Anti-TMPRSS2 antibodies and antigen-binding fragments

Abstract

The present disclosure provides antibodies and antigen-binding fragments thereof that specifically bind to TMPRSS2 and methods of using such antibodies and fragments to treat or prevent viral infection (e.g., coronavirus infection or influenza virus infection).

Description

[Technical Field]

[0001] Government Licensing Rights This invention was made with government support under Contract HHSO100201700020C awarded by the U.S. Department of Health and Human Services. The government has certain rights in this invention.

[0002] Sequence Listing An official copy of the Sequence Listing has been submitted electronically via EFS-Web contemporaneously with the present specification as an ASCII formatted Sequence Listing, with the filename "10737WO01_Sequence_Listing_ST25.txt", created on February 9, 2021, and approximately 64 KB in size. The Sequence Listing contained in this ASCII formatted document is a part of the present specification and is incorporated herein by reference in its entirety.

[0003] FIELD OF THE INVENTION The present invention relates to antibodies and antigen-binding fragments that specifically bind to TMPRSS2, and methods of treating or preventing viral infections using such antibodies and fragments. [Background technology]

[0004] Background of the Invention Viruses, such as influenza viruses, have developed resistance to currently used drugs that target the viral neuraminidase (NA) or the ion channel protein matrix protein 2 (M2). Furthermore, newly identified viruses (e.g., coronaviruses) can be difficult to treat because they are not well characterized. The emergence of drug resistance and newly identified viruses highlights the need for the development of novel antiviral strategies. Host cell targeting can reduce or avoid the emergence of escape mutants, but widespread expression can lead to "sinking" and raise concerns about toxicity. Influenza (Zmora et al., Non-human primate orthologues of TMPRSS2 cleave and activate the influenza virus hemagglutinin. PLoS ONE. 12, e0176597 (2017) (Non-Patent Document 1); Bottcher-Friebertshauser et al., Inhibition of influenza virus infection in human airway cell cultures by an antisense peptide-conjugated morpholino oligomer targeting the hemagglutinin-activating protease TMPRSS2. Journal of Virology. 85, 1554-1562 (2011) (Non-Patent Document 2); Bertram et al., TMPRSS2 and TMPRSS4 facilitate trypsin-independent spread of influenza virus in Caco-2 cells. Journal of Virology. 84, 10016-10025 (2010) (Non-Patent Document 3); Tarnow et al., TMPRSS2 is a host factor that is essential for pneumotropism and pathogenicity of H7N9 influenza A virus in mice.It has been shown that numerous respiratory viral fusion proteins require cleavage by host proteases for activation, including those from viruses such as coronaviruses (Heurich et al., TMPRSS2 and ADAM17 cleave ACE2 differentially and only proteolysis by TMPRSS2 augments entry driven by the severe acute respiratory syndrome coronavirus spike protein. Journal of Virology. 88, 2202-2013 (2014) (Non-Patent Document 5)) and viruses (Shirato et al., Clinical Isolates of Human Coronavirus 229E Bypass the Endosome for Cell Entry. Journal of Virology. 91, e01387-16 (2017) (Non-Patent Document 6); Reinke et al., Different residues in the SARS-CoV spike protein determine cleavage and activation by the host cell protease TMPRSS2. PLoS ONE.12,e0179177(2017)(Non-patent document 7);Zhou et al.,Protease inhibitors targeting coronavirus and filovirus entry.Antiviral Research.116,76-84(2015)(Non-patent document 8);Zmora et al.TMPRSS2 Isoform 1 Activates Respiratory Viruses and Is Expressed in Viral Target Cells.PLoS ONE.10, e0138380 (2015) (Non-patent Document 9)). .

[0005] The influenza A hemagglutinin precursor (HA0) requires cleavage by a host serine protease into HA1 and HA2 for activation. For example, the transmembrane proteases serine 2; TMPRSS2, TMPRSS4, and TMPRSS11D, as well as human airway trypsin-like proteases (HATs), are involved in HA cleavage (Bertram et al., TMPRSS2 and TMPRSS4 facilitate trypsin-independent spread of influenza virus in Caco-2 cells. Journal of Virology. 84, 10016-10025 (2010) (Non-Patent Document 10); Bottcher et al., Proteolytic Activation of Influenza Viruses by Serine Proteases TMPRSS2 and HAT from Human Airway Epithelium. Journal of Virology. 2006 October; 80(19):9896-8 (Non-Patent Document 11); International Patent Application Publication No. WO2017 / 151453 (Patent Document 1)). TMPRSS2 also cleaves and activates the spike protein in coronaviruses, such as severe acute respiratory syndrome coronavirus (SARS-CoV) (Heurich et al., TMPRSS2 and ADAM17 cleave ACE2 differentially and only proteolysis by TMPRSS2 augments entry driven by the severe acute respiratory syndrome coronavirus spike protein. Journal of Virology. 88, 2202-2013 (2014) (Non-Patent Document 12)). Furthermore, TMPRSS2 is a target for anti-cancer therapy. See, for example, International Patent Application Publication No. WO2008127347 (Patent Document 2) and International Patent Application Publication No. WO2002004953 (Patent Document 3).The fusion between TMPRSS2 and ERG (TMPRSS2:ERG) is a gene fusion known to be a major driver of prostate carcinogenesis, triggered by ERα and suppressed by ERβ. Bonkhoff, Estrogen receptor signaling in prostate cancer: Implications for carcinogenesis and tumor progression, Prostate 78(1):2-10(2018) (Non-Patent Document 13). [Prior art documents] [Patent documents]

[0006] [Patent Document 1] International Patent Application Publication No. WO2017 / 151453 [Patent Document 2] International Patent Application Publication No. WO2008127347 [Patent Document 3] International Patent Application Publication No. WO2002004953 [Non-patent literature]

[0007] [Non-Patent Document 1] Zmora et al.,Non-human primate orthologues of TMPRSS2 cleave and activate the influenza virus hemagglutinin.PLoS ONE.12,e0176597(2017) [Non-patent document 2] Bottcher-Friebertshauser et al.,Inhibition of influenza virus infection in human airway cell cultures by an antisense peptide-conjugated morpholino oligomer targeting the hemagglutinin-activating protease TMPRSS2.Journal of Virology.85,1554-1562(2011);

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[0008] For example, although there are small molecule inhibitors of TMPRSS2 and research antibodies that are useful for immunohistochemistry, there is a need in the art for neutralizing therapeutic anti-TMPRSS2 antibodies and their use for treating or preventing viral infections.See, for example, Shen et al.Biochimie 142:1-10(2017), International Patent Application Publication No. 2008127347; International Patent Application Publication No. WO2002004953; U.S. Patent No. 9498529; antibody ab92323 available from Abcam (Cambridge, MA), or antibodies sc-515727 and sc-101847 available from Santa Cruz Biotech (Dallas, TX). The present invention addresses this need, in part, by providing human anti-human TMPRSS2 antibodies, such as mAb8021, mAb8029, and mAb8028, and combinations thereof, including combinations with other therapeutic agents, such as, for example, anti-influenza HA antibodies (e.g., Group I HA or Group II HA) or anti-coronavirus spike protein antibodies (e.g., Severe Acute Respiratory Syndrome (SARS) coronavirus (SARS-CoV), Middle East Respiratory Syndrome (MERS) coronavirus (MERS-CoV), and SARS-CoV-2 (the coronavirus responsible for COVID-19, formerly referred to as 2019-nCoV)), and methods of use thereof to treat viral infections.

[0009] The present invention provides neutralizing human antigen-binding proteins, e.g., antibodies or antigen-binding fragments thereof, that specifically bind to human TMPRSS2. In some aspects, the antibodies or antigen-binding fragments thereof are isolated recombinant antibodies or antigen-binding fragments thereof. In some embodiments, the antigen-binding protein comprises (a) CDR-H1, CDR-H2, and CDR-H3 of an immunoglobulin heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 2, 22, or 42; and / or (b) CDR-L1, CDR-L2, and CDR-L3 of an immunoglobulin light chain comprising the amino acid sequence set forth in SEQ ID NO: 10, 30, or 50. In embodiments of the invention, the antigen-binding protein comprises (a) a light chain immunoglobulin variable region comprising an amino acid sequence having at least 90% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO: 10, 30, or 50; and / or (b) a heavy chain immunoglobulin variable region comprising an amino acid sequence having at least 90% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO: 2, 22, or 42. In embodiments of the invention, the invention provides an antigen binding protein comprising: (a) a light chain immunoglobulin CDR-L1, CDR-L2 and CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 10, 30 or 50 and at least 90% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO: 10, 30 or 50; and / or (b) a heavy chain immunoglobulin CDR-H1, CDR-H2 and CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 2, 22 or 42 and at least 90% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO: 2, 22 or 42.For example, in embodiments of the present invention, an antigen binding protein comprises a heavy chain immunoglobulin variable region comprising: (a) a CDR-H1 comprising the amino acid sequence FTFRSYD (SEQ ID NO:4); (b) a CDR-H2 comprising the amino acid sequence GSAGDT (SEQ ID NO:6); (c) a CDR-H3 comprising the amino acid sequence RVGDWGSGYLDY (SEQ ID NO:8); and a light chain immunoglobulin variable region comprising: (a) a CDR-L1 comprising the amino acid sequence SISIY (SEQ ID NO:12); (b) a CDR-L2 comprising the amino acid sequence AS (SEQ ID NO:14); and / or (c) a CDR-L3 comprising the amino acid sequence QSYGTPFT (SEQ ID NO:16). The present invention also provides antigen binding proteins comprising: (a) a heavy chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:18, 38, or 58; and / or (b) a light chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:20, 40, or 60.

[0010] In some aspects, the antigen binding protein, e.g., an isolated recombinant antibody or antigen-binding fragment thereof, that specifically binds to human transmembrane protease, serine 2 (TMPRSS2) has one or more of the following characteristics: (a) Approximately 10 -9 EC less than M 50 binds to TMPRSS2; (b) demonstrating an increase in survival in animals infected with the coronavirus after administration to the coronavirus-infected animals compared to comparable coronavirus-infected animals that have not been administered; and / or (c) (i) three heavy chain complementarity determining regions (CDRs) (CDR-H1, CDR-H2, and CDR-H3) contained within a heavy chain variable region (HCVR) comprising an amino acid sequence having at least about 90% sequence identity with the HCVR set forth in SEQ ID NO:2; and three light chain CDRs (CDR-L1, CDR-L2, and CDR-L3) contained within a light chain variable region (LCVR) comprising an amino acid sequence having at least about 90% sequence identity with the LCVR set forth in SEQ ID NO:10; or (ii) three heavy chain CDRs (CDR-H1, CDR-H2, and CDR-H3) contained within an HCVR comprising an amino acid sequence having at least about 90% sequence identity to the HCVR set forth in SEQ ID NO: 22; and three light chain CDRs (CDR-L1, CDR-L2, and CDR-L3) contained within an LCVR comprising an amino acid sequence having at least about 90% sequence identity to the LCVR set forth in SEQ ID NO: 30; or (iii) three heavy chain CDRs (CDR-H1, CDR-H2, and CDR-H3) contained within an HCVR comprising an amino acid sequence having at least about 90% sequence identity with the HCVR set forth in SEQ ID NO: 42; and three light chain CDRs (CDR-L1, CDR-L2, and CDR-L3) contained within an LCVR comprising an amino acid sequence having at least about 90% sequence identity with the LCVR set forth in SEQ ID NO: 50. Includes.

[0011] In some embodiments, the antigen binding protein, e.g., an antibody or antigen-binding fragment thereof, comprises three heavy chain CDRs (CDR-H1, CDR-H2, and CDR-H3) contained within an HCVR comprising an amino acid sequence having at least about 90% sequence identity to the HCVR set forth in SEQ ID NO:2; and three light chain CDRs (CDR-L1, CDR-L2, and CDR-L3) contained within an LCVR comprising an amino acid sequence having at least about 90% sequence identity to the LCVR set forth in SEQ ID NO:10.

[0012] In some embodiments, the antigen binding protein, e.g., an antibody or antigen-binding fragment thereof, comprises: (a) an immunoglobulin heavy chain variable region comprising CDR-H1, CDR-H2, and CDR-H3 of the immunoglobulin heavy chain comprising the amino acid sequence set forth in SEQ ID NO:2; and / or (b) an immunoglobulin light chain variable region comprising CDR-L1, CDR-L2, and CDR-L3 of the immunoglobulin light chain comprising the amino acid sequence set forth in SEQ ID NO:10.

[0013] In some embodiments, the antigen binding protein, e.g., an antibody or antigen-binding fragment thereof, comprises (a) a heavy chain immunoglobulin variable region comprising an amino acid sequence having at least 90% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO:2; and / or (b) a light chain immunoglobulin variable region comprising an amino acid sequence having at least 90% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO:10.

[0014] In some aspects, the antigen binding protein, e.g., an antibody or antigen-binding fragment thereof, (a) CDR-H1 comprising the amino acid sequence of SEQ ID NO: 4; (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 6, and / or (c) CDR-H3 comprising the amino acid sequence of SEQ ID NO: 8 a heavy chain immunoglobulin variable region comprising: (a) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 12; (b) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 14, and / or (c) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 16 a light chain immunoglobulin variable region comprising Includes.

[0015] In some embodiments, the antigen binding protein, e.g., an antibody or antigen-binding fragment thereof, comprises (a) a heavy chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO: 18, or an immunoglobulin heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 2; and / or (b) a light chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO: 20, or an immunoglobulin light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 10.

[0016] In some embodiments, the antigen binding protein, e.g., an antibody or antigen-binding fragment thereof, comprises three heavy chain CDRs (CDR-H1, CDR-H2, and CDR-H3) contained within an HCVR comprising an amino acid sequence having at least about 90% sequence identity to the HCVR set forth in SEQ ID NO: 22; and three light chain CDRs (CDR-L1, CDR-L2, and CDR-L3) contained within an LCVR comprising an amino acid sequence having at least about 90% sequence identity to the LCVR set forth in SEQ ID NO: 30.

[0017] In some embodiments, the antigen binding protein, e.g., an antibody or antigen-binding fragment thereof, comprises: (a) an immunoglobulin heavy chain variable region comprising CDR-H1, CDR-H2, and CDR-H3 of the immunoglobulin heavy chain comprising the amino acid sequence set forth in SEQ ID NO:22; and / or (b) an immunoglobulin light chain variable region comprising CDR-L1, CDR-L2, and CDR-L3 of the immunoglobulin light chain comprising the amino acid sequence set forth in SEQ ID NO:30.

[0018] In some embodiments, the antigen binding protein, e.g., an antibody or antigen-binding fragment thereof, comprises (a) a heavy chain immunoglobulin variable region comprising an amino acid sequence having at least 90% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO:22; and / or (b) a light chain immunoglobulin variable region comprising an amino acid sequence having at least 90% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO:30.

[0019] In some aspects, the antigen binding protein, e.g., an antibody or antigen-binding fragment thereof, (a) CDR-H1 comprising the amino acid sequence of SEQ ID NO: 24; (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 26, and / or (c) CDR-H3 comprising the amino acid sequence of SEQ ID NO: 28 a heavy chain immunoglobulin variable region comprising: (a) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 32; (b) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 34, and / or (c) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 36 a light chain immunoglobulin variable region comprising Includes.

[0020] In some embodiments, the antigen binding protein, e.g., an antibody or antigen-binding fragment thereof, comprises (a) a heavy chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:38, or an immunoglobulin heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO:22; and / or (b) a light chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:40, or an immunoglobulin light chain variable region comprising the amino acid sequence set forth in SEQ ID NO:30.

[0021] In some embodiments, the antigen binding protein, e.g., an antibody or antigen-binding fragment thereof, comprises three heavy chain CDRs (CDR-H1, CDR-H2, and CDR-H3) contained within an HCVR comprising an amino acid sequence having at least about 90% sequence identity to the HCVR set forth in SEQ ID NO:42; and three light chain CDRs (CDR-L1, CDR-L2, and CDR-L3) contained within an LCVR comprising an amino acid sequence having at least about 90% sequence identity to the LCVR set forth in SEQ ID NO:50.

[0022] In some embodiments, the antigen binding protein, e.g., an antibody or antigen-binding fragment thereof, comprises: (a) an immunoglobulin heavy chain variable region comprising CDR-H1, CDR-H2, and CDR-H3 of the immunoglobulin heavy chain comprising the amino acid sequence set forth in SEQ ID NO:42; and / or (b) an immunoglobulin light chain variable region comprising CDR-L1, CDR-L2, and CDR-L3 of the immunoglobulin light chain comprising the amino acid sequence set forth in SEQ ID NO:50.

[0023] In some embodiments, the antigen binding protein, e.g., an antibody or antigen-binding fragment thereof, comprises (a) a heavy chain immunoglobulin variable region comprising an amino acid sequence having at least 90% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO:42; and / or (b) a light chain immunoglobulin variable region comprising an amino acid sequence having at least 90% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO:30.

[0024] In some aspects, the antigen binding protein, e.g., an antibody or antigen-binding fragment thereof, (a) CDR-H1 comprising the amino acid sequence of SEQ ID NO: 44; (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 46, and / or (c) a CDR-H3 comprising the amino acid sequence of SEQ ID NO: 48; and / or (a) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 52; (b) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 54, and / or (c) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 56 a light chain immunoglobulin variable region comprising Includes.

[0025] In some aspects, the antigen binding protein, e.g., an antibody or antigen-binding fragment thereof, (a) a heavy chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO: 58, or an immunoglobulin heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 42; and / or (b) a light chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO: 60, or an immunoglobulin light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 50 Includes.

[0026] The present invention also provides any anti-TMPRSS2 antigen binding protein that competes with any of the antigen binding proteins described herein for binding to TMPRSS2 (e.g., as determined by use of a real-time label-free biolayer interferometry assay, e.g., on an Octet RED384 biosensor (Pall ForteBio Corp.)); or that binds to the same or overlapping epitope on TMPRSS2 (or a fragment thereof) as any of the antigen binding proteins described herein.

[0027] The present invention also provides multispecific antigen-binding proteins that bind to TMPRSS2 and another antigen, or a different epitope of TMPRSS2. For example, the multispecific molecule comprises (a) a first antigen-binding domain that specifically binds to TMPRSS2; and (b) a second antigen-binding domain that specifically binds to another antigen or TMPRSS2, or an epitope different from that of the first antigen-binding domain. In some embodiments, the multispecific antigen-binding protein is an antibody or antigen-binding fragment thereof.

[0028] The present invention also provides any anti-TMPRSS2 antigen binding protein (e.g., an antibody or antigen-binding fragment, e.g., an antibody or antigen-binding fragment comprising a sequence described herein) comprising one or more of the following properties: Inhibiting the growth of influenza virus (e.g., A / Puerto Rico / 08 / 1934(H1N1)) or coronavirus (e.g., SARS-CoV, MERS-CoV, or SARS-CoV-2) in TMPRSS2-expressing cells (e.g., Calu-3 cells); · Binds to the surface of TMPRSS-expressing cells; No significant binding to MDCK / Tet-on cells that do not express TMPRSS2; limit the spread of influenza virus or coronavirus infection of cells in vitro; and / or Mice engineered to express the human TMPRSS2 protein are protected from death and / or weight loss caused by influenza virus or coronavirus infection.

[0029] The invention also provides a complex comprising any of the antigen binding proteins described herein that binds to a TMPRSS2 polypeptide, for example, in vitro or in the body of a subject.

[0030] The present invention also provides methods for producing an anti-TMPRSS2 antigen binding protein (e.g., mAb8021, mAb8028, or mAb8029) or immunoglobulin chains thereof described herein, comprising: (a) introducing one or more polynucleotides encoding the immunoglobulin light chain and / or immunoglobulin heavy chain of the antigen binding protein; (b) culturing host cells (e.g., CHO cells, Pichia cells, or Pichia pastoris cells) under conditions favorable for expression of the polynucleotides; and (c) optionally isolating the antigen binding protein or immunoglobulin chain from the host cells and / or the medium in which the host cells are grown. The antigen binding proteins or immunoglobulin chains that are the products of such methods are part of the present invention.

[0031] (a) a V of an antibody or antigen-binding fragment comprising the amino acid sequence set forth in any one of SEQ ID NOs: 2, 22, or 42; H (b) the CDR-H1, CDR-H2, and CDR-H3 domains; or (c) the V of an immunoglobulin chain comprising the amino acid sequence set forth in SEQ ID NO: 10, 30, or 50. L Polypeptides comprising domains CDR-L1, CDR-L2, and CDR-L3 also form part of the present invention. In some embodiments, polynucleotides encoding such polypeptides are provided herein.

[0032] (a) an immunoglobulin chain V comprising the amino acid sequence set forth in SEQ ID NO: 2; H(b) the CDR1, CDR2, and CDR3 of the V domain of an immunoglobulin chain comprising the amino acid sequence set forth in SEQ ID NO: 10 L A polypeptide (e.g., an immunoglobulin) comprising CDR1, CDR2, and CDR3 of the domains (e.g., the polypeptide is in a host cell) also forms part of the invention. Similarly, the invention provides a polypeptide (e.g., an immunoglobulin) comprising (a) the V of an immunoglobulin chain comprising the amino acid sequence set forth in SEQ ID NO:22. H (b) the CDR1, CDR2, and CDR3 of the V domain of an immunoglobulin chain comprising the amino acid sequence set forth in SEQ ID NO: 30 L Similarly, the present invention provides a polypeptide (e.g., the polypeptide is in a host cell) comprising CDR1, CDR2, and CDR3 of the V domain of an immunoglobulin chain comprising the amino acid sequence set forth in SEQ ID NO: 42. H (b) the CDR1, CDR2, and CDR3 of the V domain of an immunoglobulin chain comprising the amino acid sequence set forth in SEQ ID NO: 50 L A polypeptide comprising CDR1, CDR2, and CDR3 of the domain (e.g., the polypeptide is in a host cell) is provided. In some embodiments, a polynucleotide encoding the polypeptide is provided herein.

