Viral treatment

Abstract

The disclosure provides molecules for reducing the severity and / or number of symptoms of viral infections. In particular, the disclosure provides glycan binding molecules for use in reducing the severity and / or number of symptoms of viral infections.

Description

[0001] VIRAL TREATMENT

[0002] FIELD

[0003] The disclosure provides molecules for reducing the severity and / or number of symptoms of viral infections.

[0004] BACKGROUND

[0005] Viral pathogens can cause an array of diseases and / or conditions in humans and while vaccines and antiviral therapies are available for use against some pathogens, for others there are no useful therapeutic options. In some cases, the treatment of a viral infection is limited to patient isolation (to minimise the risk of further spread) and treatment of the symptoms. In some cases a viral pathogen, in particular a respiratory viral pathogen, can lead to secondary complications such as shortness of breath, pneumonia, bronchitis and / or bronchiolitis. The young, old, immunocompromised, pregnant and individuals with underlying health conditions are often most at risk from viral pathogens.

[0006] The pathology of any given viral pathogen will vary, but those causing respiratory illness will most often be transmitted by aerosolised droplets generated by, for example, coughs and sneezes and fluids / secretions from / to mucus membranes (including the respiratory airways, the lungs, nose, mouth and eyes).

[0007] Vaccines are often difficult to develop, taking a considerable time to prepare and test. Also, antiviral drugs are variably effective and, in many cases, the virus infection can resolve before the drug takes effect.

[0008] Accordingly, there is a need for new and effective treatments to combat the symptoms and / or severity of a viral infection and the diseases and / or conditions they cause or contribute to.

[0009] SUMMARY

[0010] The present disclosure is based on the finding that the administration regime for molecules with an affinity for sialic acid (so called ‘glycan-binding’ molecules, ‘sialic-acid binding’ molecules or the carbohydrate binding module (CBM) thereof) has an effect on the severity and / or number of symptoms of a viral infection in a subject. Without wishing to be bound by theory, the inventors have further noted that the effect on the severity and / or number of symptoms is independent of the viral load. In other words, while the general administration of any of the molecules described herein may achieve a reduction in viral load (e.g. a reduction in peak viral load: as determined by qRT-PCT or viral culture), administration in accordance with the regimes described herein (e.g. administering the molecule once or twice a week, or at intervals between 2 to 7 days, e.g. 3 to 7 days) may reduce the severity and / or number of symptoms of a viral infection in a subject.

[0011] It should be noted that the terms “comprise”, “comprising” and / or “comprises” is / are used to denote that the various aspects and embodiments of this disclosure “comprise” a particular feature or features. It should be understood that this / these terms may also encompass aspects and / or embodiments which “consist essentially of” or “consist of” the relevant feature or features. Moreover, the entire contents of all references are to be regarded as incorporated herein by reference.

[0012] The term ‘viral infection’ (the severity and / or number of symptoms of which may be reduced) may embrace infections caused or contributed to by a respiratory virus, for example a virus selected from the group consisting of: influenza virus, respiratory syncytial virus (RSV), human parainfluenza virus, rhinovirus and coronavirus, Human metapneumo virus (HMPV), or a combination thereof. By way of further example, the term ‘viral infection’ may embrace coinfections caused by two or more viruses, such as those viruses described above. In particular examples, of the present disclosure, the viral infection may be caused or contributed by an influenza virus.

[0013] As used herein, the term ‘influenza virus’ embraces any virus classed as belonging to the Orthomyxoviridae. This would include species within the Genera Alphainfluenzavirus, Betainfluenzavirus, Gammainfluenzavirus and Deltainfluenzavirus. The term ‘influenza virus’ may include all known serotypes, subtypes and / or strains including, for example, types: H1 N1 , H1 N2, H2N2, H3N1 , H3N2, H3N8, H5N1 , H5N2, H5N3, H5N8, H5N9, H7N1 , H7N2, H7N3, H7N4, H7N7, H7N9, H9N2, H10N7. Influenza virus is known to cause the disease commonly referred to as ‘flu’ in humans (which term includes, for example, the specific disease variants referred to as swine ‘flu’ or bird ‘flu’). As such, the various uses, methods and medicaments described herein may be used to reduce the severity and / or number of symptoms of an influenza infection (or ‘flu’) in a subject.

[0014] The term ‘Respiratory Syncytial Virus’ (RSV) embraces species within the Genus Orthopneumovirus, characterised by the presence of an F protein (on the surface of the virus) which causes neighbouring cell membranes to merge and form large multinucleated syncytia. RSV causes a range of respiratory tract infections (ranging from mild upper respiratory tract infections (URTIs) to severe and potentially life-threatening lower respiratory tract infections (LRTIs)). The various uses, methods and medicaments described herein may be used to reduce the severity and / or number of symptoms of an RSV infection in a subject. The term ‘human parainfluenza virus’ (HPIV) embrace species within the Family Paramyxoviridae. HPIV causes human parainfluenza and embraced within this disclosure is human parainfluenza caused or contributed to by the four main serotypes of clades including HPIV-1 , HPIV-2, HPIV-3 and HPIV-4. The various uses, methods and medicaments described herein may be used to reduce the severity and / or number of symptoms of human parainfluenza in a subject.

[0015] The term ‘Rhinovirus’ embraces those viruses classified as belonging to the Family Picornaviridae and the Genus Enterovirus. The term ‘Rhinovirus’ includes, for example Rhinovirus A, Rhinovirus B, Rhinovirus C and Enterovirus types A through to L. The various uses, methods and medicaments described herein may be used to reduce the severity and / or number of symptoms of a Rhinovirus infection (e.g. the common cold) in a subject.

[0016] As used herein, the term “ Corona virus" embraces any virus classed as belonging to the Family Coronaviridiae and embraces the SARS Coronavirus’, the MERS Coronavirus’, and SARS- CoV-2 Coronavirus. The term “ Corona virus" also embraces all SARS-CoV-2 variants. Coronavirus cause or contribute to a range of respiratory diseases, including, for example, those diseases or conditions referred to as SARS and MERS. The phrase “disease or condition caused or contributed to by a Coronavirus” (the severity and / or number of symptoms of which may be reduced) includes the acute respiratory disease referred to as COVID-19 and which is caused by, or which is associated with, a SARS-CoV-2 infection. Accordingly, the various uses, methods and medicaments described herein may be used to reduce the severity and / or number of symptoms of a disease or condition caused or contributed to by a Coronavirus, including, for example SARS, MERS and / Covid-19.

[0017] Human metapneumo virus (HMPV) is asingle stranded RNA virus of the Family Pneumoviridae. HMPV infects airway epithelial cells in the nose and lung and is known to cause lower respiratory infections. As such, the various uses, methods and medicaments described herein may be used to reduce the severity and / or number of symptoms of a disease or condition caused or contributed to by HMPV.

[0018] The term ‘symptom’ may include any of the symptoms commonly associated with an infection caused by a virus, including any of the viruses described herein. The term ‘symptom’ may further include any of the symptoms commonly associated with a disease or condition caused or contributed to by a virus, including any of the viruses described herein. Additionally, the term ‘symptom’ may include those symptoms which are characteristic and / or typical of such any of these viruses, viral infections and / or diseases and / or conditions.

[0019] The term ‘symptom’ may include, for example, any one or more of the following, a ‘runny nose’ (brought about by excess mucus production in the nasal / respiratory passages), a blocked or ‘stuffed (stuffy’) nose (again brought about by excess mucus production), sneezing (potentially linked to excess mucus production in the respiratory / nasal passages), a sore (scratchy / dry) throat, earache, fatigue (malaise / tiredness), headache, aches (e.g. muscle and / or joint ache), chills and / or fever, cough, shortness of breath, vomiting, diarrhoea, change or loss in taste / smell and / or various combinations thereof. One of skill will appreciate that depending on the particular viral infection (and the disease or condition caused or contributed to thereby) not all of these symptoms may be present at any given time.

[0020] The term ‘subject’ may embrace mammalian subjects, including, for example human or animal subjects. The subject may be at risk of, vulnerable to, or susceptible to, a viral infection and / or a specific disease and / or condition caused or contributed to thereby. The subject may have an underlying disease or condition which is susceptible to virus-induced exacerbation. The underlying disease or condition susceptible to virus-induced exacerbation may be selected from chronic obstructive pulmonary disease (COPD), asthma, cystic fibrosis, non-cystic fibrosis bronchiectasis, pulmonary fibrosis, primary ciliary dyskinesia, diabetes, neutropenia, and haematological cancers. In some examples, the underlying disease or condition susceptible to virus-induced exacerbation may be selected from chronic obstructive pulmonary disease (COPD), asthma, cystic fibrosis, non-cystic fibrosis bronchiectasis, pulmonary fibrosis, and primary ciliary dyskinesia. The subjects may be subjects that are selected from subjects who are receiving chemotherapy (e.g. for their cancer) that is likely to induce neutropenia, subjects undergoing conditioning for haemopoietic stem cell transplant (HSCT), subjects who are recovering from an HSCT, and subjects who have had a lung transplant.

[0021] The present disclosure provides a number of molecules, all of which may be used to reduce the severity and / or number of symptoms of any of the disclosed viruses, viral infections and / or disease / conditions caused or contributed to thereby. These molecules may be administered to a subject (as defined above) once or twice a week, or at intervals between 2 to 7 days, e.g. 3 to 7 days. It should be noted that a reduction in the severity and / or number of symptoms associated with any given virus (including any one or more of the specific symptoms described herein) may be assessed relative to the symptoms exhibited by a subject:

[0022] (i) infected by the virus but not receiving any treatment;

[0023] (ii) infected by the virus but not receiving a treatment described herein; or

[0024] (iii) infected by the virus and receiving a glycan binding molecule-based treatment (or a sialic acid binding molecule or CBM-based treatment), which treatment is not administered once or twice a week, or at intervals between 2 to 7 days, e.g. 3 to 7 days.

[0025] The severity of and / or number of symptoms may be reduced in a subject in a period of at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 day, at least 8 days, at least 9 days, at least 10 days, at least 11 days, at least 12 days, at least 13 days or at least 14 days following a viral infection.

[0026] The severity of and / or number of symptoms may be reduced in a subject infected by the virus who has been administered the glycan binding molecule in comparison to a subject infected by the virus who has not been administered the glycan binding molecule. Additionally or alternatively, the severity of and / or number of symptoms may be reduced in a subject infected by the virus who has been administered the glycan binding molecule in accordance with a treatment regimen as described herein (in particular, which treatment is administered once or twice a week, or at intervals between 2 to 7 days, e.g. 3 to 7 days), in comparison to a subject who has not been administered a glycan binding molecule in accordance with a treatment regimen as described herein (in particular, which treatment is not administered once or twice a week, or at intervals between 2 to 7 days, e.g. 3 to 7 days).

[0027] A reduction in the ‘number of symptoms’ associated with a particular viral infection may mean that, as compared to a subject;

[0028] (i) infected by the virus but not receiving any treatment;

[0029] (ii) infected by the virus but not receiving a treatment described herein; or

[0030] (iii) infected by the virus and receiving a glycan binding molecule-based treatment (or a sialic acid binding molecule or CBM-based treatment), which treatment is not administered once or twice a week, or at intervals between 2 to 7 days, e.g. 3 to 7 days; a subject treated according to this disclosure, may exhibit fewer symptoms than usually or classically associated with the viral infection.

[0031] A reduction in the ‘severity of symptoms’ associated with a particular viral infection may mean that, as compared to a subject; (i) infected by the virus but not receiving any treatment;

[0032] (ii) infected by the virus but not receiving a treatment described herein; or

[0033] (iii) infected by the virus and receiving a glycan binding molecule-based treatment (or a sialic acid binding molecule or CBM-based treatment), which treatment is not administered once or twice a week, or at intervals between 2 to 7 days, e.g. 3 to 7 days; a subject treated according to this disclosure, may exhibit a lower total symptom score (TSS).

[0034] A total symptom score (TSS) may be derived by grading the severity of each symptom classically associated with the relevant viral infection or disease or condition caused or contributed to thereby. The grades allocated to each symptom may then be summed to provide a TSS. By way of non-limiting example, each symptom may be given a score, from 0 (not present as a symptom) through 1 (just noticeable as a symptom) and 2 (it’s clearly bothersome from time to time, but it doesn’t interfere with me doing my normal daily activities) to 3 (the symptom is bothersome most or all of the time and it stops me from participating in activities). These scores may be summed to give a TSS.

[0035] Where a subject has been administered a treatment according to this disclosure and has become infected with a specific virus (e.g. any of the viruses described herein), the computation of a TSS may be based on a single or repeated assessment of the severity of one or more symptoms known to be associated with that viral infection. For example, the severity of the symptoms and / or computation of a TSS, may be determined 1 , 2 or 3 times a day and for multiple days after an infection (or suspected infection). For example a TSS, may be determined 1 , 2 or 3 times a day and for 1 , 2, 3, 4, 5, 6, 7, 8, 9, 10, 11 , 12, 13 or 14 days after an infection (or suspected infection).

[0036] A TSS determined after an infection or suspected infection, may be compared with a TSS obtained prior to the infection or suspected infection. Additionally or alternatively, a TSS determined after an infection or suspected infection may be compared with a TSS determined prior to administration of any of the described molecules as set out herein.

[0037] It is suggested that by administering any of molecules disclosed herein to a subject (suffering from any of the viral infections described herein) once or twice a week, or at intervals between 2 to 7 days, e.g. 3 to 7 days, it is possible to reduce the total symptom score.

[0038] As stated, a TSS may be calculated daily and at multiple points through the day. Where the TSS is plotted on a graph (of TSS against time), the area under the curve (AUG) for any patient receiving a treatment according to this disclosure may be reduced (as compared to a subject

[0039] (i) infected by the virus but not receiving any treatment; (ii) infected by the virus but not receiving a treatment described herein; or (iii) infected by the virus and receiving a glycan binding molecule-based treatment (or a sialic acid binding molecule or CBM-based treatment), which treatment is not administered once or twice a week, or at intervals between 2 to 7 days, e.g. 3 to 7 days). By way of further example, the AUC for the TSS over a time period of one day, two days, three days, four days, five days, six days, seven days, eight days, nine days, ten days, eleven days, twelve days, thirteen days, or fourteen days, following a viral infection may be reduced for any patient receiving a treatment in accordance with this disclosure (as compared to a subject as described in (i) to (iii) above).

