Treatment of viral infections

Abstract

Molecules useful in the treatment or prevention of viral infections in humans and animals and / or the treatment or prevention of associated symptoms, diseases, and / or conditions are disclosed. The present disclosure provides molecules, including sugar (carbohydrate / polysaccharide / sialic acid / glycan) binding proteins, that are useful in the treatment or prevention of coronavirus infection and / or the diseases, symptoms, and / or conditions caused or contributed to thereby.

Description

Technical Field

[0001] The present disclosure provides molecules for use in compositions, medicaments and methods for the treatment or prevention of viral diseases and / or conditions.

Background Art

[0002] Viral pathogens can cause a range of diseases and / or conditions in humans, and while vaccines and antiviral therapies are available for some pathogens, there are others for which there are no useful treatment options. In some cases, the treatment of viral infections is limited to patient isolation (to minimize the risk of further spread of infection) and the treatment of symptoms. In some cases, viral pathogens, particularly respiratory viral pathogens, can cause secondary complications such as dyspnea, pneumonia, bronchitis and / or bronchiolitis. The young, the elderly, immunocompromised patients, pregnant women, and individuals with underlying diseases are often most at risk from viral pathogens. The pathology of any given viral pathogen is diverse, but pathogens that cause respiratory disease will most often be transmitted by aerosolized droplets produced by, for example, coughing and sneezing and body fluids / secretions from / to the mucosa (including the respiratory tract, lungs, nose, mouth and eyes). Occasionally, an outbreak of a virus is associated with a species and / or strain of pathogen that is new to science. Often there is no effective treatment for such infections, and it is necessary to halt and control the outbreak to avoid uncontrolled spread. Severe acute respiratory syndrome (SARS) is a viral respiratory disease of zoonotic origin. The disease is caused by the SARS coronavirus and the symptoms can include fever and / or a "flu"-like illness with muscle pain, cough, sore throat, and other non-specific symptoms (including malaise / lassitude). Infection with the SARS coronavirus can also result in dyspnea, viral pneumonia and / or secondary bacterial pneumonia. In many cases, vaccines are difficult to develop and take considerable time to manufacture and test. Furthermore, antiviral drugs have uncertain efficacy, and in many cases, the viral infection may subside before the drug can take effect. Therefore, there is a need for novel and effective treatments to combat viral infections, and especially viral respiratory infections. [Overview of the Initiative]

[0003] Molecules that can be used in the treatment or prevention of viral infections in humans and animals, and / or in the treatment or prevention of related diseases and / or conditions are disclosed herein. The molecules described may find specific applications in the treatment or prevention of viral respiratory pathogens, including, for example, pathogens belonging to the Coronaviridae family. [Modes for carrying out the invention]

[0004] Please note that the terms “comprise,” “comprising,” and / or “comprises” are used to indicate that various aspects and embodiments of this disclosure “comprise” certain characteristics or characteristics. It should also be understood that these terms may also encompass aspects and / or embodiments that “consist essentially of” or “consist of” the relevant characteristics or characteristics.

[0005] The Coronaviridae family is a group of enveloped, single-stranded positive-sense RNA viruses. The Coronaviridae family includes the genus Coronavirus. The name "Coronaviridae" or "coronavirus" derives from the characteristic shape of the virus, which contains several crown-shaped protrusions ("peplomers" or "spikes"). Coronaviruses cause respiratory infections and fatal pneumonia outbreaks in humans worldwide. Coronaviruses are large, enveloped, positive-sense RNA viruses classified into four genera: alpha-coronaviruses, beta-coronaviruses, gamma-coronaviruses, and delta-coronaviruses. They possess the largest genomes (27–32 kb) of all RNA viruses and are packaged within a helical capsid surrounded by an envelope. There are at least three structural proteins associated with the viral envelope: membrane proteins (M), envelope proteins (E), and spike (glyco) proteins (S). Some coronaviruses encode hemagglutinin-esterase proteins (HE).

[0006] The S glycoprotein forms a large protrusion on the viral surface (forming a "corona" or "crown") and is involved in viral entry into host cells. The S protein has a large external domain that can be divided into the following domains / regions: S1 (receptor-binding domain); S2 (membrane fusion domain); transmembrane anchor; and a short intracellular tail. The S1 domain can be divided into two major domains: (i) the N-terminal domain (S1-NTD) - involved in sugar binding; and (ii) the C-terminal domain (S1-CTD) - involved in the recognition of protein receptors ACE2, APN, and DPP4.

[0007] As used herein, the term "coronavirus" encompasses any virus classified as belonging to the Coronaviridae family, including SARS coronavirus, MERS coronavirus, and SARS-CoV-2 coronavirus. The term "coronavirus" also encompasses all SARS-CoV-2 variants. For example, the term "coronavirus" includes variants called B.1.1.7, B.1525, B.1.351, and P1. These variants can be characterized by mutations (e.g., addition, substitution, and / or deletion of amino acids) within the spike protein. For example, the term "coronavirus" can encompass any variant having one or more of the following mutations within the spike protein: (i) HV69-70 deletion; and / or (ii) N501Y; and / or (iii) E484K. In any case, the variant spike protein sequence may contain one or more mutations compared to the reference sequence. A suitable reference sequence can be the Wuhan-Hu-1 strain, S protein sequence, which is available under accession code QHD43416.1 / YP_009724390.1.

[0008] SARS-CoV-2 is classified as belonging to the same genus as both SARS and MERS (Betacoronavirus); it is in the same subgenus as SARS and shares approximately 80% nucleotide identity across its entire genome. Both SARS and SARS-CoV-2 use glycosylated ACE2 protein expressed on the host cell surface for cell entry. Glycosylation of 90 ACE2 residues has been shown to significantly inhibit the SARS virus. The SARS S protein has 21 predicted glycosylation sites, and at least 18 of these residues are conserved in the SARS-CoV-2 sequence.

[0009] While we do not wish to be bound by any particular theory, antibodies that neutralize coronaviruses (e.g., MERS coronavirus) can target the receptor-binding domain (RBD) of the spike glycoprotein, blocking its binding to the cell receptor dipeptidyl peptidase 4 (DPP4). Other (anti-S protein N-terminal domain (NTD)) antibodies have been shown to bind to the N-terminal domain (NTD) of the spike glycoprotein and inhibit host cell entry at high titers. Therefore, it may be possible to inhibit host cell entry in an RBD (S1-CTD) site-independent manner by targeting the NTD portion of the S protein; such inhibition may affect the conformational state of the S glycoprotein. It has also been suggested that, like certain types of influenza (e.g., influenza C), coronaviruses can bind to sialic acid-containing receptors on host cells—in particular siaroglycans, including cell surface components containing, for example, 9-O-acetylated sialic acid. Furthermore (and again, without wanting to be constrained by theory), coronaviruses possess glycoprotein profiles that may be recognized by a variety of CBMs, including those classified as belonging to CBM families 32, 40, 47, 67, and 70.

[0010] Therefore, proteins with affinity for certain carbohydrates (polysaccharides or glycans), including sialic acid (e.g., any of the CBMs described herein), may disrupt the infectivity of coronaviruses, including SARS-CoV-2, by targeting glycans on the host and the virus. Several molecules containing (or essentially composed of) sugar (carbohydrate / polysaccharide / glycan) binding proteins are disclosed. These types of proteins exhibit a specific affinity for glycans and / or sialic acid and are useful in the treatment or prevention of coronavirus infection and / or diseases and / or conditions caused or resulting therefrom.

[0011] The phrase “a disease or condition caused by or resulting from coronavirus” may include such diseases or conditions known as SARS and MERS, as well as other respiratory illnesses and / or conditions associated with coronavirus infection. In addition, the phrase “a disease or condition caused by or resulting from coronavirus” may include the acute respiratory illness known as COVID-19, as well as acute respiratory illnesses caused by or associated with SARS-CoV-2 infection. As used herein, the term "treatment" may encompass the reduction of one or more symptoms associated with the disease / condition caused or resulting from coronavirus infection. Such symptoms may include, for example, a (persistent) cough, fever, and alteration / loss of taste / smell. Accordingly, any of the glycan-binding molecules described herein can be used in compositions and / or methods for treating one or more symptoms of coronavirus infection. For example, any one of the glycan-binding molecules of the present invention can be used to reduce a persistent cough that may occur as a result of coronavirus infection.

[0012] In view of the foregoing, this disclosure provides glycan-binding molecules for use in the treatment or prevention of coronavirus infection and / or diseases or conditions caused by or resulting from coronavirus. Also disclosed are methods for treating or preventing coronavirus infection and / or diseases or conditions caused by or resulting from coronavirus, the methods comprising administering a glycan-binding molecule to a subject in need thereof. This specification provides for the use of glycan-binding molecules in the manufacture of pharmaceuticals for the treatment or prevention of coronavirus infection and / or diseases or conditions caused by or resulting from coronavirus. This disclosure also provides sialic acid-binding molecules for use in the treatment or prevention of coronavirus infection and / or diseases or conditions caused by or resulting from coronavirus. Also disclosed are methods for treating or preventing coronavirus infection and / or diseases or conditions caused by or resulting from coronavirus, the methods comprising administering a sialic acid-binding molecule to a subject in need thereof. This specification provides for the use of sialic acid-binding molecules in the manufacture of pharmaceuticals for the treatment or prevention of coronavirus infection and / or diseases or conditions caused by or resulting from coronavirus.

[0013] A particularly useful molecule is the carbohydrate-binding module (CBM). Therefore, each carbohydrate, glycan, and / or sialic acid-binding molecule described herein for use, method, and pharmaceutical purposes may be a CBM or contain a CBM. Therefore, this disclosure further provides: (i) CBM for use in the treatment or prevention of coronavirus infection, or any disease or condition caused by or resulting from coronavirus; (ii) A method for treating or preventing coronavirus infection and / or disease or condition caused by or resulting from coronavirus, comprising administering CBM to a subject in need thereof; (iii) Use of CBM in the manufacture of medicines for the treatment or prevention of coronavirus infection and / or diseases or conditions caused by or resulting from coronavirus.

[0014] Based on the foregoing, this disclosure provides a CBM for use in the treatment or prevention of diseases or conditions caused by or resulting from COVID-19 infection and / or SARS-CoV-2. This disclosure also provides methods for treating or preventing diseases or conditions caused by or resulting from COVID-19 infection and / or SARS-CoV-2, the methods comprising administering CBM to subjects in need thereof. This specification describes the use of CBM in the manufacture of pharmaceuticals for the treatment or prevention of diseases or conditions caused by or resulting from COVID-19 infection and / or SARS-CoV-2.

[0015] Examples of useful CBMs are provided below, but for completeness, please understand that the term "CBM" includes, for example, CBMs classified as belonging to CBM families 32, 40, 47, 67, and 70. Those skilled in the art will understand that there are a vast variety of CBMs, and that members of the above families can originate from a wide variety of bacterial species. More detailed information on CBMs, particularly those of families 32, 40, 47, 67, and 70, can be found in the public CAZY database (available at http: / / www.cazy.org). Some particularly useful CBMs can originate from bacterial species within the genera Streptococcus, Vibrio, and Clostridium. It should also be understood that this disclosure encompasses the use of molecules containing one or more CBMs and any modified forms of any of the CBMs described herein (as described in more detail below). A modified CBM is a CBM that contains one or more mutant residues relative to the wild-type CBM sequence.

[0016] The CBMs for the various uses, pharmaceuticals, and methods described herein are all types of carbohydrate, glycan, and / or sialic acid binding molecules. The term "sialic acid" encompasses all forms of N- or O-substituted neuraminic acid, including all synthetic, naturally occurring and / or modified forms thereof. Sialic acid can be found as a component of cell surface molecules, glycoproteins and glycolipids. In most cases, sialic acid is present at the termini (terminal regions) of sugar chains attached to cell membranes and / or proteins. For example, some cells of the human upper respiratory tract contain α-2,6-linked sialic acid receptors, and other cells of the upper and lower respiratory tracts contain α-2,3-linked sialic acid receptors. The sialic acid family encompasses several (approximately 50) derivatives that can result from acetylation, glycolylation, lactonization and methylation at C4, C5, C7, C8 and C9. All such derivatives are to be encompassed by the term "sialic acid". Sialic acid may be found where α(2,3) or α(2,6) is linked to Gal and GalNAc, or where α(2,8) or α(2,9) is linked to another sialic acid. Thus, the term "sialic acid" is used throughout this specification, but it is important to understand that it encompasses all anomers, derivatives, analogs or variants of sialic acid (whether naturally occurring or synthetically produced), as well as all monomers, dimers, trimers, oligomers, polymers or concatemers containing it.