[0033] The present invention also provides polynucleotides (e.g., DNA or RNA) encoding the polypeptides of the present invention. In embodiments of the invention, the polynucleotides encode two different immunoglobulin chains (e.g., heavy and light chains). In embodiments of the invention, one polynucleotide encodes an immunoglobulin light chain and another polynucleotide encodes an immunoglobulin heavy chain, e.g., the chains are in a host cell or in a container. For example, the polynucleotides are in a vector (e.g., a plasmid) and / or integrated into a host cell chromosome.

[0034] The present invention also provides vectors comprising the polynucleotides described herein.

[0035] A host cell of the invention (e.g., a CHO cell, a Pichia cell, or a Pichia pastoris cell) can contain an anti-TMPRSS2 antigen binding protein (e.g., mAb8021, mAb8028, or mAb8029), a polypeptide thereof, or a polynucleotide encoding such a polypeptide and / or a vector containing such a polynucleotide.

[0036] The present invention also provides compositions or kits comprising an anti-TMPRSS2 antigen binding protein described herein (e.g., mAb8021, mAb8028, or mAb8029) in association with an additional therapeutic agent (e.g., an antiviral agent and / or a vaccine). For example, the composition may be a pharmaceutical composition comprising the antigen binding protein and a pharmaceutically acceptable carrier, and, optionally, an additional therapeutic agent. The additional therapeutic agent may be remdesivir, chloroquine, lopinavir, ritonavir, ribavirin, ledipasvir, sofosbuvir, a combination of ledipasvir and sofosbuvir, oseltamivir, zanamivir, ribavirin, and interferon-alpha 2b, interferon-alpha 2a, and / or an antibody or antigen-binding fragment thereof that specifically binds to influenza HA or coronavirus spike protein. In embodiments of the present invention, the additional therapeutic agent is a medicament for the treatment of TMPRSS2, as described in International Patent Application Publication No. WO2016 / 100807. TIFF0007772702000001.tif59143TIFF0007772702000002.tif230134TIFF0007772702000003.tif175146, or an antigen-binding fragment thereof. In an embodiment of the invention, the additional therapeutic agent is an antibody or antigen-binding fragment thereof selected from the group consisting of H4sH15188P; H1H15188P; H1H15211P; H1H15177P; H4sH15211P; H1H15260P2; H1H15259P2; H1H15203P; H4sH15260P2; H4sH15231P2; H1H15237P2; H1H15208P; H1H15228P2; H1H15233P2; H1H15264P2; H1H15231P2; H1H15253P2; H1H15215P; and H1H15249P2, as described in International Patent Application Publication No. WO / 2015 / 179535.

[0037] In embodiments of the invention, an additional therapeutic agent provided in conjunction with an anti-TMPRSS2 antigen binding protein is an antibody or antigen-binding fragment that binds to an influenza group II HA protein, such as H1H14611N2; or the V of H1H14611N2 H and V L or a heavy chain immunoglobulin comprising CDR-H1, CDR-H2 and CDR-H3 of H1H14611N2 and a light chain immunoglobulin comprising CDR-L1, CDR-L2 and CDR-L3 of H1H14611N2.

[0038] In embodiments of the invention, an additional therapeutic agent provided in conjunction with an anti-TMPRSS2 antigen binding protein is an antibody or antigen-binding fragment that binds to an influenza group II HA protein, such as H1H14612N2; or the V of H1H14612N2. H and V L or a heavy chain immunoglobulin comprising CDR-H1, CDR-H2 and CDR-H3 of H1H14612N2 and a light chain immunoglobulin comprising CDR-L1, CDR-L2 and CDR-L3 of H1H14612N2.

[0039] In embodiments of the invention, an additional therapeutic agent provided in conjunction with an anti-TMPRSS2 antigen binding protein is an antibody or antigen-binding fragment that binds to an influenza group I HA protein, such as H1H11729P; or the V of H1H11729P. H and V L or a heavy chain immunoglobulin comprising CDR-H1, CDR-H2 and CDR-H3 of H1H11729P and a light chain immunoglobulin comprising CDR-L1, CDR-L2 and CDR-L3 of H1H11729P.

[0040] The invention also provides a container or injection device comprising an anti-TMPRSS2 antigen binding protein (e.g., mAb8021, mAb8028, or mAb8029) or a composition (e.g., a pharmaceutical composition) thereof.

[0041] The present invention also provides methods for treating or preventing a viral infection other than an influenza virus infection in a subject (e.g., a human) in need thereof, comprising administering a therapeutically effective amount of an anti-TMPRSS2 antigen binding protein described herein (e.g., mAb8021, mAb8028, or mAb8029).

[0042] The present invention also provides methods for treating or preventing cancer (e.g., prostate cancer, colon cancer, lung cancer, pancreatic cancer, urinary tract cancer, breast cancer, ovarian cancer, prostate adenocarcinoma, renal cell carcinoma, colorectal adenocarcinoma, lung adenocarcinoma, lung squamous cell carcinoma, and / or pleural mesothelioma) or infection, e.g., a viral infection, e.g., influenza virus, coronavirus, SARS-CoV, MERS-CoV, SARS-CoV-2, parainfluenza virus, human metapneumovirus, or hepatitis C virus (HCV) infection, in a subject (e.g., human) in need thereof, comprising administering a therapeutically effective amount of an anti-TMPRSS2 antigen binding protein described herein (e.g., mAb8021, mAb8028, or mAb8029).

[0043] In some aspects, the methods comprise administering the antigen binding protein in association with one or more additional therapeutic agents (e.g., antiviral agents and / or vaccines). In embodiments of the invention, the additional therapeutic agent is a member selected from the group consisting of remdesivir, chloroquine, lopinavir, ritonavir, ribavirin, ledipasvir, sofosbuvir, a combination of ledipasvir and sofosbuvir, oseltamivir, zanamivir, ribavirin, and interferon-alpha 2b, interferon-alpha 2a, and an antibody or antigen-binding fragment thereof that specifically binds to influenza HA. ​​In embodiments of the invention, the additional therapeutic agent is a member selected from the group consisting of: remdesivir, chloroquine, lopinavir, ritonavir, ribavirin, ledipasvir, sofosbuvir, a combination of ledipasvir and sofosbuvir, oseltamivir, zanamivir, ribavirin, and interferon-alpha 2b, interferon-alpha 2a, and an antibody or antigen-binding fragment thereof that specifically binds to influenza HA. ​​In embodiments of the invention, the additional therapeutic agent is a TIFF0007772702000004.tif189143TIFF0007772702000005.tif230134TIFF0007772702000006.tif38134, or an antigen-binding fragment thereof. In an embodiment of the invention, the additional therapeutic agent is an antibody or antigen-binding fragment thereof selected from the group consisting of H4sH15188P; H1H15188P; H1H15211P; H1H15177P; H4sH15211P; H1H15260P2; H1H15259P2; H1H15203P; H4sH15260P2; H4sH15231P2; H1H15237P2; H1H15208P; H1H15228P2; H1H15233P2; H1H15264P2; H1H15231P2; H1H15253P2; H1H15215P; and H1H15249P2, as described in International Patent Application Publication No. WO / 2015 / 179535.

[0044] The present invention also provides methods of administering an anti-TMRPSS2 antigen binding protein (e.g., mAb8021, mAb8028, or mAb8029) described herein to the body of a subject (e.g., a human), comprising injecting the antigen binding protein into the body of the subject. In some embodiments, the antigen binding protein is injected into the body of the subject parenterally (e.g., subcutaneously, intravenously, or intramuscularly). [The present invention 1001] 1. An isolated, recombinant antibody or antigen-binding fragment thereof that specifically binds to human transmembrane protease serine 2 (TMPRSS2), wherein the antibody has the following characteristics: (a) Approximately 10 -9 EC less than M 50 binding to TMPRSS2; (b) demonstrating prolonged survival in coronavirus-infected animals following administration to said coronavirus-infected animals compared to comparable coronavirus-infected animals that have not been administered; and / or (c) (i) three heavy chain complementarity determining regions (CDRs) (CDR-H1, CDR-H2, and CDR-H3) contained within a heavy chain variable region (HCVR) comprising an amino acid sequence having at least about 90% sequence identity with the HCVR set forth in SEQ ID NO:2; and three light chain CDRs (CDR-L1, CDR-L2, and CDR-L3) contained within a light chain variable region (LCVR) comprising an amino acid sequence having at least about 90% sequence identity with the LCVR set forth in SEQ ID NO:10; or (ii) three heavy chain CDRs (CDR-H1, CDR-H2, and CDR-H3) contained within an HCVR comprising an amino acid sequence having at least about 90% sequence identity to the HCVR set forth in SEQ ID NO: 22; and three light chain CDRs (CDR-L1, CDR-L2, and CDR-L3) contained within an LCVR comprising an amino acid sequence having at least about 90% sequence identity to the LCVR set forth in SEQ ID NO: 30; or (iii) three heavy chain CDRs (CDR-H1, CDR-H2, and CDR-H3) contained within an HCVR comprising an amino acid sequence having at least about 90% sequence identity with the HCVR set forth in SEQ ID NO: 42; and three light chain CDRs (CDR-L1, CDR-L2, and CDR-L3) contained within an LCVR comprising an amino acid sequence having at least about 90% sequence identity with the LCVR set forth in SEQ ID NO: 50. Contains An isolated recombinant antibody or antigen-binding fragment thereof, having one or more of: [The present invention 1002] An antibody or antigen-binding fragment thereof of the present invention, comprising: the three heavy chain CDRs (CDR-H1, CDR-H2, and CDR-H3) contained within an HCVR having an amino acid sequence having at least about 90% sequence identity with the HCVR set forth in SEQ ID NO: 2; and the three light chain CDRs (CDR-L1, CDR-L2, and CDR-L3) contained within an LCVR having an amino acid sequence having at least about 90% sequence identity with the LCVR set forth in SEQ ID NO: 10. [The present invention 1003] (a) an immunoglobulin heavy chain variable region comprising the CDR-H1, CDR-H2, and CDR-H3 of the immunoglobulin heavy chain comprising the amino acid sequence set forth in SEQ ID NO:2; and / or (b) an immunoglobulin light chain variable region comprising the CDR-L1, CDR-L2, and CDR-L3 of an immunoglobulin light chain comprising the amino acid sequence set forth in SEQ ID NO: 10; The antibody or antigen-binding fragment of the present invention 1001 or 1002, comprising: [The present invention 1004] (a) a heavy chain immunoglobulin variable region comprising an amino acid sequence having at least 90% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO:2; and / or (b) a light chain immunoglobulin variable region comprising an amino acid sequence having at least 90% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO: 10; The antibody or antigen-binding fragment of any one of 1001 to 1003 of the present invention, comprising: [The present invention 1005] (a) CDR-H1 comprising the amino acid sequence of SEQ ID NO: 4; (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 6, and / or (c) CDR-H3 comprising the amino acid sequence of SEQ ID NO: 8 a heavy chain immunoglobulin variable region comprising: (a) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 12; (b) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 14, and / or (c) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 16 a light chain immunoglobulin variable region comprising The antibody or antigen-binding fragment of any one of 1001 to 1004 of the present invention, comprising: [The present invention 1006] (a) a heavy chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO: 18, or an immunoglobulin heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 2; and / or (b) a light chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO: 20, or an immunoglobulin light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 10 The antibody or antigen-binding fragment of any one of 1001 to 1005 of the present invention, comprising: [The present invention 1007] The three heavy chain CDRs (CDR-H1, CDR-H2, and CDR-H3) contained within an HCVR comprising an amino acid sequence having at least about 90% sequence identity with the HCVR set forth in SEQ ID NO: 22; and the three light chain CDRs (CDR-L1, CDR-L2, and CDR-L3) contained within an LCVR comprising an amino acid sequence having at least about 90% sequence identity with the LCVR set forth in SEQ ID NO: 30. 1001. The antibody or antigen-binding fragment thereof of the present invention, comprising: [The present invention 1008] (a) an immunoglobulin heavy chain variable region comprising the CDR-H1, CDR-H2, and CDR-H3 of the immunoglobulin heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 22; and / or (b) an immunoglobulin light chain variable region comprising the CDR-L1, CDR-L2, and CDR-L3 of the immunoglobulin light chain comprising the amino acid sequence set forth in SEQ ID NO: 30; 1001 or 1007. The antibody or antigen-binding fragment of the present invention, comprising: [The present invention 1009] (a) a heavy chain immunoglobulin variable region comprising an amino acid sequence having at least 90% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO: 22; and / or (b) a light chain immunoglobulin variable region comprising an amino acid sequence having at least 90% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO: 30; The antibody or antigen-binding fragment of any one of 1001 and 1007 to 1008 of the present invention, comprising: [The present invention 1010] (a) CDR-H1 comprising the amino acid sequence of SEQ ID NO: 24; (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 26, and / or (c) CDR-H3 comprising the amino acid sequence of SEQ ID NO: 28 a heavy chain immunoglobulin variable region comprising: (a) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 32; (b) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 34, and / or (c) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 36 a light chain immunoglobulin variable region comprising The antibody or antigen-binding fragment of any one of 1001 and 1007 to 1009 of the present invention, comprising: [The present invention 1011] (a) a heavy chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO: 38, or an immunoglobulin heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 22; and / or (b) a light chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO: 40, or an immunoglobulin light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 30. The antibody or antigen-binding fragment of any one of 1001 or 1007 to 1010 of the present invention, comprising: [The present invention 1012] The three heavy chain CDRs (CDR-H1, CDR-H2, and CDR-H3) contained within an HCVR comprising an amino acid sequence having at least about 90% sequence identity with the HCVR set forth in SEQ ID NO: 42; and the three light chain CDRs (CDR-L1, CDR-L2, and CDR-L3) contained within an LCVR comprising an amino acid sequence having at least about 90% sequence identity with the LCVR set forth in SEQ ID NO: 50. 1001. The antibody or antigen-binding fragment thereof of the present invention, comprising: [The present invention 1013] (a) an immunoglobulin heavy chain variable region comprising the CDR-H1, CDR-H2, and CDR-H3 of the immunoglobulin heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 42; and / or (b) an immunoglobulin light chain variable region comprising the CDR-L1, CDR-L2, and CDR-L3 of the immunoglobulin light chain comprising the amino acid sequence set forth in SEQ ID NO: 50; 1001 or 1012. The antibody or antigen-binding fragment of the present invention, comprising: [The present invention 1014] (a) a heavy chain immunoglobulin variable region comprising an amino acid sequence having at least 90% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO: 42; and / or (b) a light chain immunoglobulin variable region comprising an amino acid sequence having at least 90% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO: 30; The antibody or antigen-binding fragment of any one of 1001 or 1012 to 1013 of the present invention, comprising: [The present invention 1015] (a) CDR-H1 comprising the amino acid sequence of SEQ ID NO: 44; (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 46, and / or (c) CDR-H3 comprising the amino acid sequence of SEQ ID NO: 48 a heavy chain immunoglobulin variable region comprising: (a) CDR-L1 comprising the amino acid sequence of SEQ ID NO: 52; (b) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 54, and / or (c) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 56 a light chain immunoglobulin variable region comprising The antibody or antigen-binding fragment of any one of 1001 or 1012 to 1014 of the present invention, comprising: [The present invention 1016] (a) a heavy chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO: 58, or an immunoglobulin heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 42; and / or (b) a light chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO: 60, or an immunoglobulin light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 50 The antibody or antigen-binding fragment of any one of 1001 or 1012 to 1015 of the present invention, comprising: [The present invention 1017] An antigen-binding protein that competes with any of the antibodies or antigen-binding fragments of the present invention for binding to TMPRSS2. [The present invention 1018] An antigen-binding protein that binds to the same epitope as or an overlapping epitope on TMPRSS2 as any of the antibodies or antigen-binding fragments of the present invention. [The present invention 1019] The antibody or antigen-binding fragment of any one of claims 1001 to 1018, which is polyspecific. [The present invention 1020] The following characteristics: a) inhibiting coronavirus growth in TMPRSS2-expressing cells; b) inhibiting influenza virus growth in TMPRSS2-expressing cells; c) binding to the surface of TMPRSS-expressing cells; d) no significant binding to MDCK / Tet-on cells that do not express TMPRSS2; e) limiting the spread of coronavirus infection of cells in vitro; f) limiting the spread of influenza virus infection of cells in vitro; g) protecting mice engineered to express the human TMPRSS2 protein from death and / or weight loss caused by coronavirus infection; and h) Protect mice engineered to express the human TMPRSS2 protein from death caused by influenza virus infection. The antibody or antigen-binding fragment of any one of 1001 to 1019 of the present invention, comprising one or more of the following: [The present invention 1021] A complex comprising the antibody or antigen-binding fragment of any of 1001 to 1019 of the present invention bound to a TMPRSS2 polypeptide. [The present invention 1022] (a) introducing into a host cell one or more polynucleotides encoding the antibody or antigen-binding fragment; (b) culturing the host cell under conditions favorable for expression of the one or more polynucleotides; and (c) optionally isolating the antibody or antigen-binding fragment from the host cell and / or the medium in which the host cell is grown. A method for producing the antibody or antigen-binding fragment of any of claims 1001 to 1020, comprising: [The present invention 1023] 1023. The method of claim 1022, wherein said host cell is a Chinese hamster ovary cell. [The present invention 1024] An antibody or antigen-binding fragment, which is the product of any of the methods of 1022 to 1023 of the present invention. [The present invention 1025] (a) a V of an antibody or antigen-binding fragment comprising the amino acid sequence set forth in any one of SEQ ID NOs: 2, 22, or 42; H domains CDR-H1, CDR-H2, and CDR-H3; or (b) an immunoglobulin chain V comprising the amino acid sequence set forth in SEQ ID NO: 10, 30, or 50; L Domains CDR-L1, CDR-L2, and CDR-L3 A polypeptide comprising: [The present invention 1026] A polynucleotide encoding the polypeptide of the present invention. [The present invention 1027] A vector comprising the polynucleotide of the present invention. [The present invention 1028] A host cell comprising the antibody or antigen-binding fragment, or polypeptide, or polynucleotide, or vector of any of the present inventions 1001 to 1020 and 1024 to 1027. [The present invention 1029] A composition or kit comprising an antigen-binding protein of any of the inventions 1001 to 1020 and 1024 in association with a further therapeutic agent. [The present invention 1030] A pharmaceutical composition comprising an antigen-binding protein of any of the present inventions 1001 to 1020 and 1024, and a pharmaceutically acceptable carrier, and optionally, a further therapeutic agent. [The present invention 1031] The composition or kit of any of claims 1029 to 1030, associated with a further therapeutic agent which is an antiviral agent or a vaccine. [The present invention 1032] the additional therapeutic agent is a member selected from the group consisting of remdesivir, chloroquine, lopinavir, ritonavir, ribavirin, ledipasvir, sofosbuvir, a combination of ledipasvir and sofosbuvir, oseltamivir, zanamivir, ribavirin, and interferon-alpha 2b, interferon-alpha 2a, an anti-cancer agent, and an antibody or antigen-binding fragment thereof that specifically binds to influenza HA or coronavirus spike protein; and / or TIFF0007772702000007.tif172164TIFF0007772702000008.tif223142TIFF0007772702000009.tif113153 an antibody or antigen-binding fragment thereof selected from the group consisting of The composition or kit of any one of 1029 to 1030 of the present invention. [The present invention 1033] A container or injection device comprising the antigen-binding protein or composition of any one of 1001 to 1020, 1024 and 1029 to 1032 of the present invention. [The present invention 1034] A method for treating or preventing cancer, or infection with influenza virus, coronavirus, SARS-CoV, MERS-CoV, SARS-CoV-2, parainfluenza virus, human metapneumovirus, or hepatitis C virus (HCV) in a subject in need thereof, comprising administering a therapeutically effective amount of any of the antigen-binding proteins of the present inventions 1001 to 1020 and 1024. [This invention 1035] 1034. The method of claim 1034 for treating or preventing cancer, wherein said cancer is prostate cancer, colon cancer, lung cancer, pancreatic cancer, urinary tract cancer, breast cancer, ovarian cancer, prostate adenocarcinoma, renal cell carcinoma, colorectal adenocarcinoma, lung adenocarcinoma, lung squamous cell carcinoma, and / or pleural mesothelioma. [The present invention 1036] The method of any of claims 1034 to 1035, wherein said subject is administered one or more additional therapeutic agents. [This invention 1037] The method of claim 1036, wherein said subject is administered one or more additional therapeutic agents which are antiviral agents or vaccines. [The present invention 1038] one or more additional therapeutic agents that are members selected from the group consisting of remdesivir, chloroquine, lopinavir, ritonavir, ribavirin, ledipasvir, sofosbuvir, a combination of ledipasvir and sofosbuvir, oseltamivir, zanamivir, ribavirin, and interferon-alpha 2b, interferon-alpha 2a, and an antibody or antigen-binding fragment thereof that specifically binds to influenza HA or coronavirus spike protein; and / or TIFF0007772702000010.tif135164TIFF0007772702000011.tif223142TIFF0007772702000012.tif150152 an antibody or antigen-binding fragment thereof selected from the group consisting of The method of any one of claims 1034 to 1035, wherein the compound is administered to the subject. [This invention 1039] A method for administering the antigen-binding protein of any of claims 1001 to 1020 and 1024 to the body of a subject, the method comprising injecting the antigen-binding protein into the body of the subject. [The present invention 1040] 1039. The method of claim 1039, wherein said antigen binding protein is injected into the body of said subject subcutaneously, intravenously, or intramuscularly. DETAILED DESCRIPTION OF THE INVENTION

[0045] Detailed Description of the Invention Before describing the present methods, it is to be understood that this invention is not limited to the particular methods and experimental conditions described, as methods and conditions may vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present invention will be limited only by the appended claims.

