Anti-tmprss2 antibodies and antigen-binding fragments

Abstract

To provide an antigen-binding protein that specifically binds to human TMPRSS2.SOLUTION: An antigen-binding protein comprises: (a) an immunoglobulin heavy chain variable region comprising the CDR-H1, CDR-H2, and CDR-H3 of an immunoglobulin heavy chain that comprises a specific amino acid sequence; and / or (b) an immunoglobulin light chain variable region comprising the CDR-L1, CDR-L2, and CDR-L3 of an immunoglobulin light chain that comprises a specific amino acid sequence.SELECTED DRAWING: Figure 2

Description

[Technical field]

[0001] This application claims the benefit of U.S. Provisional Patent Application No. 62 / 622,292, filed Jan. 26, 2018, which is incorporated by reference in its entirety.

[0002] The present invention relates to antibodies and antigen-binding fragments that specifically bind to TMPRSS2, and methods of treating or preventing viral infections with said antibodies and fragments. [Background technology]

[0003] Influenza viruses have acquired resistance to currently used drugs that target the viral neuraminidase (NA) or the ion channel protein matrix protein 2 (M2). The emergence of drug resistance highlights the need for the development of new antiviral strategies. Targeting the host cell may reduce or avoid the emergence of escape mutants, but may create a "sink" for widespread expression and raise concerns of toxicity. Numerous respiratory viral fusion proteins have been shown to require cleavage by host protease(s) for activation (Non-Patent Document 1; Non-Patent Document 2; Non-Patent Document 3; Non-Patent Document 4), including influenza (Non-Patent Document 5; Non-Patent Document 6; Non-Patent Document 7; Non-Patent Document 8).

[0004] Influenza A hemagglutinin precursor (HA0) requires cleavage into HA1 and HA2 by host serine proteases for activation. For example, transmembrane proteases, serine 2; TMPRSS2, TMPRSS4 and TMPRSS11D, as well as human airway trypsin-like proteases (HATs) have been implicated in HA cleavage (Non-Patent Document 7; Non-Patent Document 9; Patent Document 1). TMPRSS2 is also a target for anti-cancer therapy. See, for example, Patent Document 2 and Patent Document 3. The fusion of TMPRSS2 and ERG (TMPRSS2:ERG) is a gene fusion known to be a major driver of prostate carcinogenesis induced by ERα and suppressed by ERβ. Non-Patent Document 10. [Prior art documents] [Patent documents]

[0005] [Patent Document 1] International Publication No. WO2017 / 151453 [Patent Document 2] International Publication No. WO2008 / 127347 [Patent Document 3] International Publication No. WO2002 / 004953 [Non-patent literature]

[0006] [Non-Patent Document 1] 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 2] 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 3] Zhou et al., "Protease inhibitors targeting coronavirus and filovirus entry," Antiviral Research, Vol. 116, pp. 76-84 (2015) [Non-Patent Document 4] Zmora et al., “TMPRSS2 Isoform 1 Activates Respiratory Viruses and Is Expressed in Viral Target Cells,” PLoS ONE, 10, e0138380 (2015) [Non-Patent Document 5] Zmora et al., “Non-human primate orthologues of TMPRSS2 cleave and activate the influenza virus hemagglutinin,” PLoS ONE, 12, e0176597 (2017) [Non-Patent Document 6] 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, vol. 85, pp. 1554-1562 (2011) [Non-Patent Document 7] Bertram et al., "TMPRSS2 and TMPRSS4 facilitate trypsin-independent spread of influenza virus in Caco-2 cells," Journal of Virology, Vol. 84, pp. 10016-10025 (2010) [Non-Patent Document 8] Tarnow et al., "TMPRSS2 is a host factor that is essential for pneumotropism and pathogenicity of H7N9 influenza A virus in mice," Journal of Virology (2014), May;88(9):4744-51 [Non-Patent Document 9] Bottcher et al., "Proteolytic Activation of Influenza Viruses by Serine Proteases TMPRSS2 and HAT from Human Airway Epithelium," Journal of Virology, October 2006; Vol. 80(19): 9896-8. [Non-Patent Document 10] Bonkhoff, “Estrogen receptor signaling in prostate cancer: Implications for carcinogenesis and tumor progression,” Prostate, Vol. 78 (Issue 1): pp. 2-10 (2018) Summary of the Invention [Means for solving the problem]

[0007] Although there are small molecule inhibitors and research antibodies of TMPRSS2 that are useful, for example, for immunohistochemistry, there is a need in the art for neutralizing therapeutic anti-TMPRSS2 antibodies and their use to treat or prevent viral infections.See, for example, Shen et al., Biochimie vol. 142:1-10 (2017); International Publication No. WO2008 / 127347; International Publication No. WO2002 / 004953; 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 H1H7017N, and combinations thereof including, for example, anti-influenza HA antibodies (e.g., Group I HA or Group II HA), and methods of using them to treat viral infections.

[0008] The present invention provides neutralizing human antigen-binding proteins, e.g., antibodies or antigen-binding fragments thereof, that specifically bind to human TMPRSS2. For example, in one embodiment of the present invention, The fusion 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, 17, or 19; 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: 4 or 18. In one embodiment 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: 4 or 18; 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, 17, or 19. In one embodiment 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: 4 or 18, and at least 90% amino acid sequence identity with the amino acid sequence set forth in SEQ ID NO: 4 or 18; 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, 17 or 19, and at least 90% amino acid sequence identity with the amino acid sequence set forth in SEQ ID NO: 2, 17 or 19. For example, in one embodiment of the invention, an antigen binding protein comprises a light chain immunoglobulin variable region comprising: (a) a CDR-H1 comprising the amino acid sequence: GFTFSSYG (SEQ ID NO:6); (b) a CDR-H2 comprising the amino acid sequence: IWNDGSYV (SEQ ID NO:8); (c) a CDR-H3 comprising the amino acid sequence: AREGEWVLYYFDY (SEQ ID NO:10); and a heavy chain immunoglobulin variable region comprising: (a) a CDR-L1 comprising the amino acid sequence: QSISSW (SEQ ID NO:12); (b) a CDR-L2 comprising the amino acid sequence: KAS (SEQ ID NO:14); and / or (c) a CDR-L3 comprising the amino acid sequence: QQYNSYSYT (SEQ ID NO:16). The invention also provides antigen binding proteins comprising: (a) a heavy chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:17 or 19; and / or (b) a light chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO:18.

[0009] The 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 interference assay on an Octet RED384 biosensor (Pall ForteBio Corp.)); or that binds to the same or overlapping epitope (or fragment thereof) on TMPRSS2 as any of the antigen binding proteins described herein.

[0010] The present invention also provides multispecific antigen binding proteins that bind to TMPRSS2 and another antigen, or to TMPRSS2 at a different epitope. For example, the multispecific molecule includes (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 epitope different from TMPRSS2 or the first antigen binding domain.