[0040] The total symptom score may be reduced such that the likelihood of the subject presenting a total symptom score of grade 2 or higher is reduced.

[0041] By way of non-limiting example, where the viral infection is an influenza virus infection, there may be 11 symptoms classically associated with that infection; these symptoms may be broadly classified as : (i) runny nose; (ii) stuffy nose, (iii) sneezing, (iv) sore throat, (v) earache, (vi) malaise / tiredness, (vii) headache, (viii) muscle and / or joint ache, (ix) chilliness / feverishness, (x) cough, (xi) shortness of breath. As such, and as compared to a subject

[0042] (i) infected with influenza virus but not receiving any treatment;

[0043] (ii) infected with influenza virus but not receiving a treatment described herein; or

[0044] (iii) infected with influenza virus and receiving a glycan binding molecule-based treatment (or a sialic acid binding molecule or CBM-based treatment), which treatment is not administered once or twice a week, or at intervals between 2 to 7 days, e.g. 3 to 7 days; a subject infected with influenza virus, may

[0045] (a) exhibit fewer than all 11 of the abovementioned symptoms (for example only 10, 9, 8, 7, 6, 5, 4, 3, 2, 1 or none of the 11 symptoms).; and / or

[0046] (b) report a lower TSS - wherein the TSS associated with an influenza infection carries a maximum score of 33 (a grading of 3 points for each of the 11 symptoms).

[0047] A ‘glycan binding molecule’ for use in a method of reducing the severity and / or number of symptoms of a viral infection in a subject may comprise (consist of, or consist essentially of) sugar (carbohydrate / polysaccharide / glycan)-binding protein(s). Proteins of this type exhibit a particular affinity for glycans and / or sialic acid. A ‘glycan-binding molecule’ may be a sialic acid binding molecule and / or comprise one or more carbohydrate binding module(s) (CBMs). By way of example, a glycan binding molecule of this disclosure may comprise at least two carbohydrate binding modules (CBMs).

[0048] Examples of CBMs which may be used to reduce the severity and / or number of symptoms of a viral infection in a subject are provided below, but for completeness it should be understood that the term “CBM” includes, for example, CBMs classified as belonging to CBM families 32, 40, 47, 67 and 70.

[0049] As such, the disclosure provides a CBM32, a CBM40, a CBM47, a CBM67 and / or a CBM70 for use in a method of reducing the severity of and / or number of symptoms of a viral infection in a subject comprising: administering the CBM32, the CBM40, the CBM47, the CBM67 and / or the CBM70 to the subject once or twice a week, or at intervals between 2 to 7 days, e.g. 3 to 7 days.

[0050] In another teaching, the disclosure may provide a CBM40 for use in a method of reducing the severity of and / or number of symptoms of a viral infection in a subject comprising: administering the CBM40 to the subject once or twice a week, or at intervals between 2 to 7 days, e.g. 3 to 7 days.

[0051] One of skill will appreciate that there is a vast array of different CBMs and members of the abovementioned families that can be derived from many different bacterial species. Further information regarding CBMs, in particular the family 32, 40, 47, 67 and 70 CBMs, can be found at the public CAZY database (available at: http: / / www.cazy.org). Some particularly useful CBMs may be derived from bacterial species within the Genera: Streptococcus, Vibrio and Clostridium.

[0052] It should also be understood (and as is set out in more detail below) the disclosure embraces the use of molecules which comprise one or more CBMs and modified forms of any of the CBMs described herein. A modified CBM is a CBM which includes one or more mutated residues relative to the wild-type CBM sequence.

[0053] As such, the disclosure provides a modified CBM32, a modified CBM40, a modified CBM47, a modified CBM67 and / or a modified CBM70 for use in a method of reducing the severity of and / or number of symptoms of a viral infection in a subject comprising: administering the modified CBM32, the modified CBM40, the modified CBM47, the modified CBM67 and / or the modified CBM70 to the subject once or twice a week, or at intervals between 2 to 7 days, e.g. 3 to 7 days. In another teaching, the disclosure may provide a modified CBM40 for use in a method of reducing the severity of and / or number of symptoms of a viral infection in a subject comprising: administering the modified CBM40 to the subject once or twice a week, or at intervals between 2 to 7 days, e.g. 3 to 7 days.

[0054] The CBMs for the various uses, medicaments and methods described herein, including uses, medicaments and methods administered to a subject once or twice a week, or at intervals between 2 to 7 days, e.g. 3 to 7 days, are all types of carbohydrate, glycan and / or sialic acid binding molecule.

[0055] The term “sialic acid” embraces all forms of N- or O-substituted neuraminic acid and includes all synthetic, naturally occurring and / or modified forms thereof. Sialic acids may be found as components of cell surface molecules, glycoproteins and glycolipids. Most often, sialic acids are present at the end (terminal regions) of sugar chains connected to cell membranes and / or proteins. For example, some cells of the human upper respiratory tract comprise a-2,6-linked sialic acid receptors and other cells of the upper and lower respiratory tracts comprise a-2,3- linked sialic acid receptors. The sialic acid family encompasses a number (approximately 50) of derivatives that may result from acetylation, glycosylation, lactonisation and methylation at C4, C5, C7, C8 and C9. All such derivatives are to be embraced by the term “sialic acid”.

[0056] Sialic acids may be found linked a(2,3) or a(2,6) to Gal and GalNAc or a(2,8) or a(2,9) to another sialic acid. Accordingly, it is important to understand that while the term “sialic acid” is used throughout this specification, it encompasses all anomers, derivatives, analogues or variants (either naturally occurring or synthetically generated) thereof as well as all monomers, dimers, trimers, oligomers, polymers or concatamers comprising the same.

[0057] Accordingly, a sialic acid binding molecule or CBM for use in a method of reducing the severity and / or number of symptoms of a viral infection in a subject by administration once or twice a week, or at intervals between 2 to 7 days, e.g. 3 to 7 days, may comprise a moiety with affinity for sialic acid in any of its various forms as described above and / or which may bind / couple to and / or associate with sialic acid molecules as may be present in or on, a viral particle and / or a (mammalian) cell surface receptor.

[0058] Further detail concerning each CBM family is provided below - all of these CBMs (their subtypes, variants, orthologues etc.) are for use in a method of reducing the severity and / or number of symptoms of a viral infection in a subject and may be administered to that subject once or twice a week, or at intervals between 2 to 7 days, e.g. 3 to 7 days. CBM32

[0059] A useful CBM32 (i.e. a CBM32 for use in a method of reducing the severity and / or number of symptoms of a viral infection) may be derived from any suitable source. For example, CBM32s for use may be obtained from microorganisms, including, for example, bacteria of the genera Cellvibrio, Yersinia, Micromonospora, Streptococcus, Bifidobacteria and Clostridium. For example, useful CBM32s may be obtained or derived from, for example, Cellvibrio mixtus, Yersinia enterolitica, Clostridium perfringens, Clostridium thermocellum, Streptococcus pneumoniae, Bifidobacterium longum and Micromonospora viridifaciens. Further details concerning the source, structure and function of the CBM32 family can be found within the Carbohydrate Active Enzymes database (freely available on the internet at: http: / / www.cazy.org / CBM32.html).

[0060] An exemplary CBM32 sequence is provided by SEQ ID NO: 1 below:

[0061] SEQ ID NO: 1

[0062] AI IETAI PQSEMTASATSEEGQDPASSAIDGNTNTMWHTKWNGSDALPQSLSVNLGSSRKVSS IAITP RTSGNNGFITKYE IHAINNGVETLVAEGTWEENNLVKTVT FDSPIDAEEIKITAIQGVGGFAS IAELN VYE

[0063] Accordingly, a CBM for use for use in a method of reducing the severity and / or number of symptoms of a viral infection, may comprise, consist essentially or consist of a CBM having the sequence of SEQ ID NO: 1 or a carbohydrate binding portion thereof. A carbohydrate binding fragment of SEQ ID NO: 1 may comprise anywhere between about 5, 6, 7, 8, 9 or 10 (consecutive or contiguous) amino acids to about 138 (consecutive or contiguous) amino acids from SEQ ID NO: 1. Suitable fragments may comprise about 11 , about 15, about 20, about 25, about 30, about 35, about 40, about 45, about 50, about 55, about 60, about 65, about 70, about 75, about 80, about 85, about 90, about 95, about 100, about 105, about 110, about 115, about 120, about 125, about 130 or about 135 (consecutive or contiguous) amino acids from SEQ ID NO: 1.

[0064] CBM32 or a protein comprising, consisting essentially or consisting of SEQ ID NO: 1 may bind, for example, galactose, / V-acetylgalactosamine (GalNAc), / V-acetylglucosamine (GIcNAc) and / or lactose. Accordingly, any fragment for use may also bind galactose, N- acetylgalactosamine (GalNAc), / V-acetylglucosamine (GIcNAc) and / or lactose. One of skill will appreciate that the binding affinity of any given CBM32 molecule may depend on the precise CBM32 subtype; by way of further examples, some CBM32s have shown affinity for a variety of ligands (examples include type II blood group H-trisaccharide (Fuca1-2Gal pi-4GlcNAc), / V-acetyl-D-lactosamine (LacNAc), galactose, lacto- / V-biose, disaccharide GlcNAc-a-1,4-Gal (which may be referred to as an / V-acetylglucosamine linked alpha 1,4 to galactose), and / or GIcNAc). It should also be noted that multiple CBM32 subtypes may be derived from a single organism; these different CBM subtypes may exhibit the same, similar or different binding specificities. For example, Clostridium perfringens contains two sialidases NanJ and NanH; NanJ contains one galactose-specific CBM32; NanH contains four putative CBM32s with different binding selectivity - for example, the CBM32 encoded by NanH binds GIcNAc. As used herein, the term CBM32 embraces all CBM32 variants, derivatives and sub-types.

[0065] SEQ ID NO: 1 is derived from the sequence deposited in the UniProt database under ID No: A0A2X2YJF2. This sequence is reproduced as SEQ ID NO: 2 below (SEQ ID NO: 1 appears as residues 42-180 - shown in bold in the sequence below).

[0066] SEQ ID NO: 2

[0067] MKSKKI IATL VASLVI SNMG GYLVKANPNV NHKAVI IEDR QAIIETAIPQ SEMTASATSE

[0068] EGQDPASSAI DGNTNTMWHT KWNGSDALPQ SLSVNLGSSR KVSSIAITPR TSGNNGFITK

[0069] YEIHAINNGV ETLVAEGTWE ENNLVKTVTF DSPIDAEEIK ITAIQGVGGF ASIAELNVYE

[0070] IKGEVDEIAN YGNLKITKEE ERLNITRDLE KFSSLDEGTI VTRFNMNDTS IQSLIGLSDG

[0071] NKANNYFSLY VSGGKVGYEL RRQEGNGDFN VHHSADVTFN KGINTLALKI EKGVGAKI FL

[0072] NGSLVKTVSD PNIKFLNAIN LNSGFIGKTD RANGYNEYLF RGNIDFMNIY DKPVSDNYLL

[0073] RKTGETKAPS EDSLLPDDVY KTQPVELFYP GYLESRGYRI PALETTKKGT VLASIDVRNN

[0074] GDHDAPNNNI DVGIRRKEVN GEWEEGKVIL DYPGKSAAID TSLMSATIEE NGIEKERI FL

[0075] IVTHFPEGYG FPNTEGGSGY KEIDGKYYFI LKDAQNNEYT VREDGIVYNS EGNETDYVMK

[0076] NDKTLIQNGE EVGNALLSNS PLKAVGTAHI EMIYSDDDGN TWSEPEDLNP GLKKEWMKFF

[0077] GTAPGKGIQI KNGEHKGRLV FPIYYTNQNN FQSSAVIYSD DFGETWKLGE SPIDTASVSS

[0078] ETVSSGTQLT ECQWEMPNG QLKLFMRNTG SYTRIATSFD GGATWHDEVP EDTSLREPYC

[0079] QLSVINYSGK INGKDAI I FS NPDASSRVNG SVKVGLINEN GTYENGQPRY EFDWIYNKTV

[0080] KPGSFAYSCL TELPDGNLGL FYEGEGAGRM AYTEFDLNYL KFNASEDSPA ATVQSIESLD

[0081] EDLIYNAGDE VSIKVNFNQL VSLIGDRKIT LDIGGVDVPL NMVNYEGKSS AI FKGTI PEG

[0082] INPGNYEIK LKENNALELNT VYNKVSTLNG LDNTGINVQI GELKTTVGNS TIKVNEEVQV

[0083] GSAFEAILGI KGLNGDTEVY SAEYLFEYNA EAFKLNEITS FSDSLFVKSK EVEPGKVRIL

[0084] VASLGNEIEK DSELVKVNLT PKI SSELEVL GLTTALVGAG DGNTHDLELS SKEVKINEEA

[0085] SGEIWNPVQ NFEI PEINKK NVKLTWNAPI TTEGLEGYVI YKDGKKLSEV PAESTEFWS

[0086] KLNRHTIYNF KVAAKYSNGE LSAKESKTIR TAR

[0087] SEQ ID NOS 1 and 2 are derived from Clostridium perfringens. A CBM for use in the various aspects of this disclosure may comprise one, two, three, four or more CBM32s. A CBM for use may comprise one, two, three, four or more proteins comprising SEQ ID NO: 1 or a carbohydrate binding fragment thereof. One of skill will appreciate that useful CBM32s may comprise sequences which exhibit some degree (for example, 99%, 95%, 90%, 85%, 80%, 75%, 70%, 65% or 60%) of sequence identity or homology with the CBM32 sequences of SEQ ID NOS: 1 and 2. All such variant or divergent sequences are to be embraced within the scope of this disclosure and by the term “CBM32”. Identical and / or homologous CBM32 sequences may have carbohydrate binding function.