[0046] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. Although any methods and materials similar or equivalent to those described herein can be used in the practice or testing of the present invention, the preferred methods and materials are described herein. All publications mentioned herein are incorporated herein by reference in their entirety.

[0047] The term "coronavirus" or "CoV" refers to any virus in the coronavirus family, including, but not limited to, MERS-CoV, SARS-CoV, and SARS-CoV-2. SARS-CoV-2 refers to the newly emerged coronavirus that causes COVID-19, which was initially identified as the cause of a significant outbreak that began in Wuhan, China, and has spread rapidly worldwide. It binds to the human host cell receptor angiotensin-converting enzyme 2 (ACE2) via the viral spike protein. The spike protein also binds to and is cleaved by TMPRSS2, activating the spike protein for viral membrane fusion.

[0048] The term "CoV-S," also referred to as "S protein," refers to the spike protein of coronaviruses and can refer to specific S proteins, such as MERS-CoV-S, SARS-CoV-S, and SARS-CoV-2-S. The SARS-CoV-2 spike protein is a 1,273-amino acid type I membrane glycoprotein assembled into trimers that constitute spikes, or peplomers, on the surface of enveloped coronavirus particles. The protein has two essential functions: host receptor binding and membrane fusion, which are attributed to the N-terminal (S1) and C-terminal (S2) halves of the S protein. CoV-S binds to its cognate receptor through a receptor-binding domain (RBD) present in the S1 subunit. The amino acid sequence of the full-length SARS-CoV-2 spike protein is exemplified by the amino acid sequence provided in SEQ ID NO: 61. The term "CoV-S" includes protein variants of CoV spike proteins isolated from different CoV isolates, as well as recombinant CoV spike proteins or fragments thereof. The term also encompasses CoV spike proteins or fragments thereof linked to a signal sequence, such as, for example, a histidine tag, mouse or human Fc, or ROR1.

[0049] As used herein, the terms "coronavirus infection" or "CoV infection" refer to infection with a coronavirus, such as MERS-CoV, SARS-CoV, or SARS-CoV-2. This term often includes a respiratory tract infection in the lower respiratory tract. Symptoms include high fever, cough, shortness of breath, pneumonia, gastrointestinal symptoms such as diarrhea, organ failure (renal failure and dysfunction), septic shock, and, in severe cases, death. SARS-CoV-2 infection can cause coronavirus disease 2019 (COVID-19), which can cause symptoms such as fever, chills, shortness of breath, congestion, cough, fatigue, body / muscle aches, and loss of taste and / or smell.

[0050] The term "influenza hemagglutinin," also known as "influenza HA," is a trimeric glycoprotein found on the surface of influenza virus particles that mediates viral attachment to host cells (via HA1 binding to α-2,3 sialic acid and α-2,6 sialic acid) and entry (through conformational changes). HA consists of two structural domains: a globular head domain (subject to frequent antigenic variation) containing the receptor-binding site and a stem region (more conserved among various influenza virus strains). Influenza HA is synthesized as a precursor (HA0) and undergoes proteolytic processing to generate two subunits (HA1 and HA2), which associate with each other to form the stem / globular head structure. While viral HA is the most variable antigen within the virus, the stem (HA2) is highly conserved within each group.

[0051] The term "influenza neuraminidase," also referred to as "influenza NA," is an exosialidase (EC 3.2.1.18) that cleaves the α-ketosidic bond between sialic acid (N-acetylneuraminic acid) and the adjacent sugar residue.

[0052] The amino acid sequence of a full-length influenza HA is exemplified by the amino acid sequence of influenza isolate H1N1 A / California / 04 / 2009, provided in GenBank under accession number FJ966082.1. The term "influenza-HA" also includes protein variants of influenza HA isolated from different influenza isolates, such as GQ149237.1, NC_002017, KM972981.1, etc. The term "influenza-HA" also includes recombinant influenza HA or fragments thereof. The term also encompasses influenza HA or fragments thereof linked, for example, to a histidine tag, mouse or human Fc, or signal sequence.

[0053] An anti-TMPRSS2 "antigen binding protein" is a polypeptide or a complex of one or more polypeptides (e.g., a tetrameric IgG antibody) that specifically binds to a TMPRSS2 polypeptide, e.g., an anti-TMPRSS2 antibody or antigen-binding fragment, whether monospecific or multispecific.

[0054] TMPRSS2 TMPRSS2 (transmembrane spanning proteinase serine 2) is a protein located on human chromosome 21 that belongs to a family of serine proteases (type II transmembrane serine proteases (TTSPs)) that are important for influenza virus infectivity. TMPRSS2 has been shown to mediate the cleavage of influenza virus HA0 into HA1 and HA2.

[0055] The human TMPRSS2 gene encodes a predicted protein of 492 amino acids that is anchored in the cell membrane. The protein is converted to its mature form via autocatalytic cleavage between Arg255 and Ile256. After cleavage, the mature protease is primarily membrane-bound, although a portion may be released into the extracellular environment.

[0056] In an embodiment of the invention, human TMPRSS2(V160M) has the amino acid sequence TIFF0007772702000013.tif59150 (SEQ ID NO: 63; methionine 160 in bold). In an embodiment of the invention, the TMPRSS2 polypeptide does not comprise the V160M mutation. See also NCBI Accession No. NM_005656.3.

[0057] In an embodiment of the invention, rhesus TMPRSS2 (S129L, N251S, I415V, R431Q, D492G) has the amino acid sequence TIFF0007772702000014.tif59150 (SEQ ID NO: 64). In an embodiment of the invention, the TMPRSS2 polypeptide does not comprise the S129L, N251S, I415V, R431Q and / or D492G mutations.

[0058] In an embodiment of the invention, the Mus musculus TMPRSS2 mRNA comprises the nucleotide sequence set forth in NCBI accession number NM_015775.2.

[0059] virus The present invention includes methods for treating or preventing viral infection in a subject. The term "virus" includes any virus whose infection in a subject's body is treatable or preventable by administration of an anti-TMPRSS2 antibody or antigen-binding fragment thereof (e.g., the infectivity of the virus depends, at least in part, on TMPRSS2). In embodiments of the present invention, a "virus" is any virus that expresses HA0, a spike protein (e.g., CoV-S), or another substrate of TMPRSS2 whose proteolytic cleavage is required for full infectivity of the virus to cells in the host. The term "virus" also includes TMPRSS2-dependent respiratory viruses, which are viruses that infect respiratory tissues (e.g., upper and / or lower respiratory tract, trachea, bronchi, lungs) of a subject and that are treatable or preventable by administration of an anti-TMPRSS2 antibody. For example, in embodiments of the invention, viruses include influenza virus, coronavirus, SARS-CoV (severe acute respiratory syndrome coronavirus), MERS-CoV (Middle East respiratory syndrome (MERS) coronavirus), SARS-CoV-2, parainfluenza virus, Sendai virus (SeV), human metapneumovirus, and / or hepatitis C virus (HCV). "Viral infection" refers to the entry and proliferation of a virus in a subject's body. The invention includes embodiments in which the term "virus" excludes influenza virus, for example, where viral infection excludes influenza virus infection.

[0060] Coronavirus virions are spherical, approximately 125 nm in diameter. The most striking feature of coronaviruses is the club-shaped spikes that protrude from the surface of the virion. These spikes are the defining characteristic of the virion, giving it the appearance of a solar corona and evoking the name coronavirus. Within the virion envelope is a nucleocapsid. Coronaviruses possess a helically symmetric nucleocapsid, which is rare in positive-sense RNA viruses but much more common in negative-sense RNA viruses. MERS-CoV (Middle East Respiratory Syndrome Coronavirus), SARS-CoV (Severe Acute Respiratory Syndrome Coronavirus), and SARS-CoV-2 belong to the coronavirus family. Initial binding of the virion to the host cell is initiated by the interaction between the S protein and its receptor. The location of the receptor-binding domain (RBD) within the S1 region of the coronavirus S protein varies depending on the virus; some have the RBD at the C-terminus of S1. The S protein / receptor interaction is a major determinant for coronaviruses to infect host species and also governs the tissue tropism of the virus. Many coronaviruses utilize peptidases as their cellular receptors. After receptor binding, the virus must then access the host cell cytosol. This is generally achieved by acid-dependent proteolytic cleavage of the S protein by a cathepsin, TMPRRS2, or another protease, followed by fusion of the viral and cellular membranes.

[0061] Influenza viruses are members of the Orthomyxoviridae family. This family represents enveloped viruses whose genomes contain segmented, negative-sense, single-stranded RNA segments. This family includes four genera: A, B, C, and Thogotovirus. Influenza virus classes A, B, and C are further divided into subtypes based on the core protein and determined by the viral envelope glycoproteins hemagglutinin (HA) and neuraminidase (NA) (e.g., subtype A / H1N1). There are at least 18 influenza hemagglutinin ("HA") protein subtypes (H1-H18 or HA1-HA18) and at least 11 influenza neuraminidase (NA) protein subtypes (N1-N11 or NA1-NA11) that are used to define influenza subtypes. Group 1 influenza viruses include H1, H2, H5, H6, H8, H9, H11, H12, H13, H16, H17, and H18 subtypes, as well as NA8, NA5, Na4, and NA1 subtypes. Group 2 viruses include H3, H4, H7, H10, H14, and H15 subtypes, as well as NA6, NA9, NA7, NA2, and NA3 subtypes. Influenza A viruses infect a variety of mammalian and avian species, while infection with influenza B and C viruses is primarily limited to humans. The eight genome segments of influenza A and B viruses are loosely enclosed by nucleoproteins.

[0062] Currently, there are two genera of human parainfluenza viruses (HPIVs): respiroviruses (HPIV-1 and HPIV-3) and rubulaviruses (HPIV-2 and HPIV-4). Both genera (paramyxoviruses) are morphologically distinct from influenza viruses.

[0063] Sendai virus, also known as murine parainfluenza virus, is a type species of the Respirovirus genus, which also includes human parainfluenza virus 3, bovine parainfluenza virus 3, and human parainfluenza virus 1. TMPRSS2 is an activating protease for respiratory parainfluenza viruses, such as parainfluenza virus, and Sendai virus (SeV). See, for example, Abe et al., J. Virol. 87(21):11930-11935(2013).

[0064] Human metapneumovirus (HMPV) is classified as the first human member of the Metapneumovirus genus in the Pneumovirinae subfamily within the Paramyxoviridae family. It is an enveloped, negative-sense, single-stranded RNA virus. The RNA genome contains eight genes encoding nine different proteins. HMPV has the same gene order as avian pneumovirus (AMPV), which also belongs to the Metapneumovirus genus. TMPRSS2 is expressed in human lung epithelium, efficiently cleaves the HMPV F protein, supports HMPV proliferation, and may be involved in the development of lower respiratory tract disease in HMPV-infected patients. See, for example, Shirogane et al. J Virol. 82(17):8942-8946 (2008).

[0065] Hepatitis C virus (HCV) is a small, enveloped, positive-sense, single-stranded RNA virus in the family Flaviviridae. With at least six genotypes and numerous subtypes, HCV is a member of the genus Hepacivirus. TMPRSS2 can activate HCV infection after the binding and entry stages. See Esumi et al., Hepatology 61(2):437-446(2015).

[0066] Anti-TMPRSS2 antibodies and antigen-binding fragments The present invention provides antigen-binding proteins, such as antibodies and antigen-binding fragments thereof, that specifically bind to a TMPRSS2 protein or an antigen-binding fragment thereof.

[0067] As used herein, the term "antibody" refers to an immunoglobulin molecule comprising four polypeptide chains, two heavy chains (HC) and two light chains (LC) interconnected by disulfide bonds (i.e., a "full antibody molecule"), and multimers thereof (e.g., IgM). Exemplary antibodies include, for example, mAb8021, mAb8028, and mAb8029. Each heavy chain comprises a heavy chain variable region ("HCVR" or "V"). H ") (e.g., SEQ ID NO: 2) and a heavy chain constant region (domain C H 1. C H 2, and C H Each light chain comprises a light chain variable region ("LCVR" or "VVR"). L ") (e.g., SEQ ID NO: 4) and a light chain constant region (C L ) V H and V L The regions can be further subdivided into regions of hypervariability called complementarity determining regions (CDRs) interspersed with more conserved regions called framework regions (FRs). H and V L comprises three CDRs and four FRs arranged from amino terminus to carboxy terminus in the following order: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. In certain embodiments of the invention, the FRs of an antibody (or antigen-binding fragment thereof) are identical to human germline sequences or are naturally or artificially modified.

[0068] Typically, the variable domains of both heavy and light immunoglobulin chains contain three hypervariable regions, also called complementarity-determining regions (CDRs), located within relatively conserved framework regions (FRs). Generally, from N- to C-terminus, both light and heavy chain variable domains contain FR1, CDR1, FR2, CDR2, FR3, CDR3, and FR4. In an embodiment of the invention, the assignment of amino acids to each domain follows the immunoglobulin definition of the protein of interest, Kabat et al.; National Institutes of Health, Bethesda, Md.; 5th ed.; NIH Publication No. 91-3242 (1991); Kabat (1978) Adv. Prot. Chem. 32:1-75; Kabat et al., (1977) J. Biol. Chem. 252:6609-6616; Chothia et al., (1987) J Mol. Biol. 196:901-917 or Chothia et al., (1989) Nature 342:878-883.

[0069] The present invention includes monoclonal anti-TMPRSS2 antigen-binding proteins, e.g., antibodies and antigen-binding fragments thereof, as well as monoclonal compositions comprising a plurality of isolated monoclonal antigen-binding proteins. As used herein, the term "monoclonal antibody" refers to a population of substantially homogeneous antibodies, i.e., the antibody molecules comprising the population are identical in amino acid sequence, except for possible naturally occurring mutations that may be present in minor amounts. The "plurality" of such monoclonal antibodies and fragments in a composition refers to a concentration of identical (i.e., as discussed above in amino acid sequence, except for possible naturally occurring mutations that may be present in minor amounts) antibodies and fragments in excess of that which normally occurs in nature in the blood of a host organism, e.g., a mouse or human.

[0070] In embodiments of the invention, the anti-TMPRSS2 antigen-binding protein, e.g., antibody or antigen-binding fragment, comprises a heavy chain constant domain, e.g., of the IgA (e.g., IgA1 or IgA2), IgD, IgE, IgG (e.g., IgG1, IgG2, IgG3 and IgG4) or IgM type. In embodiments of the invention, the antigen-binding protein, e.g., antibody or antigen-binding fragment, comprises a light chain constant domain, e.g., of the kappa or lambda type.

[0071] As used herein, the term "human" antigen-binding protein, such as an antibody, includes antibodies having variable and constant regions derived from human germline immunoglobulin sequences, whether in human cells or grafted into non-human cells, e.g., mouse cells. See, e.g., U.S. Patent No. 8502018, U.S. Patent No. 6596541, or U.S. Patent No. 5789215. Human mAbs of the invention may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced in vitro by random or site-directed mutagenesis or in vivo by somatic mutation), e.g., in the CDRs, particularly CDR3. However, as used herein, the term "human antibody" is not intended to include mAbs in which CDR sequences derived from the germline of another mammalian species (e.g., a mouse) are grafted onto human FR sequences. The term includes antibodies recombinantly produced in non-human mammals or in the cells of non-human mammals. The term is not intended to include antibodies isolated from or generated in a human subject. See below.

[0072] The present invention includes anti-TMPRSS2 chimeric antigen-binding proteins, e.g., antibodies and antigen-binding fragments thereof, and methods of using same. As used herein, a "chimeric antibody" is an antibody having a variable domain from a first antibody and a constant domain from a second antibody, wherein the first and second antibodies are from different species. (See U.S. Pat. No. 4,816,567; and Morrison et al., (1984) Proc. Natl. Acad. Sci. USA 81:6851-6855.)

[0073] The term "recombinant" antigen-binding protein, such as an antibody or antigen-binding fragment, refers to such molecules that are produced, expressed, isolated, or obtained by technologies or methods known in the art as recombinant DNA technology, including, for example, DNA splicing and transgenic expression. This term includes antibodies expressed in non-human mammalian (including, for example, transgenic non-human mammalian, such as transgenic mice) or cellular (e.g., CHO cell) expression systems or isolated from recombinant combinatorial human antibody libraries.

[0074] Recombinant anti-TMPRSS2 antigen-binding proteins, e.g., antibodies and antigen-binding fragments, disclosed herein can also be produced in an E. coli / T7 expression system. In this embodiment, a nucleic acid encoding an anti-TMPRSS2 antibody immunoglobulin molecule of the invention (e.g., mAb8021, mAb8028, or mAb8029) can be inserted into a pET-based plasmid and expressed in an E. coli / T7 system. For example, the invention includes a method of expressing an antibody or antigen-binding fragment thereof in a host cell (e.g., a bacterial host cell such as E. coli, e.g., BL21 or BL21DE3), comprising expressing T7 RNA polymerase in the cell, which also contains a polynucleotide encoding an immunoglobulin chain operably linked to a T7 promoter. For example, in an embodiment of the invention, the bacterial host cell, e.g., E. coli, contains a polynucleotide encoding a T7 RNA polymerase gene operably linked to a lac promoter, and expression of the polymerase and chains is induced by incubation of the host cell with IPTG (isopropyl-beta-D-thiogalactopyranoside). See U.S. Pat. Nos. 4,952,496 and 5,693,489, or Studier and Moffatt, Use of bacteriophage T7 RNA polymerase to direct selective high-level expression of cloned genes, J. Mol. Biol. 1986 May 5;189(1):113-30.

[0075] There are several methods for producing recombinant antibodies known in the art. One example of a method for recombinant production of antibodies is disclosed in US4816567.

[0076] Transformation can be by any known method of introducing polynucleotides into host cells.Methods for introducing heterologous polynucleotides into mammalian cells are well known in the art, including dextran-mediated transfection, calcium phosphate precipitation, polybrene-mediated transfection, protoplast fusion, electroporation, encapsulation of polynucleotides in liposomes, biolistic injection, and direct microinjection of DNA into the nucleus.In addition, nucleic acid molecules can be introduced into mammalian cells by viral vectors.Methods for transforming cells are well known in the art.See, for example, U.S. Patent Nos. 4,399,216, 4,912,040, 4,740,461, and 4,959,455.

[0077] Accordingly, the present invention provides a method for producing an antigen-binding protein, e.g., a method for producing an antigen-binding protein, comprising: (i) introducing one or more polynucleotides (e.g., comprising the nucleotide sequence in any one or more of SEQ ID NOs: 1, 3, 5, 7, 9, 11, 13, 15, 17, 19, 21, 23, 25, 27, 29, 31, 33, 35, 37, 39, 41, 43, 45, 47, 49, 51, 53, 55, 57, or 59) encoding an immunoglobulin light chain and / or an immunoglobulin heavy chain, or CDRs, of mAb8021, mAb8028, or mAb8029, e.g., the polynucleotides are introduced into a vector and / or integrated into a host cell chromosome and / or operably linked to a promoter; (ii) culturing a host cell (e.g., CHO or Pichia or Pichia pastoris) under conditions favorable for expression of the polynucleotide, and (iii) optionally isolating the antigen-binding protein (e.g., antibody or fragment) or chain from the host cell and / or the medium in which the host cell is grown. When producing an antigen-binding protein (e.g., antibody or antigen-binding fragment) that comprises more than one immunoglobulin chain, e.g., an antibody that comprises two heavy immunoglobulin chains and two light immunoglobulin chains, co-expression of the chains in a single host cell leads to pairing of the chains to form the antigen-binding protein (e.g., antibody or antigen-binding fragment), e.g., in the cell or on the cell surface, or extracellularly if such chains are secreted. Methods include those in which only an immunoglobulin heavy chain or only an immunoglobulin light chain (e.g., any of those disclosed herein, including mature fragments and / or variable domains thereof) is expressed. Such chains are useful, for example, as intermediates in the expression of antibodies or antigen-binding fragments comprising such chains.For example, the present invention also includes anti-TMPRSS2 antigen-binding proteins, such as antibodies and antigen-binding fragments thereof, comprising a heavy chain immunoglobulin (or a variable domain including its CDRs) encoded by a polynucleotide comprising the nucleotide sequence set forth in SEQ ID NO: 1 and a light chain immunoglobulin (or a variable domain including its CDRs) encoded by the nucleotide sequence set forth in SEQ ID NO: 9, that are products of such production methods and, optionally, the purification methods described herein. The present invention also includes anti-TMPRSS2 antigen-binding proteins, such as antibodies and antigen-binding fragments thereof, comprising a heavy chain immunoglobulin (or a variable domain including its CDRs) encoded by a polynucleotide comprising the nucleotide sequence set forth in SEQ ID NO: 21 and a light chain immunoglobulin (or a variable domain including its CDRs) encoded by the nucleotide sequence set forth in SEQ ID NO: 29, that are products of such production methods and, optionally, the purification methods described herein. The present invention also includes anti-TMPRSS2 antigen binding proteins, such as antibodies and antigen-binding fragments thereof, comprising a heavy chain immunoglobulin (or a variable domain thereof including CDRs) encoded by a polynucleotide comprising the nucleotide sequence set forth in SEQ ID NO: 41 and a light chain immunoglobulin (or a variable domain thereof including CDRs) encoded by the nucleotide sequence set forth in SEQ ID NO: 49, that are the products of such production methods, and optionally, the purification methods described herein. For example, in embodiments of the present invention, the product of the method comprises the amino acid sequence set forth in SEQ ID NO: 2. H and V comprising the amino acid sequence set forth in SEQ ID NO: 10 L or an antibody or fragment comprising an HC comprising the amino acid sequence set forth in SEQ ID NO: 18 and an LC comprising the amino acid sequence set forth in SEQ ID NO: 20. In another embodiment of the invention, the product of the method is an anti-TMPRSS2 antigen binding protein that is an antibody or fragment comprising a V comprising the amino acid sequence set forth in SEQ ID NO: 22. H and V comprising the amino acid sequence set forth in SEQ ID NO: 30 Lor an antibody or fragment comprising an HC comprising the amino acid sequence set forth in SEQ ID NO: 38 and an LC comprising the amino acid sequence set forth in SEQ ID NO: 40. In another embodiment of the invention, the product of the method is an anti-TMPRSS2 antigen binding protein that is an antibody or fragment comprising a V comprising the amino acid sequence set forth in SEQ ID NO: 42. H and V comprising the amino acid sequence set forth in SEQ ID NO: 50 L or an antibody or fragment comprising an HC comprising the amino acid sequence set forth in SEQ ID NO:58 and an LC comprising the amino acid sequence set forth in SEQ ID NO:60.