[0011] The present invention also provides a method for producing a composition comprising the steps of: Inhibits the growth of influenza viruses (e.g., A / Puerto Rico / 08 / 1934(H1N1)) in TMPRSS2-expressing cells (e.g., Calu-3 cells); EC of e.g. 440pM or 1.06nM 50 values, binding to the surface of TMPRSS-expressing cells (e.g., MDCK / Tet-on); · No significant binding to MDCK / Tet-on cells that do not express TMPRSS2; Approximately 2.81 x 10 at 25°C -9 K of M D binds to human TMPRSS2; At 37°C, it is about 9.31 x 10 -9 K of M D binds to human TMPRSS2; Approximately 5.60 x 10 at 25°C -8 K of M D binds to cynomolgus TMPRSS2 at approximately 1.40 × 10 at 37 °C; -7 K of M Dbinds to cynomolgus TMPRSS2; limit the spread of influenza virus infection of cells in vitro; and / or protect mice engineered to express the human TMPRSS2 protein from death caused by influenza virus infection. The present invention provides any anti-TMPRSS2 antigen binding protein (e.g., an antibody or antigen-binding fragment comprising a sequence described herein) that comprises one or more of the following:

[0012] The invention also provides a complex comprising any of the antigen binding proteins described herein bound to a TMPRSS2 polypeptide, eg, in vitro or in the body of a subject.

[0013] The present invention also relates to a method of making an anti-TMPRSS2 antigen binding protein (e.g., H1H7017N) or an immunoglobulin chain thereof described herein, comprising the steps of: (a) introducing one or more polynucleotides encoding light and / or heavy immunoglobulin chains of said antigen binding protein; (b) culturing a host cell (e.g., CHO cells, Pichia cells or Pichia pastoris) under conditions favorable for expression of the polynucleotides; and (c) optionally isolating the antigen binding protein or immunoglobulin chain from the host cell and / or the medium in which the host cell is grown. The antigen binding protein or immunoglobulin chain that is the product of such a method is part of the invention.

[0014] (a) an immunoglobulin chain V comprising the amino acid sequence shown 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:4; L A polypeptide (eg, an immunoglobulin) comprising the CDR1, CDR2 and CDR3 domains (eg, the polypeptide is within a host cell) also forms part of the present invention.

[0015] The invention also provides polynucleotides (e.g., DNA or RNA) encoding the polypeptides of the invention. In one embodiment of the invention, the polynucleotides encode two different immunoglobulin chains (e.g., heavy and light chains). In one embodiment of the invention, one polynucleotide encodes the light immunoglobulin chain and another polynucleotide encodes the heavy immunoglobulin chain, e.g., the chains are in a host cell or vessel. For example, the polynucleotides are in a vector (e.g., a plasmid) and / or integrated into a host cell chromosome.

[0016] A host cell of the invention (e.g., a CHO cell, a Pichia cell or a Pichia pastoris cell) may contain an anti-TMPRSS2 antigen binding protein (e.g., H1H7017N), a polypeptide thereof, or a polynucleotide encoding such a polypeptide, and / or a vector containing such a polynucleotide.

[0017] The present invention also provides compositions or kits comprising the anti-TMPRSS2 antigen binding proteins (e.g., H1H7017N) described herein with additional therapeutic agents (e.g., antiviral drugs and / or vaccines). For example, the composition can be a pharmaceutical composition comprising the antigen binding protein and a pharma- ceutically acceptable carrier, and optionally an additional therapeutic agent. The additional therapeutic agent can be ledipasvir, sovosbuvir, a combination of ledipasvir and sovosbuvir, 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. In one embodiment of the invention, the additional therapeutic agent is H1H14611N2; H1H14612N2; H1H11723P; H1H11729P; H1H11820N; H1H11829N; H1H11829N2; H2aM11829N; H2M11830N; H1H11830N2; H1H11903N <h2 