeks, optionally for between 6 to 10 weeks.6 A composition for use according to claim 4, wherein the multidose regimen comprises administering first and second unit doses of the composition according to the regimen of claim4 for 1 to 7 days, optionally for 2, 3, 4, 5, 6 or 7 days, followed by a single dose administration regimen, wherein a unit dose is administered once every 24 hours.7 A composition for use according to claim 6, wherein the single dose regimen comprises administering 0.5 - lOOmg of the antibody , such as administering 2-5 mg / 100 pL, preferably5 mg / 100 pL, of the composition to each nostril, resulting in a total unit dose of 4-10 mg / 200 pL, preferably 10 mg / 200 pL.8 A composition for use according to any preceding claim, wherein the monoclonal IgG antibody is CR9114.9 A composition for use according to any preceding claim, wherein the composition is administered as a nasal spray.10 A composition for use according to any preceding claim, wherein the influenza virus is influenza A or B.11 A composition for use according to any preceding claim, wherein the composition is administered prior to infection and / or during period of exposure to the virus and / or during a period when there is an increased risk of exposure to the virus.12 A composition for use according to any preceding claim, wherein the mammal is a human.13 A composition formulated for intranasal or inhalation administration and effective for treatment or prophylaxis of influenza viral infection in a mammal comprising monoclonalantibody CR9114 in a single unit dose of lOmg or at least lOmg , 15mg or at least 15mg, 20mg or at least 20 mg, such as 10mg / 200pL or least 10 mg / 200 pL, 15mg / pl or at least 15 mg / 200 pL, or 20 mg / pL or at least 20mg / 200 pL.14 A composition comprising monoclonal antibody CR9114 for use in the prophylaxis or treatment of influenza virus infection in a mammal, wherein the composition is administered intranasally to said mammal as a single unit dose sufficient to achieve a threshold concentration of antibody in the nasal mucosa of at least 20 mg / L.15 A composition according to claims 13 or 14, wherein the mammal is a human.16 A method for treatment or prophylaxis of an influenza virus infection in a mammal comprising administering a monoclonal IgG antibody intranasally to said mammal according to a daily multidose regimen.17 A method according to claim 16, wherein the daily multi dose regimen consists of administering two unit doses of the composition within a 24 hour period; or administering three unit doses of the composition within a 24 hour period; or administering four unit doses of the composition within a 24 hour period;18 A method according to claim 17, wherein each of the first and second unit doses comprise or consist of 0.5 - 100 mg of monoclonal IgG antibody such as 1-20 mg of monoclonal IgG antibody, such as 1, 2, 3, 4, 5, 10, 15 or 20 mg, for example 1-20 mg / lOOpl, such as 2-5mg / 100pl, preferably 5mg / 100pl, of the composition administered each of the left and right nostrils, resulting in a total unit dose of 2- 40 mg / pl, such as 4-10 mg / 200 pl, preferably 10 mg / 200 pl.19 A method according to claims 17 or 18, wherein two unit doses are administered within a 24 hour period and the first and second unit doses are administered 2-18 hours apart, optionally 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17 or 18 hours apart, optionally 12 hours apart.20 A method according to any of claims 17-19, wherein the composition is administered according to the multidose regimen for a period of 1 to 24 weeks, optionally for 6 weeks, optionally for 12 weeks, optionally for between 6 to 10 weeks.21 A method according to claim 19, wherein the multidose regimen comprises administering first and second unit doses of the composition according to the regimen of claim 19 for 1 to 7 days, optionally for 2, 3, 4, 5, 6 or 7 days, followed by a single dose administration regimen, wherein a unit dose is administered once every 24 hours.22 A method according to claim 21, wherein the single dose regimen comprises administering 2-5 mg / 100 pL, preferably 5 mg / 100 pL, of the composition to each nostril, resulting in a total unit dose of 4-10 mg / 200 pL, preferably 10 mg / 200 pL.23 A method according to any of claims 16-22, wherein the monoclonal IgG antibody is CR9114.24 A method according to any of claims 16-23, wherein the composition is administered as a nasal spray.25 A method according to any of claims 16-24, wherein the influenza virus is influenza A or B.26 A method according to any of claims 16-25, wherein the composition is administered prior to infection and / or during period of exposure to the virus and / or during a period when there is an increased risk of exposure to the virus. 27 A method according to any of claims 16-26, wherein the mammal is a human.28 A nasal delivery device for delivering a composition to the human nasal cavity, said device comprising a composition comprising monoclonal antibody CR9114, wherein the device is metered such that a single unit dose is administered as two 5 mg / 100 pL doses applied sequentially to each nostril as a nasal spray.29 A delivery device for delivering a composition according to anyone of claims 1-15 to the human nasal cavity, wherein said device comprises said composition.

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