[0017] Thus, sialic acid-binding molecules for the various methods, uses and medicaments of the present disclosure can include a moiety having an affinity for sialic acid in any of its various forms as described above. Indeed, a CBM for use, i.e., a CBM for use in a medicament or method described herein, can exhibit an affinity for sialic acid in any of its various forms as described above, and / or can bind / couple to and / or associate with sialic acid molecules such that they can be present in or on a coronavirus, (mammalian) cell surface and / or (mammalian) cell surface receptor.

[0018] Useful CBMs can take any form and / or belong to any class or type of CBM. It can be assumed that CBMs from any one or more of the following CBM families are useful. (i) Family 32 CBM (CBM32); (ii) Family 40 CBM (CBM40); (iii) Family 47 CBM (CBM47); (iv) Family 67 CBM (CBM67); and (v) Family 70 CBM (CBM70).

[0019] For example, the present disclosure provides: (i) Coronavirus infection and / or a disease or condition caused by or resulting from coronavirus; or (ii) COVID-19 infection and / or a disease or condition caused by or resulting from SARS-CoV-2; CBM32 for use in the treatment or prevention of. Also: (i) Coronavirus infection and / or a disease or condition caused by or resulting from coronavirus; or (ii) COVID-19 infection and / or a disease or condition caused by or resulting from SARS-CoV-2; A method of treating or preventing is also disclosed, the method comprising administering one or more Family 32 CBM32 to a subject that needs it. In addition, the use of CBM32 in the manufacture of a medicament for the treatment or prevention of coronavirus infection and / or a disease or condition caused by or resulting from coronavirus is also disclosed.

[0020] The present disclosure also provides: (i) Coronavirus infection and / or a disease or condition caused by or resulting from coronavirus; or (ii) COVID-19 infection and / or a disease or condition caused by or resulting from SARS-CoV-2, We also provide CBM40, or CBM47, or CBM67, or CBM70 for use in the treatment or prevention of [unspecified disease].

[0021] This disclosure will be furthered, (i) coronavirus infection, and / or disease or condition caused by or resulting from coronavirus; or (ii) A disease or condition caused by or resulting from COVID-19 infection and / or SARS-CoV-2; A method for treating or preventing, This includes administering one or more Family 40 CBMs, Family 47 CBMs, Family 67 CBMs, or Family 70 CBMs to subjects who require them. The above method also relates to further information. In addition, this disclosure provides the use of CBM40, or CBM47, or CBM67, or CBM70 in the manufacture of a pharmaceutical product for the treatment or prevention of coronavirus infection and / or diseases or conditions caused by or resulting from coronavirus.

[0022] It should be noted that the use of any one of the disclosed CBM32, CBM40, CBM47, CBM67, or CBM70 can be combined with any other carbohydrate-binding protein, glycan-binding protein, sialic acid-binding molecule, and / or CBM. In fact, the use of CBM32, CBM40, CBM47, CBM67, or CBM70 can be combined with the use of any one of the other CBM32, CBM40, CBM47, CBM67, or CBM70. Further details regarding each CBM family are provided below - all of these CBMs (including their subtypes, variants, orthologs, etc.) are intended for use in the treatment and / or prevention of coronavirus infection and / or coronavirus-related illnesses and / or conditions described herein.

[0023] CBM32 Useful CBM32 (i.e., CBM32 for the various methods, pharmaceuticals, and uses described herein) can be derived from any suitable source. For example, CBM32 for use can be obtained from microorganisms, including, for example, bacteria of the genera Cellvibrio, Yersinia, Micromonospora, Streptococcus, Bifidobacterium, and Clostridium. For example, useful CBM32s can be obtained from or derived from, for instance, Cellvibrio mixtus, Yersinia enterolitica, Clostridium perfringens, Clostridium thermocellum, Streptococcus pneumoniae, Bifidobacterium longum, and Micromonospora viridifaciens. Further details regarding the sources, structure, and function of the CBM32 family can be found in the Carbohydrate Active Enzymes database (freely available online at http: / / www.cazy.org / CBM32.html).

[0024] An exemplary CBM32 sequence is provided by Sequence ID 1 below: Sequence ID 1 AIIETAIPQSEMTASATSEEGQDPASSAIDGNTNTMWHTKWNGSDALPQSLSVNLGSSRKVSSIAITP RTSGNNGFITKYEIHAINNGVETLVAEGTWEENNLVKTVTFDSPIDAEEIKITAIQGVGGFASIAELNVYE

[0025] Therefore, a CBM for use is essentially a CBM having the sequence of SEQ ID NO: 1 or containing a carbohydrate-binding portion thereof, or can be made from it. The carbohydrate-binding fragment of SEQ ID NO: 1 may contain anywhere between approximately 5, 6, 7, 8, 9, or 10 amino acids (consecutive or adjacent) and approximately 138 amino acids (consecutive or adjacent) from SEQ ID NO: 1. A suitable fragment may contain approximately 11, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, or 135 (consecutive or adjacent) amino acids from SEQ ID NO: 1.

[0026] Proteins containing, essentially derived from, or composed of CBM32 or SEQ ID NO: 1 can bind, for example, galactose, N-acetylgalactosamine (GalNAc), N-acetylglucosamine (GlcNAc), and / or lactose. Therefore, any fragment for use can bind galactose, N-acetylgalactosamine (GalNAc), N-acetylglucosamine (GlcNAc), and / or lactose. Those skilled in the art will understand that the binding affinity of any given CBM32 molecule can depend on the exact CBM32 subtype; as a further example, some CBM32s have shown affinity for various ligands. (Examples include type II blood group H-trisaccharide (Fucα1-2Galβ1-4GlcNAc), N-acetyl-D-lactosamine (LacNAc), galactose, lacto-N-biose, disaccharide GlcNAc-α-1,4-Gal (sometimes called N-acetylglucosamine linked to galactose at alpha-1,4), and / or GlcNAc). It should also be noted that multiple CBM32 subtypes can originate from a single organism; these various CBM subtypes can 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 to GlcNAc. As used herein, the term CBM32 encompasses all variants, derivatives, and subtypes of CBM32.

[0027] Sequence ID 1 originates from a sequence deposited in the UniProt database with ID number A0A2X2YJF2. This sequence is reproduced below as Sequence ID 2 (Sequence ID 1 appears as residues 42-180 - shown in bold in the sequence below). Sequence ID 2 MKSKKIIATL VASLVISNMG GYLVKANPNV NHKAVIIEDR QAIIETAIPQ SEMTASATSE EGQDPASSAI DGNTNTMWHT KWNGSDALPQ SLSVNLGSSR KVSSIAITPR TSGNNGFITK YEIHAINNGV ETLVAAEGTWE ENNLVKTVTF DSPIDAEEIK ITAIQGVGGF ASIAELNVYE IKGEVDEIAN YGNLKITKEE ERLNITRDLE KFSSLDEGTI VTRFNMNDTS IQSLIGLSDG NKANNYFSLY VSGGKVGYEL RRQEGNGDFN VHHSADVTFN KGINTLALKI EKGVGAKIFL NGSLVKTVSD PNIKFLNAIN LNSGFIGKTD RANGYNEYLF RGNIDFMNIY DKPVSDNYLL RKTGETKAPS EDSLLPDDVY KTQPVELFYP GYLESRGYRI PALETTKKGT VLASIDVRNN GDHDAPNNNI DVGIRRKEVN GEWEEGKVIL DYPGKSAAID TSLMSATIEE NGIEKERIFL IVTHFPEGYG FPNTEGGSGY KEIDGKYYFI LKDAQNNEYT VREDGIVYNS EGNETDYVMK NDKTLIQNGE EVGNALLSNS PLKAVGTAHI EMIYSDDDGN TWSEPEDLNP GLKKEWMKFF GTAPGKGIQI KNGEHKGRLV FPIYYTNQNN FQSSAVIYSD DFGETWKLGE SPIDTASVSS ETVSSGTQLT ECQVVEMPNG QLKLFMRNTG SYTRIATSFD GGATWHDEVP EDTSLREPYC QLSVINYSGK INGKDAIIFS NPDASSRVNG SVKVGLINEN GTYENGQPRY EFDWIYNKTV KPGSFAYSCL TELPDGNLGL FYEGEGAGRM AYTEFDLNYL KFNASEDSPA ATVQSIESLD EDLIYNAGDE VSIKVNFNQL VSLIGDRKIT LDIGGVDVPL NMVNYEGKSS AIFKGTIPEG INPGNYEIK LKENNALELNT VYNKVSTLNG LDNTGINVQI GELKTTVGNS TIKVNEEVQV GSAFEAILGI KGLNGDTEVY SAEYLFEYNA EAFKLNEITS FSDSLFVKSK EVEPGKVRIL VASLGNEIEK DSELVKVNLT PKISSELEVL GLTTALVGAG DGNTHDLELS SKEVKINEEA SGEIVVNPVQ NFEIPEINKK NVKLTWNAPI TTEGLEGYVI YKDGKKLSEV PAESTEFVVS KLNRHTIYNF KVAAKYSNGE LSAKESKTIR TAR

[0028] Sequence IDs 1 and 2 are derived from Clostridium perfringens. CBMs for use in various embodiments of this disclosure may include one, two, three, four or more CBM32 sequences. CBMs for use may include one, two, three, four or more proteins or their carbohydrate-binding fragments, including Sequence ID 1. Those skilled in the art will understand that useful CBM32 sequences may include sequences that exhibit some degree of sequence identity or homology (e.g., 99%, 95%, 90%, 85%, 80%, 75%, 70%, 65%, or 60%) to the CBM32 sequences of Sequence IDs 1 and 2. All such variant or divergent sequences will be encompassed within the scope of this disclosure by the term "CBM32". Identical and / or homologous CBM32 sequences may have carbohydrate-binding function.

[0029] CBM40 Useful CBM40s (i.e., CBM40s for the various methods, pharmaceuticals, and uses described herein) can be derived from any suitable source. For example, CBM40s for use can be obtained from microorganisms, including, for example, bacteria of the genera Clostridium, Enterococcus, Staphylococcus, Streptococcus, and Vibrio. For example, useful CBM40s can be obtained from or derived from, for example, Clostridium perfringens, Streptococcus pneumoniae, and Vibrio cholerae. Further details regarding the sources, structure, and function of the CBM40 family can be found in the carbohydrate-related enzyme database (freely available on the internet at http: / / www.cazy.org / CBM40.html).

[0030] Family 40 CBMs encompass molecules of approximately 200 residues and are often found at the N-terminus of GH33 sialidase. They may also be found inserted into the β-propeller of GH33 sialidase. At least Vibrio cholerae CBM40s bind alpha-anomers of sialic acid and, for example, α(2,3)-, α(2,6)-, and α(2,8)-linked sialosides.

[0031] Exemplary CBM40s for use may include the sialic acid-binding domain of Vibrio cholerae NanH sialidase (VcCBM: CBM40) and / or an equivalent (or homologous) domain derived from Streptococcus pneumoniae NanA sialidase (SpCBM: also CBM40). Naturally, similar or homologous sialic acid-binding modules present in other organisms would be included within the scope of the terms “CBM” and “CBM40”.