style=";text-align:left;direction:ltr">;H1H14571N;H2a14571N;H1H11704P;H1H11711P;H1H11714P;H1H11717P;H 1H11724P;H1H11727P;H1H11730P2;H1H11731P2;H1H11734P2;H1H11736P2; H1H11742P2;H1H11744P2;H1H11745P2;H1H11747P2;H1H11748P2;H1H17952B;H1H17953B;H1H17954B;H1H17955B;H1H17956B;H1H17957B;H1H17958B; H1H17959B;H1H17960B;H1H17961B;H1H17962B;H1H17963B;H1H17964B;H1H17965B;H1H17966B;H1H17967B;H1H17968B;H1H17969B;H1H17970B;H1H17 971B;H1H17972B;H1H17973B;H1H17974B;H1H17975B;H1H17976B;H1H17977B;H1H17978B;H1H17979B;H1H17980B;H1H17981B;H1H17982B;H1H17983B; H1H17984B;H1H17985B;H1H17986B;H1H17987B;H1H17988B;H1H17989B;H1H17990B;H1H17991B;H1H17992B;H1H17993B;H1H17994B;H1H17995B;H1H17 996B;H1H17997B;H1H17998B;H1H17999B;H1H18000B;H1H18001B;H1H1800 2B;H1H18003B;H1H18004B;H1H18005B;H1H18006B;H1H18007B;H1H18008B; H1H18009B;H1H18010B;H1H18011B;H1H18012B;H1H18013B;H1H18014B;H1 H18015B;H1H18016B;H1H18017B;H1H18018B;H1H18019B;H1H18020B;H1H18 021B;H1H18022B;H1H18023B;H1H18024B;H1H18025B;H1H18026B;H1H1802 7B;H1H18028B;H1H18029B;H1H18030B;H1H18031B;H1H18032B;H1H18033B;H1H18034B;H1H18035B;H1H18037B;H1H18038B;H1H18039B;H1H18040B;H1H18041B;H1H18042B;H1H18043B;H1H18044B;H1H18045B;H1H18046B;H1H18047B;H1H18048B;H1H18049B;H1H18051B;H1H18052B;H1H18053B;H1H18054B;H1H18055B;H1H18056B;H1H18057B;H1H18058B;H1H18059B;H1H18060B;H1H18061B;H1H18062B;H1H18063B;H1H18064B;H1H18065B;H1H18066B;H1H18067B;H1H18068B;H1H18069B;H1H18070B;H1H18071B;H1H18072B;H1H18073B;H1H18074B;H1H18075B;H1H18076B;H1H18077B;H1H18078B;H1H18079B;H1H18080B;H1H18081B;H1H18082B;H1H18083B;H1H18084B;H1H18085B;H1H18086B;H1H18087B;H1H18088B;H1H18089B;H1H18090B;H1H18091B;H1H18092B;H1H18093B;H1H18094B;H1H18095B;H1H18096B;H1H18097B;H1H18098B;H1H18099B;H1H18100B;H1H18101B;H1H18102B;H1H18103B;H1H18104B;H1H18105B;H1H18107B;H1H18108B;H1H18109B;H1H18110B;H1H18111B;H1H18112B;H1H18113B;H1H18114B;H1H18115B;H1H18116B;H1H18117B;H1H18118B;H1H18119B;H1H18120B;H1H18121B;H1H18122B;H1H18123B;H1H18124B;H1H18125B;H1H18126B;H1H18127B;H1H18128B;H1H18129B;H1H18130B;H1H18131B;H1H18132B;H1H18133B;H1H18134B;H1H18135B;H1H18136B;H1H18137B;H1H18138B;H1H18139B;H1H18140B;H1H18141B;H1H18142B;H1H18143B;H1H18144B;H1H18145B;H1H18146B;H1H18147B;H1H18148B;H1H18149B;H1H18150B;H1H18151B;H1H18152B;H1H18153B;H1H18154B;H1H18155B;H1H18156B;H1H18157B;H1H18158B;H1H18159B;H1H18160B;H1H18161B;H1H18162B;H1H18163B;H1H18164B;H1H18165B;H1H18166B;H1H18167B;H1H18168B;H1H18169B;H1H18170B;H1H18171B;H1H18172B;H1H18173B;H1H18174B;H1H18175B;H1H18176B;H1H18177B;H1H18178B;H1H18179B;H1H18180B;H1H18181B;H1H18182B;H1H18183B;H1H18184B;H1H18185B;H1H18186B;H1H18187B;H1H18188B;H1H18189B;H1H18190B;H1H18191B;H1H18192B;H1H18193B;H1H18194B;H1H18195B;H1H18196B;H1H18197B;H1H18198B;H1H18199B;H1H18200B;H1H18201B;H1H18202B;H1H18203B;H1H18204B;H1H18205B;H1H18206B;H1H18207B;H1H18208B;H1H18209B;H1H18210B;H1H18211B;H1H18212B;H1H18213B;H1H18214B;H1H18216B;H1H18217B;H1H18218B;H1H18219B;H1H18220B;H1H18221B;H1H18222B;H1H18223B;H1H18224B;H1H18225B;H1H18226B;H1H18227B;H1H18228B;H1H18229B;H1H18230B;H1H18231B;H1H18232B;H1H18233B;H1H18234B;H1H18235B;H1H18236B;H1H18237B;H1H18238B;H1H18239B;H1H18240B;H1H18241B;H1H18242B;H1H18243B;H 1H18244B;H1H18245B;H1H18246B;H1H18247B;H1H18248B;H1H18249B;H1 H18250B;H1H18251B;H1H18252B;H1H18253B;H1H18254B;H1H18255B;H1H 18256B;H1H18257B;H1H18258B;H1H18259B;H1H18261B;H1H18262B;H1H18 263B;H1H18264B;H1H18265B;H1H18266B;H1H18267B;H1H18268B;H1H182 69B;H1H18270B;H1H18271B;H1H18272B;H1H18274B;H1H18275B;H1H1827 6B;H1H18277B;H1H18278B;H1H18279B;H1H18280B;H1H18281B;H1H18282 B;H1H18283B;H1H18284B;H1H18285B;H1H18286B;H1H18287B;H1H18288B; H1H18289B;H1H18290B;H1H18291B;H1H18292B;H1H18293B;H1H18294B;H 1H18295B;H1H18297B;H1H18298B;H1H18299B;H1H18300B;H1H18301B;H1 H18302B;H1H18303B;H1H18304B;H1H18305B;H1H18306B;H1H18307B;H1H 18308B;H1H18309B;H1H18310B;H1H18311B;H1H18312B;H1H18313B;H1H18 314B; H1H18315B; H1H18316B; H1H18317B; H1H18318B; H1H18319B; H1H18320B; H1H18321B; H1H18322B; H1H18323B; H1H18324B; H1H18325B; H1H18326B; H1H18327B; H1H18328B; H1H18329B; H1H18330B; H1H18331B; H1H18332B; H1H18333B; H1H18334B; and H1H18335B.