[0088] CBM40

[0089] A useful CBM40 (i.e. a CBM40 for use in a method of reducing the severity and / or number of symptoms of a viral infection) may be derived from any suitable source. For example, CBM40s for use may be obtained from microorganisms, including, for example, bacteria of the genera Clostridium, Enterococcus, Staphylococcus, Streptococcus and Vibrio. For example, useful CBM40s may be obtained or derived from, for example Clostridium perfringens, Streptococcus pneumoniae and Vibrio cholerae. Further details concerning the source, structure and function of the CBM40 family can be found within the Carbohydrate Active Enzymes database (freely available on the internet at: http: / / www.cazy.org / CBM40.html).

[0090] The Family 40 CBMs embrace molecules of approximately 200 residues and are often found at the N-terminus of GH33 sialidases. They may also be found inserted in the p-propeller of GH33 sialidases. At least the CBM40 of Vibrio cholerae binds the alpha-anomer of sialic acid and, for example, a(2,3)-, a(2,6)-, and a(2,8)-linked sialosides.

[0091] Exemplary CBM40s for use in a method of reducing the severity and / or number of symptoms of a viral infection may comprise the sialic acid binding domain of Vibrio cholerae NanH sialidase (\ / cCBM: a CBM40) and / or the equivalent (or homologous) domain from Streptococcus pneumoniae NanA sialidase (SpCBM: also a CBM40). Of course, similar or homologous sialic acid binding modules present in other organisms are to be encompassed within the scope of the terms “CBM” and “CBM40”.

[0092] An exemplary Vibrio cholerae NanH sialidase amino acid sequence is deposited under accession number A5F7A4 and is reproduced below as SEQ ID NO: 3 (781 amino acids).

[0093] SEQ ID NO: 3

[0094] MRFKNVKKTA LMLAMFGMAT SSNAALFDYN ATGDTEFDSP AKQGWQDNT NNGSGVLTNA DGMPAWLVQG IGGRAQWTYS LSTNQHAQAS SFGWRMTTEM KVLSGGMITN YYANGTQRVL PIISLDSSGN LWEFEGQTG RTVLATGTAA TEYHKFELVF LPGSNPSASF YFDGKLIRDN IQPTASKQNM IVWGNGSSNT DGVAAYRDIK FEIQGDVI FR GPDRI PSIVA SSVTPGWTA FAEKRVGGGD PGALSNTNDI ITRTSRDGGI TWDTELNLTE QINVSDEFDF SDPRPIYDPS SNTVLVSYAR WPTDAAQNGD RIKPWMPNGI FYSVYDVASG NWQAPIDVTD QVKERSFQIA GWGGSELYRR NTSLNSQQDW QSNAKIRIVD GAANQIQVAD GSRKYWTLS IDESGGLVAN LNGVSAPI IL QSEHAKVHSF HDYELQYSAL NHTTTLFVDG QQITTWAGEV SQENNIQFGN ADAQIDGRLH VQKIVLTQQG HNLVEFDAFY LAQQTPEVEK DLEKLGWTKI KTGNTMSLYG NASVNPGPGH GITLTRQQNI SGSQNGRLIY PAIVLDRFFL NVMSIYSDDG GSNWQTGSTL PI PFRWKSSS ILETLEPSEA DMVELQNGDL LLTARLDFNQ IVNGVNYSPR QQFLSKDGGI TWSLLEANNA NVFSNI STGT VDASITRFEQ SDGSHFLLFT NPQGNPAGTN GRQNLGLWFS FDEGVTWKGP IQLVNGASAY SDIYQLDSEN AIVIVETDNS NMRILRMPIT LLKQKLTLSQ N

[0095] The CBM region of SEQ ID NO: 3 is from amino acid residue 25 to 216 (sequence shown in bold) - this sequence may be SEQ ID NO: 4.

[0096] An exemplary Streptococcus pneumoniae NanA sialidase amino acid sequence has been deposited under accession number P62575 and is reproduced below as SEQ ID NO: 5 (1035 amino acids).

[0097] SEQ ID NO: 5

[0098] MSYFRNRDID IERNSMNRSV QERKCRYSIR KLSVGAVSMI VGAWFGTSP VLAQEGASEQ PLANETQLSG

[0099] ESSTLTDTEK SQPSSETELS GNKQEQERKD KQEEKIPRDY YARDLENVET VIEKEDVETN ASNGQRVDLS

[0100] SELDKLKKLE NATVHMEFKP DAKAPAFYNL FSVSSATKKD EYFTMAVYNN TATLEGRGSD GKQFYNNYND APLKVKPGQW NSVTFTVEKP TAELPKGRVR LYVNGVLSRT SLRSGNFIKD MPDVTHVQIG ATKRANNTVW

[0101] GSNLQIRNLT VYNRALTPEE VQKRSQLFKR SDLEKKLPEG AALTEKTDI F ESGRNGKPNK DGIKSYRI PA

[0102] LLKTDKGTLI AGADERRLHS SDWGDIGMVI RRSEDNGKTW GDRVTITNLR DNPKASDPSI GSPVNIDMVL

[0103] VQDPETKRI F SIYDMFPEGK GI FGMSSQKE EAYKKIDGKT YQILYREGEK GAYTIRENGT VYTPDGKATD

[0104] YRVWDPVKP AYSDKGDLYK GNQLLGNIYF TTNKTSPFRI AKDSYLWMSY SDDDGKTWSA PQDITPMVKA DWMKFLGVGP GTGIVLRNGP HKGRILI PVY TTNNVSHLNG SQSSRI IYSD DHGKTWHAGE AVNDNRQVDG

[0105] QKIHSSTMNN RRAQNTESTV VQLNNGDVKL FMRGLTGDLQ VATSKDGGVT WEKDIKRYPQ VKDVYVQMSA

[0106] IHTMHEGKEY I ILSNAGGPK RENGMVHLAR VEENGELTWL KHNPIQKGEF AYNSLQELGN GEYGILYEHT

[0107] EKGQNAYTLS FRKFNWDFLS KDLI SPTEAK VKRTREMGKG VIGLEFDSEV LVNKAPTLQL ANGKTARFMT

[0108] QYDTKTLLFT VDSEDMGQKV TGLAEGAIES MHNLPVSVAG TKLSNGMNGS EAAVHEVPEY TGPLGTSGEE PAPTVEKPEY TGPLGTSGEE PAPTVEKPEY TGPLGTAGEE AAPTVEKPEF TGGVNGTEPA VHEIAEYKGS

[0109] DSLVTLTTKE DYTYKAPLAQ QALPETGNKE SDLLASLGLT AFFLGLFTLG KKREQ

[0110] The CBM region of SEQ ID NO: 5 is from amino acid residue 121 to 305 (sequence shown in bold) - this sequence may be SEQ ID NO: 6. CBMs for use for use in a method of reducing the severity and / or number of symptoms of a viral infection may comprise a protein or peptide having the sequence of SEQ ID NO: 3, 4, 5 or 6 or a carbohydrate binding fragment of any of these. For example, a useful molecule (namely a molecule for the uses, methods and medicaments described herein) may comprise a proteinaceous moiety encoded by the sialic acid binding domain of the nanH gene (encoding sialidase) of V. cholerae (as provided by SEQ ID NO: 3) or an equivalent or homologous gene present in another organism (for example the equivalent / homologous nanA sialidase gene of S. pneumoniae’, see SEQ ID NO: 5).

[0111] A molecule for the uses, methods and medicaments of this disclosure may comprise from about residue 1 , 5, 10, 15, 25 or 30 (i.e. from 1-30 or from any amino acid residue there between) to about residue 150, 175, 200, 210, 216, 220-781 (to any residue from 150 to 781 including any residue therebetween) of the V. cholerae sialidase molecule of SEQ ID NOS: 3 and 4.

[0112] For example, a use, method or medicament of this disclosure may comprise a peptide having a sequence corresponding to residue 25 to about residue 216 of SEQ ID NO: 3 above.

[0113] A carbohydrate binding fragment of SEQ ID NOS: 3 may comprise anywhere between about 5, 6, 7, 8, 9 or 10 (consecutive or contiguous) amino acids to about 191 (consecutive or contiguous) amino acids from SEQ ID NO: 3. Suitable fragments may comprise about 11 , about 15, about 20, about 25, about 30, about 35, about 40, about 45, about 50, about 55, about 60, about 65, about 70, about 75, about 80, about 85, about 90, about 95, about 100, about 105, about 110, about 115, about 120, about 125, about 130 or about 135, about 140, about 150, about 155, about 160, about 165, about 170, about 180, about 185, about 186, about 187, about 188, about 189 or about 190 (consecutive or contiguous) amino acids from SEQ ID NO: 3.

[0114] A molecule for a use, method or medicament of this disclosure may comprise from about residue 1 , 5, 10, 15, 25 or 30 (i.e. from 1-30 or from any amino acid residue there between) to about residue 150, 175, 200, 210, 216, 220-781 (to any residue from 150 to 781 including any residue therebetween) of the V. cholerae sialidase molecule of SEQ ID NOS: 3 and 4. For example, a sialic acid binding molecule for use may comprise a peptide having a sequence corresponding to residue 25 to about residue 216 of SEQ ID NO: 3 above.

[0115] A further suitable molecule may comprise a protein or peptide having the sequence of SEQ ID NO: 5 or 6 or a carbohydrate binding fragment thereof. For example, a useful sialic acid binding molecule may comprise a proteinaceous moiety encoded by the sialic acid binding domain of the Streptococcus pneumoniae nanA gene (encoding sialidase).

[0116] A sialic acid binding molecule for use may comprise from about residue 80, 90, 100, 110, 120, 121 to 130 (i.e. from any of about residues 80 to 130 including any residue therebetween) to about residue 250, 275, 300, 305, 310, 320-1035 (i.e. to any residue from about 250-1035 including to about any residue therebetween) of the S. pneumoniae sialidase molecule of SEQ ID NOS: 5 and 6.

[0117] For example, a sialic acid binding molecule for use may comprise a peptide having a sequence corresponding to residue 121 to about residue 305 of SEQ ID NO: 5 above.

[0118] A carbohydrate binding fragment may comprise anywhere between about 5, 6, 7, 8, 9 or 10 (consecutive or contiguous) amino acids to about 185 (consecutive or contiguous) amino acids from SEQ ID NO: 5. Suitable fragments may comprise about 11 , about 15, about 20, about 25, about 30, about 35, about 40, about 45, about 50, about 55, about 60, about 65, about 70, about 75, about 80, about 85, about 90, about 95, about 100, about 105, about 110, about 115, about 120, about 125, about 130, about 135, about 140, about 145, about 150, about 155, about 160, about 165, about 170, about 175, 180, 181 , 182, 183 or 184 (consecutive or contiguous) amino acids from SEQ ID NO: 5.

[0119] SEQ ID NOS 3 and 4 are derived from Vibrio cholerae and SEQ ID NOS 5 and 6 are derived from Streptococcus pneumoniae. A CBM for use in the various aspects of this disclosure may comprise one, two, three, four or more CBM40s. A CBM for use may comprise one, two, three, four or more proteins comprising SEQ ID NO: 3, 4, 5 or 6 or a carbohydrate binding fragment any of these. One of skill will appreciate that useful CBM40s may comprise sequences which exhibit some degree (for example, 99%, 95%, 90%, 85%, 80%, 75%, 70%, 65% or 60%) of sequence identity or homology with the CBM40 sequences of SEQ ID NOS: 3, 4, 5 and / or 6. All such variant or divergent sequences are to be embraced within the scope of this disclosure and by the term “CBM40”. Identical and / or homologous CBM40 sequences may have carbohydrate / sialic acid binding function.

[0120] CBM47

[0121] A useful CBM47 (i.e. a CBM47 for the various methods, medicaments and uses described herein) may be obtained from microorganisms, including, for example, bacteria of the genera Acinetobacter, Bathymodiolus, Campylobacter, Planctomycetes, Streptococcus and Streptomyces. For example, useful CBM47s may be obtained or derived from, for example, Streptococcus mitis or Streptococcus pneumoniae. Further details concerning potential sources and the structure and function of the CBM47 family can be found within the Carbohydrate Active Enzymes database (freely available on the internet at: http: / / www.cazy.org / CBM47.html).

[0122] An exemplary CBM47 sequence is provided by SEQ ID NO: 7 below:

[0123] SEQ ID NO: 7

[0124] TPDKFNDGNLNIAYAKPTTQSSVDYNGDPNRAVDGNRNGNFNSGSVTHTRADNPSWWEVDLKKMDKVG LVKIYNRTDAETQRLSNFDVILYDNNRNEVAKKHVNNLSGESVSLDFKEKGARYIKVKLLTSGVPLSL AEVEVFRES

[0125] Accordingly, a CBM for use may comprise, consist essentially or consist of a CBM having the sequence of SEQ ID NO: 7 or a carbohydrate binding portion thereof.

[0126] A carbohydrate binding fragment of SEQ ID NO: 7 may comprise anywhere between about 5, 6, 7, 8, 9 or 10 (consecutive or contiguous) amino acids to about 144 (consecutive or contiguous) amino acids from SEQ ID NO: 7. Suitable fragments may comprise about 11 , about 15, about 20, about 25, about 30, about 35, about 40, about 45, about 50, about 55, about 60, about 65, about 70, about 75, about 80, about 85, about 90, about 95, about 100, about 105, about 110, about 115, about 120, about 125, about 130 or about 135, about 140 or about 143 (consecutive or contiguous) amino acids from SEQ ID NO: 7.

[0127] CBM47 or a protein comprising, consisting essentially or consisting of SEQ ID NO: 7 may bind L-fucose, fucosyllactose, H-trisaccharide and / or Lewisyantigen. Accordingly, any fragment for use may also bind L-fucose, fucosyllactose, H-trisaccharide and / or Lewisyantigen.