[0032] The exemplary NanH sialidase amino acid sequence of Vibrio cholerae is deposited under accession number A5F7A4 and is reproduced below as Sequence ID No. 3 (781 amino acids). Sequence ID 3 MRFKNVKKTA LMLAMFGMAT SSNAALFDYN ATGDTEFDSP AKQGWMQDNT NNGSGVLTNA DGMPAWLVQG IGGRAQWTYS LSTNQHAQAS SFGWRMTTEM KVLSGGMITN YYANGTQRVL PIISLDSSGN LVVEFEGQTG RTVLATGTAA TEYHKFELVF LPGSNPSASF YFDGKLIRDN IQPTASKQNM IVWGNGSSNT DGVAAYRDIK FEIQGDVIFR GPDRIPSIVA SSVTPGVVTA FAEKRVGGGD PGALSNTNDI ITRTSRDGGI TWDTELNLTE QINVSDEFDF SDPRPIYDPS SNTVLVSYAR WPTDAAQNGD RIKPWMPNGI FYSVYDVASG NWQAPIDVTD QVKERSFQIA GWGGSELYRR NTSLNSQQDW QSNAKIRIVD GAANQIQVAD GSRKYVVTLS IDESGGLVAN LNGVSAPIIL QSEHAKVHSF HDYELQYSAL NHTTTLFVDG QQITTWAGEV SQENNIQFGN ADAQIDGRLH VQKIVLTQQG HNLVEFDAFY LAQQTPEVEK DLEKLGWTKI KTGNTMSLYG NASVNPGPGH GITLTRQQNI SGSQNGRLIY PAIVLDRFFL NVMSIYSDDG GSNWQTGSTL PIPFRWKSSS ILETLEPSEA DMVELQNGDL LLTARLDFNQ IVNGVNYSPR QQFLSKDGGI TWSLLEANNA NVFSNISTGT VDASITRFEQ SDGSHFLLFT NPQGNPAGTN GRQNLGLWFS FDEVTWKGP IQLVNGASAY SDIYQLDSEN AIVIVETDNS NMRILRMPIT LLKQKLTLSQ N

[0033] The CBM region of Sequence ID No. 3 consists of amino acid residues 25-216 (the sequence shown in bold) - this sequence can be designated as Sequence ID No. 4. An exemplary amino acid sequence of Streptococcus pneumoniae NanA sialidase is deposited under accession number P62575 and is reproduced below as Sequence ID No. 5 (1035 amino acids). Sequence ID 5 MSYFRNRDID IERNSMNRSV QERKCRYSIR KLSVGAVSMI VGAVVFGTSP VLAQEGASEQ PLANETQLSG ESSTLTDTEK SQPSSETELS GNKQEQERKD KQEEKIPRDY YARDLENVET VIEKEDVETN ASNGQRVDLS SELDKLKKLE NATVHMEFKP DAKAPAFYNL FSVSSATKKD EYFTMAVYNN TATLEGRGSD GKQFYNNYND APLKVKPGQW NSVTFTVEKP TAELPKGRVR LYVNGVLSRT SLRSGNFIKD MPDVTHVQIG ATKRANNTVW GSNLQIRNLT VYNRALTPEE VQKRSQLFKR SDLEKKLPEG AALTEKTDIF ESGRNGKPNK DGIKSYRIPA LLKTDKGTLI AGADERRLHS SDWGDIGMVI RRSEDNGKTW GDRVTITNLR DNPKASDPSI GSPVNIDMVL VQDPETKRIF SIYDMFPEGK GIFGMSSQKE EAYKKIDGKT YQILYREGEK GAYTIRENGT VYTPDGKATD YRVVVDPVKP AYSDKGDLYK GNQLLGNIYF TTNKTSPFRI AKDSYLWMSY SDDDGKTWSA PQDITPMVKA DWMKFLGVGP GTGIVLRNGP HKGRILIPVY TTNNVSHLNG SQSSRIIYSD DHGKTWHAGE AVNDNRQVDG QKIHSSTMNN RRAQNTESTV VQLNNGDVKL FMRGLTGDLQ VATSKDGGVT WEKDIKRYPQ VKDVYVQMSA IHTMHEGKEY IILSNAGGPK RENGMVHLAR VEENGELTWL KHNPIQKGEF AYNSLQELGN GEYGILYEHT EKGQNAYTLS FRKFNWDFLS KDLISPTEAK VKRTREMGKG VIGLEFDSEV LVNKAPTLQL ANGKTARFMT QYDTKTLLFT VDSEDMGQKV TGLAEGAIES MHNLPVSVAG TKLSNGMNGS EAAVHEVPEY TGPLGTSGEE PAPTVEKPEY TGPLGTSGEE PAPTVEKPEY TGPLGTAGEE AAPTVEKPEF TGGVNGTEPA VHEIAEYKGS DSLVTLTTKE DYTYKAPLAQ QALPETGNKE SDLLASLGLT AFFLGLFTLG KKREQ

[0034] The CBM region of SEQ ID NO: 5 consists of amino acid residues 121-305 (the sequence shown in bold) - this sequence can be designated as SEQ ID NO: 6. CBMs for use in various aspects and embodiments of this disclosure may include proteins or peptides having the sequence of SEQ ID NOs: 3, 4, 5, or 6, or any carbohydrate-binding fragment thereof. For example, a useful molecule (i.e., a molecule for the uses, methods, and pharmaceuticals described herein) may include a protein moiety encoded by the sialic acid-binding domain of the Vibrio cholerae nanH gene (encoding sialidase) (provided by SEQ ID NO: 3) or an equivalent or homologous gene present in another organism (e.g., the equivalent / homologous nanA sialidase gene of Streptococcus pneumoniae; see SEQ ID NO: 5).

[0035] The molecules for use, methods, and pharmaceuticals of the Vibrio cholerae sialidase molecule of SEQ ID NOs: 3 and 4 may include approximately 1, 5, 10, 15, 25, or 30 residues (i.e., residues 1 to 30 or any amino acid residues between them) to approximately 150, 175, 200, 210, 216, 220 to 781 residues (including any residues between 150 and 781, and any residues between them). For example, the use, method, or pharmaceutical of the present disclosure may include a peptide having a sequence corresponding to residues 25 to approximately 216 of SEQ ID NO: 3 above. The carbohydrate binding fragment of SEQ ID NO: 3 may contain anywhere between approximately 5, 6, 7, 8, 9, or 10 amino acids (consecutive or adjacent) and approximately 191 amino acids (consecutive or adjacent) from SEQ ID NO: 3. A suitable fragment may contain approximately 11, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 150, 155, 160, 165, 170, 180, 185, 186, 187, 188, 189, or 190 (consecutive or adjacent) amino acids from Sequence ID No. 3.

[0036] Molecules for use, method or pharmaceutical purposes of this disclosure may include approximately 1, 5, 10, 15, 25, or 30 residues of the Vibrio cholerae sialidase molecule of SEQ ID NOs: 3 and 4 (i.e., residues 1-30 or any amino acid residues between them) to approximately 150, 175, 200, 210, 216, 220-781 residues (including any residues 150-781, and any residues between them). For example, a sialic acid-binding molecule for use may include a peptide having a sequence corresponding to residues 25-216 of SEQ ID NO: 3 above. Further suitable molecules may include proteins or peptides having the sequence of SEQ ID NO: 5 or 6, or their carbohydrate-binding fragments. For example, a useful sialic acid-binding molecule may include a protein portion encoded by the sialic acid-binding domain of the nanA gene (encoding sialidase) of Streptococcus pneumoniae.

[0037] The sialic acid-binding molecule for use may include residues of approximately 80, 90, 100, 110, 120, 121-130 (i.e., any residue between approximately 80-130, including any residue between them) to approximately 250, 275, 300, 305, 310, 320-1035 (i.e., any residue between approximately 250-1035, including any residue between them). For example, a sialic acid-binding molecule for use may include a peptide having a sequence corresponding to residues 121 to approximately 305 of SEQ ID NO: 5 above.

[0038] The carbohydrate bond fragment can contain anywhere between approximately 5, 6, 7, 8, 9, or 10 amino acids (consecutive or adjacent) and approximately 185 amino acids (consecutive or adjacent) from SEQ ID NO: 5. A suitable fragment may contain approximately 11, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, 180, 181, 182, 183, or 184 (consecutive or adjacent) amino acids from Sequence ID No. 5.

[0039] Sequence IDs 3 and 4 are derived from Vibrio cholerae, and Sequence IDs 5 and 6 are derived from Streptococcus pneumoniae. CBMs for use in various embodiments of this disclosure may include one, two, three, four or more CBM40s. CBMs for use may include one, two, three, four or more proteins or any of these carbohydrate-binding fragments, including Sequence IDs 3, 4, 5, or 6. Those skilled in the art will understand that useful CBM40s may include sequences that exhibit some degree of sequence identity or homology (e.g., 99%, 95%, 90%, 85%, 80%, 75%, 70%, 65%, or 60%) to the CBM40 sequences of Sequence IDs 3, 4, 5, or 6. All such variant or divergent sequences will be encompassed within the scope of this disclosure by the term "CBM40". Identical and / or homologous CBM40 sequences may have carbohydrate / sialic acid binding function.

[0040] CBM47 Useful CBM47s (i.e., CBM47s for the various methods, pharmaceuticals, and uses described herein) can be obtained from microorganisms, including, for example, bacteria of the genera Acinetobacter, Bathymodiolus, Campylobacter, Planctomycetes, Streptococcus, and Streptomyces. For example, useful CBM47s can be obtained or derived from, for example, Streptococcus mitis or Streptococcus pneumoniae. Further details regarding the possible sources, structure, and function of the CBM47 family can be found in the carbohydrate-related enzyme database (freely available on the internet at http: / / www.cazy.org / CBM47.html).

[0041] An exemplary CBM47 sequence is provided by sequence number 7 below: Sequence ID 7 TPDKFNDGNLNIAYAKPTTQSSVDYNGDPNRAVDGNRNGNFNSGSVTHTRADNPSWWEVDLKKMDKVGLVKIYNRTDAETQRLSNFDVILYDNNRNEVAKKHVNNLSGESVSLDFKEKGARYIKVKLLTSGVPLSLAEVEVFRES Therefore, CBM for use is essentially or can be essentially a CBM having the sequence of SEQ ID NO: 7 or a CBM containing the carbohydrate binding portion thereof. The carbohydrate-binding fragment of SEQ ID NO: 7 can contain anywhere between approximately 5, 6, 7, 8, 9, or 10 amino acids (consecutive or adjacent) from SEQ ID NO: 7 and approximately 144 amino acids (consecutive or adjacent). A suitable fragment can contain approximately 11, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, or 135, 140, or 143 amino acids (consecutive or adjacent) from SEQ ID NO: 7. Proteins containing, essentially derived from, or consisting of CBM47 or Sequence ID No. 7 are L-fucose, fucosyl lactose, H-trisaccharides and / or Lewis proteins. y Antigens can be bound to them. Therefore, any fragment for use contains L-fucose, fucosyl lactose, H-trisaccharide and / or Lewis. y It can bind antigens.

[0042] Sequence ID 7 originates from a sequence deposited in the UniProt database with ID number A0A1Q2T229. This sequence is reproduced as Sequence ID 8 below (Sequence ID 7 appears as residues 601-745 - shown in bold in the sequence below). Sequence ID 8 MNKEKIKRKL ITILFVCIGM LCFGLLAGVK ADNRVQMRTT INNESPLLLS PLYGNDNGNG LWWGNTLKGA WEAIPEDVKP YAAIELHPAK VCKPTSCIPR DTKELREWYV KMLEEAQSLN IPVFLVIMSA GERNTVPPEW LDEQFQKYSV LKGVLNIENY WIYNNQLAPH SAKYLEVCAK YGAHFIWHDH EKWFWETIMN DPTFFEASQK YHKNLVLATK NTPIRDDAGT DSIVSGFWLS GLCDNWGSST DTWKWWEKHY TNTFETGRAR DMRSYASEPE SMIAMEMMNV YTGGGTVYNF ECAAYTFMTN DVPTPAFTKG IIPFFRHAIQ NPAPSKEEVV NRTKAVFWNG EGRISSLNGF YQGLYSNDET MPLYNNGRYH ILPVIHEKID KEKISSIFPN AKILTKNSEE LSSKVNYLNS LYPKLYEGDG YAQRVGNSWY IYNSNANINK NQQVMLPMYT NNTKSLSLDL TPHTYAVVKE NPNNLHILLN NYRTDKTAMW ALSGNFDASK SWKKEELELA NWISKNYSIN PVDNDFRTTT LTLKGHTGHK PQINISGDKN HYTYTENWDE NTHVYTITVN HNGMVEMSIN TEGTGPVSFP TPDKFNDGNL NIAYAKPTTQ SSVDYNGDPN RAVDGNRNGN FNSGSVTHTR ADNPSWWEVD LKKMDKVGLV KIYNRTDAET QRLSNFDVIL YDNNRNEVAK KHVNNLSGES VSLDFKEKGA RYIKVKLLTS GVPLSLAEVE VFRESDGKQS EEDIDKITED KVVSTNKVAT QSSTNYEGVA ALAVDGNKDG DYGHHSVTHT KEDSPSWWEI DLAQTEELEK LIIYNRTDAE IQRLSNFDII IYDSNDYEVF TQHIDSLESN NLSIDLKGLK GKKVRISLRN AGIPLSLAEV EVYTYK

[0043] Sequence IDs 7 and 8 are derived from Streptococcus pneumoniae. CBMs for use in various embodiments of this disclosure may include one, two, three, four or more CBMs. CBMs for use may include one, two, three, four or more proteins or their carbohydrate-binding fragments, including SEQ ID NO: 8. Those skilled in the art will understand that useful CBM47s may include sequences that exhibit some degree of sequence identity or homology (e.g., 99%, 95%, 90%, 85%, 80%, 75%, 70%, 65%, or 60%) to the CBM47 sequences of SEQ ID NOs. All such variant or divergent sequences are encompassed within the scope of this disclosure by the term "CBM47". Identical and / or homologous CBM47 sequences may have carbohydrate-binding function.