[0018] In one embodiment of the invention, the additional therapeutic agent provided in conjunction with the 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 (e.g., SEQ ID NOs: 25-27) and a light chain immunoglobulin comprising CDR-L1, CDR-L2 and CDR-L3 of H1H14611N2 (e.g., SEQ ID NOs: 29-31).

[0019] In one embodiment of the invention, the additional therapeutic agent provided in conjunction with the 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 (e.g., SEQ ID NOs: 41-43) and a light chain immunoglobulin comprising CDR-L1, CDR-L2 and CDR-L3 of H1H14612N2 (e.g., SEQ ID NOs: 45-47).

[0020] In one embodiment of the invention, the additional therapeutic agent provided in conjunction with the 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 (e.g., SEQ ID NOs: 33-35) and a light chain immunoglobulin comprising CDR-L1, CDR-L2 and CDR-L3 of H1H11729P (e.g., SEQ ID NOs: 37-39).

[0021] The invention also provides a container or injection device comprising an anti-TMPRSS2 antigen binding protein (eg, H1H7017N) or a composition thereof (eg, a pharmaceutical composition).

[0022] The present invention also provides a method 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 (e.g., H1H7017N) described herein.

[0023] The present invention also provides a method of treating or preventing cancer (e.g., prostate cancer) or infection, such as infection by influenza virus, coronavirus, SARS-Co virus, MERS-Co virus, parainfluenza virus, human metapneumovirus, or hepatitis C virus (HCV), in a subject (e.g., a human) in need thereof, comprising administering a therapeutically effective amount of an anti-TMPRSS2 antigen binding protein (e.g., H1H7017N) as described herein. For example, the antigen binding protein is administered with one or more additional therapeutic agents (e.g., antiviral drugs and / or vaccines). In one embodiment of the present invention, the additional therapeutic agent is a member selected from the group consisting of ledipasvir, sovosbuvir, a combination of ledipasvir and sovosbuvir, oseltamivir, zanamivir, ribavirin, and interferon-alpha 2b, interferon-alpha 2a, and an antibody or antigen-binding fragment that specifically binds to influenza HA. In one embodiment of the invention, the additional therapeutic agent is H1H14611N2;H1H14612N2;H1H11723P;H1H11729P;H1H11820N;H1H11829N;H1H11829N2;H2aM11829N;H2M11830N;H1H11830N2;H1H11903N;H1H14571N;H2a14571N;H1H11704P;H1H11711P;H1H11714P;H1H11717P;H1H11724P;H1H11727P;H1H11730P2;H1H11 <h2 style=";text-align:left;direction:ltr">731P2;H1H11734P2;H1H11736P2;H1H11742P2;H1H11744P2;H1H11745P2;H 1H11747P2;H1H11748P2;H1H17952B;H1H17953B;H1H17954B;H1H17955B;H 1H17956B;H1H17957B;H1H17958B;H1H17959B;H1H17960B;H1H17961B;H1H17962B;H1H17963B;H1H17964B;H1H17965B;H1H17966B;H1H17967B;H1H17 968B;H1H17969B;H1H17970B;H1H17971B;H1H17972B;H1H17973B;H1H17974B;H1H17975B;H1H17976B;H1H17977B;H1H17978B;H1H17979B;H1H17980B ;H1H17981B;H1H17982B;H1H17983B;H1H17984B;H1H17985B;H1H17986B;H1H17987B;H1H17988B;H1H17989B;H1H17990B;H1H17991B;H1H17992B;H1H 17993B;H1H17994B;H1H17995B;H1H17996B;H1H17997B;H1H17998B;H1H17999B;H1H18000B;H1H18001B;H1H18002B;H1H18003B;H1H18004B;H1H1800 5B;H1H18006B;H1H18007B;H1H18008B;H1H18009B;H1H18010B;H1H18011B ;H1H18012B;H1H18013B;H1H18014B;H1H18015B;H1H18016B;H1H18017B;H 1H18018B;H1H18019B;H1H18020B;H1H18021B;H1H18022B;H1H18023B;H1H 18024B;H1H18025B;H1H18026B;H1H18027B;H1H18028B;H1H18029B;H1H18 030B;H1H18031B;H1H18032B;H1H18033B;H1H18034B;H1H18035B;H1H1803 7B;H1H18038B;H1H18039B;H1H18040B;H1H18041B;H1H18042B;H1H18043B;H1H18044B;H1H18045B;H1H18046B;H1H18047B;H1H18048B;H1H18049B;H1H18051B;H1H18052B;H1H18053B;H1H18054B;H1H18055B;H1H18056B;H1H18057B;H1H18058B;H1H18059B;H1H18060B;H1H18061B;H1H18062B;H1H18063B;H1H18064B;H1H18065B;H1H18066B;H1H18067B;H1H18068B;H1H18069B;H1H18070B;H1H18071B;H1H18072B;H1H18073B;H1H18074B;H1H18075B;H1H18076B;H1H18077B;H1H18078B;H1H18079B;H1H18080B;H1H18081B;H1H18082B;H1H18083B;H1H18084B;H1H18085B;H1H18086B;H1H18087B;H1H18088B;H1H18089B;H1H18090B;H1H18091B;H1H18092B;H1H18093B;H1H18094B;H1H18095B;H1H18096B;H1H18097B;H1H18098B;H1H18099B;H1H18100B;H1H18101B;H1H18102B;H1H18103B;H1H18104B;H1H18105B;H1H18107B;H1H18108B;H1H18109B;H1H18110B;H1H18111B;H1H18112B;H1H18113B;H1H18114B;H1H18115B;H1H18116B;H1H18117B;H1H18118B;H1H18119B;H1H18120B;H1H18121B;H1H18122B;H1H18123B;H1H18124B;H1H18125B;H1H18126B;H1H18127B;H1H18128B;H1H18129B;H1H18130B;H1H18131B;H1H18132B;H1H18133B;H1H18134B;H1H18135B;H1H18136B;H1H18137B;H1H18138B;H1H18139B;H1H18140B;H1H18141B;H1H18142B;H1H18143B;H1H18144B;H1H18145B;H1H1814; <h2 style=";text-align:left;direction:ltr">6B;H1H18147B;H1H18148B;H1H18149B;H1H18150B;H1H18151B;H1H18152B;H1H18153B;H1H18154B;H1H18155B;H1H18156B;H1H18157B;H1H18158B;H 1H18159B;H1H18160B;H1H18161B;H1H18162B;H1H18163B;H1H18164B;H1H 18165B;H1H18166B;H1H18167B;H1H18168B;H1H18169B;H1H18170B;H1H18 171B;H1H18172B;H1H18173B;H1H18174B;H1H18175B;H1H18176B;H1H1817 7B;H1H18178B;H1H18179B;H1H18180B;H1H18181B;H1H18182B;H1H18183B ;H1H18184B;H1H18185B;H1H18186B;H1H18187B;H1H18188B;H1H18189B;H 1H18190B;H1H18191B;H1H18192B;H1H18193B;H1H18194B;H1H18195B;H1H 18196B;H1H18197B;H1H18198B;H1H18199B;H1H18200B;H1H18201B;H1H18 202B;H1H18203B;H1H18204B;H1H18205B;H1H18206B;H1H18207B;H1H1820 8B;H1H18209B;H1H18210B;H1H18211B;H1H18212B;H1H18213B;H1H18214B ;H1H18216B;H1H18217B;H1H18218B;H1H18219B;H1H18220B;H1H18221B;H 1H18222B;H1H18223B;H1H18224B;H1H18225B;H1H18226B;H1H18227B;H1H18228B;H1H18229B;H1H18230B;H1H18231B;H1H18232B;H1H18233B;H1H18 234B;H1H18235B;H1H18236B;H1H18237B;H1H18238B;H1H18239B;H1H18240B;H1H18241B;H1H18242B;H1H18243B;H1H18244B;H1H18245B;H1H18246B;H1H18247B;H1H18248B;H1H18249B;H1H18250B;H1H18251B;H1H18252B;H1H18253B;H1H18254B;H1H18255B;H1H18256B;H1H18257B;H 1H18258B;H1H18259B;H1H18261B;H1H18262B;H1H18263B;H1H18264B;H1H18265B;H1H18266B;H1H18267B;H1H18268B;H1H18269B;H1 H18270B;H1H18271B;H1H18272B;H1H18274B;H1H18275B;H1H18276B;H1H18277B;H1H18278B;H1H18279B;H1H18280B;H1H18281B;H1H 18282B;H1H18283B;H1H18284B;H1H18285B;H1H18286B;H1H18287B;H1H18288B;H1H18289B;H1H18290B;H1H18291B;H1H18292B;H1H18 293B;H1H18294B;H1H18295B;H1H18297B;H1H18298B;H1H18299B;H1H18300B;H1H18301B;H1H18302B;H1H18303B;H1H18304B;H1H183 05B;H1H18306B;H1H18307B;H1H18308B;H1H18309B;H1H18310B;H1H18311B;H1H18312B;H1H18313B;H1H18314B;H1H18315B;H1H18316 B; H1H18317B; H1H18318B; H1H18319B; H1H18320B; H1H18321B; H1H18322B; H1H18323B; H1H18324B; H1H18325B; H1H18326B; H1H18327B; H1H18328B; H1H18329B; H1H18330B; H1H18331B; H1H18332B; H1H18333B; H1H18334B; and H1H18335B, or an antigen-binding fragment thereof;

[0024] The invention also relates to the anti-TMRPSS2 antigen binding proteins described herein (e.g., The present invention provides a method of administering an antigen-binding protein (H1H7017N) to the body of a subject (e.g., a human) comprising parenterally (e.g., subcutaneously, intravenously, or intramuscularly) injecting the antigen-binding protein into the body of the subject. [Brief description of the drawings]