[0078] Eukaryotic and prokaryotic host cells, including mammalian cells, may be used as hosts for expression of anti-TMPRSS2 antigen-binding proteins. Such host cells are well known in the art, and many are available from the American Type Culture Collection (ATCC). These host cells include, among others, Chinese hamster ovary (CHO) cells, NSO, SP2 cells, HeLa cells, baby hamster kidney (BHK) cells, monkey kidney cells (COS), human hepatocellular carcinoma cells (e.g., Hep G2), A549 cells, 3T3 cells, HEK-293 cells, and numerous other cell lines. Mammalian host cells include human, mouse, rat, dog, monkey, pig, goat, cow, horse, and hamster cells. Other cell lines that can be used include insect cell lines (e.g., Spodoptera frugiperda or Trichoplusia ni), amphibian cells, bacterial cells, plant cells, and fungal cells.Fungal cells include, for example, Pichia pastoris, Pichia finlandica, Pichia trehalophila, Pichia koclamae, Pichia membranaefaciens, Pichia minuta (Ogataea minuta, Pichia lindneri), Pichia opuntiae, Pichia thermotolerans, Pichia salictaria, Pichia guercuum, Pichia piperi, and the like. pijperi, Pichia stiptis, Pichia methanolica, Pichia species, Saccharomyces cerevisiae, Saccharomyces species, Hansenula polymorpha, Kluyveromyces species, Kluyveromyces lactis, Candida albicans, Aspergillus nidulans, Aspergillus niger, Aspergillus oryzae, Trichoderma reesei, Chrysosporium rockenown lucknowense, Fusarium species, Fusarium gramineum, Fusarium venenatum, Physcomitrella patens, and Neurospora crassa, as well as yeast and filamentous fungal cells.The present invention includes an isolated host cell (eg, a CHO cell) comprising an antigen binding protein, such as mAb8021, mAb8028, or mAb8029; or a polynucleotide encoding such a polypeptide.

[0079] The term "specifically binds" refers to an affinity of at least about 10 as measured, for example, on an Octet® HTX biosensor at 25°C or 37°C by real-time label-free biolayer interferometry assay, or surface plasmon resonance, e.g., BIACORE™, or solution affinity ELISA. -8 M (e.g., 2.81 × 10 -9 M;9.31×10 -9 M;10 -9 M;10 -10 M, 10 -11 M, or 10 -12 M)'s K D The term "antigen-binding protein" refers to an antigen-binding protein (e.g., a mAb) that has binding affinity to an antigen, such as a TMPRSS2 protein (e.g., human TMPRSS2), represented as: The present invention includes antigen-binding proteins that specifically bind to a TMPRSS2 protein.

[0080] As used herein, the terms "antigen-binding portion" or "antigen-binding fragment" of an antibody or antigen-binding protein include any naturally occurring, enzymatically obtainable, synthetic, or genetically engineered polypeptide or glycoprotein that specifically binds to an antigen to form a complex. Non-limiting examples of antigen-binding fragments include (i) Fab fragments, (ii) F(ab')2 fragments, (iii) Fd fragments, (iv) Fv fragments, (v) single-chain Fv (scFv) molecules, (vi) dAb fragments, and (vii) minimal recognition units consisting of amino acid residues mimicking the hypervariable regions of an antibody (e.g., isolated complementarity-determining regions (CDRs), such as CDR3 peptides), or a constrained FR3-CDR3-FR4 peptide. Other engineered molecules, such as domain-specific antibodies, single-domain antibodies, domain-deleted antibodies, chimeric antibodies, CDR-grafted antibodies, diabodies, triabodies, tetrabodies, minibodies, nanobodies (e.g., as defined in International Patent Application Publication No. WO08 / 020079 or International Patent Application Publication No. WO09 / 138519) (e.g., monovalent nanobodies, bivalent nanobodies, etc.), small modular immunopharmaceuticals (SMIPs), and shark variable IgNAR domains, are also encompassed by the term "antigen-binding fragment" as used herein. In embodiments of the invention, the antigen-binding fragment comprises three or more CDRs (e.g., CDR-H1, CDR-H2, and CDR-H3; or CDR-L1, CDR-L2, and CDR-L3) of mAb8021, mAb8028, or mAb8029.

[0081] Antigen-binding fragments of antibodies, in embodiments of the present invention, comprise at least one variable domain. A variable domain may be of any size or amino acid composition and generally comprises at least one CDR adjacent to, or in frame with, one or more framework sequences. L V bound to the domain H In an antigen-binding fragment having a domain, V H Domains and V L The domains can be positioned relative to each other in any suitable arrangement. For example, the variable region can be a dimer, with the VH -V H , V H -V L , or V L -V L Alternatively, the antigen-binding fragment of an antibody may contain a dimer of monomeric V H or V L It may contain domains.

[0082] In certain embodiments, an antigen-binding fragment of an antibody may contain at least one variable domain covalently linked to at least one constant domain. Non-limiting exemplary configurations of variable and constant domains that may be found in an antigen-binding fragment of an antibody of the invention include: (i) a V H -C H 1, (ii) V H -C H 2, (iii) V H -C H 3, (iv) V H -C H 1-C H 2. (v) V H -C H 1-C H 2-C H 3. (vi) V H -C H 2-C H 3, (vii)V H -C L、 (viii)V L -C H 1, (ix)V L -C H 2. (x)V L -C H 3. (xi) V L -C H 1-C H 2, (xii)V L -C H 1-C H 2-C H 3, (xiii)V L -C H 2-C H 3, and (xiv) V L -C LIn any configuration of variable and constant domains, including any of the exemplary configurations listed above, the variable and constant domains may be directly linked to each other or may be linked by a full or partial hinge or linker region. The hinge region may consist of at least two (e.g., 5, 10, 15, 20, 40, 60, or more) amino acids that provide a flexible or semi-flexible connection between adjacent variable and / or constant domains in a single polypeptide molecule. Furthermore, antigen-binding fragments of antibodies of the present invention may be linked to each other and / or to one or more monomeric V H Or V L It may comprise homodimers or heterodimers (or other multimers) of any of the variable and constant domain configurations listed above in non-covalent association with the domains (e.g., via disulfide bonds).

[0083] Antigen-binding proteins (e.g., antibodies and antigen-binding fragments) can be monospecific or multispecific (e.g., bispecific). Multispecific antigen-binding proteins are discussed further herein.

[0084] In certain embodiments, the antibodies or antibody fragments of the invention may be conjugated to a moiety, such as a ligand or therapeutic moiety (an "immunoconjugate"), such as an antiviral agent, a second anti-influenza antibody, or any other therapeutic moiety useful for treating a viral infection, e.g., influenza virus infection. See below.

[0085] The present invention also provides complexes comprising an anti-TMPRSS2 antigen-binding protein, e.g., an antibody or antigen-binding fragment, as discussed herein, complexed with a TMPRSS2 polypeptide or antigenic fragment thereof and / or a secondary antibody or antigen-binding fragment thereof (e.g., a directly labeled secondary antibody) that specifically binds to the anti-TMPRSS2 antibody or fragment. In embodiments of the invention, the antibody or fragment is in vitro (e.g., immobilized on a solid substrate) or within a subject's body. In embodiments of the invention, TMPRSS2 is in vitro (e.g., immobilized on a solid substrate), on the surface of a cell, or within a subject's body. Immobilized anti-TMPRSS2 antibodies and antigen-binding fragments thereof covalently bound to an insoluble substrate material (e.g., glass or polysaccharides such as agarose or Sepharose, e.g., beads or other particles thereof) are also part of the invention, and optionally the immobilized antibody is complexed with TMPRSS2 or an antigenic fragment thereof or a secondary antibody or fragment thereof.

[0086] "Isolated" antigen-binding proteins, antibodies or antigen-binding fragments thereof, polypeptides, polynucleotides, and vectors are at least partially free from other biological molecules from the cell or cell culture in which they are produced. Such biological molecules include nucleic acids, proteins, other antibodies or antigen-binding fragments, lipids, carbohydrates, or other substances such as cellular debris and growth medium. Isolated antibodies or antigen-binding fragments may further be at least partially free from expression system components such as biological molecules of the host cell or its growth medium. In general, the term "isolated" is not intended to refer to the complete absence of such biological molecules, or the absence of water, buffers, or salts, or the absence of components of a pharmaceutical formulation that includes the antibody or fragment.

[0087] The term "epitope" refers to a specific antigen-binding site of an antigen-binding protein, known as a paratope, e.g., an antigenic determinant (e.g., on a TMPRSS2 polypeptide) that interacts with the variable region of an antibody molecule. A single antigen may have more than two epitopes. Thus, different antibodies may bind to different regions on an antigen and have different biological effects. The term "epitope" also refers to the site on an antigen to which B cells and / or T cells respond. The term also refers to the region of an antigen to which an antibody binds. Epitopes may be defined as structural or functional. Functional epitopes are generally a subset of structural epitopes and contain residues directly involved in the affinity of the interaction. Epitopes may be linear or steric, i.e., composed of nonlinear amino acids. In certain embodiments, epitopes may include determinants that are chemically active surface groups of molecules such as amino acids, sugar side chains, phosphoryl groups, or sulfonyl groups, and in certain embodiments, may have specific three-dimensional structural characteristics, and / or specific charge characteristics.

[0088] Methods for determining the epitope of an antigen-binding protein, e.g., an antibody or fragment or polypeptide, include alanine scanning mutation analysis, peptide blot analysis (Reineke (2004) Methods Mol. Biol. 248:443-63), peptide truncation analysis, crystallographic studies, and NMR analysis. Additionally, methods such as epitope excision, epitope extraction, and chemical modification of antigens can be used (Tomer (2000) Prot. Sci. 9:487-496). Another method that can be used to identify amino acids within a polypeptide with which an antigen-binding protein (e.g., an antibody or fragment or polypeptide) (e.g., conversin) interacts is hydrogen / deuterium exchange detected by mass spectrometry. In general terms, the hydrogen / deuterium exchange method involves labeling a protein of interest with deuterium, followed by binding the antigen-binding protein, e.g., an antibody or fragment or polypeptide, to the deuterium-labeled protein. The TMPRSS2 protein / antigen-binding protein complex is then transferred to water, and exchangeable protons in amino acids protected by the antibody complex undergo back-exchange from deuterium to hydrogen at a slower rate than exchangeable protons in amino acids that are not part of the interface. As a result, amino acids that form part of the protein / antigen-binding protein may retain deuterium and therefore exhibit a relatively high mass compared to amino acids that are not included in the interface. After dissociation of the antigen-binding protein (e.g., antibody or fragment or polypeptide), the target protein is subjected to protease cleavage and mass spectrometry analysis, thereby revealing deuterium-labeled residues corresponding to the specific amino acids with which the antigen-binding protein interacts. See, for example, Ehring (1999) Analytical Biochemistry 267:252-259; Engen and Smith (2001) Anal. Chem. 73:256A-265A.

[0089] As used herein, the term "compete" refers to an antigen-binding protein (e.g., an antibody or antigen-binding fragment thereof) that binds to an antigen (e.g., TMPRSS2) and inhibits or blocks the binding of another antigen-binding protein (e.g., an antibody or antigen-binding fragment thereof) to the antigen. The term also includes bidirectional competition between two antigen-binding proteins, e.g., antibodies, i.e., a first antibody that binds to and blocks the binding of a second antibody, and vice versa. In certain embodiments, a first antigen-binding protein (e.g., an antibody) and a second antigen-binding protein (e.g., an antibody) may bind to the same epitope. Alternatively, the first and second antigen-binding proteins (e.g., antibodies) may bind to different, but overlapping, epitopes, such that the binding of one inhibits or blocks the binding of the second antibody, e.g., through steric hindrance. Competition between antigen-binding proteins (e.g., antibodies) may be measured by methods known in the art, such as real-time label-free biolayer interferometry assays. In embodiments of the invention, competition between a first and second anti-TMPRSS2 antigen-binding protein (e.g., antibody) is determined by measuring the ability of an immobilized first anti-TMPRSS2 antigen-binding protein (e.g., antibody) (initially uncomplexed with TMPRSS2 protein) to bind to soluble TMPRSS2 protein complexed with a second anti-TMPRSS2 antigen-binding protein (e.g., antibody). A reduction in the ability of the first anti-TMPRSS2 antigen-binding protein (e.g., antibody) to bind to the complexed TMPRSS2 protein compared to the uncomplexed TMPRSS2 protein indicates that the first and second anti-TMPRSS2 antigen-binding proteins (e.g., antibodies) compete. The degree of competition can be expressed as a percentage of reduced binding. Such competition can be measured using a real-time label-free biolayer interferometry assay, such as an Octet RED384 biosensor (Pall ForteBio Corp.), enzyme-linked immunosorbent assay (ELISA), or surface plasmon resonance (SPR).

[0090] Binding competition between anti-TMPRSS2 antigen-binding proteins (e.g., monoclonal antibodies (mAbs)) can be determined using a real-time label-free biolayer interferometry assay on an Octet RED384 biosensor (Pall ForteBio Corp.). For example, to determine competition between two anti-human TMPRSS2 monoclonal antibodies, the anti-TMPRSS2 mAb can be first captured onto an Octet biosensor chip (Pall ForteBio Corp., product number 18-5060) coated with an anti-hFc antibody by immersing the chip in a solution of anti-human TMPRSS2 mAb (hereinafter referred to as "mAb1"). As a positive control for blocking, the biosensor chip with the captured antibody can then be saturated with a known blocking isotype control mAb (hereinafter referred to as "blocking mAb") by immersing it in a blocking mAb solution. To determine whether mAb2 competes with mAb1, the biosensor chip can then be immersed in a co-complex solution of pre-incubated human TMPRSS2 polypeptide and a second anti-human TMPRSS2 mAb (hereinafter referred to as "mAb2") for a certain period of time, and the binding of mAb1 to the TMPRSS2 polypeptide can be determined. The biosensor chip can be washed with buffer between all steps of the experiment. Real-time binding responses can be monitored throughout the course of the experiment, and the binding responses can be recorded at the end of each step.

[0091] For example, in embodiments of the invention, the competitive assay is carried out at 25° C., at a pH of about 7, eg, 7.4, in the presence of, eg, buffer, salt, detergent, and non-specific protein (eg, bovine serum albumin).

[0092] Typically, antibodies or antigen-binding fragments of the invention, modified in some way, retain the ability to specifically bind to TMPRSS2, e.g., retain at least 10% of their TMPRSS2-binding activity (compared to the parent antibody) when that activity is expressed on a molar basis. Preferably, antibodies or antigen-binding fragments of the invention retain at least 20%, 50%, 70%, 80%, 90%, 95%, or 100% or more of the TMPRSS2-binding affinity of the parent antibody. It is also intended that antibodies or antigen-binding fragments of the invention may include conservative or non-conservative amino acid substitutions (referred to as "conservative variants" or "functionally conservative variants" of antibodies) that do not substantially alter their biological activity.

[0093] "Variants" of polypeptides such as immunoglobulin chains (e.g., mAb8021 V H , V L , HC, or LC, mAb8028 V H , V L , HC, or LC, or mAb8029 V H , V L , HC, or LC) to a reference amino acid sequence described herein (e.g., SEQ ID NOs: 2, 10, 18, 20, 22, 30, 38, 40, 42, 50, 58, or 60) when compared using the BLAST algorithm, with the algorithm parameters selected to give the largest match between each sequence over the entire length of each reference sequence (e.g., expectation threshold: 10, word size: 3, maximum matches in query range: 0, BLOSUM62 matrix, gap cost: extent 11, extension 1, conditional composition score matrix adjusted), refers to a polypeptide containing an amino acid sequence that is at least about 70 to 99.9% (e.g., 70, 72, 74, 75, 76, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 99.5, 99.9%) identical or similar.

[0094] A "variant" of a polynucleotide refers to a polynucleotide comprising a nucleotide sequence that is at least about 70-99.9% (e.g., at least about 70, 72, 74, 75, 76, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 99.5, or 99.9%) identical to a reference nucleotide sequence described herein (e.g., SEQ ID NOs: 1, 9, 17, 19, 21, 29, 37, 39, 41, 49, 57, or 59) when the comparison is performed using the BLAST algorithm and the algorithm parameters are selected to give the largest match between each sequence over the entire length of each reference sequence (e.g., expectation threshold: 10, word size: 28, maximum match in query range: 0, match / mismatch score: 1, -2, gap cost: linear).

[0095] Anti-TMPRSS2 antigen binding proteins, e.g., antibodies and antigen-binding fragments thereof, of the present disclosure, in embodiments of the invention, comprise a heavy chain immunoglobulin variable region having at least 70% (e.g., 80%, 85%, 90%, 95%, 99%) amino acid sequence identity to the amino acids set forth in SEQ ID NO: 2, 18, 22, 38, 42, or 58, and / or a light chain immunoglobulin variable region having at least 70% (e.g., 80%, 85%, 90%, 95%, 99%) amino acid sequence identity to the amino acids set forth in SEQ ID NO: 10, 20, 30, 40, 50, or 60.

[0096] Additionally, variants of anti-TMPRSS2 antigen binding proteins can include polypeptides comprising an amino acid sequence as described herein, except for one or more (e.g., 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10) mutations, such as, for example, missense mutations (e.g., conservative substitutions), nonsense mutations, deletions, or insertions. For example, the present invention includes antigen binding proteins comprising immunoglobulin light chain variants comprising the amino acid sequence set forth in SEQ ID NO: 10, 20, 30, 40, 50, or 60, but with one or more such mutations, and / or immunoglobulin heavy chain variants comprising the amino acid sequence set forth in SEQ ID NO: 2, 18, 22, 38, 42, or 58, but with one or more such mutations. In embodiments of the invention, the variant anti-TMPRSS2 antigen binding proteins comprise immunoglobulin light chain variants comprising CDR-L1, CDR-L2, and CDR-L3, wherein one or more (e.g., one or two or three) of such CDRs have one or more of such mutations (e.g., conservative substitutions), and / or immunoglobulin heavy chain variants comprising CDR-H1, CDR-H2, and CDR-H3, wherein one or more (e.g., one or two or three) of such CDRs have one or more of such mutations (e.g., conservative substitutions).

[0097] The present invention further provides variant anti-TMPRSS2 antigen binding proteins, e.g., antibodies or antigen-binding fragments thereof, comprising one or more variant CDRs described herein (e.g., any one or more of CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2, and / or CDR-H3), with at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99%, or 99.9% sequence identity or similarity to, e.g., SEQ ID NOs: 4, 6, 8, 12, 14, and / or 16; or 24, 26, 28, 32, 34, and / or 36; or 44, 46, 48, 52, 54, and / or 56.

[0098] Embodiments of the present invention also include the corresponding V specifically described herein. H , V Limmunoglobulin V, comprising an amino acid sequence having 70% or more (e.g., 80%, 85%, 90%, 95%, 97%, or 99%) overall amino acid sequence identity or similarity to the amino acid sequence of HC or LC. H and V L or variants of antigen binding proteins, such as anti-TMPRSS2 antibodies and antigen-binding fragments thereof, comprising the HC and LC, provided that CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2, and CDR-H3 of the immunoglobulin are not variant and comprise the amino acid sequences set forth in SEQ ID NOs: 4, 6, 8, 12, 14, and 16; or 24, 26, 28, 32, 34, and 36; or 44, 46, 48, 52, 54, and 56, respectively. Thus, in such embodiments, the CDRs within the variant antigen binding protein are not themselves variant.

[0099] Conservatively modified variants of anti-TMPRSS2 antibodies and antigen-binding fragments thereof are also part of the present invention. "Conservatively modified variants" or "conservative substitutions" refer to variants in which one or more amino acids in a polypeptide are replaced with other amino acids having similar properties (e.g., charge, side chain size, hydrophobicity / hydrophilicity, backbone structure, and rigidity). Such changes can often be made without significantly impairing the biological activity of the antibody or fragment. Those skilled in the art generally recognize that single amino acid substitutions in non-essential regions of a polypeptide do not substantially alter biological activity (see, e.g., Watson et al. (1987) Molecular Biology of the Gene, The Benjamin / Cummings Pub. Co., p. 224 (4th ed.)). Furthermore, substitutions of structurally or functionally similar amino acids are unlikely to significantly destroy biological activity.