[0128] SEQ ID NO: 7 is derived from the sequence deposited in the UniProt database under ID No: A0A1Q2T229. This sequence is reproduced as SEQ ID NO: 8 below (SEQ ID NO: 7 appears as residues 601-745 - shown in bold in the sequence below):

[0129] SEQ ID NO: 8

[0130] MNKEKIKRKL ITILFVCIGM LCFGLLAGVK ADNRVQMRTT INNESPLLLS PLYGNDNGNG

[0131] LWWGNTLKGA WEAIPEDVKP YAAIELHPAK VCKPTSCIPR DTKELREWYV KMLEEAQSLN

[0132] IPVFLVIMSA GERNTVPPEW LDEQFQKYSV LKGVLNIENY WIYNNQLAPH SAKYLEVCAK YGAHFIWHDH EKWFWETIMN DPTFFEASQK YHKNLVLATK NTPIRDDAGT DSIVSGFWLS

[0133] GLCDNWGSST DTWKWWEKHY TNTFETGRAR DMRSYASEPE SMIAMEMMNV YTGGGTVYNF

[0134] ECAAYTFMTN DVPTPAFTKG IIPFFRHAIQ NPAPSKEEVV NRTKAVFWNG EGRISSLNGF

[0135] YQGLYSNDET MPLYNNGRYH ILPVIHEKID KEKISSIFPN AKILTKNSEE LSSKVNYLNS

[0136] LYPKLYEGDG YAQRVGNSWY IYNSNANINK NQQVMLPMYT NNTKSLSLDL TPHTYAVVKE

[0137] NPNNLHILLN NYRTDKTAMW ALSGNFDASK SWKKEELELA NWISKNYSIN PVDNDFRTTT

[0138] LTLKGHTGHK PQINISGDKN HYTYTENWDE NTHVYTITVN HNGMVEMSIN TEGTGPVSFP

[0139] TPDKFNDGNL NIAYAKPTTQ SSVDYNGDPN RAVDGNRNGN FNSGSVTHTR ADNPSWWEVD

[0140] LKKMDKVGLV KIYNRTDAET QRLSNFDVIL YDNNRNEVAK KHVNNLSGES VSLDFKEKGA

[0141] RYIKVKLLTS GVPLSLAEVE VFRESDGKQS EEDIDKITED KVVSTNKVAT QSSTNYEGVA

[0142] ALAVDGNKDG DYGHHSVTHT KEDSPSWWEI DLAQTEELEK LI IYNRTDAE IQRLSNFDI I

[0143] IYDSNDYEVF TQHIDSLESN NLSIDLKGLK GKKVRISLRN AGIPLSLAEV EVYTYK

[0144] SEQ ID NOS 7 and 8 are derived from Streptococcus pneumoniae.

[0145] A CBM for use in the various aspects of this disclosure may comprise one, two, three, four or more CBM47s. A CBM for use may comprise one, two, three, four or more proteins comprising SEQ ID NO: 8 or a carbohydrate binding fragment thereof.

[0146] One of skill will appreciate that useful CBM47s may comprise sequences which exhibit some degree (for example, 99%, 95%, 90%, 85%, 80%, 75%, 70%, 65% or 60%) of sequence identity or homology with the CBM47 Sequences of SEQ ID NOS: 7 and 8 All such variant or divergent sequences are to be embraced within the scope of this disclosure and by the term “CBM47”. Identical and / or homologous CBM47 sequences may have carbohydrate binding function.

[0147] CBM67

[0148] A useful CBM67 (i.e. a CBM67 for the various methods, medicaments and uses described herein) may be derived from any suitable source. For example, CBM67s for use may be obtained from microorganisms, including, for example, bacteria of the genera Bacillus, Paenibacillus, Planctomycetes and Streptomyces. For example, a useful CBM67 may be obtained or derived from, for example, Streptomyces avermitilis. Further details concerning potential sources and the structure and function of the CBM67 family can be found within the Carbohydrate Active Enzymes database (freely available on the internet at: http: / / www.cazy.org / CBM67.html). An exemplary CBM67 sequence is provided by SEQ ID NO: 9 below:

[0149] SEQ ID NO: 9

[0150] APSLEGSSWIWFPEGEPANSAPAATRWFRRTVDLPDDITGATLAISADNVYAVSVDGAEVARTDLEAD

[0151] NEGWRRPAVIDVLDHVHSGNNTLAVSASNASVGPAGWICVLVLTTASGEKKI FSDASWKSTDHEPADG

[0152] WREPDFDDSGWPAAKVAAAWGAGPWGRVA

[0153] Accordingly, a CBM for use may comprise, consist essentially or consist of a CBM having the sequence of SEQ ID NO: 9 or a carbohydrate binding portion thereof.

[0154] A carbohydrate binding fragment of SEQ ID NO: 9 may comprise anywhere between about 5, 6, 7, 8, 9 or 10 (consecutive or contiguous) amino acids to about 164 (consecutive or contiguous) amino acids from SEQ ID NO: 9. Suitable fragments may comprise about 11 , about 15, about 20, about 25, about 30, about 35, about 40, about 45, about 50, about 55, about 60, about 65, about 70, about 75, about 80, about 85, about 90, about 95, about 100, about 105, about 110, about 115, about 120, about 125, about 130, about 135 about 140, about 145, about 150, about 155, about 160 or about 163 (consecutive or contiguous) amino acids from SEQ ID NO: 9.

[0155] CBM67 or a protein comprising, consisting essentially or consisting of SEQ ID NO: 9 may bind L-rhamnose. Accordingly, any fragment for use may also bind L-rhamnose. SEQ ID NO: 9 is derived from the sequence deposited in the UniProt database under ID No: Q82PP4. This sequence is reproduced as SEQ ID NO: 10 below (SEQ ID NO: 9 appears as residues 132- 296 - shown in bold in the sequence below):

[0156] SEQ ID NO: 10

[0157] MSALRVTSPS VEYVQRPLGL DAAHPRLSWP MASAAPGRRQ SAYQVRVASS AAGLSHPDVW DSGKVVSDDS VLVPYAGPPL KPRTRYFWSV RVWDADGGAS EWSAPSWWET GLMGASQWSA KWISAPAPLT EAPSLEGSSW IWFPEGEPAN SAPAATRWFR RTVDLPDDIT GATLAISADN VYAVSVDGAE VARTDLEADN EGWRRPAVID VLDHVHSGNN TLAVSASNAS VGPAGWICVL VLTTASGEKK IFSDASWKST DHEPADGWRE PDFDDSGWPA AKVAAAWGAG PWGRVAPVAS AANQLRHEFR LPHKKVSRAR LYATALGLYE AHLNGRRVGR DQLAPGWTDY RKRVQYQTYD VTSSVRPGAN ALAAYVAPGW YAGNVGMFGP HQYGERPALL AQLEVEYADG TSERITSGPD WRAASGPIVS ADLLSGETYD ARKETAGWTS PGFDDRAWLA VRGADNDVPE QIVAQVDGPV RIAKELPARK VTEPKPGVFV LDLGQNMVGS VRLRVSGDAG TTVRLRHAEV LNPDGTIYTA NLRSAAATDT YTLKGQGEET YEPRFTFHGF RYVEVTGFPG KPSTTSVTGR VMHTSAPFTF EFETNVPMLN KLHSNITWGQ RGNFLSVPTD TPARDERLGW TGDINVFAPT AAYTMESARF LTKWLVDLRD AQTSDGAFTD VAPAVGNLGN GVAGWGDAGV TVPWALYQAY GDRQVLADAL PSVHAWLRYL EKHSDGLLRP ADGYGDWLNV SDETPKDVIA TAYFAHSADL AARMATELGK DAAPYTDLFT RIRKAFQTAY VASDGKVKGD TQSAYVLTLS MNLVPDALRK AAADRLVALI EAKDWHLSTG FLGTPRLLPV LTDTGHTDVA YRLLHQRTFP SWGYPIDKGS TTMWERWDSI QPDGGFQTPE MNSFNHYAYG SVGEWMYANI AGIAPGRAGY RQVVIRPRPG GEVTSARATF ASLHGPVSTR WQQRSGGFVL TCSVPPNTTA EVWIPADHPD RVQHTHGTFV RAEDGCAVFE VGSGSHRFTV

[0158] SEQ ID NOS 9 and 10 are derived from Streptomyces avermitilis.

[0159] A CBM for use in the various aspects of this disclosure may comprise one, two, three, four or more CBM67s.

[0160] A CBM for use may comprise one, two, three, four or more proteins comprising SEQ ID NO: 9 or a carbohydrate binding fragment thereof.

[0161] One of skill will appreciate that useful CBM67s may comprise sequences which exhibit some degree (for example 99%, 95%, 90%, 85%, 80%, 75%, 70%, 65% or 60%) of sequence identity or homology with the CBM67 Sequences of SEQ ID NOS: 9 and 10. All such variant or divergent sequences are to be embraced within the scope of this disclosure and by the term “CBM67”. Identical and / or homologous CBM67 sequences may have carbohydrate binding function.

[0162] CBM70

[0163] A useful CBM70 (i.e. a CBM70 for the various methods, medicaments and uses described herein) may be derived from any suitable source. For example, CBM70s for use may be obtained from microorganisms, including, for example, bacteria of the genera Bacillus, Paenibacillus, Planctomycetes and Streptococcus. For example, a useful CBM70s may be obtained or derived from, for example, Streptomyces pneumoniae. Further details concerning potential sources and the structure and function of the CBM70 family can be found within the Carbohydrate Active Enzymes database (freely available on the internet at: http: / / www.cazy.org / CBM70.html)

[0164] An exemplary CBM70 sequence is provided by SEQ ID NO: 11 below: SEQ ID NO: 11

[0165] NLVENGDFGQTEDGSSPWTGSKAQGWSAWVDQKNSADASTRVIEAKDGAITISSHEKLRAALHRMVPI EAKKKYKLRFKIKTDNKIGIAKVRI IEESGKDKRLWNSATTSGTKDWQTIEADYSPTLDVDKIKLELF YETGTGTVSFKDIELVEVADQLS

[0166] A carbohydrate binding fragment of SEQ ID NO: 11 may comprise anywhere between about 5, 6, 7, 8, 9 or 10 (consecutive or contiguous) amino acids to about 158 (consecutive or contiguous) amino acids from SEQ ID NO: 11. Suitable fragments may comprise about 11 , about 15, about 20, about 25, about 30, about 35, about 40, about 45, about 50, about 55, about 60, about 65, about 70, about 75, about 80, about 85, about 90, about 95, about 100, about 105, about 110, about 115, about 120, about 125, about 130, about 135, about 140, about 145, about 150, about 155 or about 157 (consecutive or contiguous) amino acids from SEQ ID NO: 11.

[0167] CBM70 or a protein comprising, consisting essentially or consisting of SEQ ID NO: 11 may bind hyaluronan. Accordingly, any fragment for use may also bind hyaluronan.

[0168] SEQ ID NO: 11 is derived from the sequence deposited in the UniProt database under ID No: Q54873. This sequence is reproduced as SEQ ID NO: 12 below (SEQ ID NO: 11 appears as residues 54-212 - shown in bold in the sequence below):

[0169] SEQ ID NO: 12

[0170] MQTKTKKLIV SLSSLVLSGF LLNHYMTIGA EETTTNTIQQ SQKEVQYQQR DTKNLVENGD FGQTEDGSSP WTGSKAQGWS AWVDQKNSAD ASTRVIEAKD GAITISSHEK LRAALHRMVP IEAKKKYKLR FKIKTDNKIG IAKVRIIEES GKDKRLWNSA TTSGTKDWQT IEADYSPTLD VDKIKLELFY ETGTGTVSFK DIELVEVADQ LSEDSQTDKQ LEEKIDLPIG KKHVFSLADY TYKVENPDVA SVKNGILEPL KEGTTNVIVS KDGKEVKKIP LKILASVKDA YTDRLDDWNG I IAGNQYYDS KNEQMAKLNQ ELEGKVADSL SSISSQADRT YLWEKFSNYK TSANLTATYR KLEEMAKQVT NPSSRYYQDE TVVRTVRDSM EWMHKHVYNS EKSIVGNWWD YEIGTPRAIN NTLSLMKEYF SDEEIKKYTD VIEKFVPDPE HFRKTTDNPF KALGGNLVDM GRVKVIAGLL RKDDQEISST IRSIEQVFKL VDQGEGFYQD GSYIDHTNVA YTGAYGNVLI DGLSQLLPVI QKTKNPIDKD KMQTMYHWID KSFAPLLVNG ELMDMSRGRS ISRANSEGHV AAVEVLRGIH RIADMSEGET KQCLQSLVKT IVQSDSYYDV FKNLKTYKDI SLMQSLLSDA GVASVPRPSY LSAFNKMDKT AMYNAEKGFG FGLSLFSSRT LNYEHMNKEN KRGWYTSDGM FYLYNGDLSH YSDGYWPTVN PYKMPGTTET DAKRADSDTG KVLPSAFVGT SKLDDANATA TMDFTNWNQT LTAHKSWFML KDKIAFLGSN IQNTSTDTAA TTIDQRKLES GNPYKVYVND KEASLTEQEK DYPETQSVFL ESFDSKKNIG YFFFKKSSIS MSKALQKGAW KDINEGQSDK EVENEFLTIS QAHKQNRDSY GYMLIPNVDR ATFNQMIKEL ESSLIENNET LQSVYDAKQG VWGIVKYDDS VSTISNQFQV LKRGVYTIRK EGDEYKIAYY NPETQESAPD QEVFKKLEQA AQPQVQNSKE KEKSEEEKNH SDQKNLPQTG EGQSILASLG FLLLGAFYLF RRGKNN

[0171] SEQ ID NOS 11 and 12 are derived from Streptococcus pneumoniae.

[0172] A CBM for use in the various aspects of this disclosure may comprise one, two, three, four or more CBM70s.

[0173] A CBM for use may comprise one, two, three, four or more proteins comprising SEQ ID NO: 11 or a carbohydrate binding fragment thereof.