[0025] [Figure 1] Progression of A / Puerto Rico / 08 / 1934 (H1N1)-GFP virus spreading in different cell lines at an initial multiplicity of infection of 0.01 (A) or 0.001 (B) in the absence of exogenous trypsin: Calu3 (circle), A549 (square), MDCK (triangle) and HepG2 (inverted triangle) cells. [Diagram 2] Application of H1H7017N during the infection cycle reduces the number of A / Puerto Rico / 08 / 1934 (H1N1) fluorescent focus units (FFU) at 72 hours post-infection compared to isotype control antibody, no antibody, anti-HA antibody and uninfected controls. [Diagram 3] Anti-TMPRSS2, H1H7017N, binds to human and cynomolgus TMPRSS2 expressed on cells. (A) H1H7017N binds to MDCK / Tet-on / hTMPRSS2 and MDCK / Tet-on / mfTMPRSS2 with EC50 values ​​of 460 pM and 1.06 nM, respectively, and showed no significant binding to MDCK / Tet-on cells. (B) Control mAb1, an irrelevant isotype control antibody, showed no binding to any of the cell lines tested. [Figure 4] Survival curves of mice engineered to express human TMPRSS2 protein treated with 5 mg / kg H1H7017N on day -1 PI (inverted triangles, dashed line) or day 0 PI (circles, solid line) demonstrated protection against H1N1 in a prophylactic model. Mice treated with the isotype control H1H1238N (triangles, solid line) demonstrated no protection. [Diagram 5]Survival curves of mice engineered to express the human TMPRSS2 protein infected with H1N1 and treated with 10 mg / kg H1H7017N showed protection. Mice were treated on days 0 (diamonds, dotted line), 1 (circles, solid line), 2 (inverted triangles, solid line), or 3 (squares, dashed line) PI. The isotype control H1H1238N (triangles, solid line) had partial protection with 25% survival. [Figure 6] Survival curves of hTPMRSS2 mice treated with 10 mg / kg H1H7017N on day 1 PI (triangles) or day 2 PI (circles) showed protection against H3N2. Untreated mice (squares) showed no protection. [Figure 7] Survival curves of wild-type mice (A) or mice engineered to express the human TMPRSS2 protein (B) infected with 150 PFU (triangles), 750 PFU (squares), or 1,500 PFU (circles) of A / Puerto Rico / 08 / 1934 (H1N1). Mice were weighed daily until day 14 PI. [Figure 8] Survival curves for mice engineered to express human TMPRSS2 protein infected with A / Aichi / 2 / 68 (HA, NA) × A / PR / 8 / 34 (H3N2) on day 0 and treated with a combination of H1H7017N and H1H14611N2 at 2.5 mg / kg each (diamonds), H1H1H7017N at 10 mg / kg (triangles), H1H14611N2 at 10 mg / kg each (squares), H1H7017N and H1H14611N2 at 5 mg / kg each, or hIgG1 isotype control at 10 mg / kg (circles). Mice were weighed daily until day 14 PI. [Figure 9]Survival curves for mice engineered to express human TMPRSS2 protein infected with A / Puerto Rico / 08 / 1934 (H1N1) on day 1 PI and treated with a combination of 1 mg / kg H1H7017N and 2 mg / kg H1H11729P (circles), 2.5 mg / kg each of H1H7017N and H1H11729P (inverted triangles), 5 mg / kg H1H11729P (diamonds), 5 mg / kg H1H7017N (squares), or 5 mg / kg hIgG1 isotype control (triangles). Mice were weighed daily until day 14 PI. DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS

[0026] Before describing the method, it should be understood that the invention is not limited to the particular methods and experimental conditions described, for example, the methods and conditions may vary. It should also be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, and that the scope of the invention will be limited only by the appended claims.

[0027] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by those skilled in the art to which this invention belongs.Although any method and material similar or equivalent to those described herein can be used in the practice or testing of this invention, the preferred method and material are described herein.All publications described herein are incorporated herein by reference in their entirety.

[0028] The term "influenza hemagglutinin", also referred to as "influenza HA", is a trimeric glycoprotein found on the surface of influenza virions that mediates viral attachment (through binding of HA1 to α-2,3- and α-2,6-sialic acids) and entry into host cells (through conformational changes). HA is composed of two structural domains: a globular head domain (which undergoes high frequency antigenic mutations) containing the receptor binding site and a stem region (which is more conserved among various influenza virus strains). Influenza HA is synthesized as a precursor (HA0), which undergoes proteolytic processing to generate two subunits (HA1 and HA2), which associate with each other to form the stem / globular head structure. The viral HA is the most variable antigen on the virus, while the stem (HA2) is highly conserved within each group.

[0029] The term "influenza neuraminidase," also referred to as "influenza NA," is an exosialidase (EC 3.2.1.18) that cleaves the α-ketosidic linkage between sialic acid (N-acetylneuraminic acid) and adjacent sugar residues.

[0030] The amino acid sequence of 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 coupled to, for example, a histidine tag, a mouse or human Fc, or a signal sequence.

[0031] An anti-TMPRSS2 "antigen binding protein" is a polypeptide, or a complex of more than one polypeptide (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.

[0032] TMPRSS2 TMPRSS2 (transmembrane protease 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 demonstrated to mediate the cleavage of influenza virus HA0 into HA1 and HA2.

[0033] The human TMPRSS2 gene encodes a putative protein of 492 amino acids that anchors to the cell membrane. This protein is converted to a mature form via autocatalytic cleavage between Arg255 and Ile256. After cleavage, the mature protease is mostly membrane-bound, but some of them are released into the extracellular environment.

[0034] In one embodiment of the invention, human TMPRSS2(V160M) has the amino acid sequence: [ka] (SEQ ID NO:22; methionine 160 is in bold). In one embodiment of the invention, the TMPRSS2 polypeptide does not comprise a V160M mutation. See also NM_005656.3.

[0035] In one embodiment of the invention, Macaca mulatta TMPRSS2 (S129L, N251S, I415V, R431Q, D492G) has the amino acid sequence: [ka] (SEQ ID NO: 23). In one embodiment of the invention, the TMPRSS2 polypeptide does not comprise a S129L, N251S, I415V, R431Q and / or D492G mutation.

[0036] In one embodiment of the present invention, Mus musculus TMPRSS2 The mRNA comprises the nucleotide sequence shown in NM_015775.2.

[0037] virus The present invention includes methods of treating or preventing a viral infection in a subject. The term "virus" includes any virus whose infection in a subject's organism is treatable or preventable by administration of an anti-TMPRSS2 antibody or antigen-binding fragment thereof (e.g., the infectivity of the virus is at least partially dependent on TMPRSS2). In one embodiment of the invention, the "virus" is any virus that expresses HA0 or another substrate of TMPRSS2, the proteolytic cleavage of which is required for full infectivity of the virus to cells in a 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, bronchi, lungs) of a subject and are treatable or preventable by administration of an anti-TMPRSS2. For example, in one embodiment of the present invention, the virus includes influenza virus, coronavirus, SARS-Co virus (Severe Acute Respiratory Syndrome Coronavirus), MERS-Co virus (Middle East Respiratory Syndrome (MERS) CoV), parainfluenza virus, Sendai virus (SeV), human metapneumovirus, and / or hepatitis C virus (HCV). "Viral infection" refers to the entry and reproduction of a virus in the living body of a subject. The present invention includes embodiments, with the proviso that "virus" excludes influenza virus, e.g., viral infection excludes influenza virus infection.

[0038] 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) can be morphologically separated from influenza viruses.

[0039] Sendai virus, also known as mouse parainfluenza virus, is the type species of the genus Respirovirus, which also contains human parainfluenza virus 3, bovine parainfluenza virus 3, and human parainfluenza virus 1. TMPRSS2 is the activating protease of respiratory parainfluenza viruses, such as parainfluenza virus and Sendai virus (SeV). See Abe et al., J. Virol., 87(21):11930-11935 (2013).

[0040] Human metapneumovirus (HMPV) is classified as the first human member of the genus Metapneumovirus in the subfamily Pneumovirinae within the family Paramyxoviridae. It is an enveloped, negative-sense, single-stranded RNA virus. The RNA genome contains eight genes encoding nine proteins. HMPV has the same gene order as avian pneumovirus (AMPV), which belongs to the genus Metapneumovirus. TMPRSS2 is expressed in human lung epithelium, efficiently cleaves HMPV F protein, and supports HMPV reproduction, as well as may be involved in the development of lower respiratory tract disease in HMPV-infected patients. See Shirogane et al., J Virol., 82(17):8942-8946 (2008).

[0041] Hepatitis C virus (HCV) is a small, enveloped, positive-sense, single-stranded RNA virus in the Flaviviridae family. HCV has at least six genotypes and numerous subtypes and is a member of the hepacivirus genus. TMPRSS2 can activate HCV infection post-binding and during the entry phase. Esumi et al., Hepatology, 61(2):437-446 (2015).