[0100] Examples of groups of amino acids having side chains with similar chemical properties include: 1) aliphatic side chains: glycine, alanine, valine, leucine, and isoleucine, 2) aliphatic-hydroxyl side chains: serine and threonine, 3) amide-containing side chains: asparagine and glutamine, 4) aromatic side chains: phenylalanine, tyrosine, and tryptophan, 5) basic side chains: lysine, arginine, and histidine, 6) acidic side chains: aspartic acid and glutamic acid, and 7) sulfur-containing side chains: cysteine ​​and methionine. Preferred conservative amino acid substitution groups are valine-leucine-isoleucine, phenylalanine-tyrosine, lysine-arginine, alanine-valine, glutamic acid-aspartic acid, and asparagine-glutamine. Alternatively, a conservative substitution is any change that has a positive value in the PAM250 log-likelihood matrix disclosed in Gonnet et al. (1992) Science 256:1443-1445.

[0101] Function-conservative variants of anti-TMPRSS2 antibodies and antigen-binding fragments thereof are also part of the present invention. Any variant of an anti-TMPRSS2 antibody and antigen-binding fragment thereof (discussed herein) may be a "function-conservative variant." Such function-conservative variants may, in some instances, also be characterized as conservatively modified variants. As used herein, "function-conservative variant" refers to a variant of an anti-TMPRSS2 antibody or antigen-binding fragment thereof in which one or more amino acid residues have been altered without significantly altering one or more functional properties of the antibody or fragment. In certain embodiments of the present invention, a function-conservative variant of an anti-TMPRSS2 antibody or antigen-binding fragment thereof of the present invention comprises a variant amino acid sequence and exhibits one or more of the following functional properties: Inhibiting the growth of influenza viruses (e.g., A / Puerto Rico / 08 / 1934(H1N1)) or coronaviruses (e.g., SARS-CoV-2) in TMPRSS2-expressing cells (e.g., Calu-3 cells); No significant binding to MDCK / Tet-on cells that do not express TMPRSS2; limiting the spread of coronavirus infection (e.g., by SARS-CoV-2) or influenza virus infection (e.g., by H1_PR34; H1_CA09; H1_Bris; H9N2 or H3N2 influenza viruses) of cells, e.g., Calu-3, in vitro; and / or Mice engineered to express the human TMPRSS2 protein are protected from death caused by influenza virus infection (e.g., H1N1 or H3N2) or coronavirus infection (e.g., SARS-CoV-2), optionally when combined with anti-HA or anti-spike protein antibodies, e.g., by infecting the mice with an otherwise lethal dose of the virus. Mice engineered to express the human TMPRSS2 protein are protected from weight loss caused by influenza virus infection (e.g., H1N1 or H3N2) or coronavirus infection (e.g., SARS-CoV-2), optionally when combined with anti-HA or anti-spike protein antibodies, e.g., by infecting the mice with a dose of the virus that would otherwise cause weight loss.

[0102] The present invention includes mice engineered to express human TMPRSS2 protein in the mouse body, including anti-TMPRSS2 antigen-binding proteins (e.g., antibodies or antigen-binding fragments) such as mAb 8021, mAb 8028, or mAb 8029. See International Patent Application Publication No. WO 2017 / 151453.

[0103] A "neutralizing" or "antagonist" anti-TMPRSS2 antigen-binding protein, e.g., an antibody or antigen-binding fragment, refers to a molecule that inhibits the activity of TMPRSS2 to any detectable extent, e.g., inhibits the proteolytic enzymatic activity of TMPRSS2, e.g., a substrate such as HA; Cbz-Gly-Gly-Arg-AMC (Sigma) (where Cbz is benzyloxycarbonyl and AMC is 7-amino-4-methylcoumarin); influenza virus HA0; coronavirus S protein; or a TMPRSS2 precursor that is autocatalytically cleaved between Arg255 and Ile256, and / or inhibits influenza virus entry into cells and / or inhibits influenza virus reproduction in the body of a subject.

[0104] mAb8021, mAb8028, and mAb8029 have heavy or V chains as described below. H (or a variant thereof) and light chain or V L (or a variant thereof); or V comprising its CDRs (CDR-H1 (or a variant thereof), CDR-H2 (or a variant thereof) and CDR-H3 (or a variant thereof)). H and V comprising its CDRs (CDR-L1 (or a variant thereof), CDR-L2 (or a variant thereof), and CDR-L3 (or a variant thereof)). L

[0033] The term "antigen-binding protein" refers to an antigen-binding protein, such as an antibody and antigen-binding fragment thereof, comprising, for example, the immunoglobulin chains, variable regions and / or CDRs, comprising the specific amino acid sequences described below.

[0105] In an embodiment of the present invention, mAb8028 refers to an antibody or antigen-binding fragment thereof comprising CDR-H1, CDR-H2, and CDR-H3 of an immunoglobulin heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 2 or 18, and CDR-L1, CDR-L2, and CDR-L3 of an immunoglobulin light chain comprising the amino acid sequence set forth in SEQ ID NO: 10 or 20.

[0106] In an embodiment of the present invention, mAb8021 refers to an antibody or antigen-binding fragment thereof comprising CDR-H1, CDR-H2, and CDR-H3 of an immunoglobulin heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 22 or 38, and CDR-L1, CDR-L2, and CDR-L3 of an immunoglobulin light chain comprising the amino acid sequence set forth in SEQ ID NO: 30 or 40.

[0107] In an embodiment of the present invention, mAb8029 refers to an antibody or antigen-binding fragment thereof comprising CDR-H1, CDR-H2, and CDR-H3 of an immunoglobulin heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 42 or 58, and CDR-L1, CDR-L2, and CDR-L3 of an immunoglobulin light chain comprising the amino acid sequence set forth in SEQ ID NO: 50 or 60.

[0108] In an embodiment of the invention, mAb8028 comprises a V H and V comprising the amino acid sequence set forth in SEQ ID NO: 10 L It refers to an antibody or antigen-binding fragment thereof comprising:

[0109] In an embodiment of the invention, mAb8021 comprises a V H and V comprising the amino acid sequence set forth in SEQ ID NO: 30 L It refers to an antibody or antigen-binding fragment thereof comprising:

[0110] In an embodiment of the invention, mAb8029 comprises a V H and V comprising the amino acid sequence set forth in SEQ ID NO: 50 L It refers to an antibody or antigen-binding fragment thereof comprising:

[0111] In an embodiment of the present invention, mAb8028 refers to an antibody or antigen-binding fragment comprising a heavy chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO: 18; and a light chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO: 20.

[0112] In an embodiment of the present invention, mAb8021 refers to an antibody or antigen-binding fragment comprising a heavy chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:38; and a light chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:40.

[0113] In an embodiment of the present invention, mAb8029 refers to an antibody or antigen-binding fragment comprising a heavy chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:58; and a light chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:60.

[0114] The antibodies described herein also include V H is fused to wild-type IgG4 (e.g., residue 108 is S) or an IgG4 variant (e.g., residue 108 is P).

[0115] The antibodies and antigen-binding fragments of the invention include immunoglobulin chains comprising the amino acid sequences described herein, as well as cellular and in vitro post-translational modifications to the antibodies. For example, the invention includes antibodies and antigen-binding fragments thereof that specifically bind to TMPRSS2, comprising the heavy and / or light chain amino acid sequences described herein (e.g., CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and / or CDR-L3), as well as antibodies and fragments in which one or more amino acid residues are glycosylated, one or more Asn residues are deamidated, one or more residues (e.g., Met, Trp, and / or His) are oxidized, the N-terminal Gln is pyroglutamate (PyroE), and / or the C-terminal lysine is missing.

[0116] Antibody administration The invention provides methods of administering an anti-TMPRSS2 antigen binding protein of the invention, e.g., mAb 8021, mAb 8028, or mAb 8029, comprising introducing the antigen binding protein into the body of a subject (e.g., a human). For example, the method may comprise puncturing the subject's body with a syringe needle and injecting the antigen binding protein into the subject's body, e.g., into a vein, artery, tumor, muscle tissue, or subcutaneously within the subject.

[0117] The invention provides a container (e.g., a plastic or glass vial, a hollow bore or syringe cylinder, e.g., with a cap or chromatography column) containing an anti-TMPRSS2 antigen binding protein of the invention, e.g., mAb8021, mAb8028, or mAb8029.

[0118] The present invention also provides an injection device comprising one or more antigen-binding proteins (e.g., antibodies or antigen-binding fragments) that specifically bind to TMPRSS2, e.g., mAb8021, mAb8028, or mAb8029, or a pharmaceutical composition thereof. The injection device may be packaged in a kit. An injection device is a device for introducing a substance into a subject's body via a parenteral route, e.g., intramuscularly, subcutaneously, or intravenously. For example, the injection device may comprise, for example, a cylinder or barrel for holding the fluid to be injected (e.g., comprising an antibody or fragment thereof, or a pharmaceutical composition thereof), a syringe (e.g., pre-filled with the pharmaceutical composition, such as an auto-injector) comprising a needle for piercing the skin and / or blood vessel for injection of the fluid, and a plunger for forcing the fluid from the cylinder through the needle bore. In an embodiment of the present invention, the injection device comprising an antigen-binding protein, e.g., an antibody or antigen-binding fragment thereof, or a pharmaceutical composition thereof, from the combination of the present invention is an intravenous (IV) injection device. Such devices can contain an antigen-binding protein or pharmaceutical composition thereof in a cannula or trocar / needle, which can be attached to a tube that can be attached to a bag or reservoir for holding a fluid (e.g., saline) to be introduced into the subject's body through the cannula or trocar / needle. In embodiments of the invention, the trocar and cannula are inserted into a subject's vein, and the antibody or fragment or pharmaceutical composition thereof can be introduced into the device once the trocar is removed from the inserted cannula. The IV device can be inserted, for example, into a peripheral vein (e.g., the hand or arm); into the superior or inferior vena cava or the right atrium of the heart (e.g., a central IV); or into the subclavian, internal jugular, or femoral vein, and can be advanced toward the heart until it reaches, for example, the superior or right vena cava (e.g., a central venous line). In embodiments of the invention, the injection device is an autoinjector, jet injector, or external infusion pump. Jet injectors use a high-pressure, narrow liquid jet that penetrates the epidermis to introduce the antibody or fragment or pharmaceutical composition thereof into the subject's body. An external infusion pump is a medical device that delivers an antibody or fragment or pharmaceutical composition thereof in controlled amounts to the body of a subject.External infusion pumps may be electrically or mechanically powered. Different pumps operate in different ways; for example, syringe pumps hold fluid in a syringe reservoir and a movable piston controls fluid delivery; elastomeric pumps hold fluid in a stretchable balloon reservoir and pressure from the balloon's elastic walls effects fluid delivery. In peristaltic pumps, a set of rollers squeezes a length of flexible tubing, pushing the fluid forward. In multi-channel pumps, fluid can be delivered from multiple reservoirs at multiple speeds.

[0119] Preparation of human antibodies Methods for generating human antibodies in transgenic mice are known in the art. Any such known methods can be used in the context of the present invention to generate human antibodies that specifically bind to TMPRSS2. Antibodies against TMPRSS2 can be generated using immunogens including any one of the following: In specific embodiments of the present invention, antibodies of the present invention are obtained from mice immunized with full-length native TMPRSS2, or with live attenuated or inactivated virus, or with DNA encoding the protein or a fragment thereof. Alternatively, TMPRSS2 protein or a fragment thereof can be produced and modified using standard biochemical techniques and used as an immunogen. In one embodiment of the present invention, the immunogen is a recombinant TMPRSS2 protein or a fragment thereof. In specific embodiments of the present invention, the immunogen can be a TMPRSS2 polypeptide vaccine. In certain embodiments, one or more booster injections can be administered. In certain embodiments, the immunogen can be a recombinant TMPRSS2 polypeptide expressed in Escherichia coli or any other eukaryotic or mammalian cell, such as Chinese hamster ovary (CHO) cells.

[0120] Using VELOCIMMUNE® technology (see, e.g., US Pat. No. 6,596,541, Regeneron Pharmaceuticals, Inc., VELOCIMMUNE®) or any other known method for generating monoclonal antibodies, a high-affinity chimeric antibody against TMPRSS2 having a human variable region and a mouse constant region is first isolated. VELOCIMMUNE® technology involves generating transgenic mice whose genomes contain human heavy and light chain variable regions operably linked to endogenous mouse constant region loci, such that the mice produce antibodies containing the human variable region and the mouse constant region in response to antigenic challenge. DNA encoding the heavy and light chain variable regions of the antibody is isolated and operably linked to DNA encoding the human heavy and light chain constant regions. The DNA is then expressed in cells capable of expressing fully human antibodies.

[0121] Generally, VELOCIMMUNE® mice are challenged with an antigen of interest, and lymphocytes (such as B cells) are collected from the mice that express antibodies. Lymphocytes can be fused with myeloma cell lines to prepare immortalized hybridoma cell lines, which are then screened and selected to identify hybridoma cell lines that produce antibodies specific to the antigen of interest. DNA encoding the heavy and light chain variable regions can be isolated and linked to constant regions of the desired heavy and light chain isotypes. Such antibody proteins can be produced in cells such as CHO cells. Alternatively, DNA encoding the antigen-specific chimeric antibody or the light and heavy chain variable domains can be isolated directly from antigen-specific lymphocytes.

[0122] First, a high-affinity chimeric antibody having a human variable region and a mouse constant region is isolated. As in the experimental section below, the antibody is characterized and selected for desirable characteristics, including affinity, selectivity, epitope, etc. The mouse constant region is replaced with a desired human constant region to generate a fully human antibody of the present invention, e.g., a wild-type or modified IgG1 or IgG4. While the constant region selected can vary depending on the particular application, the high-affinity antigen binding and target specificity characteristics reside in the variable region.

[0123] Anti-TMPRSS2 antibodies containing Fc variants According to certain embodiments of the present invention, there are provided anti-TMPRSS2 antigen binding proteins, e.g., antibodies or antigen-binding fragments, comprising an Fc domain comprising one or more mutations that enhance or decrease antibody binding to the FcRn receptor, e.g., at acidic pH compared to neutral pH. For example, the present invention provides an Fc domain comprising one or more mutations that enhance or decrease antibody binding to the FcRn receptor, e.g., at acidic pH compared to neutral pH. H 2 or C HThe present invention also includes anti-TMPRSS2 antibodies containing mutations in the three regions that increase the affinity of the Fc domain for FcRn in acidic environments (e.g., endosomes at pHs ranging from about 5.5 to about 6.0). Such mutations can extend the serum half-life of the antibody when administered to an animal. Non-limiting examples of such Fc modifications include, for example, modifications at positions 250 (e.g., E or Q), 250 and 428 (e.g., L or F), 252 (e.g., L / Y / F / W or T), 254 (e.g., S or T), and 256 (e.g., S / R / Q / E / D or T), or at positions 428 and / or 433 (e.g., H / L / R / S / P / Q or K) and / or 434 (e.g., A, W, H, F, or Y [N434A, N434W, N434H, N434F or N434Y]), or modifications at positions 250 and / or 428, or at positions 307 or 308 (e.g., 308F, V308F), and 434. In one embodiment, the modifications include 428L (e.g., M428L) and 434S (e.g., N434S) modifications, 428L, 259I (e.g., V259I), and 308F (e.g., V308F) modifications, 433K (e.g., H433K) and 434 (e.g., 434Y) modifications, 252, 254, and 256 (e.g., 252Y, 254T, and 256E) modifications, 250Q and 428L modifications (e.g., T250Q and M428L), and 307 and / or 308 modifications (e.g., 308F and / or 308P). In yet another embodiment, the modifications include 265A (e.g., D265A) and / or 297A (e.g., N297A) modifications.

[0124] For example, the present invention provides 250Q and 248L (e.g., T250Q and M248L), 252Y, 254T and 256E (e.g., M252Y, S254T and T256E), 428L and 434S (e.g., M428L and N434S), 257I and 311I (e.g., P257I and Q311I), 257I and 434H (e.g., P257I and N434H), 376V and 434H (e.g., D376V and N434H), 307A, 380A and 434A (e.g., T307A, E380A and N434A), and 433K and 434F (e.g., H433K and N434F).

[0125] V as described herein, including any possible combination of the aforementioned Fc domain mutations. H and / or V L Anti-TMPRSS antigen binding proteins, e.g., antibodies and antigen-binding fragments thereof, including are considered within the scope of the present invention.

[0126] The present invention also provides a method for treating a V H and chimeric heavy chain constant (C H ) region, and comprising an anti-TMPRSS2 antigen-binding protein, antibody, or antigen-binding fragment, and a chimeric C H The region contains C H For example, the antibodies of the present invention may comprise a segment derived from a C region derived from a human IgG1 molecule, a human IgG2 molecule, or a human IgG4 molecule. H C derived from a human IgG1 molecule, a human IgG2 molecule, or a human IgG4 molecule in combination with part or all of the three domains H Chimeric C containing part or all of the 2 domains H According to a particular embodiment, the antibody of the present invention may comprise a chimeric C region having a chimeric hinge region. HFor example, the chimeric hinge may comprise an "upper hinge" amino acid sequence (amino acid residues at positions 216-227 according to EU numbering) derived from a human IgG1 hinge region, a human IgG2 hinge region, or a human IgG4 hinge region combined with a "lower hinge" sequence (amino acid residues at positions 228-236 according to EU numbering) derived from a human IgG1 hinge region, a human IgG2 hinge region, or a human IgG4 hinge region. According to certain embodiments, the chimeric hinge region comprises amino acid residues derived from a human IgG1 upper hinge or a human IgG4 upper hinge and amino acid residues derived from a human IgG2 lower hinge. The chimeric C described herein H Antibodies comprising the region, in certain embodiments, exhibit modified Fc effector functions without negatively impacting the therapeutic or pharmacokinetic properties of the antibody (see, e.g., WO 2014 / 022540).

[0127] Immunoconjugates The present invention encompasses anti-TMPRSS2 antigen-binding proteins, e.g., antibodies or antigen-binding fragments, conjugated to another moiety, e.g., a therapeutic moiety ("immunoconjugate"), such as a toxoid or antiviral agent for treating influenza virus infection. In embodiments of the invention, the anti-TMPRSS2 antibody or fragment is conjugated to any of the additional therapeutic agents described herein. As used herein, the term "immunoconjugate" refers to an antigen-binding protein, e.g., an antibody or antigen-binding fragment, that is chemically or biologically bound to a radioactive agent, cytokine, interferon, target or reporter moiety, enzyme, peptide or protein, or therapeutic agent. The antigen-binding protein may be linked to the radioactive agent, cytokine, interferon, target or reporter moiety, enzyme, peptide, or therapeutic agent anywhere along the molecule, as long as it is capable of binding to its target (TMPRSS2). Examples of immunoconjugates include antibody-drug conjugates and antibody-toxin fusion proteins. In one embodiment of the present invention, the agent may be a second, different antibody that specifically binds to TMPRSS2. The type of therapeutic moiety that can be conjugated to the anti-TMPRSS2 antigen binding protein (eg, antibody or fragment) will take into account the condition being treated and the desired therapeutic effect to be achieved.See, for example, Arnon et al., “Monoclonal Antibodies For Immunotargeting Of Drugs In Cancer Therapy”, Monoclonal Antibodies And Cancer Therapy, Reisfeld et al. (eds.), pp. 243-56 (Alan R. Liss, Inc. 1985); Hellstrom et al., “Antibodies For Drug Delivery”, Controlled Drug Delivery (2nd Ed.), Robinson et al. (eds.), pp. 623-53 (Marcel Dekker, Inc. 1987); Thorpe, “Antibody Carriers Of Cytotoxic Agents In Cancer Therapy: A Review”, Monoclonal Antibodies 1984: Biological And Clinical Applications, Pinchera et al. (eds.), pp. 475-506 (1985); “Analysis, Results, And Future Prospective Of The Therapeutic Use Of Radiolabeled Antibody In Cancer Therapy”, Monoclonal Antibodies For Cancer Detection And Therapy, Baldwin et al. (eds.), pp. 303-16 (Academic Press 1985), and Thorpe et al., “The Preparation And Cytotoxic Properties Of Antibody-Toxin Conjugates”, Immunol. Rev., 62: 119-58 (1982).

[0128] Bispecific antibody The present invention includes anti-TMPRSS2 antigen-binding proteins, e.g., antibodies and antigen-binding fragments thereof, as well as methods of use and methods of making such antigen-binding proteins. The term "anti-TMPRSS2" antigen-binding protein, e.g., antibody or antigen-binding fragment, includes multispecific (e.g., bispecific or dual-paratopic) molecules comprising at least one first antigen-binding domain that specifically binds to TMPRSS2 (e.g., an antigen-binding domain from mAb8021, mAb8028, or mAb8029) and at least one second antigen-binding domain that binds to a different antigen or epitope on TMPRSS2 that is different from that of the first antigen-binding domain (e.g., an influenza HA antigen-binding domain from mAb8021, mAb8028, or mAb8029). In an embodiment of the invention, the first and second epitopes overlap. In another embodiment of the invention, the first and second epitopes do not overlap. For example, in an embodiment of the invention, the multispecific antibody is a bispecific IgG antibody (e.g., IgG1 or IgG4) comprising a first antigen-binding domain that specifically binds to TMPRSS2, comprising the immunoglobulin heavy and light chains of mAb8021, mAb8028, or mAb8029, and a second antigen-binding domain that specifically binds to influenza HA (comprising a different immunoglobulin light and heavy chain, such as mAb8021, mAb8028, or mAb8029).