[0174] One of skill will appreciate that useful CBM70s may comprise sequences which exhibit some degree (for example 99%, 95%, 90%, 85%, 80%, 75%, 70%, 65% or 60%) of sequence identity or homology with the CBM70 Sequences of SEQ ID NOS: 11 and 12. All such variant or divergent sequences are to be embraced within the scope of this disclosure and by the term “CBM70”. Identical and / or homologous CBM70 sequences may have carbohydrate binding function.

[0175] The various molecules described herein (including the various carbohydrate, sialic acid and / or glycan binding proteins / molecules) which are all for the various uses, medicaments and methods described herein, may further comprise an oligomerisation domain.

[0176] Suitable oligomerisation domains may exhibit an ability to self-associate to form multimeric structures, for example trimers. An oligomerisation domain for use may comprise any molecule with oligomerisation properties or any functional fragment thereof. For example, one or more (for example, two) sialic acid / glycan binding molecules (for example, CBMs) may be bound, coupled or fused to an oligomerisation domain - the resulting sialic acid / glycan binding molecule: :oligomerisation domain “fusion” may then be used (with one or more other such “fusions”) as a molecule for use in a method of reducing the severity and / or number of symptoms of a viral infection.

[0177] Suitable oligomerisation domains may be derived from, for example, Pseudomonas aeruginosa pseudaminidase. An exemplary Pseudomonas aeruginosa pseudaminidase sequence amino acid sequence has been deposited under accession number PAO579 and is reproduced below as SEQ ID NO: 13 (438 amino acids).

[0178] SEQ ID NO: 13

[0179] MNTYFDIPHR LVGKALYESY YDHFGQMDIL SDGSLYLIYR RATEHVGGSD GRVVFSKLEG

[0180] GIWSAPTIVA QAGGQDFRDV AGGTMPSGRI VAASTVYETG EVKVYVSDDS GVTWVHKFTL

[0181] ARGGADYNFA HGKSFQVGAR YVIPLYAATG VNYELKWLES SDGGETWGEG STIYSGNTPY

[0182] NETSYLPVGD GVILAVARVG SGAGGALRQF ISLDDGGTWT DQGNVTAQNG DSTDILVAPS

[0183] LSYIYSEGGT PHVVLLYTNR TTHFCYYRTI LLAKAVAGSS GWTERVPVYS APAASGYTSQ

[0184] VVLGGRRILG NLFRETSSTT SGAYQFEVYL GGVPDFESDW FSVSSNSLYT LSHGLQRSPR

[0185] RWVEFARSS SPSTWNIVMP SYFNDGGHKG SGAQVEVGSL NIRLGTGAAV WGTGYFGGID

[0186] NSATTRFATG YYRVRAWI

[0187] The oligomerisation domain of SEQ ID NO: 13 is from amino acid residue 333 to 438 - this sequence may be SEQ ID NO: 14 (sequence shown in bold).

[0188] Thus, an oligomerisation domain for use may comprise from about residue 250, 275, 300, 310, 320, 333, 340 to 350 (i.e. from about residue 250 to about residue 350 including from about any residue therebetween) to about residue 400, 410, 420, 430 or 438 (i.e. to about any residue from about residue 400 residue 438 including to about any residue therebetween) of the P. aeruginosa pseudaminidase trimerisation domain (PaTD) provided by SEQ ID NO: 5. For example, a useful sialic acid / glycan binding molecule may exploit an oligomerisation domain comprising residues 333 to 438 of SEQ ID NO: 13.

[0189] Sialic acid / glycan binding molecules for reducing the severity of and / or number of symptoms of a viral infection (by administration to the subject once or twice a week, or at intervals between 2 to 7 days, e.g. 3 to 7 days) may include modified forms of any of the CBMs described herein.

[0190] The term “modified” embraces molecules which contain one or more mutations relative to a reference sequence.

[0191] A “reference sequence” may be any wild type CBM sequence. For example, a reference sequence may comprise, consist essentially of or consist of a wild type family 40 CBM sequence, e.g. the wild type CBM sequences from Vibrio cholerae NanH sialidase or Streptococcus pneumoniae NanA sialidase (it should be appreciated that similar or homologous CBMs (including CBM40s) present in other organisms are to be encompassed within the scope of the term “CBM” and / or as CBM reference sequences). A reference sequence may comprise (or consist of, or consist essentially of) any of SEQ ID NOS: 1-12 above.

[0192] Accordingly, a modified CBM sequence may be derived from a specific or particular wild type CBM sequence.

[0193] A modified CBM sequence may comprise a wild type CBM sequence which has been modified to include one or more mutations. For example, a modified CBM sequence may comprise a wild type CBM sequence which has been modified to include between 1 and 30, between 1 and 20, between 1 and 10, or between 1 and 5 mutations.

[0194] The one or more mutations may be functional - that is to say they may individually (and / or independently) or collectively (for example, synergistically) modulate (alter, improve or suppress / inhibit) one or more of the physiological, biological immunological and / or pharmacological properties characteristic of a wild type CBM (for example the wild type CBM from which the modified CBM is derived). In particular, the one or more mutations may:

[0195] (i) alter the immunogenicity (or antigenicity) of the CBM; and / or

[0196] (ii) alter (for example improve) the efficacy (of the CBM or of any multimeric molecule comprising a modified CBM)’ and / or

[0197] (iii) they may modulate (for example improve) the thermostability of the CBM; and / or

[0198] (iv) they may modulate (for example improve) the solubility of the CBM; and / or

[0199] (v) they may modulate the binding affinity of the CBM for its target (for example a glycan, sialic acid, galactose, lactose, polygalacturonic acid, LacNAc, fucose, L- rhamnose; hyaluronan); and / or

[0200] (vi) they may modulate (for example improve) the in vivo half-life of the molecule.

[0201] A “mutation” may include any alteration to a wild-type CBM molecule. For example, the term “mutation” may embrace, for example:

[0202] (i) one or more amino acid substitution(s) (where one or more of the wild type amino acid(s) is / are swapped or changed for another (different) amino acid - the term “substitutions” would include conservative amino acid substitutions); and / or

[0203] (ii) one or more amino acid deletion(s) (where one or more of the wild type amino acid residue(s) are removed); and / or (iii) one or more amino acid addition(s) / insertion(s) (where additional amino acid residue(s) are added to a wild type (or reference) primary sequence); and / or

[0204] (iv) one or more amino acid / sequence inversions (usually where two or more consecutive amino acids in a primary sequence are reversed; and / or

[0205] (v) one or more amino acid / sequence duplications (where an amino acid or a part of the primary amino acid sequence (for example a stretch of 5-10 amino acids) is repeated).

[0206] Accordingly, a modified CBM according to this disclosure may comprise one or more of the mutations described herein.

[0207] An exemplary wild type CBM (in other words a reference sequence from which a useful modified CBM may be derived) is the Streptococcus pneumoniae NanA sialidase, the amino acid sequence for which has been deposited under accession number P62575 and is reproduced above as SEQ ID NO: 5 (1035 amino acids). As also noted above, the CBM region of SEQ ID NO: 5 is from amino acid residue 121 to 305 - this sequence is designated SEQ ID NO: 6.

[0208] Thus, this disclosure provides carbohydrate binding molecules with an affinity for sialic acid, which molecules comprise modified forms of SEQ ID NO: 6 (and / or SEQ ID NO: 5). A modified form of SEQ ID NO: 6 may comprise one or more mutated residues - the mutations being, for example, amino acid substitutions, additions / insertions, duplications, deletions and / or inversions made relative to the sequence of SEQ ID NO: 6. In some examples, the modified form of SEQ ID NO: 6 may comprise between 1 and 30, between 1 and 20, between 1 and 10, or between 1 and 5 mutated residues - the mutations being, for example, amino acid substitutions, additions / insertions, duplications, deletions and / or inversions made relative to the sequence of SEQ ID NO: 6. In yet further examples, the modified form of SEQ ID NO: 6 may comprise between 1 and 30, between 1 and 20, between 1 and 10, or between 1 and 5 mutated residues - the mutations being conservative amino acid substitutions made relative to the sequence of SEQ ID NO: 6.

[0209] An exemplary Vibrio cholerae NanH sialidase amino acid sequence is deposited under accession umber A5F7A4 and is reproduced above as SEQ ID NO: 7 (781 amino acids). The CBM region of SEQ ID NO: 7 is from amino acid residue 25 to 216 - this is SEQ ID NO: 8.

[0210] Thus, this disclosure provides sialic acid / glycan binding molecules which comprise modified forms of SEQ ID NO: 8 (and / or SEQ ID NO: 7). A modified form of SEQ ID NO: 8 may comprise one or more mutated residues - the mutations being, for example, amino acid substitutions, additions / insertions, duplications, deletions and / or inversions made relative to the sequence of SEQ ID NO: 8. In some examples, the modified form of SEQ ID NO: 8 may comprise between 1 and 30, between 1 and 20, between 1 and 10, or between 1 and 5 mutated residues - the mutations being, for example, amino acid substitutions, additions / insertions, duplications, deletions and / or inversions made relative to the sequence of SEQ ID NO: 8. In yet further examples, the modified form of SEQ ID NO: 8 may comprise between 1 and 30, between 1 and 20, between 1 and 10, or between 1 and 5 mutated residues - the mutations being conservative amino acid substitutions made relative to the sequence of SEQ ID NO: 8.

[0211] Thus, a useful modified CBM (i.e. a CBM for use in the medical uses and methods described herein) may comprise one or more of the mutations described herein.

[0212] By way of non-limiting example, the following represent individual units (referred to as “HEX” units) which may be used to make modified sialic acid / glycan binding molecules for the various uses, methods and medicaments described herein (for example, for use in a method of reducing the severity and / or number of symptoms of a viral infection comprising: administering the glycan binding molecule to the subject once or twice a week, or at intervals between 2 to 7 days, e.g. 3 to 7 days).

[0213] (i) HEX1

[0214] CBMX1 (L170T V239A V246G I28 6A Y292E) - CBMX2 (L170T V239A V24 6G I286A

[0215] Y292 E) - TD (S342D L348 D R403K)

[0216] (ii) HEX2

[0217] CBMX1 (V239A V246G I 28 6A Y2 92 E) - CBMX2 (V239A V24 6G I 28 6A Y2 92 E) - TD

[0218] (S342D R403K)

[0219] (iii) HEX3

[0220] CBMX1 (V239A V246G I286A)- — CBMX2 (V239A V246G I286A)- — TD (S342D R403K)

[0221] (iv) HEX4

[0222] CBMX1 (V239A V246G)- — CBMX2 (V239A V246G)- — TD (S342D)

[0223] (v) HEX5

[0224] CBMX1 (V239A V246G)- — CBMX2 (V239A V246G)- — TD (R403K)

[0225] (vi) HEX6 CBMX1 (V239A V246G) - CBMX2 (V239A V246G) - TD (S342D R403K)

[0226] (vii) HEX17

[0227] CBMX1 (V239A V246G A162 P) - CBMX2 (V239A V24 6G A162 P) - TD (S342 D R403K)

[0228] A suitable HEX unit may comprise two modified CBMs (denoted CBMX1 and CBMX2 above) with the specific mutations introduced to each CBM being identified in parenthesis. In one teaching the units CBMX1 and CBMX2 may comprise (consist of, or consist essentially of) any type of CBM (for example any member of the CBM40, any CBM32, any CBM47, any CBM67 and / or any CBM70 groups / classes). It should be noted that a symbol indicates an amino acid linker (linking one modified CBM to another modified CBM or a modified CBM to an oligomerisation domain).

[0229] In each case, the oligomerisation domain (denoted “TD”) present in each HEX unit conjugates the units together as a trimer. While any given hexamer may comprise three identical copies of one of the units described above, one of skill will appreciate that further options are available. For example, a HEX unit may be made up of two CBMs, each having different mutations (the mutations being one or more selected from the options detailed herein).

[0230] It will be noted that HEX6 and HEX17 are identical except for the additional A162P mutation. This proline mutation (a substitution for the wild type alanine at residue 162) has been shown to improve thermostability (the single CBM Tm by 3-4°C). Further information regarding the use of proline mutations may be derived from Fu 2009, 'Increasing protein stability by improving beta-turns' (DOI 10.1002 / prot.22509) which describes the general approach. The proline mutation does not affect (increase or decrease) the predicted immunogenicity of the CBM molecule, is not located near the other mutations, the N- or C-termini or the ligand binding site. Rather unexpectedly, beyond the modest improvement in thermostability, it was noted that the A162P mutation yields a molecule exhibiting a marked improvement in in vivo experiments - in particular in comparison to those same experiments conducted using a hexameric molecule comprising other (for example, HEX6) HEX units. For example, the modified molecules (in particular, a molecule comprising a HEX17 unit) exhibit modulation over pro-inflammatory cytokines, including, for example, IL-8. Indeed, the modulatory effect (specifically an inhibitory effect) on the production of IL-8 by a molecule comprising a HEX17 unit, was improved over other tested modified molecules.