[0044] CBM67 Useful CBM67 (i.e., CBM67 for the various methods, pharmaceuticals, and uses described herein) can be derived from any suitable source. For example, CBM67 for use can be obtained from microorganisms, including, for example, bacteria of the genera Bacillus, Paenibacillus, Planctomyces, and Streptomyces. For example, useful CBM67 can be obtained from or derived from, for example, Streptomyces avermitilis. Further details regarding possible sources, structure, and function of the CBM67 family can be found in the carbohydrate-related enzyme database (freely available on the internet at http: / / www.cazy.org / CBM67.html).

[0045] An exemplary CBM67 sequence is provided by sequence number 9 below: Sequence ID 9 APSLEGSSWIWFPEGEPANSAPAATRWFRRTVDLPDDITGATLAISADNVYAVSVDGAEVARTDLEADNEGWRRPAVIDVLDHVHSGNNTLAVSASNASVGPAGWICVLVLTTASGEKKIFSDASWKSTDHEPADGWREPDFDDSGWPAAKVAAAWGAGPWGRVA Therefore, CBM for use is essentially or can be essentially a CBM having the sequence of SEQ ID NO: 9 or a CBM containing the carbohydrate binding portion thereof.

[0046] The carbohydrate-binding fragment of Sequence ID No. 9 can contain anywhere between approximately 5, 6, 7, 8, 9, or 10 amino acids (consecutive or adjacent) and approximately 164 amino acids (consecutive or adjacent) from Sequence ID No. 9. A suitable fragment can contain approximately 11, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, 160, or 163 amino acids (consecutive or adjacent) from Sequence ID No. 9.

[0047] Proteins containing, essentially derived from, or composed of CBM67 or SEQ ID NO: 9 can bind L-rhamnose. Therefore, any fragment for use can bind L-rhamnose. SEQ ID NO: 9 is derived from a sequence deposited in the UniProt database with ID number Q82PP4. This sequence is reproduced below as SEQ ID NO: 10 (SEQ ID NO: 9 appears as residues 132–296 – shown in bold in the sequence below). Sequence ID 10 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

[0048] Sequence IDs 9 and 10 are derived from Streptomyces abelmytilis. A CBM for use in various embodiments of this disclosure may include one, two, three, four or more CBMs. CBMs for use may contain one, two, three, four or more proteins or their carbohydrate-binding fragments, including SEQ ID NO: 9. Those skilled in the art will understand that useful CBM67 may include sequences that exhibit some degree of sequence identity or homology (e.g., 99%, 95%, 90%, 85%, 80%, 75%, 70%, 65%, or 60%) to the CBM67 sequences of SEQ ID NOs. All such variant or divergent sequences are encompassed within the scope of this disclosure by the term "CBM67". Identical and / or homologous CBM67 sequences may have carbohydrate-binding function.

[0049] CBM70 Useful CBM70 (i.e., CBM70 for the various methods, pharmaceuticals, and uses described herein) can be derived from any suitable source. For example, CBM70 for use can be obtained from microorganisms, including, for example, bacteria of the genera Bacillus, Paenibacillus, Planctomyces, and Streptococcus. For example, useful CBM70 can be obtained from or derived from, for example, Streptococcus pneumoniae. Further details regarding possible sources, structure, and function of the CBM70 family can be found in the carbohydrate-related enzyme database (freely available on the internet at http: / / www.cazy.org / CBM70.html).

[0050] An exemplary CBM70 sequence is provided by sequence number 11 below: Sequence ID 11 NLVENGDFGQTEDGSSPWTGSKAQGWSAWVDQKNSADASTRVIEAKDGAITISSHEKLRAALHRMVPIEAKKKYKLRFKIKTDNKIGIAKVRIIEESGKDKRLWNSATTSGTKDWQTIEADYSPTLDVDKIKLELFYETGTGTVSFKDIELVEVADQLS The carbohydrate-binding fragment of SEQ ID NO: 11 can contain anywhere between approximately 5, 6, 7, 8, 9, or 10 amino acids (consecutive or adjacent) from SEQ ID NO: 11 and approximately 158 amino acids (consecutive or adjacent). A suitable fragment can contain approximately 11, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130, 135, 140, 145, 150, 155, or 157 amino acids (consecutive or adjacent) from SEQ ID NO: 11.

[0051] Proteins containing, essentially derived from, or composed of CBM70 or SEQ ID NO: 11 can bind hyaluronan. Therefore, any fragment for use can bind hyaluronan. Sequence ID 11 originates from a sequence deposited in the UniProt database with ID number Q54873. This sequence is reproduced below as Sequence ID 12 (Sequence ID 11 appears as residues 54-212 - shown in bold in the sequence below). Sequence ID 12 MQTKTKKLIV SLSSLVLSGF LLNHYMTIGA EETTTNTIQQ SQKEVQYQQR DTKNLVENGD FGQTEDGSSP WTGSKAQGWS AWVDQKNSAD ASTRVIEAKD GAITISSHEK LRAALHRMVP IEAKKKYKLR FKIKTDNKIG IAKVRIEES GKDKRLWNSA TTSGTKDWQT IEADYSPTLD VDKIKLELFY ETGTGTVSFK DIELVEVADQ LSEDSQTDKQ LEEKIDLPIG KKHVFSLADY TYKVENPDVA SVKNGILEPL KEGTTNVIVS KDGKEVKKIP LKILASVKDA YTDRLDDWNG IIAGNQYYDS 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

[0052] Sequence IDs 11 and 12 are derived from Streptococcus pneumoniae. A CBM for use in various embodiments of this disclosure may include one, two, three, four or more CBMs. CBMs for use may contain one, two, three, four or more proteins or their carbohydrate-binding fragments, including SEQ ID NO: 11. Those skilled in the art will understand that useful CBM70s may include sequences that exhibit some degree of sequence identity or homology (e.g., 99%, 95%, 90%, 85%, 80%, 75%, 70%, 65%, or 60%) to the CBM70 sequences of SEQ ID NOs. All such variant or divergent sequences are encompassed within the scope of this disclosure by the term "CBM70". Identical and / or homologous CBM70 sequences may have carbohydrate-binding function.

[0053] The various molecules described herein (including various carbohydrate, sialic acid, and / or glycan-binding proteins / molecules) for all of the various uses, pharmaceuticals, and methods described herein may further include oligomerized domains. A suitable oligomerization domain can exhibit the ability to self-associate to form a multimeric construct, for example, a trimer. An oligomerization domain for use can include any molecule or any functional fragment thereof possessing oligomerization properties. For example, one or more (e.g., two) sialic acid / glycan binding molecules (e.g., CBMs) can be bound, coupled, or fused to the oligomerization domain – the resulting sialic acid / glycan binding molecule::oligomerization domain "fusion" can then be used (together with one or more other such "fusions") as a molecule for treating or preventing coronavirus infection and / or associated diseases or conditions.

[0054] A suitable oligomerization domain can be derived, for example, from Pseudomonas aeruginosa pseudoaminidase. The amino acid sequence of an exemplary Pseudomonas aeruginosa pseudoaminidase sequence is deposited under accession number PA0579 and is reproduced below as SEQ ID NO: 13 (438 amino acids). Sequence ID 13 MNTYFDIPHR LVGKALYESY YDHFGQMDIL SDGSLYLIYR RATEHVGGSD GRVVFSKLEG GIWSAPTIVA QAGGQDFRDV AGGTMPSGRI VAASTVYETG EVKVYVSDDS GVTWVHKFTL ARGGADYNFA HGKSFQVGAR YVIPLYAATG VNYELKWLES SDGGETWGEG STIYSGNTPY NETSYLPVGD GVILAVARVG SGAGGALRQF ISLDDGGTWT DQGNVTAQNG DSTDILVAPS LSYIYSEGGT PHVVLLYTNR TTHFCYYRTI LLAKAVAGSS GWTERVPVYS APAASGYTSQ VVLGGRRILG NLFRETSSTT SGAYQFEVYL GGVPDFESDW FSVSSNSLYT LSHGLQRSPR RVVVEFARSS SPSTWNIVMP SYFNDGGHKG SGAQVEVGSL NIRLGTGAAV WGTGYFGGID NSATTRFATG YYRVRAWI

[0055] The oligomerized domain of sequence number 13 is amino acid residues 333-438 - this sequence can be represented as sequence number 14 (shown in bold). Therefore, the oligomerization domain for use may include residues approximately 250, 275, 300, 310, 320, 333, 340-350 (i.e., approximately residues 250-350, including approximately any residues between them) to approximately 400, 410, 420, 430, or 438 (i.e., approximately any residues between approximately residues 400-438, including approximately any residues between them) of the trimerization domain (PaTD) of Pseudomonas aeruginosa pseudoaminidase provided by SEQ ID NO: 5. For example, useful sialic acid / glycan binding molecules can utilize the oligomerization domain containing residues 333-438 of SEQ ID NO: 13.

[0056] The sialic acid / glycan binding molecules for any of the methods, uses, and / or pharmaceuticals described herein may include any modified form of any of the CBMs described herein. The term "modified" encompasses molecules that contain one or more mutations compared to the reference sequence. The “reference sequence” can be any wild-type CBM sequence. For example, the reference sequence may include, be essentially, or be essentially a wild-type CBM sequence derived from, for example, Vibrio cholerae NanH sialidase or Streptococcus pneumoniae NanA sialidase, a wild-type CBM sequence (note that similar or homologous CBMs present in other organisms (including CBM40) will be included within the scope of the term “CBM” and / or as CBM reference sequences). The reference sequence may include (or consist of, or be essentially) any of the above sequence numbers 1-12. Therefore, modified CBM sequences can be derived from specific or particular wild-type CBM sequences.

[0057] A modified CBM sequence may contain a wild-type CBM sequence that has been modified to include one or more mutations. One or more mutations can be functional – that is, a mutation can modulate (alter, improve, or suppress / inhibit) one or more physiological, biological, immunological, and / or pharmacological properties that are characteristic of wild-type CBM (e.g., wild-type CBM from which the modified CBM originates), individually (and / or independently) or collectively (e.g., synergistically). In particular, one or more mutations: (i) The immunogenicity (or antigenicity) of CBM can be altered; and / or (ii) The effectiveness of (CBM or any multimeric molecule including modified CBM) can be altered (e.g., improved); and / or (iii) Mutations can modulate (e.g., improve) the thermal stability of CBM; and / or (iv) Mutations can modulate (e.g., improve) the solubility of CBM; and / or (v) Mutations can modulate the binding affinity of CBM to its targets (e.g., glycans, sialic acid, galactose, lactose, polygalacturonic acid, LacNAc, fucose, L-rhamnose; hyaluronan); and / or (vi) Mutations can modulate (e.g., improve) the in vivo half-life of a molecule.

[0058] "Mutation" can include any modification to the wild-type CBM molecule. For example, the term "mutation" can encompass, for instance, the following: (i) one or more amino acid substitutions (in which case one or more wild-type amino acids are replaced or changed by another (different) amino acid - the term “substitution” will include conservative amino acid substitutions); and / or (ii) One or more amino acid deletions (in which case one or more wild-type amino acid residues are removed); and / or (iii) Addition / insertion of one or more amino acids (in which case further amino acid residues are added to the wild-type (or reference) primary sequence); and / or (iv) Inversion of one or more amino acids / sequences (usually this involves inversion of two or more consecutive amino acids in the primary sequence); and / or (v) Duplication of one or more amino acids / sequences (in this case, a repeating amino acid or part of a primary amino acid sequence (e.g., a stretch of 5 to 10 amino acids)). Therefore, the modified CBMs provided for in this disclosure may include one or more of the mutations described herein.