[0129] mAb8021, mAb8028, and mAb8029 are CDR-H and CDR-L, V of mAb8021, mAb8028, or mAb8029, respectively. H and V L or a multispecific molecule, such as an antibody or antigen-binding fragment (including variants thereof described herein), comprising an HC and an LC.

[0130] In embodiments of the invention, an antigen binding domain that specifically binds to TMPRSS that may be included in a multispecific molecule comprises: (1) (i) a heavy chain variable domain sequence comprising CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 4, CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 6, and CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 8; and (ii) a light chain variable domain sequence comprising CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 12, CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 14, and CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 16; or (2) (i) a heavy chain variable domain sequence comprising the amino acid sequence set forth in SEQ ID NO:2, and (ii) a light chain variable domain sequence comprising the amino acid sequence set forth in SEQ ID NO: 10; or (3) (i) a heavy chain immunoglobulin sequence comprising the amino acid sequence set forth in SEQ ID NO: 18, and (ii) a light chain immunoglobulin sequence comprising the amino acid sequence set forth in SEQ ID NO: 20.

[0131] In embodiments of the invention, an antigen binding domain that specifically binds to TMPRSS that may be included in a multispecific molecule comprises: (1) (i) a heavy chain variable domain sequence comprising CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 24, CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 26, and CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 28; and (ii) a light chain variable domain sequence comprising CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 32, CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 34, and CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 36; or (2) (i) a heavy chain variable domain sequence comprising the amino acid sequence set forth in SEQ ID NO: 22; and (ii) a light chain variable domain sequence comprising the amino acid sequence set forth in SEQ ID NO: 30; or (3) (i) a heavy chain immunoglobulin sequence comprising the amino acid sequence set forth in SEQ ID NO: 38, and (ii) a light chain immunoglobulin sequence comprising the amino acid sequence set forth in SEQ ID NO: 40.

[0132] In embodiments of the invention, an antigen binding domain that specifically binds to TMPRSS that may be included in a multispecific molecule comprises: (1) (i) a heavy chain variable domain sequence comprising CDR-H1 comprising the amino acid sequence set forth in SEQ ID NO: 44, CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO: 46, and CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 48; and (ii) a light chain variable domain sequence comprising CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 52, CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 54, and CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 56; or (2) (i) a heavy chain variable domain sequence comprising the amino acid sequence set forth in SEQ ID NO: 42, and (ii) a light chain variable domain sequence comprising the amino acid sequence set forth in SEQ ID NO: 50; or (3) (i) a heavy chain immunoglobulin sequence comprising the amino acid sequence set forth in SEQ ID NO: 58, and (ii) a light chain immunoglobulin sequence comprising the amino acid sequence set forth in SEQ ID NO: 60.

[0133] In embodiments of the invention, the multispecific antibody or fragment comprises more than two different binding specificities (e.g., a trispecific molecule), e.g., one or more additional antigen-binding domains, which are the same as or different from the first and / or second antigen-binding domains.

[0134] In embodiments of the invention, the multispecific molecule comprises, in addition to an antigen binding site that specifically binds TMPRSS2, the H4sH15188P; H1H15188P; H1H15211P; H1H15177P; H4sH15211P; H1H15260P2; H1H15259P2; H1H15203P; H4s ... and H1H15249P2.

[0135] In an embodiment of the invention, the multispecific molecule comprises, in addition to an antigen binding site that specifically binds to TMPRSS2, a polypeptide comprising: TIFF0007772702000015.tif161142TIFF0007772702000016.tif230134TIFF0007772702000017.tif65134(e.g., CDR-H, V H or heavy chain; and its CDR-L, V L and a light chain).

[0136] In embodiments of the invention, the multispecific molecule comprises an antigen binding site that specifically binds to TMPRSS2, as well as an antigen binding site that specifically binds to the V H and V L or an antigen-binding site that specifically binds to an influenza group II HA protein, comprising a heavy chain immunoglobulin comprising CDR-H1, CDR-H2 and CDR-H3 of H1H14611N2 and a light chain immunoglobulin comprising CDR-L1, CDR-L2 and CDR-L3 of H1H14611N2.

[0137] In embodiments of the invention, the multispecific molecule comprises an antigen binding site that specifically binds to TMPRSS2, as well as an antigen binding site that specifically binds to the V H and V L or an antigen-binding site that specifically binds to an influenza group II HA protein, comprising a heavy chain immunoglobulin comprising CDR-H1, CDR-H2 and CDR-H3 of H1H14612N2 and a light chain immunoglobulin comprising CDR-L1, CDR-L2 and CDR-L3 of H1H14612N2.

[0138] In embodiments of the invention, the multispecific molecule comprises, in addition to an antigen binding site that specifically binds TMPRSS2, e.g., the V H and V L or an antigen-binding site that specifically binds to an influenza group I HA protein, comprising a heavy chain immunoglobulin comprising CDR-H1, CDR-H2 and CDR-H3 of H1H11729P and a light chain immunoglobulin comprising CDR-L1, CDR-L2 and CDR-L3 of H1H11729P.

[0139] In one embodiment of the present invention, the bispecific antigen-binding fragment comprises a first scFv (e.g., the V of mAb8021, mAb8028, or mAb8029) that has binding specificity for a first epitope (e.g., TMPRSS2). H and V L and a second scFv (e.g., the V of an anti-influenza HA antibody) having binding specificity for a second, different epitope. H and V L For example, in embodiments of the invention, the first and second scFvs may comprise a linker, such as a peptide linker (e.g., (GGGGS) n(SEQ ID NO: 62), where n is, for example, 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10. Other bispecific antigen-binding fragments include F(ab)2 of a bispecific IgG antibody that contains the heavy and light chain CDRs of mAb8021 or mAb8028 or mAb8029 and the heavy and light chain CDRs of another antibody that binds to a different epitope.

[0140] Treatment method The present invention provides methods for treating or preventing viral infection or cancer (e.g., prostate cancer) by administering a therapeutically effective amount of an anti-TMPRSS2 antigen-binding protein, e.g., an antibody or antigen-binding fragment (e.g., mAb8021, mAb8028, or mAb8029), to a subject (e.g., a human) in need of such treatment or prevention.

[0141] Coronavirus or influenza virus infection can be treated or prevented in a subject by administering an anti-TMPRSS2 antigen binding protein of the present invention to the subject. Influenza viruses are classified into types A, B, and C based on their core proteins. Influenza A virus subtypes are determined by envelope glycoproteins that have either hemagglutinin (HA) or neuraminidase (NA) activity. There are several HA subtypes (e.g., HA1, HA2, HA3, HA4, HA5, HA6, HA7, HA8, HA9, HA10, HA11, HA12, HA13, HA14, HA15, HA16, HA17, or HA18—these subtypes may be named H1, H2, H3, etc.) and NA subtypes (e.g., NA1, NA2, NA3, NA4, NA5, NA6, NA7, NA8, NA9, NA10, or NA11—these subtypes may be named N1, N2, N3, etc.) of influenza A viruses, which are used to designate influenza A subtypes. For example, influenza A viruses H1N1 and H3N2 are commonly known human pathogens. Humans are generally infected with viruses of subtype H1, H2, or H3, and N1 or N2. The present invention includes methods of treating or preventing infection with the influenza virus subtypes discussed herein. In embodiments of the present invention, multispecific antibodies and antigen-binding fragments thereof that bind to TMPRSS2 also bind to the spike protein of a coronavirus (e.g., SARS-CoV-2, MERS-CoV, or SARS-CoV) or the HA and / or NA of an influenza virus (e.g., an influenza virus of a subtype described herein).

[0142] An effective or therapeutically effective dose of an anti-TMPRSS2 antigen-binding protein, e.g., an antibody or antigen-binding fragment (e.g., mAb8021, mAb8028, or mAb8029), for treating or preventing a viral infection refers to the amount of the antibody or fragment sufficient to alleviate one or more signs and / or symptoms of infection in a treated subject, by inducing regression or elimination of such signs and / or symptoms, or by inhibiting the progression of such signs and / or symptoms. The dosage may vary depending on the age and size of the subject to be administered, the target disease, condition, route of administration, etc. In embodiments of the invention, an effective or therapeutically effective dose of an antibody or antigen-binding fragment of the invention for treating or preventing a viral infection, e.g., in an adult human subject, is about 0.01 to about 200 mg / kg, e.g., up to about 150 mg / kg. In embodiments of the invention, the dosage is up to about 10.8 or 11 grams (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, or 11 grams). The frequency and duration of treatment can be adjusted depending on the severity of the infection. In certain embodiments, the antigen-binding protein of the present invention is administered in an initial dose, followed by one or more secondary doses. In certain embodiments, the initial dose may be followed by administration of a subsequent dose or multiple subsequent doses of the antibody or antigen-binding fragment thereof, in an amount that may be approximately the same as or less than the initial dose, wherein the subsequent doses are separated by at least 1 to 3 days, at least 1 week, at least 2 weeks, at least 3 weeks, at least 4 weeks, at least 5 weeks, at least 6 weeks, at least 7 weeks, at least 8 weeks, at least 9 weeks, at least 10 weeks, at least 12 weeks, or at least 14 weeks.

[0143] As used herein, the term "subject" refers to a mammal (e.g., rat, mouse, cat, dog, cow, sheep, horse, goat, rabbit), preferably a human, for example, a human in need of prevention and / or treatment of a disease or disorder such as a viral infection or cancer. The subject may have or be susceptible to a viral infection, such as an influenza infection. Subjects susceptible to infection or who may be at increased risk of contracting an infection (e.g., coronavirus or influenza virus) include subjects with an immunocompromised immune system due to an autoimmune disease, subjects undergoing immunosuppressive therapy (e.g., after organ transplantation), subjects suffering from human immunodeficiency syndrome (HIV) or acquired immunodeficiency syndrome (AIDS), subjects with a form of anemia that depletes or destroys white blood cells, subjects undergoing radiation or chemotherapy, or subjects suffering from an inflammatory disease. In addition, very young subjects (e.g., under the age of 5) or elderly subjects (e.g., over the age of 65) are at increased risk. Additionally, subjects may be at risk of contracting a viral infection due to proximity to a disease outbreak, e.g., living in a densely populated city or in close proximity to a subject with confirmed or suspected viral infection, or due to occupational choices, e.g., hospital workers, pharmaceutical researchers, travelers to affected areas, or frequent flyers.

[0144] "Treat" or "treating" means administering an anti-TMPRSS2 antigen binding protein of the invention, e.g., an antibody or antigen-binding fragment (e.g., mAb8021 or mAb8028 or mAb8029), to a subject having one or more signs or symptoms of a disease or infection, e.g., a viral infection, against which the antigen binding protein is effective when administered to the subject in an effective or therapeutically effective amount or dose (discussed herein).

[0145] The present invention also encompasses the prophylactic administration of an anti-TMPRSS2 antigen-binding protein of the present invention, e.g., an antibody or antigen-binding fragment thereof (e.g., mAb8021, mAb8028, or mAb8029), to a subject at risk of viral infection to prevent such infection. Passive antibody-based immunoprophylaxis has proven to be an effective strategy for preventing subjects from viral infection. For example, Berry et al.,Passive broad-spectrum influenza immunoprophylaxis Influenza Res Treat.2014 ;2014:267594.Epub 2014 Sep 22;and Jianqiang et al.,Passive immune neutralization strategies for prevention and control of influenza A infections,Immunotherapy.2012 February;4(2):175-186;Prabhu et al. See al., Antivir Ther 2009;14(7):911-21, Prophylactic and therapeutic efficacy of a chimeric monoclonal antibody specific for H5 hemagglutinin against lethal H5N1 influenza. "Prevent" or "preventing" means administering to a subject an anti-TMPRSS2 antigen binding protein, e.g., an antibody or antigen-binding fragment (e.g., mAb8021 or mAb8028 or mAb8029) of the invention, where the antigen binding protein is effective when administered to a subject in an effective or therapeutically effective amount or dose (discussed herein), to inhibit the appearance of disease or infection (e.g., viral infection) in the subject's body.

[0146] In an embodiment of the present invention, a sign or symptom of a viral infection in a subject is the survival or proliferation of the virus in the subject's body, for example, as determined by a viral titer assay (e.g., growth of coronavirus or influenza virus in embryonated eggs, a coronavirus spike protein assay, or an influenza virus hemagglutination assay). Other signs and symptoms of viral infection are discussed herein.

[0147] The present invention provides a method for treating or preventing, or inducing the regression or clearance of, a viral infection (e.g., influenza virus or coronavirus infection) in a subject (e.g., a human) in need thereof by administering a therapeutically effective amount of an anti-TMPRSS2 antigen binding protein (e.g., mAb8021 or mAb8028 or mAb8029) to the subject, e.g., by injection of the protein into the subject's body; · Fever or feeling feverish / chills; cough; · Sore throat; · Runny or stuffy nose; sneezing; · Muscle or body pain; headaches; · Fatigue (tiredness); · Vomiting; · diarrhea; Respiratory tract infections; · Chest discomfort; Shortness of breath; bronchitis; and / or · Pneumonia The present invention provides a method for inhibiting the progression of at least one sign or symptom of a viral infection, such as a viral infection, wherein the sign or symptom is secondary to the viral infection.

[0148] The present invention also includes methods of treating or preventing cancer in a subject, e.g., metastatic cancer, e.g., prostate cancer (e.g., characterized by expression of a TMPRSS2:ERG fusion), colon cancer, lung cancer, pancreatic cancer, urinary tract cancer, breast cancer, ovarian cancer, prostate adenocarcinoma, renal cell carcinoma, colorectal adenocarcinoma, lung adenocarcinoma, lung squamous cell carcinoma, and / or pleural mesothelioma, by administering a therapeutically effective amount of a TMPRSS2 antigen binding protein (e.g., mAb8021 or mAb8028 or mAb8029) to the subject, e.g., by injecting the protein into the subject's body. In embodiments of the invention, the subject is also administered the TMPRSS2 antigen binding protein in conjunction with an additional therapeutic agent, e.g., an anti-cancer therapeutic agent. In embodiments of the invention, the cancer is a tumor whose cells express TMPRSS2 or a variant thereof.

[0149] Combinations and Pharmaceutical Compositions To prepare a pharmaceutical composition of an anti-TMPRSS2 antigen binding protein, for example, antibodies and antigen binding fragments thereof (e.g., mAb8021 or mAb8028 or mAb8029), the antigen binding protein is mixed with a pharmaceutically acceptable carrier or excipient. For example, Remington's Pharmaceutical Sciences and USPharmacopeia: National Formulary, Mack Publishing Company, Easton, Pa. (1984); Hardman, et al. (2001) Goodman and Gilman's The Pharmacological Basis of Therapeutics, McGraw-Hill, New York, NY; Gennaro (2000) Remington: The Science and Practice of Pharmacy, Lippincott, Williams, and Wilkins, New York, NY; Avis, et al. (eds.) (1993) Pharmaceutical Dosage Forms: Parenteral Medications, Marcel Dekker, NY; Lieberman, et al. (eds.) (1990) Pharmaceutical Dosage Forms: Tablets, Marcel Dekker, NY; Lieberman, et al. (eds.) (1990) Pharmaceutical Dosage Forms: Disperse Systems,Marcel See Dekker, NY; Weiner and Kotkoskie (2000) Excipient Toxicity and Safety, Marcel Dekker, Inc., New York, NY. In an embodiment of the invention, the pharmaceutical composition is sterile. Such compositions are part of the present invention.

[0150] The scope of the present invention includes a dried composition, e.g., a lyophilized composition, comprising an anti-TMPRSS2 antigen-binding protein, e.g., an antibody or antigen-binding fragment thereof (e.g., mAb8021 or mAb8028 or mAb8029), or a pharmaceutical composition thereof, comprising a pharmaceutically acceptable carrier but substantially lacking water.

[0151] In a further embodiment of the invention, the additional therapeutic agent administered to a subject in association with an anti-TMPRSS2 antigen binding protein, e.g., an antibody or antigen-binding fragment thereof (e.g., mAb8021 or mAb8028 or mAb8029) disclosed herein is selected from the group consisting of antibodies, antibodies, and antibodies to TMPRSS2, as described in the Physicians' Desk Reference 2003 (Thomson Healthcare; 57 th The subject is administered the FDA-approved ...

[0152] The mode of administration can vary, including oral, rectal, transmucosal, intestinal, parenteral, intramuscular, subcutaneous, intradermal, intramedullary, intrathecal, direct intraventricular, intravenous, intraperitoneal, intranasal, intraocular, inhalation, insufflation, topical, dermal, transdermal, or intra-arterial.

[0153] The present invention provides methods of administering an anti-TMPRSS2 antigen-binding protein, e.g., an antibody or antigen-binding fragment thereof (e.g., mAb8021, mAb8028, or mAb8029), comprising introducing the protein into a subject's body. For example, the method includes puncturing the subject's body with a syringe needle and injecting the antigen-binding protein into the subject's body, e.g., into a vein, artery, tumor, muscle tissue, or subcutaneously within the subject.

[0154] The present invention relates to anti-TMPRSS2 antigen-binding proteins, e.g., antibodies or antigen-binding fragments thereof (e.g., mAb8021, mAb8028, or mAb8029), polypeptides (e.g., HC, LC, V of mAb8021, mAb8028, or mAb8029), and the like, as described herein. H Or V L) or a container (e.g., a plastic or glass vial, e.g., a hollow, bored or syringe cylinder with a cap or a chromatography column) containing either the polynucleotide or vector or a pharmaceutical composition thereof including a pharmaceutically acceptable carrier.

[0155] In embodiments of the invention, an anti-TMPRSS2 antigen-binding protein of the invention, e.g., an antibody or antigen-binding fragment thereof (e.g., mAb8021, mAb8028, or mAb8029), is associated with one or more additional therapeutic agents. For example, in embodiments of the invention, the additional therapeutic agent is an antiviral agent and / or a vaccine. As used herein, the term "antiviral agent" refers to any anti-infective drug or therapy used to treat, prevent, or ameliorate a viral infection in a subject. The term "antiviral agent" includes, but is not limited to, cationic steroid antibacterial agents, leupeptin, aprotinin, amantadine, rimantadine, oseltamivir, zanamivir, ribavirin, or interferon-alpha 2b. Methods of treating or preventing a viral (e.g., coronavirus or influenza virus) infection in a subject in need of such treatment or prevention by administering mAb8021, mAb8028, or mAb8029 in association with an additional therapeutic agent are part of the present invention.

[0156] For example, in embodiments of the invention, the additional therapeutic agent is a vaccine, e.g., a coronavirus vaccine or an influenza vaccine. In embodiments of the invention, the vaccine is an inactivated / killed virus vaccine, a live attenuated virus vaccine, or a viral subunit vaccine.

[0157] For example, in embodiments of the invention, the additional therapeutic agent is TIFF0007772702000018.tif37128(Camostat mesilate); TIFF0007772702000019.tif38128 (Nafamostat mesylate); TIFF0007772702000020.tif32128(Bromhexine hydrochloride (BHH)); TIFF0007772702000021.tif18128(4-(2-aminomethyl)benzenesulfonyl fluoride hydrochloride (AEBSF)); TIFF0007772702000022.tif164136 (polyamide). See Shen et al. Biochimie 142:1-10 (2017).

[0158] In embodiments of the invention, the antiviral agent is an antibody or antigen-binding fragment that specifically binds to a coronavirus, e.g., CoV-S. For example, in embodiments of the invention, the anti-CoV-S antibody is one of the antibodies H4sH15188P; H1H15188P; H1H15211P; H1H15177P; H4sH15211P; H1H15260P2; H1H15259P2; H1H15203P; H4sH15260P2; H4sH15231 ... and H1H15249P2, or an antigen-binding fragment thereof, e.g., the antibody or fragment is any one of CDR-L1, CDR-L2, and CDR-L3 (e.g., V, VL ... L or light chain); and CDR-H1, CDR-H2, and CDR-H3 (e.g., V ... H or heavy chain).

[0159] In embodiments of the invention, the antiviral agent is an antibody or antigen-binding fragment that specifically binds to an influenza virus, e.g., influenza HA. ​​For example, in embodiments of the invention, the anti-HA antibody is an antibody or antigen-binding fragment described in International Patent Application Publication No. WO2016 / 100807. or an antigen-binding fragment thereof, e.g., the antibody or fragment comprises CDR-L1, CDR-L2, and CDR-L3 (e.g., V, VL ... L or light chain); and a light chain immunoglobulin comprising CDR-H1, CDR-H2, and CDR-H3 (e.g., the V H or heavy chain).

[0160] In an embodiment of the invention, the additional therapeutic agent is an antibody or antigen-binding fragment that binds to an influenza group II HA protein, such as H1H14611N2; or H and V L or an antibody or fragment comprising a heavy chain immunoglobulin comprising CDR-H1, CDR-H2, and CDR-H3 of H1H14611N2 and a light chain immunoglobulin comprising CDR-L1, CDR-L2, and CDR-L3 of H1H14611N2. "H1H14611N2" refers to any anti-Group II HA antibody containing such sequences.

[0161] In an embodiment of the invention, the additional therapeutic agent is an antibody or antigen-binding fragment that binds to an influenza group II HA protein, such as H1H14612N2; or H and V L or an antibody or fragment comprising a heavy chain immunoglobulin comprising CDR-H1, CDR-H2, and CDR-H3 of H1H14612N2 and a light chain immunoglobulin comprising CDR-L1, CDR-L2, and CDR-L3 of H1H14612N2. "H1H14612N2" refers to any anti-Group II HA antibody containing such sequences.