[0042] Influenza viruses belong to the family Orthomyxoviridae. This family refers to enveloped viruses whose genome contains a segmented negative-sense single-stranded RNA segment. There are four genera in this family: types A, B, C and Thogotovirus. Influenza virus classes A, B and C are based on the core protein and are further divided into subtypes (e.g., subtype A / H1N1) determined by the viral envelope glycoproteins hemagglutinin (HA) and neuraminidase (NA). 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) are used to define influenza subtypes. Group 1 influenza includes the H1, H2, H5, H6, H8, H9, H11, H12, H13, H16, H17, and H18 subtypes, as well as the NA8, NA5, Na4, and NA1 subtypes. Group 2 includes the H3, H4, H7, H10, H14, and H15 subtypes, as well as the NA6, NA9, NA7, NA2, and NA3 subtypes. Influenza A viruses infect a variety of mammalian and avian species, while types B and C infections are primarily restricted to humans. The eight genome segments of influenza A and B viruses are loosely encapsidated by nucleoprotein.

[0043] Coronavirus virions are spherical with a diameter of approximately 125 nm. The most prominent feature of coronaviruses are the club-like spiked projections that emerge from the surface of the virion. These spikes are the defining feature of the virion, giving it the appearance of a solar corona and prompting the name coronavirus. Within the virion envelope is the nucleocapsid. Coronaviruses have a helically symmetrical nucleocapsid, which is uncommon in positive-sense RNA viruses but much more common in negative-sense RNA viruses. Both MERS-CoV (Middle East Respiratory Syndrome coronavirus) and SARS-CoV (Severe Acute Respiratory Syndrome coronavirus) belong to the Coronaviridae family. The initial binding of the virion to the host cell is initiated by the interaction of the S protein with its receptor. The site of the receptor binding domain (RBD) within the S1 region of the coronavirus S protein varies from virus to virus, with some having the RBD at the C-terminus of S1. The S protein / receptor interaction is the major determinant of coronavirus infection of 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 gain access to the host cell cytoplasm. This is generally accomplished by acid-dependent proteolytic cleavage of the S protein by a cathepsin, TMPRS2 or another protease, followed by fusion of the viral and cellular membranes.

[0044] 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 antigenic fragment thereof.

[0045] The term "antibody," as used herein, refers to an immunoglobulin molecule comprising four polypeptide chains, two heavy chains (HC) and two light chains (LC), interconnected by disulfide bonds (i.e., "intact antibody molecules"), as well as multimers thereof (e.g., IgM), e.g., H1H701N. Each heavy chain comprises a heavy chain variable region ("HCVR" or "VH ") (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 "V L ") (e.g., SEQ ID NO: 4) and a light chain constant region (C L ) is composed of V H and V L The regions are further subdivided into hypervariable regions 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 the antibody (or antigen-binding fragment thereof) are identical to human germline sequences or are naturally or artificially modified.

[0046] 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-terminus to C-terminus, both light and heavy chain variable domains contain FR1, CDR1, FR2, CDR2, FR3, CDR3 and FR4. In one embodiment of the invention, the assignment of amino acids to each domain is based on the Sequences of Proteins of Immunological Interest, Kabat et al.; National Institutes of Health, Bethesda, Md.; 5th ed.; NIH Publ. 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 according to the definitions in Chothia et al., (1989) Nature 342:878-883.

[0047] 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. The term "monoclonal antibody" as used herein 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 small amounts. A "plurality" of such monoclonal antibodies and fragments in a composition refers to a concentration of identical (i.e., in amino acid sequence, except for possible naturally occurring mutations that may be present in small amounts, as discussed above) antibodies and fragments, which typically exceeds the concentration naturally present in the blood of a host organism, e.g., a mouse or human.

[0048] In one embodiment of the invention, the anti-TMPRSS2 antigen binding protein, e.g., an antibody or antigen binding fragment, comprises a heavy chain constant domain, e.g., of type IgA (e.g., IgA1 or IgA2), IgD, IgE, IgG (e.g., IgG1, IgG2, IgG3 and IgG4), or IgM. In one embodiment of the invention, the antigen binding protein, e.g., an antibody or antigen binding fragment, comprises a light chain constant domain, e.g., of type kappa or lambda.

[0049] The term "human" antigen-binding protein, e.g., antibody, as used herein, 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. Pat. No. 8502018, U.S. Pat. No. 6596541, or U.S. Pat. No. 5789215. The human mAbs of the present invention may include, e.g., in the CDRs and especially in CDR3, amino acid residues that are not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by in vitro random or site-specific mutagenesis or by in vivo somatic mutation). However, the term "human antibody" as used herein is not intended to include mAbs in which CDR sequences derived from the germline of another mammalian species (e.g., mouse) are grafted onto human FR sequences. The term includes antibodies recombinantly produced in non-human mammals or in cells of non-human mammals. The term is not intended to include antibodies isolated from or produced in a human subject. See below.

[0050] The present invention includes anti-TMPRSS2 chimeric antigen binding proteins, such as antibodies and antigen-binding fragments thereof, and methods of using them. 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, where the first antibody and the second antibody are from different species (U.S. Pat. No. 4,816,567; and Morrison et al., (1984) Proc. Natl. Acad. Sci. USA, vol. 81: 6851-6855).

[0051] The term "recombinant" antigen binding protein, e.g., an antibody or antigen binding fragment thereof, refers to such molecules made, expressed, isolated or obtained by techniques or methods known in the art as recombinant DNA technology, including, for example, DNA splicing and transgenic expression. The term also applies to non-human mammals (transgenic non-human mammals, For example, antibodies expressed in human (including transgenic mouse) or cellular (eg, CHO cell) expression systems or isolated from a recombinant combinatorial human antibody library.

[0052] Recombinant anti-TMPRSS2 antigen binding proteins, e.g., antibodies and antigen binding fragments, disclosed herein are also 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., H1H7017N) 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, or an immunoglobulin chain 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 that also comprises a polynucleotide encoding an immunoglobulin chain operably linked to a T7 promoter. For example, in one embodiment of the invention, a bacterial host cell, e.g., E. coli, comprises a polynucleotide encoding a T7 RNA polymerase gene operably linked to a lac promoter, and expression of the polymerase and chains is induced by incubating the host cell with IPTG (isopropyl-beta-D-thiogalactopyranoside). No. 4,952,496 and U.S. Pat. No. 5,693,489, or Studier & Moffatt, "Use of bacteriophage T7 RNA polymerase to direct See, "selective high-level expression of cloned genes," J. Mol. Biol., 1986, May 5;189(1):113-30.

[0053] 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 US Patent No. 4,816,567.

[0054] Transformation can be carried out by any known method for introducing polynucleotides into host cells. Methods for introducing heterologous polynucleotides into mammalian cells are well known in the art, and include dextran-mediated transfection, calcium phosphate precipitation, polybrene-mediated transfection, protoplast fusion, electroporation, encapsulation of polynucleotide(s) in liposomes, biolistec injection, and direct microinjection of DNA into nuclei. 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 No. 4,399,216, U.S. Patent No. 4,912,040, U.S. Patent No. 4,740,461, and U.S. Patent No. 4,959,455.