[0231] Relative to the amino acid sequences of Sp2CBMTD (aka “SpOrig”) the amino acid sequence of the HEX6 and HEX17 molecules is:

[0232] SpOrig GAMVIEKEDVETNASNGQRVDLSSELDKLKKLENATVHMEFKPDAKAPAFYNLFSVSSAT HEX 6 GAMVIEKEDVETNASNGQRVDLSSELDKLKKLENATVHMEFKPDAKAPAFYNLFSVSSAT

[0233] HEX17 GAMVIEKEDVETNASNGQRVDLSSELDKLKKLENATVHMEFKPDPKAPAFYNLFSVSSAT

[0234] SpOrig KKDEYFTMAVYNNTATLEGRGSDGKQFYNNYNDAPLKVKPGQWNSVTFTVEKPTAELPKG

[0235] HEX6 KKDEYFTMAVYNNTATLEGRGSDGKQFYNNYNDAPLKVKPGQWNSVTFTVEKPTAELPKG

[0236] HEX17 KKDEYFTMAVYNNTATLEGRGSDGKQFYNNYNDAPLKVKPGQWNSVTFTVEKPTAELPKG

[0237] SpOrig RVRLYVNGVLSRTSLRSGNFIKDMPDVTHVQIGATKRANNTVWGSNLQIRNLTVYNRALT

[0238] HEX6 RARLYVNGGLSRTSLRSGNFIKDMPDVTHVQIGATKRANNTVWGSNLQIRNLTVYNRALT

[0239] HEX17 RARLYVNGGLSRTSLRSGNFIKDMPDVTHVQIGATKRANNTVWGSNLQIRNLTVYNRALT

[0240] SpOrig PEEVQKRSGGGSGVIEKEDVETNASNGQRVDLSSELDKLKKLENATVHMEFKPDAKAPAF

[0241] HEX6 PEEVQKRSGGGSGVIEKEDVETNASNGQRVDLSSELDKLKKLENATVHMEFKPDAKAPAF

[0242] HEX17 PEEVQKRSGGGSGVIEKEDVETNASNGQRVDLSSELDKLKKLENATVHMEFKPDPKAPAF

[0243] SpOrig YNLFSVSSATKKDEYFTMAVYNNTATLEGRGSDGKQFYNNYNDAPLKVKPGQWNSVTFTV

[0244] HEX6 YNLFSVSSATKKDEYFTMAVYNNTATLEGRGSDGKQFYNNYNDAPLKVKPGQWNSVTFTV

[0245] HEX17 YNLFSVSSATKKDEYFTMAVYNNTATLEGRGSDGKQFYNNYNDAPLKVKPGQWNSVTFTV

[0246] SpOrig EKPTAELPKGRVRLYVNGVLSRTSLRSGNFIKDMPDVTHVQIGATKRANNTVWGSNLQIR

[0247] HEX6 EKPTAELPKGRARLYVNGGLSRTSLRSGNFIKDMPDVTHVQIGATKRANNTVWGSNLQIR

[0248] HEX17 EKPTAELPKGRARLYVNGGLSRTSLRSGNFIKDMPDVTHVQIGATKRANNTVWGSNLQIR

[0249] SpOrig NLTVYNRALTPEEVQKRSGGALGVPDFESDWFSVSSNSLYTLSHGLQRSPRRVVVEFARS

[0250] HEX6 NLTVYNRALTPEEVQKRSGGSLGVPDFESDWFDVSSNSLYTLSHGLQRSPRRVVVEFARS

[0251] HEX17 NLTVYNRALTPEEVQKRSGGSLGVPDFESDWFDVSSNSLYTLSHGLQRSPRRVVVEFARS SpOrig SSPSTWNIVMPSYFNDGGHKGSGAQVEVGSLNIRLGTGAAVWGTGYFGGIDNSATTRFAT

[0252] HEX6 SSPSTWNIVMPSYFNDGGHKGSGAQVEVGSLNIKLGTGAAVWGTGYFGGIDNSATTRFAT

[0253] HEX17 SSPSTWNIVMPSYFNDGGHKGSGAQVEVGSLNIKLGTGAAVWGTGYFGGIDNSATTRFAT

[0254] SpOrig GYYRVRAWI

[0255] HEX 6 GYYRVRAWI

[0256] HEX17 GYYRVRAWI

[0257] [SpOrig (SEQ ID NO: 15); HEX6 (SEQ ID NO: 16) and HEX17 (SEQ ID NO: 17)]

[0258] SpOrig may be modified to include the following mutation (R274Q). This mutation modulates the binding affinity of the CBM for sialic acid.

[0259] A modified CBM comprising a sequence of SEQ ID NO: 17 may find particular application in the methods and uses described herein. Thus, the present disclosure further extends to variants thereof. In some examples, a variant of SEQ ID NO: 17 may encompass molecules comprising the general structure as shown below:

[0260] (vii) HEX17

[0261] CBMX1 (V239A V246G A162P)- — CBMX2 (V239A V246G A162P)- — TD (S342D R403K); which further comprise one or more additional amino acid mutations (as described herein) relative to the wild-type sequence SpOrig (SEQ ID NO: 15). By way of example the variant may comprise between 1 and 30, between 1 and 20, between 1 and 10, or between 1 and 5 additional amino acid mutations (such as conservative amino acid substitutions) relative to SpOrig.

[0262] In view of the above, the disclosure provides a modified CBM, for use in a method of reducing the severity of and / or number of symptoms of a viral infection in a subject comprising: administering the modified CBM to the subject once or twice a week, or at intervals between 2 to 7 days, e.g. 3 to 7 days.

[0263] Additionally disclosed, is the use of a modified CBM in the manufacture of a medicament for for use in a method of reducing the severity of and / or number of symptoms of a viral infection in a subject comprising: administering the modified CBM to the subject once or twice a week, or at intervals between 2 to 7 days, e.g. 3 to 7 days.

[0264] The disclosure provides a method of reducing the severity of and / or number of symptoms of a viral infection in a subject, said method comprising administering a subject in need thereof a modified CBM once or twice a week, or at intervals between 2 to 7 days, e.g. 3 to 7 days.

[0265] The disclosure further provides a modified CBM, for use in a method of reducing the severity of and / or number of symptoms of an influenza infection in a subject comprising: administering the modified CBM to the subject once or twice a week, or at intervals between 2 to 7 days, e.g. 3 to 7 days.

[0266] Further disclosed, is the use of a modified CBM in the manufacture of a medicament for reducing the severity of and / or number of symptoms of influenza (or ‘flu’) in a subject comprising: administering the modified CBM to the subject once or twice a week, or at intervals between 2 to 7 days, e.g. 3 to 7 days.

[0267] In a further teaching, the disclosure provides a method of reducing the severity of and / or number of symptoms of influenza (or ‘flu’) in a subject, said method comprising administering a subject in need thereof a modified CBM once or twice a week, or at intervals between 2 to 7 days, e.g. 3 to 7 days. In view of the above, the disclosure provides:

[0268] (i) HEX17; or

[0269] (ii) a sialic acid binding molecule or modified CBM comprising the following sequence: GAMVIEKEDVETNASNGQRVDLSSELDKLKKLENATVHMEFKPDPKAPAFYNLFSVSSATKKDEYFTM AVYNNTATLEGRGSDGKQFYNNYNDAPLKVKPGQWNSVTFTVEKPTAELPKGRARLYVNGGLSRTSLR SGNFIKDMPDVTHVQIGATKRANNTVWGSNLQIRNLTVYNRALTPEEVQKRSGGGSGVIEKEDVETNA SNGQRVDLSSELDKLKKLENATVHMEFKPDPKAPAFYNLFSVSSATKKDEYFTMAVYNNTATLEGRGS DGKQFYNNYNDAPLKVKPGQWNSVTFTVEKPTAELPKGRARLYVNGGLSRTSLRSGNFIKDMPDVTHV QIGATKRANNTVWGSNLQIRNLTVYNRALTPEEVQKRSGGSLGVPDFESDWFDVSSNSLYTLSHGLQR SPRRVVVEFARSSSPSTWNIVMPSYFNDGGHKGSGAQVEVGSLNIKLGTGAAVWGTGYFGGIDNSATT RFATGYYRVRAWI for use in a method of reducing the severity of and / or number of symptoms of a viral infection in a subject comprising: administering the modified CBM to the subject once or twice a week, or at intervals between 2 to 7 days, e.g. 3 to 7 days. Additionally disclosed, is the use of

[0270] (i) HEX17; or

[0271] (ii) a sialic acid binding molecule or modified CBM comprising the following sequence: GAMVIEKEDVETNASNGQRVDLSSELDKLKKLENATVHMEFKPDPKAPAFYNLFSVSSATKKDEYFTM AVYNNTATLEGRGSDGKQFYNNYNDAPLKVKPGQWNSVTFTVEKPTAELPKGRARLYVNGGLSRTSLR SGNFIKDMPDVTHVQIGATKRANNTVWGSNLQIRNLTVYNRALTPEEVQKRSGGGSGVIEKEDVETNA SNGQRVDLSSELDKLKKLENATVHMEFKPDPKAPAFYNLFSVSSATKKDEYFTMAVYNNTATLEGRGS DGKQFYNNYNDAPLKVKPGQWNSVTFTVEKPTAELPKGRARLYVNGGLSRTSLRSGNFIKDMPDVTHV QIGATKRANNTVWGSNLQIRNLTVYNRALTPEEVQKRSGGSLGVPDFESDWFDVSSNSLYTLSHGLQR SPRRVVVEFARSSSPSTWNIVMPSYFNDGGHKGSGAQVEVGSLNIKLGTGAAVWGTGYFGGIDNSATT RFATGYYRVRAWI in the manufacture of a medicament for use in a method of reducing the severity of and / or number of symptoms of a viral infection in a subject comprising: administering the modified CBM to the subject once or twice a week, or at intervals between 2 to 7 days, e.g. 3 to 7 days.

[0272] The disclosure provides a method of reducing the severity of and / or number of symptoms of a viral infection in a subject, said method comprising administering a subject in need thereof :

[0273] (i) HEX17; or

[0274] (ii) a sialic acid binding molecule or modified CBM comprising the following sequence: GAMVIEKEDVETNASNGQRVDLSSELDKLKKLENATVHMEFKPDPKAPAFYNLFSVSSATKKDEYFTM AVYNNTATLEGRGSDGKQFYNNYNDAPLKVKPGQWNSVTFTVEKPTAELPKGRARLYVNGGLSRTSLR SGNFIKDMPDVTHVQIGATKRANNTVWGSNLQIRNLTVYNRALTPEEVQKRSGGGSGVIEKEDVETNA SNGQRVDLSSELDKLKKLENATVHMEFKPDPKAPAFYNLFSVSSATKKDEYFTMAVYNNTATLEGRGS DGKQFYNNYNDAPLKVKPGQWNSVTFTVEKPTAELPKGRARLYVNGGLSRTSLRSGNFIKDMPDVTHV QIGATKRANNTVWGSNLQIRNLTVYNRALTPEEVQKRSGGSLGVPDFESDWFDVSSNSLYTLSHGLQR SPRRVVVEFARSSSPSTWNIVMPSYFNDGGHKGSGAQVEVGSLNIKLGTGAAVWGTGYFGGIDNSATT RFATGYYRVRAWI once or twice a week, or at intervals between 2 to 7 days, e.g. 3 to 7 days.

[0275] A molecule for reducing the severity of and / or number of symptoms of a viral infection in a subject, may comprise any one or more of the CBM molecules (including the modified CBMs) described herein. Molecules of this type may further comprise the abovementioned trimerisation domain allowing the formation of sialic acid / glycan binding molecules which are multivalent CBMs. For example, a molecule for use may comprise a plurality or multiple (i.e. two, three, four or more) CBMs. Molecules which comprise a plurality of CBMs may be termed “multivalent glycan or sialic acid binding molecules” or “multivalent CBMs”.

[0276] Multivalent CBM molecules, may be prepared as constructs comprising multiple (identical or different) CBMs linked by amino acid / peptide linkers. Each CBM (for example, VcCBM, SpCBM or modified CBM) may be linked to another (for example, VcCBM, SpCBM or modified CBM) or to a trimerisation domain (TD) by, for example, peptides comprising 5, 10 or 15 amino acids.

[0277] By way of example any one or more of the following peptides may be used to link two or more CBMs or a CBM to a trimerisation domain, to produce a multivalent CBM:

[0278] (i) 5 amino acid linkers: LNGS

[0279] LQ LG GGNSG GGGSG GGALG

[0280] (ii) 10 amino acid linkers: ALNGSGGGSG

[0281] LQALGGGGSL

[0282] (iii) 15 amino acid linkers: ALNGSGGGSGGGGSG

[0283] A multivalent CBM for reducing the severity of and / or number of symptoms of a viral infection in a subject may, for example, comprise two or more Family 32 CBMs, two or more Family 40 CBMs, two or more Family 47 CBMs, two or more Family 67 CBMs, two or more Family 70 CBMs or two or more of any of the modified CBMs described herein. A multivalent CBM may comprise a mix of different CBM types, for example CBMs from different CBM families, different modified CBMs, or repeats of the same CBM.

[0284] Various examples of useful mono- and multivalent glycan / sialic acid binding molecules are described herein.

[0285] For example, the molecules presented in Figure 1 may be used to reduce the severity of and / or number of symptoms of a viral infection in a subject. A molecule for reducing the severity of and / or number of symptoms of a viral infection in a subject may comprise (consist essentially of, or consist of) one or more (for example, two, three, four or more) VcCBMs (that is a CBM40 derived from Vibrio cholerae). The CBM (for example the VcCBM) may be fused, bound or conjugated to an oligomerisation domain (such as a PaTD or oligomerisation fragment thereof). A glycan / sialic acid binding molecule may comprise, consist or consist essentially of two fused (or bound) CBMs which are in turn fused to an oligomerisation domain (see, for example, molecule Vc2CBMTD shown in Figure 1).

[0286] Every one of the disclosed molecules alone or in combination with another disclosed molecule, may be for use in reducing the severity of and / or number of symptoms of a viral infection in a subject.

[0287] Any of the disclosed molecules - especially the modified CBM molecules (for reducing the severity of and / or number of symptoms of a viral infection in a subject), may be generated using PCR-based cloning techniques and a suitable method for the generation of multivalent molecules of this type is described in, for example, Connaris et al, 2009 (Enhancing the Receptor Affinity of the Sialic Acid-Binding Domain of Vibrio cholerae Sialidase through Multivalency; J. Biol. Chem; Vol. 284(11); pp 7339-7351). For example, multivalent CBM molecules, including the likes of HEX17, Vc2CBM, Vc4CBM and Sp2CBM as disclosed herein, may be prepared as constructs comprising multiple CBMs linked by amino acid / peptide linkers - such as those described above.

[0288] In the context of this disclosure, molecules (carbohydrate binding molecules, sialic acid binding molecules, glycan binding proteins / molecules and / or CBMs) for use in reducing the severity of and / or number of symptoms of a viral infection in a subject, may comprise one or more CBMs selected from the group consisting of:

[0289] (i) one or more (for example, 2, 3, 4 or more) Family 32 CBMs;

[0290] (ii) one or more (for example, 2, 3, 4 or more) Family 40 CBMs;

[0291] (iii) one or more (for example, 2, 3, 4 or more) Family 47 CBMs;

[0292] (iv) one or more (for example, 2, 3, 4 or more) Family 67 CBMs;

[0293] (v) one or more (for example, 2, 3, 4 or more) Family 70 CBMs; and

[0294] (vi) a modified CBM (as described herein).