[0059] An exemplary wild-type CBM (in other words, a reference sequence from which useful modified CBMs can be derived) is Streptococcus pneumoniae NanA sialidase, whose amino acid sequence is deposited under accession number P62575 and reproduced above as SEQ ID NO: 5 (1035 amino acids). Also, as mentioned above, the CBM region of SEQ ID NO: 5 consists of amino acid residues 121-305 - this sequence is designated as SEQ ID NO: 6. Accordingly, the present disclosure provides a carbohydrate-binding molecule having affinity for sialic acid, the molecule including modified forms of SEQ ID NO: 6 (and / or SEQ ID NO: 5). Modified forms of SEQ ID NO: 6 may include one or more mutant residues - the mutations are, for example, amino acid substitutions, additions / insertions, duplications, deletions and / or inversions made to the sequence of SEQ ID NO: 6.

[0060] The exemplary Vibrio cholerae NanH sialidase amino acid sequence is deposited at accession number A5F7A4 and reproduced above as SEQ ID NO: 7 (781 amino acids). The CBM region of SEQ ID NO: 7 consists of amino acid residues 25-216 - this is SEQ ID NO: 8. Accordingly, this disclosure provides a sialic acid / glycan binding molecule including modified forms of SEQ ID NO: 8 (and / or SEQ ID NO: 7). Modified forms of SEQ ID NO: 8 may include one or more mutant residues - the mutations are, for example, amino acid substitutions, additions / insertions, duplications, deletions and / or inversions made to the sequence of SEQ ID NO: 8. Therefore, a useful modified CBM (i.e., a CBM for use in the medical uses and methods described herein) may contain one or more of the mutations described herein.

[0061] As a non-limiting example, the following represent individual units (referred to as "HEX" units) that can be used to construct modified sialic acid / glycan conjugate molecules for the various uses, methods, and pharmaceuticals described herein (for example, for use in methods of treating or preventing coronavirus infection). (i)HEX1 CBMX1 (L170T V239A V246G I286A Y292E)-----CBMX2 (L170T V239A V246G I286A Y292E)-----TD (S342D L348D R403K) (ii)HEX2 CBMX1 (V239A V246G I286A Y292E)----CBMX2 (V239A V246G I286A Y292E)----TD (S342D R403K) (iii)HEX3 CBMX1 (V239A V246G I286A)-----CBMX2 (V239A V246G I286A)-----TD (S342D R403K) (iv)HEX4 CBMX1 (V239A V246G)-----CBMX2 (V239A V246G)-----TD (S342D) (v)HEX5 CBMX1 (V239A V246G)-----CBMX2 (V239A V246G)-----TD (R403K) (vi)HEX6 CBMX1 (V239A V246G)-----CBMX2 (V239A V246G)-----TD (S342D R403K) (vii)HEX17 CBMX1 (V239A V246G A162P)-----CBMX2 (V239A V246G A162P)-----TD (S342D R403K)

[0062] A suitable HEX unit can contain two modified CBMs (indicated above as CBMX1 and CBMX2), with the specific mutations introduced into each CBM identified in parentheses. In one teaching, units CBMX1 and CBMX2 can contain (consist of, or essentially consist of) any type of CBM (e.g., any member of the group / class of CBM40, any CBM32, any CBM47, any CBM67 and / or any CBM70). Note that the "----" symbol indicates an amino acid linker (linking one modified CBM to another, or linking one modified CBM to an oligomeric domain).

[0063] In each case, the oligomerization domain (indicated as "TD") present in each HEX unit conjugates the unit together as a trimer. While any given hexamer may contain three identical copies of one of the above units, those skilled in the art will understand that further options are available. For example, a HEX unit may consist of two CBMs, each having a different mutation (one or more mutations selected from the options detailed herein).

[0064] It should be noted that HEX6 and HEX17 are identical except for a further A162P mutation. This proline mutation (substitution of wild-type alanine at residue 162) has been shown to improve thermal stability (single CBM Tm at 3–4°C). Further information on the use of proline mutations can be found in Fu 2009, 'Increasing protein stability by improving beta-turns' (DOI 10.1002 / prot.22509), which describes a general approach. The proline mutation does not affect the predicted immunogenicity of the CBM molecule (neither increasing nor decreasing it) and is not located near other mutations, the N-terminus or C-terminus, or ligand-binding sites. Rather unexpectedly, it has been noted that beyond a moderate improvement in thermal stability, the A162P mutation results in molecules that exhibit significant improvements in in vivo experiments—particularly compared to such same experiments performed using hexamer molecules containing other (e.g., HEX6) HEX units. For example, modified molecules (particularly those containing the HEX17 unit) exhibit modulation of pro-inflammatory cytokines, including IL-8. In fact, the modulation effect (particularly the inhibitory effect) on IL-8 production by molecules containing the HEX17 unit was significantly improved compared to other modified molecules tested.

[0065] The amino acid sequences of HEX6 and HEX17 molecules, compared to the amino acid sequence of Sp2CBMTD (also known as "SpOrig"), are as follows: SpOrig GAMVIEKEDVETNASNGQRVDLSSELDKLKKLENATVHMEFKPDAKAPAFYNLFSVSSAT HEX6 GAMVIEKEDVETNASNGQRVDLSSELDKLKKLENATVHMEFKPDAKAPAFYNLFSVSSAT HEX17 GAMVIEKEDVETNASNGQRVDLSSELDKLKKLENATVHMEFKPDPKAPAFYNLFSVSSAT Question KKDEYFTMAVYNNTATLEGRGSDGKQFYNNYNDAPLKVKPGQWNSVTFTVEKPTAELPKG HEX6 KKKDEYFTMAVYNNTATLEGRGSDGKQFYNNYNDAPLKVKPGQWNSVTFTVEKPTAELPKG HEX17 KKKDEYFTMAVYNNTATLEGRGSDGKQFYNNYDPLKVKPGQWNSVTFTVEKPTAELPKG SpOrig RVRLYVNGVLSRTSLRSGNFIKDMPDVTHVQIGATKRANNTVWGSNLQIRNLTVYNRALT HEX6 RARLYVNGGLSRTSLRSGNFIKDMPDVTHVQIGATKRANNTVWGSNLQIRNLTVYNRALT HEX17 RARLYVNGGLSRTSLRSGNFIKDMPDVTHVQIGATKRANNTVWGSNLQIRNLTVYNRALT Question: PEEVQKRSGGGSGVIEKEDVETNASNGQRVDLSSELDKLKKLENATVHMEFKPDAKAPAF HEX6 PEEVQKRSGGGSGVIEKEDVETNASNGQRVDLSSELDKLKKLENATVHMEFKPDAKAPAF HEX17 PEEVQKRSGGGSGVIEKEDVETNASNGQRVDLSSELDKLKKLENATVHMEFKPDPKAPAF SpOrig YNLFSVSSATKKDEYFTMAVYNNTATLEGRGSDGKQFYNNYNDAPLKVKPGQWNSVTFTV HEX6 YNLFSVSSATKKDEYFTMAVYNNTATLEGRGSDGKQFYNNYNDAPLKVKPGQWNSVTFTV HEX17 YNLFSVSSATKKDEYFTMAVYNNTATLEGRGSDGKQFYNNYNDAPLKVKPGQWNSVTFTV SpOrig EKPTAELPKGRVRLYVNGVLSRTSLRSGNFIKDMPDVTHVQIGATKRANNTVWGSNLQIR HEX6 EKPTAELPKGRARLYVNGGLSRTSLRSGNFIKDMPDVTHVQIGATKRANNTVWGSNLQIR HEX17 EKPTAELPKGRARLYVNGGLSRTSLRSGNFIKDMPDVTHVQIGATKRANNTVWGSNLQIR SpOrig NLTVYNRALTPEEVQKRSGGALGVPDFESDWFSVSSNSLYTLSHGLQRSPRRVVVEFARS HEX6 NLTVYNRALTPEEVQKRSGGSLGVPDFESDWFDVSSNSLYTLSHGLQRSPRRVVVEFARS HEX17 NLTVYNRALTPEEVQKRSGGSLGVPDFESDWFDVSSNSLYTLSHGLQRSPRRVVVEFARS SpOrig SSPSTWNIVMPSYFNDGGHKGSGAQVEVGSLNIRLGTGAAVWGTGYFGGIDNSATTRFAT HEX6 SSPSTWNIVMPSYFNDGGHKGSGAQVEVGSLNIKLGTGAAVWGTGYFGGIDNSATTRFAT HEX17 SSPSTWNIVMPSYFNDGGHKGSGAQVEVGSLNIKLGTGAAVWGTGYFGGIDNSATTRFAT SpOrig GYYRVRAWI HEX6 GYYRVRAWI HEX17 GYYRVRAWI

[0066] [SpOrig(sequence code 15);HEX6(sequence code 16) and HEX17(sequence code 17)] SpOrig can be modified to include the following mutation (R274Q), which modulates the binding affinity of CBM to sialic acid.

[0067] In light of the above, this disclosure is: (i) coronavirus infection, and / or disease or condition caused by or resulting from coronavirus; or (ii) A disease or condition caused by or resulting from COVID-19 infection and / or SARS-CoV-2; A modified CBM is provided for use in the treatment or prevention of [unspecified condition]. Also: (i) coronavirus infection, and / or disease or condition caused by or resulting from coronavirus; or (ii) A disease or condition caused by or resulting from COVID-19 infection and / or SARS-CoV-2; Methods for treating or preventing the condition are also disclosed, which include administering a modified CBM to a subject in need thereof.

[0068] In addition, the use of modified CBMs in the manufacture of pharmaceuticals for the treatment or prevention of coronavirus infection and / or diseases or conditions caused by or resulting from coronavirus is also disclosed. In addition, this disclosure states: (i) coronavirus infection, and / or disease or condition caused by or resulting from coronavirus; or (ii) A disease or condition caused by or resulting from COVID-19 infection and / or SARS-CoV-2; HEX17 is provided for use in the treatment or prevention of [unspecified condition].

[0069] Also: (i) coronavirus infection, and / or disease or condition caused by or resulting from coronavirus; or (ii) A disease or condition caused by or resulting from COVID-19 infection and / or SARS-CoV-2; Methods for treating or preventing the condition are also disclosed, the methods comprising administering HEX17 to a subject in need thereof. In addition, the use of HEX17 in the manufacture of pharmaceuticals for the treatment or prevention of coronavirus infection and / or diseases or conditions caused by or resulting from coronavirus is also disclosed.

[0070] The molecules for use, methods, and pharmaceuticals described herein may include one or more of the CBM molecules described herein. These types of molecules may further include the aforementioned trimerizing domains that enable the formation of a sialic acid / glycan binding molecule, which is a polyvalent CBM. For example, a molecule for use may contain multiple (i.e., two, three, four or more) CBMs. A molecule containing multiple CBMs can be called a "polyvalent glycan or sialic acid binding molecule" or "polyvalent CBM".

[0071] Polyvalent CBM molecules can be prepared as constructs containing multiple (identical or different) CBMs linked by amino acid / peptide linkers. Each CBM (e.g., VcCBM, SpCBM, or modified CBM) can be linked to another (e.g., VcCBM, SpCBM, or modified CBM) or to a trimerizing domain (TD) by a peptide containing, for example, 5, 10, or 15 amino acids. For example, one or more of the following peptides can be used to ligate two or more CBMs or one CBM to a trimerizing domain, thereby generating a polyvalent CBM: (i) 5-amino acid linker: ALNGS LQALG GGNSG GGGSG GGALG (ii) Linker of 10 amino acids: ALNGSGGGSG LQALGGGGSL (iii) Linker of 15 amino acids: ALNGSGGGSGGGGSG

[0072] The polyvalent CBMs described herein for use and pharmaceutical purposes may include, for example, two or more Family 32 CBMs, two or more Family 40 CBMs, two or more Family 47 CBMs, two or more Family 67 CBMs, and two or more Family 70 CBMs. The polyvalent CBMs may include mixtures of different CBM types, for example, CBMs from different CBM families, or repeats of the same CBM. Various examples of useful monovalent and polyvalent glycan / sialic acid binding molecules are described herein. For example, the molecules presented in Figure 1 may be used in the methods, uses, and pharmaceuticals of this disclosure. Suitable molecules may contain (essentially consist of) or consist of one or more (e.g., two, three, four, or more) VcCBMs (which are CBM40 derived from Vibrio cholerae). CBMs (e.g., VcCBMs) may be fused, bound, or conjugated to an oligomerized domain (e.g., PaTD or its oligomerized fragment). Glycan / sialic acid binding molecules may contain, consist of, or essentially consist of two fused (or bound) CBMs fused to an oligomerized domain (see, for example, the molecule Vc2CBMTD shown in Figure 1).