[0162] In an embodiment of the invention, the additional therapeutic agent is an antibody or antigen-binding fragment that binds to an influenza group I HA protein, such as H1H11729P; or H and V L or an antibody or fragment comprising a heavy chain immunoglobulin comprising CDR-H1, CDR-H2, and CDR-H3 of H1H11729P and a light chain immunoglobulin comprising CDR-L1, CDR-L2, and CDR-L3 of H1H11729P. "H1H11729P" refers to any anti-Group I HA antibody containing such sequences.

[0163] In certain embodiments of the invention, the additional therapeutic agent is not amantadine, rimantadine, oseltamivir, zanamivir, aprotinin, leupeptin, a cationic steroid antimicrobial, an influenza vaccine (e.g., killed, live, attenuated whole virus or subunit vaccine), or an antibody to an influenza virus (e.g., an anti-hemagglutinin antibody).

[0164] " and connection Did (did) The term "anti-TMPRSS2" indicates that a component anti-TMPRSS2 antigen-binding protein of the invention, e.g., an antibody or antigen-binding fragment thereof, along with another agent such as oseltamivir, can be formulated in a single composition or separately in two or more compositions (e.g., a kit), e.g., for simultaneous delivery. Each component can be administered to a subject at a time different from when the other components are administered, e.g., each administration may be given non-simultaneously (e.g., separately or sequentially), spaced apart over a predetermined period of time. Furthermore, the separate components may be administered to a subject by the same or a different route (e.g., an anti-TMPRSS2 antibody or antigen-binding fragment thereof).

[0165] kit Further provided are kits comprising one or more components, including, but not limited to, an anti-TMPRSS2 antigen binding protein, e.g., an antibody or antigen-binding fragment (e.g., mAb 8021, mAb 8028, or mAb 8029), as discussed herein, in association with one or more additional components, including, but not limited to, an additional therapeutic agent, as discussed herein. The antigen binding protein and / or additional therapeutic agent may be formulated separately, together with a pharmaceutically acceptable carrier, as a single composition or in two or more compositions, e.g., in a pharmaceutical composition.

[0166] In one embodiment of the invention, the kit comprises an anti-TMPRSS2 antigen binding protein of the invention, e.g., an antibody or antigen-binding fragment thereof (e.g., mAb8021, mAb8028, or mAb8029), or a pharmaceutical composition thereof, in one container (e.g., a sterile glass or plastic vial), and an additional therapeutic agent in another container (e.g., a sterile glass or plastic vial).

[0167] In another embodiment, the kit comprises, in a single common container, a combination of the invention comprising an anti-TMPRSS2 antigen binding protein of the invention, e.g., an antibody or antigen-binding fragment thereof (e.g., mAb8021, mAb8028, or mAb8029), or a pharmaceutical composition thereof, optionally combined with one or more additional therapeutic agents formulated together in a pharmaceutical composition.

[0168] Where the kit includes a pharmaceutical composition for parenteral administration to a subject, the kit can include a device (e.g., an injection device) for performing such administration. For example, the kit can include one or more hypodermic needles or other injection devices discussed herein containing an anti-TMPRSS2 antigen binding protein of the invention, e.g., an antibody or antigen-binding fragment thereof (e.g., mAb8021, mAb8028, or mAb8029).

[0169] The kit may include a package insert containing information about the pharmaceutical compositions and dosage forms in the kit. Generally, such information will assist patients and physicians in effectively and safely using the enclosed pharmaceutical compositions and dosage forms. For example, the following information about the combination of the present invention may be provided in the package insert: pharmacokinetics, pharmacodynamics, clinical studies, efficacy parameters, indications and usage, contraindications, warnings, precautions, side effects, overdose, appropriate dosage and administration, method of delivery, appropriate storage conditions, references, manufacturer / distributor information, and patent information.

[0170] Diagnostic uses of antibodies Anti-TMPRSS2 antigen binding proteins of the invention, e.g., antibodies or antigen-binding fragments (e.g., mAb8021, mAb8028, or mAb8029), may be used to detect and / or measure TMPRSS2 in a sample. An exemplary assay for TMPRSS2 may include contacting a sample with an anti-TMPRSS2 antigen binding protein of the invention, where the anti-TMPRSS2 antigen binding protein is labeled with a detectable label or reporter molecule or used as a capture ligand to selectively isolate TMPRSS2 from the sample. The presence of the anti-TMPRSS2 antigen binding protein complexed with TMPRSS2 indicates the presence of TMPRSS2 in the sample. Alternatively, an unlabeled anti-TMPRSS2 antibody may be used in combination with a secondary antibody that is itself detectably labeled. The detectable label or reporter molecule may be, for example, 3 H, 14 C. 32 P, 35 S, or 125The antigen-binding protein may be a radioisotope such as I, a fluorescent or chemiluminescent moiety such as fluorescein isothiocyanate or rhodamine, or an enzyme such as alkaline phosphatase, β-galactosidase, horseradish peroxidase, or luciferase. Specific exemplary assays that can be used to detect or measure TMPRSS2 in a sample include enzyme-linked immunosorbent assay (ELISA), radioimmunoassay (RIA), and fluorescence-activated cell sorting (FACS). Accordingly, the present invention includes methods for detecting the presence of a TMPRSS2 polypeptide in a sample, comprising contacting the sample with an anti-TMPRSS2 antigen-binding protein and detecting the presence of the TMPRSS / anti-TMPRSS2 antigen-binding protein, wherein the presence of a complex indicates the presence of TMPRSS2.

[0171] The present invention includes a cell-based ELISA method using an anti-TMPRSS2 antigen binding protein of the invention, e.g., an antibody and antigen-binding fragment thereof (e.g., mAb8021, mAb8028, or mAb8029), to detect the presence of TMPRSS2 on a cell. In an embodiment of the invention, the method comprises the following steps: (i) contacting cells fixed on a solid surface (e.g., a microplate) to be tested for the presence of TMPRSS2 with an anti-TMPRSS2 antigen binding protein of the invention; (ii) optionally, washing the mixture to remove unbound anti-TMPRSS2 antigen-binding protein; (iii) contacting the anti-TMPRSS2 antigen-binding protein with a labeled second antibody or antigen-binding fragment thereof that binds to the anti-TMPRSS2 antigen-binding protein; (iv) optionally washing the complex to remove unbound antigen-binding protein; and (v) detecting the presence of a label on the second antibody or fragment, wherein detection of the label indicates that the cells contain TMPRSS2. For example, the present invention provides a method for detecting TMPRSS2 in a sample. + Such cell-based ELISA methods for identifying cells include:

[0172] The anti-TMPRSS2 antigen binding proteins of the invention (e.g., mAb8021, mAb8028, or mAb8029) may be used in Western blot or immunoprotein blotting methods to detect the presence of TMPRSS2 or a fragment thereof in a sample. Such methods form part of the invention and include, for example, the following steps: (1) providing a membrane or other solid substrate containing a sample to be tested for the presence of TMPRSS2, e.g., optionally transferring proteins from the sample to be tested for the presence of TMPRSS2 (e.g., from PAGE or SDS-PAGE electrophoretic separation of proteins in the sample) onto the membrane or other solid substrate using methods known in the art (e.g., semi-dry blotting or tank blotting); and contacting the membrane or other solid substrate to be tested for the presence of TMPRSS2 or a fragment thereof with an anti-TMPRSS2 antigen binding protein of the invention.

[0173] Such membranes may take the form of, for example, nitrocellulose or vinyl-based (e.g., polyvinylidene fluoride (PVDF)) membranes onto which proteins to be tested for the presence of TMPRSS2 have been transferred (e.g., after electrophoretic separation in a gel) in a non-denaturing PAGE (polyacrylamide gel electrophoresis) gel or SDS-PAGE (sodium dodecyl sulfate polyacrylamide gel electrophoresis) gel. Prior to contacting the membrane with the anti-TMPRSS2 antigen-binding protein, the membrane is optionally blocked, for example, with non-fat dry milk, to bind to nonspecific protein-binding sites on the membrane. (2) washing the membrane one or more times to remove unbound anti-TMPRSS2 antigen-binding protein and other unbound substances; and (3) detecting the bound anti-TMPRSS2 antigen-binding protein.

[0174] Detection of bound antigen-binding protein indicates that TMPRSS2 protein is present on a membrane or substrate in the sample. Detection of bound antigen-binding protein may be by binding the antigen-binding protein with a directly labeled secondary antibody (anti-immunoglobulin antibody) and then detecting the presence of the secondary antibody label.

[0175] The anti-TMPRSS2 antigen binding proteins (e.g., antibodies and antigen-binding fragments (e.g., mAb8021, mAb8028, or mAb8029)) disclosed herein may be used for immunohistochemistry. Such methods form part of the invention and include, for example, the following: (1) contacting a tissue to be tested for the presence of TMPRSS2 protein with an anti-TMPRSS2 antigen binding protein of the invention; and (2) detecting antigen-binding proteins on or in tissues;

[0176] If the antigen-binding protein itself is detectably labeled, it can be detected directly. Alternatively, the antigen-binding protein may be bound by a directly labeled secondary antibody, and the label then detected. [Example]

[0177] The following examples are provided to fully disclose to those skilled in the art how to make and use the methods and compositions of this invention, and are not intended to limit the scope of what the inventors regard as their invention. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, temperatures, etc.), but some experimental error and deviation should be accounted for. Unless otherwise indicated, parts are parts by weight, molecular weight is average molecular weight, temperature is in degrees Celsius, room temperature is about 25°C, and pressure is at or near atmospheric.

[0178] Example 1: Generation of human antibodies against TMPRSS2 Human antibodies against TMPRSS2 were generated in VELOCIMMUNE® mice containing DNA encoding the human immunoglobulin heavy chain variable region and kappa light chain variable region. Mice were immunized with a vector expressing TMPRSS2, followed by a booster dose of TMPRSS2. Antibody immune responses were monitored by a TMPRSS2-specific immunoassay. Anti-TMPRSS2 antibodies were isolated directly from antigen-positive mouse B cells without fusion with myeloma cells, as described in U.S. Patent No. 7,582,298, which is expressly incorporated herein by reference in its entirety. Using this method, fully human anti-TMPRSS2 antibodies (i.e., antibodies possessing human variable and constant domains) were obtained.

[0179] The exemplary antibodies described herein are designated mAb 8028, mAb 8021, and mAb 8029. The biological properties of the exemplary antibodies produced according to the methods of this example are described in detail in the Examples set forth below.

[0180] Example 2: Amino acid and nucleotide sequences of heavy and light chain variable regions Table 1 lists the amino acid sequence identifiers for the heavy and light chain variable regions (HCVR and LCVR, respectively) and CDRs (HCDR-1, HCDR-2, HCDR-3, LCDR-1, LCDR-2, and LCDR-3), as well as the heavy chain (HC) and light chain (LC) sequences of exemplary anti-TMPRSS2 antibodies. The corresponding nucleic acid sequence identifiers are listed in Table 2.

[0181] Table 1: Amino acid sequence identifiers TIFF0007772702000026.tif37150

[0182] Table 2: Nucleic acid sequence identifiers TIFF0007772702000027.tif37150

[0183] The antibodies disclosed herein have fully human variable regions, but can have mouse constant regions (e.g., mouse IgG1 Fc or mouse IgG2 Fc (a or b isotype)) or human constant regions (e.g., human IgG1 Fc or human IgG4 Fc). As will be understood by one of skill in the art, an antibody with a particular Fc isotype can be converted to an antibody with a different Fc isotype (e.g., an antibody with mouse IgG1 Fc can be converted to an antibody with human IgG4, etc.), but in either case, the variable domains (including CDRs) indicated by the numerical identifiers shown in Tables 1 and 2 will remain constant, and the binding characteristics to the antigen are expected to be the same or substantially similar regardless of the nature of the Fc domain.

[0184] Example 3: In vitro multicycle replication The ability of influenza virus, influenza AA / Puerto Rico / 08 / 1934 (H1_PR34), to replicate in Calu3 cells after treatment with mAb8021, mAb8028, or mAb8029 was assessed.

[0185] Table 3: Reagents used TIFF0007772702000028.tif69156

[0186] Calu-3 cells were seeded at 40,000 cells / well in 96-well plates in DMEM:F12 medium with 5% FBS. The following day, influenza virus was diluted to an MOI of 0.01, and antibodies were diluted to 25 μg / mL. HA antibodies were preincubated with influenza virus in a separate plate at 37°C for 1 hour. After the preincubation period, uninfected Calu-3 cells were treated with either the anti-HA antibody / virus mixture or one of the anti-TMPRSS2 antibodies mAb8021, mAb8028, or mAb8029 in combination with influenza virus. The treated Calu-3 cells were then incubated for 1 hour. After 1 hour of infection, cells were washed three times with PBS, and new antibodies (anti-HA or anti-TMPRSS2 matching the previous antibody) were added to each well along with fresh medium. Additional antibodies were added 24 and 48 hours postinfection. At 72 hours post-infection, cells were stained with anti-NP and imaged on a CTL-ImmunoSpot® S6 Universal Analyzer (Cellular Technology Limited, Cleveland, OH).

[0187] Calu-3 is an immortalized human airway epithelial cell line that has been shown to allow multicycle replication of human influenza viruses in the absence of exogenous trypsin (Zeng et al., Highly pathogenic avian influenza H5N1 viruses elicit an attenuated type I interferon response in polarized human bronchial epithelial cells. J Virol. 81:12439-12449 (2007)). Furthermore, Calu-3 cells have been shown to express TMPRSS2, which is important for testing anti-TMPRSS2 antibodies. While these antibodies were assayed for their ability to prevent multicycle replication of influenza PR8 virus, those skilled in the art will recognize that TMPRSS2 is also involved in coronavirus infection and, therefore, blocking TMPRSS2 would have a similar effect on coronavirus infection. Furthermore, blocking TMPRSS2 may be beneficial because it may inhibit a broader range of viral infections than blocking specific viral proteins. As a positive control for inhibition of infection, an anti-HA neutralizing antibody was used. An isotype control antibody was used as a negative control. Imaging of infected cells was performed on a CTL-ImmunoSpot® S6 Universal Analyzer (Cellular Technology Limited, Cleveland, OH). TMPRSS2 mAbs demonstrated inhibition of influenza infection, as observed by examining the difference in infected cell numbers between cells treated with anti-TMPRSS2 mAbs and those treated with the isotype control. The relative degree of mAb-mediated inhibition of viral replication is reported in Table 4 below.

[0188] Table 4. Viral inhibition by mAb8021, mAb8028, and mAb8029 TIFF0007772702000029.tif47128Level of viral inhibition compared to infected untreated controls: no inhibition (-), no virus-positive cells (++++), few virus-positive cells (+++).

[0189] Example 3 : Binding kinetics of anti-TMPRSS2 antibodies The equilibrium dissociation constants (K) for TMPRSS2 binding to different TMPRSS2 monoclonal antibodies (mAbs) D ) was determined using a real-time surface plasmon resonance biosensor with a Biacore T200 instrument. All binding studies were performed at 25°C and 37°C in a running buffer of 10 mM HEPES, 150 mM NaCl, and 0.05% v / v surfactant Tween-20, pH 7.4 (HBS-P). The Biacore CM5 sensor chip surface was first derivatized by amine coupling with an anti-human Fc mAb (REGN2567) to capture different TMPRSS2 mAbs expressed with a C-terminal myc-myc-hexahistidine (MMH) tag. The ectodomains of human TMPRSS2-myc-myc-His (hTMPRSS2-MMH), cynomolgus monkey TMPRSS2-MMH (mfTMPRSS2-MMH), rat TMPRSS2-MMH (rTMPRSS2-MMH), and mouse TMPRSS2-MMH (mTMPRSS2-MMH) prepared in HBS-P running buffer at different concentrations (100–3.7 nM, 3-fold serial dilutions) were injected over the TMPRSS2 mAb capture surface for 150 seconds at a flow rate of 30 μL / min, and their dissociation in HBS-P running buffer was monitored for 10 minutes. At the end of each cycle, the TMPRSS2 mAb capture surface was regenerated using a 12-second injection of 20 mM phosphate.

[0190] Association rates (k) were calculated by fitting the real-time binding sensorgrams to a 1:1 binding model using mass transport limitation using Scrubber 2.0c curve fitting software. a ) and dissociation rate (kd The binding-dissociation equilibrium constant (K D ) and dissociation half-life (t 1 / 2 ) was calculated from the kinetic velocity as follows: TIFF0007772702000030.tif5128

[0191] The binding kinetic parameters for TMPRSS2 binding to different TMPRSS2 mAbs of the present invention at 25°C and 37°C are shown in Tables 5-12. Both mAb8021 and mAb8029 showed strong binding to TMPRSS2, with mAb8021 in particular showing preferential binding to hTMPRSS2 and mfTMPRSS2 over rTMPRSS2 and mTMPRSS2.

[0192] Table 5. Binding kinetics of anti-TMPRSS2 mAb binding to hTMPRSS2-MMH at 25°C TIFF0007772702000031.tif58164 * Positive control antibody H4H7017N, described in International Patent Application Publication No. WO / 2019 / 147831

[0193] Table 6. Binding kinetics of anti-TMPRSS2 mAb binding to hTMPRSS2-MMH at 37°C TIFF0007772702000032.tif58164 * Positive control antibody H4H7017N, described in International Patent Application Publication No. WO / 2019 / 147831

[0194] Table 7. Binding kinetics of anti-TMPRSS2 mAb binding to mfTMPRSS2-MMH at 25°C TIFF0007772702000033.tif59164 * Positive control antibody H4H7017N, described in International Patent Application Publication No. WO / 2019 / 147831

[0195] Table 8. Binding kinetics of anti-TMPRSS2 mAb binding to mfTMPRSS2-MMH at 37°C TIFF0007772702000034.tif58164 * Positive control antibody H4H7017N, described in International Patent Application Publication No. WO / 2019 / 147831

[0196] Table 9. Binding kinetics of anti-TMPRSS2 mAb binding to rTMPRSS2-MMH at 25°C TIFF0007772702000035.tif58164 * No binding was observed under the current experimental conditions ** Positive control antibody H4H7017N, described in International Patent Application Publication No. WO / 2019 / 147831

[0197] Table 10. Binding kinetics of anti-TMPRSS2 mAb binding to rTMPRSS2-MMH at 37°C TIFF0007772702000036.tif58164 * No binding was observed under the current experimental conditions. ** Positive control antibody H4H7017N, described in International Patent Application Publication No. WO / 2019 / 147831

[0198] Table 11. Binding kinetics of anti-TMPRSS2 mAb binding to mTMPRSS2-MMH at 25°C TIFF0007772702000037.tif58164 * No binding was observed under the current experimental conditions. ** Positive control antibody H4H7017N, described in International Patent Application Publication No. WO / 2019 / 147831

[0199] Table 12. Binding kinetics of anti-TMPRSS2 mAb binding to mTMPRSS2-MMH at 37°C TIFF0007772702000038.tif58164 * indicates that no binding was observed under the current experimental conditions. **Positive control antibody H4H7017N, described in International Patent Application Publication No. WO / 2019 / 147831

[0200] Example 4 pH sensitivity of anti-TMPRSS2 antibodies The equilibrium dissociation constant (kd) for TMPRSS2 binding to purified anti-TMPRSS2 monoclonal antibodies was determined using a real-time surface plasmon resonance-based Biacore T200 biosensor platform. All binding studies were performed at 37°C using two running buffers: (i) 1.9 mM NaH2PO4, 8.1 mM Na2HPO4, 2.7 mM KCl, 137 mM NaCl, 0.05% v / v surfactant Tween-20, pH 7.4 (PBS-T-pH 7.4), and (ii) 8.8 mM NaH2PO4, 1.2 mM Na2HPO4, 2.7 mM KCl, 137 mM NaCl, 0.05% v / v surfactant Tween-20, pH 6.0 (PBS-T-pH 6.0). Different anti-TMPRSS2 mAbs were captured using a CM5 Biacore sensor surface derivatized by amine coupling with an anti-human Fc-specific mouse mAb. All TMPRSS2 reagents were expressed with a C-terminal myc-myc-hexahistidine tag (hereafter referred to as TMPRSS2-MMH). Different concentrations of human TMPRSS2-MMH, cynomolgus monkey TMPRSS2-MMH, rat TMPRSS2-MMH, or mouse TMPRSS2-MMH prepared in PBS-T pH 7.4 running buffer (25 nM to 6.25 nM; 4-fold serial dilutions) were injected at a flow rate of 30 μL / min for 2.5 min. Dissociation of bound TMPRSS2-MMH protein in PBS-T pH 7.4 or PBS-T pH 6.0 running buffer was continued for 8 min.

[0201] Dissociation rate constants (kd) in the two running buffers were determined by fitting the real-time binding sensorgrams to a 1:1 binding model using Scrubber 2.0c curve fitting software.

[0202] Dissociation rate values ​​for anti-TMPRSS2 mAb binding to human TMPRSS2-MMH, monkey TMPRSS2-MMH, rat TMPRSS2-MMH, and mouse TMPRSS2-MMH in PBS-T pH 7.4 and PBS-T pH 6.0 at 37°C are shown in Tables 13 to 16. None of the antibodies exhibited pH sensitivity.