[0055] Thus, the present invention includes recombinant methods for making anti-TMPRSS2 antigen binding proteins, e.g., antibodies or antigen binding fragments thereof of the invention, or immunoglobulin chains thereof, comprising the steps of: (i) introducing one or more polynucleotides (e.g., comprising any one or more of the nucleotide sequences of SEQ ID NOs: 1, 3, 5, 7, 9, 11, 13 or 15) encoding the antigen binding protein, e.g., the light and / or heavy immunoglobulin chains of H1H7017N or H4H7017N, e.g., wherein the polynucleotide is in a vector; and / or integrated into a host cell chromosome and / or operably linked to a promoter; (ii) culturing the host cell (e.g., CHO or Pichia or Pichia pastoris) under conditions suitable 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. Antigen-binding proteins (e.g., antibodies or antigen-binding fragments) that contain more than one immunoglobulin chain, e.g., two heavy immunoglobulin chains and two light immunoglobulin chains. When making antibodies comprising TMPRSS2 chains, co-expression of the chains in a single host cell results in association of the chains, e.g., intracellularly or on the cell surface or extracellularly, when such chains are secreted, to form an antigen-binding protein (e.g., an antibody or antigen-binding fragment). The methods include those in which only heavy immunoglobulin chains, or only light immunoglobulin chains (e.g., any of those discussed herein including mature fragments and / or variable domains thereof) are expressed. Such chains are useful, e.g., 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, e.g., antibodies and antigen-binding fragments thereof, including heavy chain immunoglobulins (or variable domains or CDRs thereof) encoded by a polynucleotide comprising the nucleotide sequence set forth in SEQ ID NO:1, and light chain immunoglobulins (or variable domains or CDRs thereof) encoded by a nucleotide sequence set forth in SEQ ID NO:3, which are products of such production methods, and optionally purification methods described herein. For example, in one embodiment of the present invention, the products of the methods include V-type immunoglobulins comprising the amino acid sequence set forth in SEQ ID NO:2, and V-type immunoglobulins comprising the amino acid sequence set forth in SEQ ID NO:3, which are products of such production methods, and optionally purification methods described herein. H and V comprising the amino acid sequence shown in SEQ ID NO:4 L or an anti-TMPRSS2 antigen binding protein that is an antibody or fragment comprising an HC comprising the amino acid sequence set forth in SEQ ID NO:17 or 19, and an LC comprising the amino acid sequence set forth in SEQ ID NO:18.

[0056] Eukaryotic and prokaryotic host cells, such as mammalian cells, are used as hosts for the 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 may be used are insect cell lines (e.g. Spodoptera frugiperda or Trichoplusia ni), amphibian cells, bacterial cells, plant cells and fungal cells. Fungal cells include yeast and filamentous fungal cells, such as Pichia pastoris, Pichia finlandica, Pichia trehalophila, Pichia koclamae, Pichia membranaefaciens, Pichia minuta, Pichia ni ... minuta (Ogataea minuta, Pichia lindneri), Pichia opuntiae, Pichia thermotolerans, Pichia salictaria, Pichia guercuum, Pichia pijperi, Pichia stiptis, Pichia methanolica, Pichia sp., Saccharomyces cerevisiae, Saccharomyces sp., Hansenula polymorpha polymorpha, Kluyveromyces sp., Kluyveromyces lactis, Candida albicans, Aspergillus nidulans, Aspergillus niger, Aspergillus oryzae, Trichoderma reesei, Chrysosporium lactis, Chrysoprion spp. ... sosporium lucknowense, Fusarium sp., Fusarium gramineum, Fusarium venenatum, Physcomitrella patens, and Neurospora crassa. The present invention includes an isolated host cell (e.g., a CHO cell) containing an antigen-binding protein, such as H1H7017N; or a polynucleotide encoding such a polypeptide.

[0057] The term "specifically binds" refers to a binding affinity of at least about 10, as measured, for example, by a real-time label-free biolayer interference assay at 25°C or 37°C, for example, by an Octet® HTX biosensor, or by surface plasmon resonance, for example, BIACORE™, or by 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)K D The term "mAb" refers to an antigen binding protein (e.g., mAb) that has binding affinity for an antigen, such as a TMPRSS2 protein (e.g., human TMPRSS2), expressed as a serotype 1 (SEQ ID NO: 1). The present invention includes antigen binding proteins that specifically bind to a TMPRSS2 protein.

[0058] The term "antigen-binding portion" or "antigen-binding fragment" of an antibody or antigen-binding protein, as used herein, includes any naturally occurring, enzymatically obtained, 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 that mimic the hypervariable regions of an antibody (e.g., isolated CDRs, such as complementarity determining region (CDR) 3 peptides), or constrained FR3-CDR3-FR4 peptides. 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 Publication No. WO08 / 020079 or International Publication No. WO09 / 138519) (e.g., monovalent nanobodies, bivalent nanobodies, etc.), small modular immunopharmaceuticals (SMIPs), and shark variable IgNAR domains are also encompassed within the expression "antigen-binding fragment" as used herein. In one embodiment of the present invention, the antigen-binding fragment comprises three or more CDRs of H1H7017N (e.g., CDR-H1, CDR-H2 and CDR-H3; or CDR-L1, CDR-L2 and CDR-L3).

[0059] An antigen-binding fragment of an antibody, in one embodiment of the invention, comprises 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 Domain-associated V H For antigen-binding fragments containing domains, V H Domains and V L The domains may be arranged relative to one another in any suitable configuration. For example, the variable region may be a dimer and may have a V H -V H , V H -VL or V L -V L Alternatively, the antigen-binding fragment of the antibody may contain a monomeric V H or V L It may include a domain.

[0060] In certain embodiments, an antigen-binding fragment of an antibody may comprise at least one variable domain covalently linked to at least one constant domain. Non-limiting exemplary configurations of variable and constant domains 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;(ii)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 ;(vii)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 the variable and constant domains, including any of the exemplary configurations listed above, the variable and constant domains are directly linked to each other or 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 and provides a flexible or semi-flexible linkage 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 The domains may comprise homodimers or heterodimers (or other multimers) of any of the variable and constant domain configurations listed above, non-covalently associated (e.g., by disulfide bond(s)).

[0061] 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.

[0062] In a specific embodiment, the antibodies or antibody fragments of the invention are conjugated to a moiety such as a ligand or a therapeutic moiety (an "immunoconjugate"), such as an anti-viral drug, a second anti-influenza antibody, or any other therapeutic moiety useful for treating a viral infection, e.g., an influenza virus infection. See below.

[0063] The present invention also provides a complex comprising an antibody or antigen-binding fragment discussed herein complexed with an anti-TMPRSS2 antigen-binding protein, such as a TMPRSS2 polypeptide or antigenic fragment thereof, and / or a secondary antibody or antigen-binding fragment thereof (e.g., a detectably labeled secondary antibody) that specifically binds to a TMPRSS2 polypeptide or fragment thereof. In one embodiment of the present invention, the antibody or fragment is in vitro (e.g., immobilized on a solid substrate) or in the subject's living body. In one embodiment of the present invention, the TMPRSS2 is in vitro (e.g., immobilized on a solid substrate) or on the surface of a cell or in the subject's living body. Also part of the present invention are immobilized anti-TMRPSS2 antibodies and antigen-binding fragments thereof covalently linked to an insoluble matrix material (e.g., glass or polysaccharides, such as agarose or sepharose, such as beads or other particles thereof); optionally, the immobilized antibody is complexed with TMPRSS2 or an antigenic fragment thereof, or a secondary antibody or fragment thereof.