[0073] Any of the disclosed molecules may be used, alone or in combination with other disclosed molecules, for use in treating or preventing coronavirus infection or associated disease or condition, for use in a method for treating coronavirus infection or associated disease or condition, or for use in the manufacture of a medicine for treating or preventing coronavirus infection or associated disease or condition.

[0074] Any of the disclosed molecules (for the uses, pharmaceuticals, and methods described herein), particularly modified CBM molecules, can be generated using PCR-based cloning techniques, and suitable methods for generating this type of polyvalent molecule are described, for example, in 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). Polyvalent CBM molecules, including, for example, HEX17, Vc2CBM, Vc4CBM, and Sp2CBM, can be prepared as constructs containing multiple CBMs linked by amino acid / peptide linkers, such as those described above.

[0075] In the context of this disclosure, molecules for use in the treatment or prevention of coronavirus infection (carbohydrate-binding molecules, sialic acid-binding molecules, glycan-binding proteins / molecules and / or CBMs) may include one or more CBMs selected from the group consisting of: (i) One or more (e.g., two, three, four or more) families of 32 CBMs; (ii) One or more families of 40 CBMs (e.g., two, three, four or more); (iii) One or more (e.g., two, three, four or more) families of 47 CBMs; (iv) One or more (e.g., two, three, four or more) families of 67 CBMs; (v) one or more (e.g., two, three, four or more) families of 70 CBMs; and (vi) Modified CBM (as described herein).

[0076] In addition, molecules for use in the treatment or prevention of coronavirus infection (carbohydrate-binding molecules, sialic acid-binding molecules, glycan-binding molecules / proteins and / or CBMs) may include CBMs selected from the following group: (i) Clostridium perfringens CBM32 (CpCBM32); (ii) Streptococcus pneumoniae CBM40 (SpCBM40); (iii) Vibrio cholerae CBM40 (VcCBM40); (iv) Streptococcus pneumoniae CBM47 (SpCBM47); (v) Streptomyces abelmytilis CBM67 (SaCBM67); (vi) Streptococcus pneumoniae CBM70 (SpCBM70); (vii) Vibrio cholerae NanH sialidase CBM; (vii) Sialic acid-binding fragment of Vibrio cholerae NanH sialidase CBM: (xi) Streptococcus pneumoniae nanA sialidase CBM; and (x) Sialic acid-binding fragment of Streptococcus pneumoniae nanA sialidase CBM.

[0077] The various uses, methods, and pharmaceutical applications of polyvalent CBMs described herein may include: (i) VcCBM; or (ii) Two (or more) VcCBMs; or (iii) Three or four (or more) VcCBMs; or (iv) SpCBM; or (v) Two (or more) SpCBMs; or (vi) Three or four (or more) SpCBMs; or (vii) CpCBM; or (viii) Two (or more) CpCBMs; or (viii) Three or four (or more) CpCBMs; or (ix) SaCBM; or (x) Two (or more) SaCBMs; or (xi) Three or four (or more) SaCBMs.

[0078] Polyvalent CBMs for use, methods, or pharmaceuticals of this disclosure may include one or more CBM32 together with a mixture of different CBMs, for example, one or more other CBMs selected from the group consisting of: (i) Family 40 CBM (ii) Family 47 CBM (iii) Family 67 CBM (iv) Family 70 CBM For example, the methods, uses, or pharmaceuticals described herein can utilize combinations of CBM32 and Family 40 CBM (CBM40). Alternatively, the polyvalent CBMs for use, methods, and pharmaceuticals described herein may include one or more CBMs 40 together with a mixture of different CBMs, for example, one or more other CBMs selected from the group consisting of: (i) Family 32 CBM (ii) Family 47 CBM (iii) Family 67 CBM (iv) Family 70 CBM

[0079] For example, the methods, uses, or pharmaceuticals described herein can utilize combinations of CBM40 and Family 32 CBM (CBM32). The polyvalent CBMs for use, methods, and pharmaceuticals described herein may include one or more CBMs, along with a mixture of different CBMs, for example, one or more other CBMs selected from the group consisting of: (i) Family 32 CBM (ii) Family 40 CBM (iii) Family 67 CBM (iv) Family 70 CBM

[0080] The polyvalent CBMs for use, methods, and pharmaceuticals described herein may include one or more CBM67s together with a mixture of different CBMs, for example, one or more other CBMs selected from the group consisting of: (i) Family 32 CBM (ii) Family 40 CBM (iii) Family 47 CBM (iv) Family 70 CBM The polyvalent CBMs for use, methods, and pharmaceuticals described herein may include one or more CBMs, together with a mixture of different CBMs, for example, one or more other CBMs selected from the group consisting of: (i) Family 32 CBM (ii) Family 40 CBM (iii) Family 47 CBM (iv) Family 67 CBM

[0081] The various uses, methods, and pharmacopoeias of polyvalent CBMs disclosed herein may include, for example, molecules selected from the group consisting of: (i) Cp2CBM32TD (containing, essentially, or consisting of two CBMs (CBM32) derived from Clostridium perfringens fused to a trimerizing domain); (ii) Sp2CBM40TD (containing, essentially, or consisting of two CBMs (CBM40) derived from Streptococcus pneumoniae fused to a trimerizing domain); (iii) Vc2CBM40TD (containing, essentially, or consisting of two CBMs (CBM40) derived from Vibrio cholerae fused to a trimerizing domain); and (iv) Vc4CBM (containing, essentially consisting of, or comprising four CBMs derived from Vibrio cholerae (CBM40)); (v)Sp2CBM47TD (containing, essentially, or consisting of two CBMs (CBM47) derived from Streptococcus pneumoniae fused to a trimerizing domain); (vi)Sp2CBM67TD(containing, essentially, or consisting of two CBMs (CBM67) derived from Streptococcus abelmytilis fused to a trimerizing domain); (vii)Sp2CBM70TD (containing, essentially, or consisting of two CBMs (CBM70) derived from Streptococcus pneumoniae fused to a trimerizing domain).

[0082] This disclosure provides uses (compositions, methods, and pharmaceuticals), which include, but are not limited to, uses of one or more of the CBMs disclosed herein in isolated forms (where at least one CBM component (of the therapeutic molecule) contains or is essentially one of the CBM sequences and / or functional fragments described herein), but also uses where the CBM component of the therapeutic molecule is contained within a larger molecule. For example, the various CBMs described herein are provided and / or can be used in the form of a larger molecule containing the CBM component. A CBM component (e.g., a sialic acid binding molecule) may, for example, contain (consist of or be essentially composed of) one of the CBMs described herein (including, for example, CBM32, CBM40, CBM47, CBM67, and CBM70). As a (non-limiting) example, a molecule of this disclosure (e.g., a CBM and / or glycan / sialic acid binding molecule) may have one or more other functions in addition to exhibiting the ability to bind glycans (or components thereof) or sialic acid. For example, molecules can possess enzymatic activity. For instance, useful molecules include CBMs (as described herein) and can exhibit some sialidase activity.

[0083] A useful molecule can be a fusion protein comprising an enzyme moiety and a glycan / sialic acid binding moiety – in this case, the glycan / sialic acid binding moiety comprises a CBM as described herein. In such a case, the enzyme moiety can be fused to the glycan / sialic acid binding moiety. As stated above, the enzyme moiety of any useful fusion protein may contain (or have or exhibit) sialidase activity.

[0084] In one embodiment, the sialic acid-binding molecule, glycan-binding molecule, or CBM for the various uses described herein may not be provided as part of a molecule (e.g., a fusion protein) possessing enzymatic (e.g., sialidase) activity, nor may it be contained within such molecule. Additionally or alternatively, the sialic acid / glycan-binding molecule may (i) not bind heparin or heparin sulfate, and / or (ii) not contain a GAG-binding domain of a protein that binds the heparin or heparin sulfate moiety. Constructs comprising the sialic acid / glycan-binding molecule, glycan-binding molecule, or CBM may not exhibit or present enzymatic (e.g., sialidase) activity.

[0085] This disclosure also provides pharmaceutical compositions that can be utilized, in particular, in the uses, methods and pharmaceuticals described herein. Therefore, any of the useful molecules described herein (e.g., CBMs (modified, polyvalent, or other) or glycan-binding molecules) can be formulated for subsequent use. For convenience, it should be noted that the term "CBM" encompasses all monovalent, polyvalent, and modified CBM molecules described herein. For example, sialic acid-binding molecules, glycan-binding molecules, or CBMs can be formulated as therapeutic or pharmaceutical compositions. Various compositions may include one or more of the sialic acid-binding molecules / glycan-binding molecules / CBMs described herein and one or more pharmaceutically acceptable excipients. For example, pharmaceutical formulations containing the molecules described herein may be mixed with stabilizers, wetting agents, emulsifiers, salts (for use in affecting osmotic pressure), buffers, and / or other substances that do not react adversely with the active compound. A given therapeutic use or treatment method may require administration of one or more of these compositions (together, simultaneously, or separately), and such composition may contain one or more different CBMs.

[0086] The pharmaceutical compositions described herein can be prepared for oral, mucosal, intranasal, or parenteral (intravenous) administration. Such formulations for mucosal or intranasal administration can be prepared conventionally, for example, by incorporating by reference all the information in 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), containing substances conventionally used in pharmaceuticals. Liquid dosage forms for oral and / or intranasal administration may include emulsions, liquids, suspensions, syrups, and elixirs. In addition to the compound or composition, the liquid dosage form may contain water or other solvents, solubilizers, and emulsifiers, which are commonly used in the art and may contain inert diluents.

[0087] Any appropriate amount of sialic acid-binding molecules, glycan-binding molecules, or CBMs can be used. For example, whether a composition containing sialic acid-binding molecules, glycan-binding molecules, or CBMs is administered intravenously or mucosally (e.g., intranasally), the dose of sialic acid-binding molecules / glycan-binding molecules / CBMs can be anywhere between approximately 0.1 μg and approximately 6000 μg. For example, doses of sialic acid-binding molecules / glycan-binding molecules / CBMs of approximately (e.g., + / - 0.5 μg) 0.1 μg, 0.5 μg, 1 μg, 5 μg, 10 μg, 11 μg, 12 μg, 13 μg, 14 μg, 15 μg, 20 μg, 30 μg, 40 μg, 50 μg, 100 μg, 200 μg, 300 μg, 400 μg, 500 μg, 600 μg, 700 μg, 800 μg, 900 μg, 950 μg, 1000 μg, 1500 μg, 2000 μg, 2500 μg, 3000 μg, 3500 μg, 4000 μg, 4500 μg, 5000 μg, 5500 μg, or 6000 μg can be used. These amounts can be provided in any suitable volume of excipient, diluent, or buffer. For example, the amount of sialic acid-binding molecule / glycan-binding molecule / CBM can be provided somewhere between approximately 1 μl and approximately 0.5 ml of excipient, diluent, or buffer. For example, the required amount of sialic acid-binding molecules / glycan-binding molecules or CBM can be combined (or formulated) in approximately 5 μl, 10 μl, 15 μl, 20 μl, 25 μl, 30 μl, 35 μl, 40 μl, 45 μl, 50 μl, 55 μl, 60 μl, 65 μl, 70 μl, 75 μl, 80 μl, 85 μl, 90 μl, 95 μl, 100 μl, 140 μl, 200 μl, 280 μl, 300 μl, 400 μl, 500 μl, 560 μl, 600 μl, 700 μl, 800 μl, 900 μl, or 1 ml. A concentration of 0.1–15 mg (e.g., 0.5 mg, 1 mg, 2 mg, 3 mg, 4 mg, 5 mg, 6 mg, 7 mg, 8 mg, 9 mg, 10 mg, 11 mg, 12 mg, 13 mg, or 14 mg) (sialic acid / glycan binding protein) per 1 ml (excipient, diluent, or buffer) can be considered most useful. A concentration of 10 mg / ml (excipient, diluent, or buffer) can be considered extremely useful.

[0088] The compositions of this disclosure (for example, compositions comprising any of the sialic acid-binding molecules, glycan-binding molecules, or CBMs disclosed herein) can be administered to a subject (prophylactically) at fixed and / or predetermined times. For example, the compositions described herein can be administered at fixed and / or predetermined times before, during, or after a subject enters or encounters a scenario in which they may be vulnerable and / or susceptible to coronavirus infection.