[0295] Additionally, molecules (carbohydrate binding molecules, sialic acid binding molecules, glycan binding molecules / proteins and / or CBMs) for use in reducing the severity of and / or number of symptoms of a viral infection in a subject, may comprise a CBM selected from the group consisting of: (i) a Clostridium perfringens CBM32 (CpCBM32);

[0296] (ii) a Streptococcus pneumoniae CBM40 (SpCBM40);

[0297] (iii) a Vibrio cholerae CBM40 (\ / cCBM40);

[0298] (iv) a Streptococcus pneumoniae CBM47(SpCBM47);

[0299] (v) a Streptomyces avermitilis CBM67 (SaCBM67);

[0300] (vi) a Streptococcus pneumoniae CBM70 (SpCBM70);

[0301] (vii) a Vibrio cholerae NanH sialidase CBM;

[0302] (vii) a Vibrio cholerae NanH sialidase CBM sialic acid binding fragment thereof.

[0303] (xi) a Streptococcus pneumoniae nanA sialidase CBM; and

[0304] (x) a Streptococcus pneumoniae nanA sialidase CBM sialic acid binding fragment thereof.

[0305] A multivalent CBM for reducing the severity of and / or number of symptoms of a viral infection in a subject may comprise:

[0306] (i) a \ / cCBM; or

[0307] (ii) two (or more) \ / cCBM(s); or

[0308] (iii) three or four (or more) \ / cCBM(s); or

[0309] (iv) a SpCBM; or

[0310] (v) two (or more) SpCBM(s); or

[0311] (vi) three or four (or more) SpCBM(s); or

[0312] (vii) a CpCBM; or

[0313] (viiiO two (or more) CpCBM(s); or

[0314] (viii) three or four (or more) CpCBM(s); or

[0315] (ix) a SaCBM; or

[0316] (x) two (or more) SaCBM(s); or

[0317] (xi) three or four (or more) SaCBM(s).

[0318] A multivalent CBM for reducing the severity of and / or number of symptoms of a viral infection in a subject may comprise a mixture of different CBMs, for example, one or more CBM32s with one more other CBMs selected from the group consisting of:

[0319] (i) a Family 40 CBM

[0320] (ii) a Family 47 CBM

[0321] (iii) a Family 67 CBM

[0322] (iv) a Family 70 CBM For example, reducing the severity of and / or number of symptoms of a viral infection in a subject may comprise using the combination of a CBM32 with a Family 40 CBM (a CBM40).

[0323] Alternatively, a multivalent CBM for reducing the severity of and / or number of symptoms of a viral infection in a subject may comprise a mixture of different CBMs, for example one or more CBM40s with one more other CBMs selected from the group consisting of:

[0324] (i) a Family 32 CBM

[0325] (ii) a Family 47 CBM

[0326] (iii) a Family 67 CBM

[0327] (iv) a Family 70 CBM

[0328] For example, reducing the severity of and / or number of symptoms of a viral infection in a subject may use the combination of a CBM40 with a Family 32 CBM (a CBM32).

[0329] A multivalent CBM for reducing the severity of and / or number of symptoms of a viral infection in a subject may comprise a mixture of different CBMs, for example one or more CBM47s with one more other CBMs selected from the group consisting of:

[0330] (i) a Family 32 CBM

[0331] (ii) a Family 40 CBM

[0332] (iii) a Family 67 CBM

[0333] (iv) a Family 70 CBM

[0334] A multivalent CBM for reducing the severity of and / or number of symptoms of a viral infection in a subject may comprise a mixture of different CBMs, for example one or more CBM67s with one more other CBMs selected from the group consisting of:

[0335] (i) a Family 32 CBM

[0336] (ii) a Family 40 CBM

[0337] (iii) a Family 47 CBM

[0338] (iv) a Family 70 CBM

[0339] A multivalent CBM for reducing the severity of and / or number of symptoms of a viral infection in a subject may comprise a mixture of different CBMs, for example one or more CBM70s with one more other CBMs selected from the group consisting of: (i) a Family 32 CBM

[0340] (ii) a Family 40 CBM

[0341] (iii) a Family 47 CBM

[0342] (iv) a Family 67 CBM

[0343] Multivalent CBMs for reducing the severity of and / or number of symptoms of a viral infection in a subject may include, for example, molecules selected from the group consisting of:

[0344] (i) Cp2CBM32TD (comprising, consisting essentially of or consisting of: 2 CBMs (CBM32s) from Clostridium perfringens fused to a trimerisation domain);

[0345] (ii) Sp2CBM40TD (comprising, consisting essentially of or consisting of: 2 CBMs (CBM40s) from Streptococcus pneumoniae fused to a trimerisation domain;

[0346] (iii) \ / c2CBM40TD (comprising, consisting essentially of or consisting of: 2 CBMs (CBM40s) from Vibrio cholerae fused to a trimerisation domain; and

[0347] (iv) \ / c4CBM (comprising, consisting essentially of or consisting of: 4 CBMs (CBM40s) from Vibrio cholerae)

[0348] (v) Sp2CBM47TD comprising, consisting essentially of or consisting of: 2 CBMs (CBM47s) from Streptococcus pneumoniae fused to a trimerisation domain);

[0349] (vi) Sa2CBM67TD comprising, consisting essentially of or consisting of: 2 CBMs (CBM67s) from Streptococcus avermitilis fused to a trimerisation domain);

[0350] (vii) Sp2CBM70TD comprising, consisting essentially of or consisting of: 2 CBMs (CBM70s) from Streptococcus pneumoniae fused to a trimerisation domain).

[0351] It should be noted that this disclosure provides uses (compositions, methods and medicaments for reducing the severity of and / or number of symptoms of a viral infection in a subject) which comprise not only any one or more of the CBMs disclosed herein, in isolated form (that is uses in which at least the CBM component (of the therapeutic molecule) comprises or consists essentially of one or more of the CBM sequence(s) described herein and / or functional fragments thereof), but also uses in which the CBM component of the therapeutic molecule is comprised within a larger molecule. By way of example, the various CBMs described herein may be provided and / or used (to reduce the severity of and / or number of symptoms of a viral infection in a subject) in the form of large molecules comprising a CBM component. The CBM component (e.g. the sialic acid binding molecule) may itself comprise (consist of or consist essentially of), for example, any one of the CBMs described herein (including, for example CBM32, CBM40, CBM47, CBM67 and CBM70). By way of (nonlimiting) example, molecules (e.g. the CBMs and / or glycan / sialic acid binding molecules) of this disclosure may not only exhibit an ability to bind a glycan (or a component thereof) or sialic acid, but may also have one or more other functions. For example, the molecules may have enzymatic activity. For example, a useful molecule may comprise a CBM (as described herein) and exhibit some sialidase activity.

[0352] A molecule for use in reducing the severity of and / or number of symptoms of a viral infection in a subject may be a fusion protein comprising an enzymatic portion and a glycan / sialic acid binding portion - wherein the glycan / sialic acid binding portion comprises a CBM as described herein. In such cases, the enzymatic portion may be fused to the glycan / sialic acid binding portion. As stated, the enzymatic portion of any useful fusion protein may comprise (or have, or exhibit) sialidase activity.

[0353] In one embodiment, the sialic acid binding molecule, glycan binding molecule, or CBM for reducing the severity of and / or number of symptoms of a viral infection in a subject , may not be provided as part of, or comprised within, a molecule (for example a fusion protein) with enzymatic (for example sialidase) activity. Additionally or alternatively, the sialic acid / glycan binding molecule may not (i) bind heparin or heparin sulfate and / or (ii) comprise the GAG- binding domain of a protein that binds heparin or heparin sulfate moieties. A construct comprising the sialic acid / glycan binding molecule, glycan binding molecule, or CBM, may not show or exhibit enzymatic (for example sialidase) activity.

[0354] This disclosure also provides compositions, in particular pharmaceutical compositions which may be used to reduce the severity of and / or number of symptoms of a viral infection in a subject.

[0355] As such, any of the disclosed molecule(s) (for example, the CBMs (modified, multivalent or otherwise) or glycan binding molecules) for reducing the severity of and / or number of symptoms of a viral infection in a subject may be formulated for subsequent use. For convenience, it should be noted that the term “CBM” embraces, all monovalent, multivalent and modified CBM molecules described herein.

[0356] For example, a sialic acid binding molecule, a glycan binding molecule or CBM may be formulated as a therapeutic or pharmaceutical composition. The various compositions may comprise one or more of the sialic acid binding molecule(s) / glycan binding molecule(s) / CBM(s) as described herein and one or more pharmaceutically acceptable excipients. For example, the pharmaceutical preparations comprising the molecules described herein may be mixed with stabilisers, wetting agents, emulsifiers, salts (for use in influencing osmotic pressure), buffers and / or other substances that do not react deleteriously with the active compounds. The therapeutic uses of this disclosure may require the administration (together, concurrently or separately) of one or more of these compositions, which compositions may comprise one or more different CBMs.

[0357] Pharmaceutical compositions comprising any of the glycan / sialic acid-binding or CBMs described herein, may be prepared for oral, mucosal, intranasal or parenteral (intravenous) administration. Those formulations for mucosal or intranasal administration may be prepared conventionally, comprising substances that are customarily used in pharmaceuticals and as described in, for example, Remington's The Sciences and Practice of Pharmacy, 22nd Edition (Pharmaceutical Press 2012) and / or Handbook of Pharmaceutical Excipients, 7th edition (compiled by Rowe et al, Pharmaceutical Press, 2012) - the entire content of all of these documents and references being incorporated by reference.

[0358] A pharmaceutical composition of this disclosure may be sterile and may comprise any of the glycan / sialic acid-binding or CBMs described herein together with one or more pharmaceutically acceptable excipients / diluents.

[0359] A composition for administration to a subject in accordance with this disclosure, may be prepared so that is comprises the required dose of glycan-binding molecule, sialic acid-binding molecule or CBM.

[0360] Any suitable amount of a sialic acid binding molecule, glycan binding molecule or CBM may be used to reduce the severity of and / or number of symptoms of a viral infection in a subject. For example, whether a composition comprising a sialic acid binding molecule, glycan binding molecule or CBM is to be administered intravenously or mucosally (for example, intranasally e.g. by intranasal spray) the dose of sialic acid binding molecule / glycan binding molecule / CBM may comprise anywhere between about 0.05 mg and about 5 mg, between about 0.5 mg and about 4 mg, between about 1 mg and about 3 mg, or between about 2 mg and about 3 mg, such as about 2.8 mg.

[0361] Each dose may be formulated (or diluted) in a volume of excipient or diluent (for example pharmaceutic diluent or excipient), for example in about 10pl to about 500pl of diluent / excipient. Each dose may be diluted in about 50 pl to about 300 pl, or about 100 pl to about 200 pl of excipient / diluent. For example, a dose (for example between about 0.05 mg and about 5 mg, between about 0.5 mg and about 4 mg, between about 1 mg and about 3 mg, or between about 2 mg and about 3 mg, such as about 2.8 mg) of any of the disclosed glycan / sialic acid binding (or CBM containing) molecules, may be formulated in about 110 pl, 120 pl, 130 pl, 140 pl, 150 pl, 160 pl, 170 pl, 180 pl, or 190 pl.

[0362] Liquid dosage forms for oral and / or intranasal administration (e.g. by intranasal spray) may include emulsions, solutions, suspensions, syrups, and elixirs. In addition to the compound or composition, the liquid dosage forms may contain inert diluents commonly used in the art, such as water or other solvents, solubilising agents and emulsifiers.

[0363] A composition of this disclosure (for example, a composition comprising any of the sialic acid binding molecules, glycan binding molecules or CBMs disclosed herein) may be administered (prophylactically) to a subject at regular and / or predetermined times. For example, a composition described herein may be administered at least 3 days before the subject enters or encounters a scenario during which they might be vulnerable and / or susceptible to any of the viral infections described herein - including an influenza infection.

[0364] A glycan / sialic acid binding molecule, CBM or composition comprising the same may be administered (at any of the doses described herein) once or twice a week, or at intervals between 2 to 7 days, e.g. 3 to 7 days.

[0365] A glycan / sialic acid binding molecule, CBM or composition comprising the same, may be administered at intervals between 3 to 5 days, intervals between 3 and 4 days, or intervals of 3 days.

[0366] A glycan / sialic acid binding molecule, CBM or composition comprising the same, may be administered no more than once a week, or no more than twice a week on non-consecutive days.

[0367] A glycan / sialic acid binding molecule, CBM or composition comprising the same, may be administered to the subject for a period of at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks, or for a period of at least 1 month, at least 2 months, at least 3 months, or at least 4 months, at least 5 months or at least 6 months.

[0368] The present disclosure will now be described, by way of example only, with reference to the following figures which show:

[0369] Figure 1 : Building blocks of the multivalent CBM forms and their affinities for sialic acid, a, \ / cCBM, residues 25-216 of the V. cholerae sialidase (PDB:1w0p) with a-2,3-sialyllactose drawn as spheres, b, SpCBM, residues 121-305 of S. pneumoniae NanA sialidase with a-2,3- sialyllactose (PDB:4c1w). c, TD, the trimerisation domain, residues 333-438, of the P. aeruginosa pseudaminidase (PDB:2w38) in rainbow colours; the other two monomers in single colours, d, Multivalent forms: their molecular weights, valencies and binding affinities for a2,3-sialyllactose as determined by surface plasmon resonance (SPR) at 25°C (KD values for \ / cCBM, \ / c2CBM and Vc3CBM had been reported previously (Connaris et al, 2009)). Tandem repeat CBMs, and oligomeric CBMs fused to TD are linked by a 5-amino linker (details in Connaris, H. et al., (2014). PNAS 111 :6401-6406).

[0370] Figure 2: Study Schematic.