[0203] Table 13. Dissociation rate constants for anti-TMPRSS2 mAb binding to human TMPRSS2-MMH in PBS-T pH 7.4 and PBS-T pH 6.0 at 37°C. TIFF0007772702000039.tif71150 * Positive control antibody H4H7017N, described in International Patent Application Publication No. WO / 2019 / 147831

[0204] Table 14. Dissociation rate constants for anti-TMPRSS2 mAb binding to monkey TMPRSS2-MMH in PBS-T pH 7.4 and PBS-T pH 6.0 at 37°C. TIFF0007772702000040.tif69150 * Positive control antibody H4H7017N, described in International Patent Application Publication No. WO / 2019 / 147831

[0205] Table 15. Dissociation rate constants of anti-TMPRSS2 mAb binding to rat TMPRSS2-MMH in PBS-T pH 7.4 and PBS-T pH 6.0 at 37°C. TIFF0007772702000041.tif65150 * No binding was observed under the current experimental conditions ** Positive control antibody H4H7017N, described in International Patent Application Publication No. WO / 2019 / 147831

[0206] Table 16. Dissociation rate constants of anti-TMPRSS2 mAb binding to mouse TMPRSS2-MMH in PBS-T pH 7.4 and PBS-T pH 6.0 at 37°C. TIFF0007772702000042.tif65150 * No binding was observed under the current experimental conditions. ** Positive control antibody H4H7017N, described in International Patent Application Publication No. WO / 2019 / 147831

[0207] Example 5: Octet cross-competition between anti-TMPRSS2 monoclonal antibodies Binding competition of TMPRSS2 monoclonal antibodies (mAbs) was determined using a real-time, label-free biolayer interferometry (BLI) assay on an Octet HTX biosensor platform (Pall ForteBio Corp.). The entire experiment was performed at 25°C with the plate shaking at 1000 rpm in 10 mM HEPES, 150 mM NaCl, 0.05% v / v surfactant Tween-20, and 1 mg / mL BSA, 0.02% NaN3, pH 7.4 (HBS-P) buffer. To assess whether the two mAbs could compete with each other for binding to their respective epitopes on TMPRSS2, the ectodomain of human TMPRSS2-MMH (hTMPRSS2-MMH) was first captured onto an Octet biosensor chip coated with an anti-penta-His antibody (HIS1K) by immersing the biosensor chip in a well containing a 10 μg / mL solution of hTMPRSS2-MMH for 1.5 minutes. The biosensor chip with captured hTMPRSS2-MMH was then saturated with anti-TMPRSS2 mAb (hereafter referred to as mAb-1) by immersing it in a well containing a 50 μg / mL solution of mAb-1 for 5 minutes. The biosensor chip was then immersed in a well containing a 50 μg / mL solution of a second TMPRSS2 mAb (hereafter referred to as mAb-2) for 5 minutes. The biosensor chip was washed with HBS-P buffer between all steps of the experiment. Real-time binding responses were monitored throughout the course of the experiment and recorded at the end of each step. The binding responses of mAb-2 to pre-complexed hTMPRSS2-MMH with mAb-1 were compared to determine the competitive / non-competitive behavior of the different anti-TMPRSS2 mAbs, as shown in Table 17.

[0208] Dissociation rate constants (kd) in the two running buffers were determined by fitting the real-time binding sensorgrams to a 1:1 binding model using Scrubber 2.0c curve fitting software.

[0209] Table 17. Cross-competition between anti-TMPRSS2 mAbs TIFF0007772702000043.tif74128 * Positive control antibody H4H7017N, described in International Patent Application Publication No. WO / 2019 / 147831

[0210] All references cited herein are incorporated by reference to the same extent as if each individual publication, database entry (e.g., Genbank sequence or GeneID entry), patent application, or patent was specifically and individually indicated to be incorporated by reference. It is intended by the applicant that the reference to incorporate by reference relates to each and every individual publication, database entry (e.g., Genbank sequence or GeneID entry), patent application, or patent, even if such citation is not immediately adjacent to the specific reference to incorporate by reference. The inclusion of a specific reference to incorporate by reference within this specification, if any, does not in any way weaken this general reference to incorporate by reference. Citation of references herein is not intended as an admission that the references are relevant prior art, nor does it constitute any admission as to the content or date of these publications or documents.

Claims

1. 1. An isolated, recombinant antibody or antigen-binding fragment thereof that specifically binds to human transmembrane protease serine 2 (TMPRSS2), wherein the antibody comprises: (i) three heavy chain complementarity determining regions (CDRs) (CDR-H1, CDR-H2, and CDR-H3) contained within a heavy chain variable region (HCVR) comprising the amino acid sequence set forth in SEQ ID NO: 22; and three light chain CDRs (CDR-L1, CDR-L2, and CDR-L3) contained within a light chain variable region (LCVR) comprising the amino acid sequence set forth in SEQ ID NO: 30; or (ii) three heavy chain CDRs (CDR-H1, CDR-H2, and CDR-H3) contained within an HCVR comprising the amino acid sequence set forth in SEQ ID NO:2; and three light chain CDRs (CDR-L1, CDR-L2, and CDR-L3) contained within an LCVR comprising the amino acid sequence set forth in SEQ ID NO:10; or (iii) three heavy chain CDRs (CDR-H1, CDR-H2, and CDR-H3) contained within an HCVR comprising the amino acid sequence set forth in SEQ ID NO: 42; and three light chain CDRs (CDR-L1, CDR-L2, and CDR-L3) contained within an LCVR comprising the amino acid sequence set forth in SEQ ID NO:

50.

1. An isolated recombinant antibody or antigen-binding fragment thereof comprising:

2. (a) an immunoglobulin heavy chain variable region (HCVR) comprising the CDR-H1, CDR-H2, and CDR-H3 of an immunoglobulin heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 22; and / or (b) an immunoglobulin light chain variable region (LCVR) comprising the CDR-L1, CDR-L2, and CDR-L3 of an immunoglobulin light chain comprising the amino acid sequence set forth in SEQ ID NO: 30; The antibody or antigen-binding fragment thereof of claim 1, comprising:

3. (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 24; (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 26; and (c) CDR-H3 comprising the amino acid sequence of SEQ ID NO: 28 an immunoglobulin HCVR comprising: (a) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 32; (b) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 34; and (c) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 36 immunoglobulin LCVR comprising The antibody or antigen-binding fragment thereof of claim 2, comprising:

4. (a) an immunoglobulin HCVR comprising an amino acid sequence having at least 90% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO: 22; and / or (b) an immunoglobulin LCVR comprising an amino acid sequence having at least 90% amino acid sequence identity with the amino acid sequence set forth in SEQ ID NO: 30; The antibody or antigen-binding fragment thereof of claim 3, comprising:

5. (a) an immunoglobulin HCVR comprising the amino acid sequence set forth in SEQ ID NO: 22; and / or (b) an immunoglobulin LCVR comprising the amino acid sequence set forth in SEQ ID NO: 30; The antibody or antigen-binding fragment thereof of claim 1, comprising:

6. (a) an immunoglobulin heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 38; and / or (b) an immunoglobulin light chain comprising the amino acid sequence set forth in SEQ ID NO:

40. The antibody or antigen-binding fragment thereof of claim 5, comprising:

7. (a) an immunoglobulin HCVR comprising the CDR-H1, CDR-H2, and CDR-H3 of an immunoglobulin heavy chain comprising the amino acid sequence set forth in SEQ ID NO:2; and / or (b) an immunoglobulin LCVR comprising the CDR-L1, CDR-L2, and CDR-L3 of an immunoglobulin light chain comprising the amino acid sequence set forth in SEQ ID NO: 10; The antibody or antigen-binding fragment thereof of claim 1, comprising:

8. (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 4; (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 6; and (c) CDR-H3 comprising the amino acid sequence of SEQ ID NO: 8 an immunoglobulin HCVR comprising: (a) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 12; (b) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 14; and (c) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 16 immunoglobulin LCVR comprising The antibody or antigen-binding fragment thereof of claim 7, comprising:

9. (a) an immunoglobulin HCVR comprising an amino acid sequence having at least 90% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO:2; and / or (b) an immunoglobulin LCVR comprising an amino acid sequence having at least 90% amino acid sequence identity with the amino acid sequence set forth in SEQ ID NO: 10; The antibody or antigen-binding fragment thereof of claim 8, comprising:

10. (a) an immunoglobulin HCVR comprising the amino acid sequence set forth in SEQ ID NO:2; and / or (b) an immunoglobulin LCVR comprising the amino acid sequence set forth in SEQ ID NO: 10; The antibody or antigen-binding fragment thereof of claim 1, comprising:

11. (a) an immunoglobulin heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 18; and / or (b) an immunoglobulin light chain comprising the amino acid sequence set forth in SEQ ID NO: 20; The antibody or antigen-binding fragment thereof of claim 10, comprising:

12. (a) an immunoglobulin heavy chain variable region (HCVR) comprising the CDR-H1, CDR-H2, and CDR-H3 of an immunoglobulin heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 42; and / or (b) an immunoglobulin light chain variable region (LCVR) comprising the CDR-L1, CDR-L2, and CDR-L3 of an immunoglobulin light chain comprising the amino acid sequence set forth in SEQ ID NO: 50; The antibody or antigen-binding fragment thereof of claim 1, comprising:

13. (a) a CDR-H1 comprising the amino acid sequence of SEQ ID NO: 44; (b) a CDR-H2 comprising the amino acid sequence of SEQ ID NO: 46; and (c) CDR-H3 comprising the amino acid sequence of SEQ ID NO: 48 an immunoglobulin HCVR comprising: (a) a CDR-L1 comprising the amino acid sequence of SEQ ID NO: 52; (b) a CDR-L2 comprising the amino acid sequence of SEQ ID NO: 54; and (c) CDR-L3 comprising the amino acid sequence of SEQ ID NO: 56 immunoglobulin LCVR comprising The antibody or antigen-binding fragment thereof of claim 12, comprising:

14. (a) an immunoglobulin HCVR comprising an amino acid sequence having at least 90% amino acid sequence identity to the amino acid sequence set forth in SEQ ID NO: 42; and / or (b) an immunoglobulin LCVR comprising an amino acid sequence having at least 90% amino acid sequence identity with the amino acid sequence set forth in SEQ ID NO: 50; The antibody or antigen-binding fragment thereof of claim 13, comprising:

15. (a) an immunoglobulin HCVR comprising the amino acid sequence set forth in SEQ ID NO: 42; and / or (b) an immunoglobulin LCVR comprising the amino acid sequence set forth in SEQ ID NO: 50; The antibody or antigen-binding fragment thereof of claim 1, comprising:

16. (a) an immunoglobulin heavy chain comprising the amino acid sequence set forth in SEQ ID NO: 58; and / or (b) an immunoglobulin light chain comprising the amino acid sequence set forth in SEQ ID NO:

60. The antibody or antigen-binding fragment thereof of claim 15, comprising:

17. The antibody or antigen-binding fragment thereof according to any one of claims 1 to 16, which is multispecific.

18. The following characteristics: a) inhibiting coronavirus growth in TMPRSS2-expressing cells; b) inhibiting influenza virus growth in TMPRSS2-expressing cells; c) binding to the surface of TMPRSS-expressing cells; d) no significant binding to MDCK / Tet-on cells that do not express TMPRSS2; e) limiting the spread of coronavirus infection of cells in vitro; f) limiting the spread of influenza virus infection of cells in vitro; g) protecting mice engineered to express the human TMPRSS2 protein from death and / or weight loss caused by coronavirus infection; and h) Protecting mice engineered to express the human TMPRSS2 protein from death caused by influenza virus infection The antibody or antigen-binding fragment thereof according to any one of claims 1 to 17, comprising one or more of:

19. A complex comprising the antibody or antigen-binding fragment thereof of any one of claims 1 to 18 bound to a TMPRSS2 polypeptide.

20. A method for producing the antibody or antigen-binding fragment thereof of any one of claims 1 to 18, comprising: (a) introducing into a host cell one or more polynucleotides encoding the antibody or antigen-binding fragment thereof; (b) culturing the host cell under conditions favorable for expression of the one or more polynucleotides; and (c) optionally isolating the antibody or antigen-binding fragment thereof from the host cell and / or the medium in which the host cell is grown. A method comprising:

21. 21. The method of claim 20, wherein the host cell is a Chinese hamster ovary cell.

22. (a) a V of an antibody or antigen-binding fragment thereof comprising the amino acid sequence set forth in any one of SEQ ID NOs: 22, 2, or 42; Ｈ Domains CDR-H1, CDR-H2, and CDR-H3; and (b) a V of an antibody or antigen-binding fragment thereof comprising the amino acid sequence set forth in any one of SEQ ID NOs: 30, 10, or 50; Ｌ Domains CDR-L1, CDR-L2, and CDR-L3 A polypeptide comprising:

23. A polynucleotide encoding the polypeptide of claim 22.

24. A vector comprising the polynucleotide of claim 23.

25. A host cell comprising the antibody or antigen-binding fragment thereof, or polypeptide, or polynucleotide, or vector according to any one of claims 1 to 18 and 22 to 24.

26. A composition or kit comprising an antibody or antigen-binding fragment thereof according to any one of claims 1 to 18 in association with a further therapeutic agent.

27. A pharmaceutical composition comprising the antibody or antigen-binding fragment thereof of any one of claims 1 to 18, and a pharmaceutically acceptable carrier, and optionally, a further therapeutic agent.

28. 28. The composition or kit of claim 26 or 27, associated with a further therapeutic agent which is an antiviral agent or a vaccine.

29. the additional therapeutic agent is a member selected from the group consisting of remdesivir, chloroquine, lopinavir, ritonavir, ribavirin, ledipasvir, sofosbuvir, a combination of ledipasvir and sofosbuvir, oseltamivir, zanamivir, ribavirin, and interferon-alpha 2b, interferon-alpha 2a, an anti-cancer drug, and an antibody or antigen-binding fragment thereof that specifically binds to influenza HA or coronavirus spike protein; and / or an antibody or antigen-binding fragment thereof selected from the group consisting of 28. A composition or kit according to claim 26 or 27.

30. A container or injection device comprising the antibody or antigen-binding fragment thereof or composition of any one of claims 1 to 18 and 26 to 29.

31. 20. A pharmaceutical composition for use in the treatment or prevention of cancer, or infection with influenza virus, coronavirus, SARS-CoV, MERS-CoV, SARS-CoV-2, parainfluenza virus, human metapneumovirus, or hepatitis C virus (HCV) in a subject in need thereof, comprising a therapeutically effective amount of the antibody or antigen-binding fragment thereof of any one of claims 1 to 18.

32. 32. The pharmaceutical composition of claim 31 for treating or preventing cancer, wherein the cancer is prostate cancer, colon cancer, lung cancer, pancreatic cancer, urinary tract cancer, breast cancer, ovarian cancer, prostate adenocarcinoma, renal cell carcinoma, colorectal adenocarcinoma, lung adenocarcinoma, lung squamous cell carcinoma, and / or pleural mesothelioma.

33. 33. The pharmaceutical composition of claim 31 or 32, administered in combination with one or more additional therapeutic agents.

34. 34. The pharmaceutical composition of claim 33, wherein the one or more additional therapeutic agents is an antiviral agent or a vaccine.

35. the one or more additional therapeutic agents a member selected from the group consisting of remdesivir, chloroquine, lopinavir, ritonavir, ribavirin, ledipasvir, sofosbuvir, a combination of ledipasvir and sofosbuvir, oseltamivir, zanamivir, ribavirin, and interferon-alpha 2b, interferon-alpha 2a, and an antibody or antigen-binding fragment thereof that specifically binds to influenza HA or coronavirus spike protein; and / or an antibody or antigen-binding fragment thereof selected from the group consisting of 35. The pharmaceutical composition of claim 33 or 34.

36. A pharmaceutical composition comprising an antibody or antigen-binding fragment thereof according to any one of claims 1 to 18 for use in administration to the body of a subject.

37. 37. The pharmaceutical composition of claim 36, formulated for subcutaneous, intravenous, or intramuscular injection.

38. 19. The isolated recombinant antibody or antigen-binding fragment thereof according to any one of claims 1 to 18, wherein the antibody has the following characteristics: (a) about 10 －９ EC below M ５０ and / or (b) demonstrating an increase in survival in coronavirus-infected animals after administration to the coronavirus-infected animals compared to comparable coronavirus-infected animals that have not been treated; An antibody or antigen-binding fragment thereof having one or more of:

39. A polynucleotide encoding HCVR and / or LCVR of an antibody or antigen-binding fragment thereof that specifically binds to human transmembrane protease serine 2 (TMPRSS2), (a) the HCVR comprises the amino acid sequence set forth in SEQ ID NO:2 and the LCVR comprises the amino acid sequence set forth in SEQ ID NO:10; or (b) the HCVR comprises the amino acid sequence set forth in SEQ ID NO: 22 and the LCVR comprises the amino acid sequence set forth in SEQ ID NO: 30; or (c) the HCVR comprises the amino acid sequence set forth in SEQ ID NO: 42, and the LCVR comprises the amino acid sequence set forth in SEQ ID NO: 50; Polynucleotide.

40. (a) the HCVR is encoded by the nucleic acid sequence set forth in SEQ ID NO:1 and the LCVR is encoded by the nucleic acid sequence set forth in SEQ ID NO:9; or (b) the HCVR is encoded by the nucleic acid sequence set forth in SEQ ID NO:21 and the LCVR is encoded by the nucleic acid sequence set forth in SEQ ID NO:29; or (c) the HCVR is encoded by the nucleic acid sequence set forth in SEQ ID NO: 41, and the LCVR is encoded by the nucleic acid sequence set forth in SEQ ID NO: 49; 40. The polynucleotide of claim 39.

41. A vector comprising the polynucleotide of claim 39 or 40.

42. A host cell comprising a polynucleotide according to claim 39 or 40, or a vector according to claim 41.

43. (A) a first polynucleotide encoding a polypeptide comprising an HCVR of an antibody or antigen-binding fragment thereof that specifically binds to human TMPRSS2; and (B) a second polynucleotide encoding a polypeptide comprising the LCVR of the antibody or antigen-binding fragment thereof. a set of polynucleotides comprising: (a) the HCVR comprises the amino acid sequence set forth in SEQ ID NO:2 and the LCVR comprises the amino acid sequence set forth in SEQ ID NO:10; or (b) the HCVR comprises the amino acid sequence set forth in SEQ ID NO: 22 and the LCVR comprises the amino acid sequence set forth in SEQ ID NO: 30; or (c) the HCVR comprises the amino acid sequence set forth in SEQ ID NO: 42, and the LCVR comprises the amino acid sequence set forth in SEQ ID NO: 50; A set of polynucleotides.

44. (a) a first polynucleotide comprises the nucleic acid sequence of SEQ ID NO: 1 encoding the HCVR of an antibody or antigen-binding fragment thereof that specifically binds to human TMPRSS2; and the second polynucleotide comprises the nucleic acid sequence of SEQ ID NO: 9 encoding the LCVR of the antibody or antigen-binding fragment thereof; or (b) the first polynucleotide comprises the nucleic acid sequence of SEQ ID NO: 21 encoding the HCVR of an antibody or antigen-binding fragment thereof that specifically binds to human TMPRSS2; and the second polynucleotide comprises the nucleic acid sequence of SEQ ID NO: 29 encoding the LCVR of the antibody or antigen-binding fragment thereof; or (c) the first polynucleotide comprises the nucleic acid sequence of SEQ ID NO: 41 encoding the HCVR of an antibody or antigen-binding fragment thereof that specifically binds to human TMPRSS2; and the second polynucleotide comprises the nucleic acid sequence of SEQ ID NO: 49 encoding the LCVR of the antibody or antigen-binding fragment thereof; 44. The set of polynucleotides of claim 43.

45. A vector comprising the set of polynucleotides of claim 43 or 44.

46. A host cell comprising the set of polynucleotides of claim 43 or 44, or the vector of claim 45.

47. 47. The host cell of claim 46, which is a Chinese hamster ovary cell.

48. (A) a first polynucleotide encoding a polypeptide comprising an HCVR of an antibody or antigen-binding fragment thereof that specifically binds to human TMPRSS2; and (B) a second polynucleotide encoding a polypeptide comprising the LCVR of the antibody or antigen-binding fragment thereof. A host cell comprising: (a) the HCVR comprises the amino acid sequence set forth in SEQ ID NO:2 and the LCVR comprises the amino acid sequence set forth in SEQ ID NO:10; or (b) the HCVR comprises the amino acid sequence set forth in SEQ ID NO: 22 and the LCVR comprises the amino acid sequence set forth in SEQ ID NO: 30; or (c) the HCVR comprises the amino acid sequence set forth in SEQ ID NO: 42, and the LCVR comprises the amino acid sequence set forth in SEQ ID NO: 50; host cell.

49. (a) a first polynucleotide comprises the nucleic acid sequence of SEQ ID NO: 1 encoding the HCVR of an antibody or antigen-binding fragment thereof that specifically binds to human TMPRSS2; and the second polynucleotide comprises the nucleic acid sequence of SEQ ID NO: 9 encoding the LCVR of the antibody or antigen-binding fragment thereof; or (b) the first polynucleotide comprises the nucleic acid sequence of SEQ ID NO: 21 encoding the HCVR of an antibody or antigen-binding fragment thereof that specifically binds to human TMPRSS2; and the second polynucleotide comprises the nucleic acid sequence of SEQ ID NO: 29 encoding the LCVR of the antibody or antigen-binding fragment thereof; or (c) the first polynucleotide is Comprising the nucleic acid sequence of SEQ ID NO: 41 encoding the HCVR of an antibody or antigen-binding fragment thereof that specifically binds to human TMPRSS2; and the second polynucleotide comprises the nucleic acid sequence of SEQ ID NO: 49 encoding the LCVR of the antibody or antigen-binding fragment thereof; 49. The host cell of claim 48.

50. 50. The host cell of claim 48 or 49, which is a Chinese hamster ovary cell.

51. A method for producing an antibody or antigen-binding fragment thereof that specifically binds to TMPRSS2, comprising culturing a host cell described in claim 50 under conditions favorable for expression of one or more polynucleotides.

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