[0089] The compositions of this disclosure may be administered daily and / or every few days. The compositions of this disclosure can be administered multiple times throughout any given day. The compositions described herein may be administered over periods of several weeks, months, or years. The exact dosing regimen will vary depending on the subject, the subject's health condition, and the duration for which the subject is considered to be at risk of or vulnerable to coronavirus infection. The present disclosure will now be explained with reference to the following diagram: [Brief explanation of the drawing]

[0090] [Figure 1]This diagram shows the building blocks of the polyvalent CBM morphology and their affinity for sialic acid. a. VcCBM, residues 25-216 of Vibrio cholerae sialidase with α-2,3-sialyl lactose, depicted as a sphere (PDB: 1w0p). b. SpCBM, residues 121-305 of Streptococcus pneumoniae NanA sialidase with α-2,3-sialyl lactose (PDB: 4c1w). c. TD, trimerization domain, residues 333-438 of Pseudoaminidase, iridescent (PDB: 2w38); the other two monomers in their respective colors. d. Polyvalent morphology: molecular weight, binding valency, and their binding affinity to α2,3-sialyl lactose, determined by surface plasmon resonance (SPR) at 25°C (KD values ​​for VcCBM, Vc2CBM, and Vc3CBM have been previously reported (Connaris et al, 2009)). Tandem repeat CBMs and oligomeric CBMs fused to TDs are linked by 5-aminolinkers (for details, see Connaris, H. et al., (2014). PNAS 111:6401-6406). [Figure 2] This graph shows the results of the assay under condition 1. In particular, when tested at 3 mg / mL, all CBM compounds exhibited observable antiviral effects. [Figure 3] This is a diagram of the plaque assay showing the results of assay condition 1; here again, the antiviral effect was demonstrated against all three CBMs, especially when used at higher concentrations. [Figure 4] This figure shows a graph illustrating the results of the Condition 2 assay. In this case, cells were exposed to CBM before SARS-CoV-2 infection; therefore, the Condition 2 assay represents a preventive model. Antiviral effects are shown for at least CBM2 and CBM3. [Figure 5] This is a graph showing the results of assay condition 3. In this case, infected cells (with SARS-CoV-2) were treated with various CBMs. Antiviral effects were shown for all three CBMs, particularly CBM3. [Figure 6]This is a diagram of the plaque assay showing the results of assay condition 3. Here again, this shows the antiviral effect of all three CBMs - in particular CBM3. [Figure 7] Figure 7 shows the mean values ​​and SEM images of the total clinical findings for groups 2 (control) and 3 (Neumifil) from 0DPC to the day of euthanasia (7DPC). Figure 7 also shows the mean percentage change in body weight (along with SEM) for the same period (y-axis on the right). [Figure 8] This figure shows the detection of Neumifil (HEX17) binding to the SARS-CoV-2 spike S1 variant. The dotted line represents the 4PL curve fit of the data. Inset: EC50 values ​​for each variant. [Figure 9] This figure shows the detection of Neumifil (HEX17) binding to recombinant human ACE2. The dotted line represents the 4PL curve fitting of the data. Inset: EC50 values.

[0091] method Plaque reduction assay Thaw the SARS-CoV-2 titration vial on ice. Two working stocks of SARS-CoV-2 were prepared, and the virus was diluted with serum-free (SF) DMEM to 500 pfu / mL (working stock for condition 1) and 250 pfu / mL (working stock for conditions 2 and 3). Store the prepared stock on ice. For the preparation of the test reagent, a fixed amount of each CBM (100 μL per vial at 10 mg / mL) is thawed on ice, the contents are transferred to a new sterile 1.5 mL Eppendorf tube, and the mixture is centrifuged at 13,000 rpm for 5 minutes to pellet any precipitate that forms. The supernatant is then transferred to a new sterile 1.5 mL Eppendorf tube. Dilute the CBM supernatant with a 50:50 mixture of serum-free DMEM:PBS to prepare two working stock concentrations (3 mg / mL and 1 mg / mL from a 10 mg / mL master stock) for the three CBMs to be tested (see Table 1). The final CBM concentration varies depending on the conditions. Condition 1: Final CBM concentrations of 1.5 mg / mL and 0.5 mg / mL, and conditions 2 and 3: Final CBM concentrations of 3 mg / mL and 1 mg / mL.

[0092] [Table 1]

[0093] Condition 1: Virus and CBM are mixed before being added to the cells. Mix equal volumes of SARS-CoV-2 and CBM and incubate on ice for 1 hour. For a positive control, mix SARS-CoV-2 with SF diluent (a 50-50 mixture of SF DMEM and PBS) instead of CBM. For a negative control, mix 3 mg / mL of CBM 1 with SF DMEM instead of SARS-CoV-2. ○ Tube 1: 300 μl SARS-CoV-2 in 500 pfu / mL along with 300 μl CBM1 at 3 mg / mL ○ Tube 2: 300 μl of SARS-CoV-2 at 500 pfu / mL, along with 300 μl of CBM2 at 3 mg / mL. ○ Tube 3: 300 μl of SARS-CoV-2 in 500 pfu / mL along with 300 μl of CBM3 in 3 mg / mL ○ Tube 4: 300 μl of SARS-CoV-2 in 500 pfu / mL along with 300 μl of CBM1 in 1 mg / mL ○ Tube 5: 300 μl SARS-CoV-2 in 500 pfu / mL along with 300 μl CBM1 at 1 mg / mL ○ Tube 6: 300 μl SARS-CoV-2 in 500 pfu / mL along with 300 μl CBM1 at 1 mg / mL ○ Tube NC: Negative control: 300 μl SF DMEM with 300 μl CBM diluent ○ Tube PC: Positive control, 300 μl SARS-CoV-2 at 500 pfu / mL together with 300 μl CBM diluent After 1 hour, remove the serum-free medium from the cells and wash with sterile PBS (<0.5 mL / well). After washing with PBS, 200 μL of the prepared inoculum is added to appropriate wells, with tubes 1-3 (and control) placed on plate 1 and tubes 4-6 (and control) on plate 2. Place the plate in a flat-bottomed, airtight container and transfer it to an incubator. Incubate at 37°C with 5% CO2 for 1 hour. After 1 hour, remove the inoculum and add 1 mL of overlay to each well. Transfer the plate to a flat-bottomed, airtight container. Incubate at 37°C in 5% CO2 for 5 days.

[0094] Condition 2: Cells exposed to CBM before viral infection Remove serum-free DMEM and wash with sterile PBS (<0.5 mL / well). After washing with PBS, add 200 μL of CBM at a concentration of 3 mg / mL to plate 3 and 1 mg / mL to plate 4. Incubate at 37°C in 5% CO2 for 1 hour. After 1 hour, remove the CBM and wash with sterile PBS (<0.5 mL / well). After washing with PBS, add 200 μL of SARS-CoV-2 diluted to 250 pfu / mL to each well except the negative control well. Add 200 μL of SF DMEM to the negative control well. Place the plate in a flat-bottomed, airtight container and transfer it to an incubator. Incubate at 37°C with 5% CO2 for 1 hour. After 1 hour, remove the inoculum and add 1 mL of overlay to each well. Transfer the plate to a flat-bottomed, airtight container. Incubate at 37°C in 5% CO2 for 5 days.

[0095] Condition 3: Cells infected with SARS-CoV-2 and then treated with CBM. Remove serum-free DMEM and wash with sterile PBS (0.5 mL / well). After washing with PBS, add 200 μL of SARS-CoV-2 diluted to 250 pfu / mL to each well except the negative control. Add 200 μL of SF DMEM to the negative control well. Place the plate in a flat-bottomed, airtight container and transfer it to an incubator. Incubate at 37°C with 5% CO2 for 1 hour. After 1 hour, remove the inoculum and add 200 μL of CBM to each well except the negative control (3 mg / mL CBM in plate 5 and 1 mg / mL CBM in plate 6). Add 200 μL of CBM diluent (SF DMEM-PBS) to the negative control well. Place the plate in a flat-bottomed, airtight container and transfer it to an incubator. Incubate at 37°C with 5% CO2 for 1 hour. After 1 hour, remove the CBM and add 1 mL of overlay to each well. Transfer the plate to a flat-bottomed, airtight container. Incubate at 37°C in 5% CO2 for 4 days.

[0096] Specific details: CBM: • CBM1:Vc2CBM40TD (a trimer (hexavalent) form based on Vibrio cholerae CBM40) • CBM2: Neumifil (HEX17: Trimeric (hexavalent) form based on Streptococcus pneumoniae CBM40) • CBM3:Cp2CBM32TD (a trimer (hexavalent) form based on Clostridium perfringens CBM32) CBM was added at two concentrations (1 mg / mL and 3 mg / mL) and tested under three conditions (as described): Condition 1: Mix SARS-CoV-2 and CBM before adding to cells. Condition 2: Exposure of cells to CBM before SARS-CoV-2 infection (prevention model) Condition 3: Cells are infected with SARS-CoV-2, and then CBM is added (treatment model). Regarding the configuration of CBM: 3 mg / mL: 450 μL stock + 1050 μL DMEM: PBS 1 mg / mL: 150 μL stock + 1350 μL DMEM:PBS SARS-CoV-2 Vial label: SARS CoV2, England 2, P2 HCM / V / 52, 05.03.20 2.4 × 10 5 Stock in pfu / mL → 100 μL stock + 900 μL medium = 2.4 × 10 4 pfu / mL → 200 μL of the above + 1800 μL of medium = 2.4 × 10 3 pfu / mL → 1.5 mL above + 6 mL medium = 480 pfu / mL ← Used in the study VeroE6 cells P15, Split 1:2 25 / 3 / 20 (Obtained from PHE HCM Group) Reagent details → DMEM Sigma D5796, Lot RNBH6732, Exp 06 / 2020 → PBS Gibco 10010-023, Lot 2098597, Exp 30 / 6 / 2021 → Overlay (a set amount prepared, enough for two plates each): · 7.5mL 4%CMC PHE medium, MR / 19 / 1044, date of manufacture 04 / DEC / 19 · 7.5mL 2%CMC PHE medium, MR / 19 / 1043, manufacturing date 04 / DEC / 19 ·15mL 2×MEM Gibco 21935-028, Lot 2150451, Exp 31 / 12 / 2020 · 800μL FCS Gibco 10100-139, Lot 216386RP, Exp 05 / 2024 · 400μL anti-anti Sigma A5955, Lot 035M4800V

[0097] result Condition 1: Mix the virus and CBM before adding them to the cells. In the main phase of the test, the SARS-CoV-2 virus and CBM compounds were incubated for 1 hour before being added to the cells. Plate 1 contained the compounds tested at a working concentration of 3 mg / mL, and Plate 2 contained the compounds tested at 1 mg / mL. All negative control wells were intact, while positive controls yielded plaque counts between 119 and 164 per well. The results are shown in Figures 2 (graph) and 3 (plaque assay). The results indicate that all CBM compounds exhibit observable antiviral effects. 3.2. Condition 2: Exposure of cells to CBM before viral infection In the main phase of the study, VeroE6 cells were exposed to the CBM compound before the addition of the SARS-CoV-2 virus. Plate 3 contained the compound tested at a working concentration of 3 mg / mL, and Plate 4 contained the compound tested at 1 mg / mL. Plaques were counted after staining. As shown in Figure 4, when tested at a working concentration of 1 mg / mL, the CBM2 and CBM3 compounds showed observable antiviral effects. 3.3. Condition 3: Cells were infected with SARS-CoV-2 and then treated with CBM. In the main phase of the experiment, VeroE6 cells were infected with the SARS-CoV-2 virus before adding the CBM compound. Plate 5 contained the compound tested at a working concentration of 3 mg / mL, and Plate 6 contained the compound tested at 1 mg / mL. Plaques were counted after staining. All negative control wells were intact, while the positive controls gave plaque counts between 117 and 132 per well. When plotted on a graph (see Figure 5), all three CBM compounds were shown to have antiviral effects, with CBM3 showing the greatest reduction. The results from both CBM1 and CBM2 at their effective concentrations were similar. The results of the plaque assay are also shown in Figure 6.

[0098] conclusion All CBMs showed some form of antiviral activity against the SARS-CoV-2 virus in vitro. CBM2 (Neumifil: Hex17) and CBM3 (CBM32-based) are extremely promising. The results demonstrated here provide a basis for urgent in vivo studies in appropriate animal models.