[0371] Figure 3: Graph showing rate of symptomatic Infection (confirmed by qRT-PCR). Results show a clinically significant reduction of -50% in symptomatic infections based on qRT PCR test. * Denotes statistical significance; p<0.05.

[0372] Figure 4: Graph showing rate of symptomatic Infection (confirmed by viral culture). The results show a clinically significant reduction of -63% in symptomatic infections based on viral culture. * Denotes statistical significance; p<0.05.

[0373] Figure 5: Graph showing viral load based on qRT PCR test. * Denotes statistical significance; p<0.05.

[0374] Figure 6: Graph showing viral load based on viral culture. * Denotes statistical significance; p<0.05.

[0375] Figure 7: Graph showing peak viral load as determined by qRT-PCR. Results show a significant reduction in peak viral load. * Denotes statistical significance; p<0.05.

[0376] Figure 8: graph showing peak viral load as determined by viral culture. Results show a significant reduction in peak viral load. * Denotes statistical significance; p<0.05.

[0377] Figure 9: Graph showing the Mean TSS per dose over time. The results show that a single dose on D- 3 significantly reduces the symptoms and severity based on the Total Symptom Score.

[0378] Influenza Human Challenge Study A randomized, double blind, placebo controlled, Phase 2 study was carried out to assess the safety, tolerability, and prophylactic antiviral activity against influenza of Neumifil (a modified CBM: HEX17) via a human viral challenge model in healthy adult participants. The study details are outlined below.

[0379] Study Overview:

[0380] Controlled Human Infection Model (CHIM)

[0381] • Controlled study with defined timelines & population

[0382] • Healthy subjects (18-55)

[0383] Primary Objective

[0384] To evaluate the effect of Neumifil (a liquid vehicle comprising a modified CBM: HEX17 as described herein)

[0385] • In reducing the incidence of symptomatic influenza infection and / or

[0386] • In reducing the severity of symptoms after influenza viral challenge, compared to placebo

[0387] End Points: Infection & Symptomology

[0388] Cohorts: 3 cohorts; 100 subjects

[0389] Challenge Strain: Influenza (H3N2)

[0390] Dose: 2.8 mg dose active (HEX17) or Placebo (liquid vehicle only) intranasal dosing, 140 pL per nostril

[0391] The total symptom score (TSS) was calculated using the methodology outlined below.

[0392]

[0393] Results

[0394] • Neumifil delivers up to 63% relative reduction in symptomatic infection • Significant reduction of the incidence of symptomatic infection

[0395] • Neumifil significantly reduces viral load and the severity of symptoms

[0396] • Potential to reduce dosing frequency

[0397] Significant Reduction of Symptomatic Infection (qRT-PCR) * Denotes statistical significance; p<0.05

[0398] Significant Reduction of Symptomatic Infection (viral culture)

[0399] Denotes statistical significance; p<0.05

[0400] Viral load by qRT-PCT

[0401] Denotes statistical significance; p<0.05

[0402] Viral Load by viral culture

[0403] Denotes statistical significance; p<0.05

[0404] Viral Load (VLPeak) by qRT-PCR

[0405] Denotes statistical significance; p<0.05 Viral Load (VLPeak) by culture

[0406] Denotes statistical significance; p<0.05

[0407] Severity of symptoms * Denotes statistical significance; p<0.05

[0408] Symptoms & Severity

[0409] Denotes statistical significance; p<0.05

Claims

CLAIMS1. A glycan binding molecule for use in a method of reducing the severity of and / or number of symptoms of a viral infection in a subject comprising: administering the glycan binding molecule to the subject once or twice a week, or at intervals between 2 to 7 days or 3 to 7 days.

2. The glycan binding molecule for use of claim 1 , wherein the glycan binding molecule is administered at intervals between 3 to 5 days, intervals between 3 and 4 days, or intervals of 3 days.

3. The glycan binding molecule for use of claim 1 wherein the glycan binding molecule is administered no more than once a week, or no more than twice a week on non-consecutive days.

4. The glycan binding molecule for use of claim 1 , wherein the glycan binding molecule is administered to the subject for a period of at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, or at least 6 weeks, or for a period of at least 1 month, at least 2 months, at least 3 months, or at least 4 months, at least 5 months or at least 6 months.

5. The glycan binding molecule for use of any one of the preceding claims, wherein the glycan binding molecule is administered in a dosage amount of between about 0.05 mg and about 5 mg, between about 0.5 mg and about 4 mg, between about 1 mg and about 3 mg, or between about 2 mg and about 3 mg, such as about 2.8 mg.

6. The glycan binding molecule for use of any one of the preceding claims, wherein the glycan binding molecule is prophylactically administered to the subject.

7. The glycan binding molecule for use of any one of the preceding claims, wherein the viral infection is caused by a virus selected from the group consisting of: influenza virus, human parainfluenza virus, respiratory syncytial virus, rhinovirus, HMPV and coronavirus infection, or a combination thereof.

8. The glycan binding molecule for use of claim 7, wherein the virus is influenza virus.

9. The glycan binding molecule for use of any one of the preceding claims, wherein the symptoms of the viral infection are selected from the group consisting of: fever (including symptoms of chilliness and feverishness), cough, sneezing, sore throat, runny or stuffy nose,aches (e.g. muscle aches), pains (e.g. muscle and / or joint pain), headaches, fatigue (including malaise / tiredness), earache, vomiting, diarrhoea, shortness of breath, change or loss in taste and / or smell, and combinations thereof.

10. The glycan binding molecule for use of any one of the preceding claims, wherein the symptoms of the viral infection are selected from the group consisting of: runny nose, stuffy nose, sneezing, sore throat, earache, malaise / tiredness / fatigue, headache, muscle and / or joint pain, chilliness / feverishness, cough and shortness of breath.11 . The glycan binding molecule for use of any one of the preceding claims, wherein the severity of symptoms is measured and / or determined by a total symptom score (TSS).

12. The glycan binding molecule for use of any one of the preceding claims, wherein the likelihood of the subject presenting a total symptom score of grade 2 or higher is reduced.

13. The glycan binding molecule for use of any one of the preceding claims, wherein the area under curve (AUC) for the total symptom score over time is reduced for the subject.

14. The glycan binding molecule for use of any one of the preceding claims, wherein the severity of and / or number of symptoms is reduced in a subject in a period of at least 1 day, at least 2 days, at least 3 days, at least 4 days, at least 5 days, at least 6 days, at least 7 day, at least 8 days, at least 9 days, at least 10 days, at least 11 days, at least 12 days, at least 13 days or at least 14 days following a viral infection.

15. The glycan binding molecule for use of any one of the preceding claims, wherein:(i) the severity of and / or number of symptoms is reduced in a subject infected by the virus who has been administered the glycan binding molecule in comparison to a subject infected by the virus who has not been administered the glycan binding molecule; and / or(ii) the severity of and / or number of symptoms is reduced in a subject infected by the virus who has been administered the glycan binding molecule in accordance with claim 1 in comparison to a subject who has not been administered a glycan binding molecule in accordance with claim 1.

16. The glycan binding molecule for use of any one of the preceding claims, wherein at least one of: the likelihood of viral infection is reduced; and / or the likelihood of symptomatic viral infection is reduced.

17. The glycan binding molecule for use of any one of the preceding claims, wherein the subject is a mammalian subject, optionally a human subject.

18. The glycan binding molecule for use of any one of the preceding claims, wherein the subject is a subject that is deemed to be at risk of, vulnerable to, or susceptible to, a viral infection.

19. The glycan binding molecule for use of any one of the preceding claims, wherein the subject has an underlying disease or condition which is susceptible to virus-induced exacerbation.

20. The glycan binding molecule for use of 19, wherein the underlying disease or condition susceptible to virus-induced exacerbation is selected from chronic obstructive pulmonary disease (COPD), asthma, cystic fibrosis, non-cystic fibrosis bronchiectasis, pulmonary fibrosis, primary ciliary dyskinesia, diabetes, neutropenia, and haematological cancers; or wherein the subjects are selected from subjects who are receiving chemotherapy that is likely to induce neutropenia, subjects undergoing conditioning for haemopoietic stem cell transplant (HSCT), subjects who are recovering from an HSCT, and subjects who have had a lung transplant.

21. The glycan binding molecule for use of any one of the preceding claims, wherein the glycan binding molecule is a sialic acid binding molecule and / or comprises one or more carbohydrate binding module(s) (CBMs), optionally wherein the glycan binding molecule comprises at least two carbohydrate binding modules.

22. The glycan binding molecule for use of any one of the preceding claims, wherein the glycan binding molecule does not exhibit sialidase activity.

23. The glycan binding molecule for use of any one of the preceding claims, wherein the glycan binding molecule does not bind heparin or heparin sulfate and / or does not comprise the GAG-binding domain of a protein that binds heparin or heparin sulfate.

24. The glycan binding molecule for use of any one of the preceding claims, wherein the glycan binding molecule comprises a CBM selected from the group consisting of:(i) a Family 40 CBM (CBM40);(ii) a Family 32 CBM (CBM32).

25. The glycan binding molecule for the use any one of the preceding claims, wherein the glycan binding molecule comprises a CBM selected from the group consisting of:(i) a Clostridium perfringens CBM32 (CpCBM32);(ii) a Streptococcus pneumoniae CBM40 (SpCBM40);(iii) a Vibrio cholerae CBM40 (\ / cCBM40);(iv) a Vibrio cholerae NanH sialidase CBM;(v) a Vibrio cholerae NanH sialidase CBM sialic acid binding fragment thereof.(vi) a Streptococcus pneumoniae nanA sialidase CBM; and(vii) a Streptococcus pneumoniae nanA sialidase CBM sialic acid binding fragment thereof.

26. The glycan binding molecule for the use of any one of claims 1 to 23, wherein the glycan binding molecule is selected from the group consisting of:(i) Cp2CBM32TD, wherein Cp2CBM32TD comprises, consists essentially of or consists of: two CBMs (CBM32s) from Clostridium perfringens fused to a trimerisation domain;(ii) Sp2CBM40TD, wherein Sp2CBM40TD comprises, consists essentially of or consists of: 2 CBMs (CBM40s) from Streptococcus pneumoniae fused to a trimerisation domain;(iii) \ / c2CBM40TD, wherein \ / c2CBM40TD comprises, consists essentially of or consists of: 2 CBMs (CBM40s) from Vibrio cholerae fused to a trimerisation domain; and(iv) \ / c4CBM, wherein \ / c4CBM comprises, consists essentially of or consists of: 4 CBMs (CBM40s) from Vibrio cholerae.

27. The glycan binding molecule for the use of any one of claims 1 to 23, wherein the glycan binding molecule is a CBM32 and comprises the amino acid sequence of SEQ ID NO: 1 :AI IETAI PQSEMTASATSEEGQDPASSAIDGNTNTMWHTKWNGSDALPQSLSVNLGSSRKVSS IAITP RTSGNNGFITKYE IHAINNGVETLVAEGTWEENNLVKTVT FDSPIDAEEIKITAIQGVGGFAS IAELN VYE or a glycan / carbohydrate binding fragment thereof.

28. The glycan binding molecule for the use of any one of claims 1 to 23 wherein the glycan binding molecule is a CBM40 and comprises the amino acid sequence of SEQ ID NO: 4: ALFDYNATGDTEFDSPAKQGWMQDNTNNGSGVLTNADGMPAWLVQGIGGRAQWTYSLSTNQHAQASSFGWRMTTEMKVLSGGMITNYYANGTQRVLPI I SLDSSGNLWEFEGQTGRTVLATGTAATEYHKFELVFLPGSNPSASFYFDG KLIRDNIQPTASKQNMIVWGNGSSNTDGVAAYRDIKFEIQGDor SEQ ID NO: 6VIEKEDVETNASNGQRVDLSSELDKLKKLENATVHMEFKPDAKAPAFYNLFSVSSATKKDEYFTMAVYNNTATLE GRGSDGKQFYNNYNDAPLKVKPGQWNSVTFTVEKPTAELPKGRVRLYVNGVLSRTSLRSGNFIKDMPDVTHVQIG ATKRANNTVWGSNLQIRNLTVYNRALTPEEVQKRS or a glycan / sialic acid binding fragment of either.

29. The glycan binding molecule for use of any one of claims 1 to 23, wherein the glycan binding molecule is a modified CBM, wherein the modified CBM comprises a wild type CBM sequence which has been modified to include one or more mutations.

30. The glycan binding molecule for use of claim 29, wherein the modified CBM comprises the sequence of SEQ ID NO: 17:GAMVIEKEDVETNASNGQRVDLSSELDKLKKLENATVHMEFKPDPKAPAFYNLFSVSSATKKDEYFTM AVYNNTATLEGRGSDGKQFYNNYNDAPLKVKPGQWNSVTFTVEKPTAELPKGRARLYVNGGLSRTSLR SGNFIKDMPDVTHVQIGATKRANNTVWGSNLQIRNLTVYNRALTPEEVQKRSGGGSGVIEKEDVETNA SNGQRVDLSSELDKLKKLENATVHMEFKPDPKAPAFYNLFSVSSATKKDEYFTMAVYNNTATLEGRGS DGKQFYNNYNDAPLKVKPGQWNSVTFTVEKPTAELPKGRARLYVNGGLSRTSLRSGNFIKDMPDVTHV QIGATKRANNTVWGSNLQIRNLTVYNRALTPEEVQKRSGGSLGVPDFESDWFDVSSNSLYTLSHGLQR SPRRVVVEFARSSSPSTWNIVMPSYFNDGGHKGSGAQVEVGSLNIKLGTGAAVWGTGYFGGIDNSATT RFATGYYRVRAWI or a glycan acid binding fragment thereof.

31. The glycan binding molecule for use of any one of the preceding claims, wherein the glycan binding molecule is administered mucosally, optionally intranasally.

32. The glycan binding molecule for use of claim 31 , wherein the glycan binding molecule is administered as an intranasal spray33. The glycan binding molecule for use of any one of the preceding claims, wherein the glycan binding molecule is administered to the subject in a pharmaceutical composition comprising one or more additional excipients and / or carriers.

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