[0099] (Example 2): Test of CBM vs OC43 OC43 is the human coronavirus OC43-(HCoV-OC43); it is a beta-coronavirus and is associated with the development of "common cold" type illnesses. Its S protein binds to sugar-based receptor determinants, particularly 9-O-acetylated sialic acid (9-O-Ac-Sias), which is attached as a terminal residue to glycan chains on glycoproteins and lipids. A series of experiments were completed to determine whether CBM can be used to treat or prevent OC43 infection.

[0100] The following CBMs were used in each experiment: 1) Vc2CBM40TD(VC2) 2) Cp2CBM32TD(CBM32) 3) HEX17 Each CBM was used at concentrations of 1 mg / ml and 3 mg / ml. Condition 1: Prophylactic CBM treatment Cells were treated with CBM for 1 hour and then incubated with hCoV-OC43 for 1 hour. The assay showed that treatment with any of the tested CBMs prevented subsequent hCoV-OC43 infection compared to infected cells that were not pretreated with any of the CBMs. This effect was most pronounced with VC2 (at 3 mg / ml and 1 mg / ml), HEX17 (at 1 mg / ml), and CBM32 (at 3 mg / ml and 1 mg / ml). Condition 2: Simultaneous CBM processing Cells were simultaneously treated with CBM and hCoV-OC43 for 1 hour. The assay showed that simultaneous treatment with either CBM at either concentration (1 mg / ml or 3 mg / ml) reduced hCoV-OC43 infection compared to infected cells that were not treated with either CBM.

[0101] (Example 3) Animal (hamster) study testing Neumifil against SARS-CoV-2 Protocol / Research Design

[0102] [Table 2] TIFF2023516995000004.tif218155 TIFF2023516995000005.tif67144

[0103] [Table 3] result

[0104] [Table 4]

[0105] (Example 4) Angiotensin-converting enzyme 2 (ACE2) plays a major role in the recognition, binding, fusion, and entry of SARS-CoV-2 into host cells [1]. Glycans, including sialic acid, can also be important in this interaction. The ACE2 receptor has eight glycosylation sites, three of which (N90, N322, and N546) may play a crucial role in the interaction with the SARS-CoV-2 spike. Glycans on the SARS-CoV-2 spike can also modulate the conformation of the spike's receptor-binding domain (RBD), which is involved in ACE2 recognition and binding. Deficiency of such glycans significantly reduces ACE2 binding.

[0106] Novel, multiple variants of SARS-CoV-2 have emerged and are circulating globally [1, 2]. Of particular concern are the B.1.1.7 and B.1525 variants identified in the UK, the B.1.351 South African variant, and the P.1 Brazil variant. The B.1.1.7UK ("Kent") virus is characterized by HV69-70 deletions and N501Y in the spike protein, which may be associated with increased infectivity and a higher risk of death. The B.1.351 S.African variant shares some mutations with B.1.1.7, along with further differences, including the E484K mutation, which is a potential vaccine escape. The Brazil variant, P.1, is spreading to many countries and is associated with reinfection. P.1 also contains E484K, in addition to the N501Y mutation, which is associated with the increased infectivity of the UK variant. A further variant, B1525, has been identified in the UK, which contains the important E484K mutation in parallel with the Kent B.1.1.7 mutation.

[0107] the purpose: 1) To determine whether Neumifil interacts with the SARS-CoV-2 spike S1 protein and whether its affinity is affected by novel variant mutations. 2) To determine whether Neumifil interacts with human ACE2. method: Table 1 summarizes the variant S1 spike sequence information provided by the manufacturer. The spike protein and ACE2 protein were each recombinantly expressed in HEK293 cells.

[0108] [Table 5]

[0109] EC50 (50% effective concentration) values ​​were determined by ELISA. Spike protein or ACE2 protein was immobilized overnight at 4°C at a concentration of 1 μg / mL in a highly binding ELISA plate. The wells were then incubated with Neumifil (3-fold dilution series: 29160, 9720, 3240, 1080, 360, 120, 40, 0 ng / mL) in three overlapping cycles for 1.5 hours. Immunodetection of Neumifil binding was performed by incubation with rabbit anti-Neumifil (1 hour), followed by incubation with HRP-labeled anti-rabbit IgG (1 hour), and development of a TMB substrate. The binding curve was analyzed using 4-parameter logistic (4PL) curve fitting to determine the inflection point (EC50).

[0110] result: As shown in Figure 8, Neumifil binds to the original spike protein (consisting of the Wuhan-Hu-1 December 2019 isolate sequence) with an EC50 of 174 ng / mL. The variant binding profile and EC50 indicate that this affinity is not significantly affected by mutations present in the South African and UK (Kent) sequences. Figure 9 shows that Neumifil binds to ACE2 with an EC50 of 235 ng / mL.

[0111] Literature 1. Understanding variants of SARS-CoV-2 (2021) The Lancet World Report 397(102), P462 2. McNally, A. (2021) BMJ 372, 504

Claims

Claim 1: A pharmaceutical composition for use in the treatment or prevention of a coronavirus infection, a disease or condition caused or contributed to by a coronavirus, and / or symptoms of a coronavirus infection / disease, comprising a carbohydrate binding module (CBM), wherein the CBM: (i) Family 40 CBM; and (ii) Family 32 CBM; A pharmaceutical composition selected from the group consisting of:

2. The pharmaceutical composition of claim 1, wherein the disease or condition caused or contributed to by a coronavirus is COVID-19, SARS, or MERS.

3. 3. The pharmaceutical composition of claim 1 or 2, wherein the treatment of a coronavirus infection and / or a disease or condition caused or contributed to by a coronavirus includes treatment of one or more symptoms associated with the coronavirus infection, disease or condition.

4. 4. The pharmaceutical composition of claim 3, wherein the symptoms are persistent cough and / or fever and / or alteration / loss in / of taste / smell.

5. The carbohydrate binding module (CBM) comprises: (i) Clostridium perfringens CBM32 (CpCBM32); (ii) Streptococcus pneumoniae CBM40 (SpCBM40); (iii) Vibrio cholerae CBM40 (VcCBM40); (iv) Vibrio cholerae NanH sialidase CBM; (v) sialic acid-binding fragment of Vibrio cholerae NanH sialidase CBM; (vi) Streptococcus pneumoniae nanA sialidase CBM; and (vii) a sialic acid-binding fragment of Streptococcus pneumoniae nanA sialidase CBM; The pharmaceutical composition according to any one of claims 1 to 4, selected from the group consisting of:

6. The pharmaceutical composition according to any one of claims 1 to 5, wherein the coronavirus is SARS-CoV-2.

7. The pharmaceutical composition according to any one of claims 1 to 6, wherein the coronavirus is a SARS-CoV-2 variant.

8. 8. The pharmaceutical composition of claim 7, wherein the SARS-CoV-2 variant comprises a mutation in the spike protein, and the mutation is an amino acid change relative to the amino acid sequence of the spike protein of the Wuhan-Hu-1 isolate having accession code: QHD43416.1 / YP_009724390.

1.

9. The variants have the following spike protein mutations: (i) HV69-70 deletion; and / or (ii) N501Y; and / or (iii) E484K, 9. The pharmaceutical composition of claim 7 or 8, comprising one or more of:

10. 10. The pharmaceutical composition of claim 7, 8 or 9, wherein the SARS-CoV-2 variant is a B.1.1.7 variant and / or a B.1525 variant and / or a B.1.351 variant.

11. Including CBM32, (i) a coronavirus infection, a disease or condition caused or contributed to by a coronavirus, and / or symptoms of a coronavirus infection / disease; or (ii) COVID-19, a disease or condition caused or contributed to by SARS-CoV-2, and / or symptoms of COVID-19 20. A pharmaceutical composition for use in the treatment or prevention of:

12. CBM32 has the amino acid sequence of SEQ ID NO:1: AIIETAIPQSEMTASATSEEGQDPASSAIDGNTNTMWHTKWNGSDALPQSLSVNLGSSRKVSSIAITPRTSGNNGFITKYEIHAINNGVETLVAEGTWEENNLVKTVTFDSPIDAEEIKITAIQGVGGFASIAELNVYE or glycan / carbohydrate binding fragments thereof 12. The pharmaceutical composition of claim 11, comprising:

13. Including CBM40, (i) a coronavirus infection, a disease or condition caused or contributed to by a coronavirus; or (ii) COVID-19, a disease or condition caused or contributed to by SARS-CoV-2, and / or symptoms of COVID-19 20. A pharmaceutical composition for use in the treatment or prevention of:

14. CBM40 has the amino acid sequence of SEQ ID NO:4: ALFDYNATGDTEFDSPAKQGWMQDNTNNGSGVLTNADGMPAWLVQGIGGRAQWTYSLSTNQHAQASSFGWRMTTEMKVLSGGMITNYYANGTQRVLPIISLDSGNLVVEFEGQTGRTVLATGTAATEYHKFELVFLPGSNPSASFYFDGKLIRDNIQPTASKQNMIVWGNGSSNTDGVAAYRDIKFEIQGD Or the amino acid sequence of SEQ ID NO:6: VIEKEDVETNASNGQRVDLSSELDKLKKLENATVHMEFKPDAKAPAFYNLFSVSSATKKDEYFTMAVYNNTATLEGRGSDGKQFYNNYNDAPLKVKPGQWNSVTFTVEKPTAELPKGRVRLYVNGVLSRTSLRSGNFIKDMPDVTHVQIGATKRANNTVWGSNLQIRNLTVYNRALTPEEVQKRS or any glycan / sialic acid binding fragment 14. The pharmaceutical composition of claim 13, comprising:

15. containing an engineered carbohydrate binding module (CBM), (i) a coronavirus infection, a disease or condition caused or contributed to by a coronavirus, and / or symptoms of a coronavirus infection / disease; or (ii) COVID-19, a disease or condition caused or contributed to by SARS-CoV-2, and / or symptoms of COVID-19 1. A pharmaceutical composition for use in the treatment or prevention of A modified CBM comprises a wild-type CBM sequence that has been modified to include one or more mutations, said CBM comprising: (i) Family 40 CBM; and (ii) Family 32 CBM; A pharmaceutical composition selected from the group consisting of:

16. The modified CBM has the sequence of SEQ ID NO: 17: GAMVIEKEDVETNASNGQRVDLSSELDKLKKLENATVHMEFKPDPKAPAFYNLFSVSSATKKDEYFTMAVYNNTATLEGRGSDGKQFYNNYNDAPLKVKPGQWNSVTFTVEKPTAELPKGRA RLYVNGGLSRTSLRSGNFIKDMPDVTHVQIGATKRANNTVWGSNLQIRNLTVYNRALTPEEVQKRSGGGSGVIEKEDVETNASNGQRVDLSSELDKLKKLENATVHMEFKPDPKAPAFYNLF SVSSATKKDEYFTMAVYNNTATLEGRGSDGKQFYNNYNDAPLKVKPGQWNSVTFTVEKPTAELPKGRARLYVNGGLSRTSLRSGNFIKDMPDVTHVQIGATKRANNTVWGSNLQIRNLTVYN RALTPEEVQKRSGGSLGVPDFESDWFDVSSNSLYTLSHGLQRSPRRVVVEFARSSSPSTWNIVMPSYFNDGGHKGSGAQVEVGSLNIKLGTGAAVWGTGYFGGIDNSATTRFATGYYRVRAWI or a glycan-binding fragment thereof 16. The pharmaceutical composition of claim 15, comprising:

17. The method of claim 16, comprising administering to a mammalian subject the method of claim 16, further comprising administering to said ... wherein the carbohydrate binding module (CBM) comprises: (i) a coronavirus infection, a disease or condition caused or contributed to by a coronavirus, and / or symptoms of a coronavirus infection / disease; or (ii) COVID-19, a disease or condition caused or contributed to by SARS-CoV-2, and / or symptoms of COVID-19; 1. A pharmaceutical composition for use in the treatment or prevention of The carbohydrate binding module (CBM) is: (i) Cp2CBM32TD (comprising or consisting of two CBMs (CBM32) from Clostridium perfringens fused to a trimerization domain); (ii) Sp2CBM40TD (comprising or consisting of two CBMs (CBM40) from Streptococcus pneumoniae fused to a trimerization domain); (iii) Vc2CBM40TD (comprising or consisting of two CBMs (CBM40) from Vibrio cholerae fused to a trimerization domain); and (iv) Vc4CBM (comprising or consisting of four CBMs (CBM40) from Vibrio cholerae); A pharmaceutical composition selected from the group consisting of:

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