[0064] "Isolated" antigen-binding proteins, antibodies or antigen-binding fragments thereof, polypeptides, polynucleotides and vectors are at least partially free of 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 cell debris and growth medium. An isolated antibody or antigen-binding fragment may further be at least partially free of expression system components such as biological molecules from 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 components of pharmaceutical formulations that include the antibody or fragment.

[0065] The term "epitope" refers to an antigenic determinant (e.g., on a TMPRSS2 polypeptide) that interacts with a specific antigen-binding site of an antigen-binding protein, e.g., a variable region of an antibody molecule, known as a paratope. A single antigen can have more than one epitope. Thus, different antibodies can bind to different regions on an antigen and have different biological effects. The term "epitope" also refers to a specific antigen binding site of an antigen-binding protein, e.g., an antigenic determinant (e.g., on a TMPRSS2 polypeptide) that interacts with a variable region of an antibody molecule, known as a paratope. A single antigen can have more than one epitope. Thus, different antibodies can bind to different regions on an antigen and have different biological effects. The term "epitope" also refers to a specific antigen binding site of an antigen that interacts with a specific antigen binding site of an antigen, e.g., an antigenic determinant (e.g., on a TMPRSS2 polypeptide) that interacts with a specific antigen binding site of an antigen-binding protein, e.g., an antigenic determinant (e.g., on a TMPRSS2 polypeptide). It refers to the site on an antigen to which an antibody responds. It also refers to the region of an antigen to which an antibody binds. Epitopes are defined as structural or functional. Functional epitopes are generally a subset of structural epitopes and have residues that directly contribute to the affinity of the interaction. Epitopes can be linear or conformational, i.e., composed of non-linear amino acids. In certain embodiments, epitopes can 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, can have specific three-dimensional structural characteristics, and / or specific charge characteristics.

[0066] 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., vol. 248: 443-63), peptide truncation analysis, crystallographic studies and NMR analysis. In addition, methods such as epitope removal, epitope extraction and chemical modification of the antigen can be used (Tomer (2000) Prot. Sci., vol. 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., Coversin) interacts is hydrogen / deuterium exchange detected by mass spectrometry. In general terms, the hydrogen / deuterium exchange method involves deuterium labeling of the protein of interest, followed by binding of 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 deuterium-hydrogen back exchange 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 interface retain deuterium and therefore exhibit a relatively higher 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, thereby revealing deuterium-labeled residues that correspond to the specific amino acids with which the antigen-binding protein interacts. See, e.g., Ehring (1999) Analytical Biochemistry 267:252-259; Engen and Smith (2001) Anal.Chem., 73:256A-265A.

[0067] The term "compete" as used herein 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 that antigen. The term also includes competition between two antigen-binding proteins, e.g., antibodies, in both directions, i.e., a first antibody binds 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, a first and a second antigen-binding protein (e.g., an antibody) may bind to different, but overlapping, epitopes, such that the binding of one inhibits or blocks the binding of the second antibody, e.g., via steric hindrance. Competition between antigen-binding proteins (e.g., antibodies) is measured by methods known in the art, e.g., real-time label-free biolayer interference assays. In one embodiment of the invention, competition between a first and a second anti-TMPRSS2 antigen binding protein (e.g., an antibody) is determined by measuring the ability of an immobilized first anti-TMPRSS2 antigen binding protein (e.g., an antibody) (not initially complexed with TMPRSS2 protein) to bind to soluble TMPRSS2 protein complexed with a second anti-TMPRSS2 antigen binding protein (e.g., an antibody). A decrease in the ability of the first anti-TMPRSS2 antigen binding protein (e.g., an antibody) to bind to complexed TMPRSS2 protein compared to uncomplexed TMPRSS2 protein indicates a decrease in the ability of the first and second anti-TMPRSS2 antigen binding proteins to bind to soluble TMPRSS2 protein. This indicates that the quality (e.g., antibody) competes with the antibody. The degree of competition can be expressed as a percentage of the reduction in binding. Such competition can be measured using real-time label-free biolayer interference assays, such as Octet RED384 biosensor (Pall ForteBio Corp.), ELISA (enzyme-linked immunosorbent assay) or SPR (surface plasmon resonance).

[0068] Binding competition between anti-TMPRSS2 antigen-binding proteins (e.g., monoclonal antibodies (mAbs)) can be determined using a real-time label-free biolayer interference assay on an Octet RED384 biosensor (Pall ForteBio Corp.). For example, to determine competition between two anti-human TMPRSS2 monoclonal antibodies, an anti-hFc antibody-coated Octet biosensor chip (Pall ForteBio Corp.) was probed by immersing the tip in a solution of anti-human TMPRSS2 mAb (hereafter referred to as "mAb1"). Anti-TMPRSS2 mAb can be first captured on an antibody capture biosensor chip (ForteBio Corp., #18-5060). Next, as a positive control for blocking, the antibody capture biosensor chip can be saturated with a known blocking isotype control mAb (hereafter referred to as "blocking mAb") by immersion in a solution of the blocking mAb. Next, to determine whether mAb2 competes with mAb1, the biosensor chip can then be immersed in a co-complexed solution of human TMPRSS2 polypeptide and a second anti-human TMPRSS2 mAb (hereafter referred to as "mAb2"), which is pre-incubated for a period of time, and the binding of mAb1 to the TMPRSS2 polypeptide can be determined. The biosensor chip can be washed in buffer between each step of the experiment. Real-time binding responses can be monitored over the course of the experiment, and the binding responses at the end of each step can be recorded.

[0069] For example, in one embodiment of the invention, the competitive assay is carried out at 25° C. and a pH of about 7, eg, 7.4, in the presence of buffers, salts, detergents and non-specific proteins (eg, bovine serum albumin).

[0070] Typically, an antibody or antigen-binding fragment of the invention, modified in any way, retains the ability to specifically bind to TMPRSS2, e.g., retains at least 10% of its TMPRSS2 binding activity (compared to the parent antibody) when that activity is expressed on a molar basis. Preferably, an antibody or antigen-binding fragment of the invention retains at least 20%, 50%, 70%, 80%, 90%, 95% or 100% or more of the TMPRSS2 binding affinity of the parent antibody. It is also contemplated that an antibody or antigen-binding fragment of the invention may include conservative or non-conservative amino acid substitutions (referred to as "conservative variants" or "functionally conserved variants" of an antibody) that do not substantially alter its biological activity.

[0071] Immunoglobulin chains (e.g., H1H7017N, V H , V L A "variant" of a polypeptide, such as a polypeptide sequence (e.g., SEQ ID NO: 2, 4, 17, 18 or 19) is a polypeptide that contains an amino acid sequence that is at least about 70-99.9% (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 to a reference amino acid sequence described herein (e.g., SEQ ID NO: 2, 4, 17, 18 or 19); when the comparison is performed by the BLAST algorithm, the parameters of the algorithm are selected to give maximum matching between each sequence over the entire length of the respective reference sequences (e.g., expectation threshold: 10; word size: 3; maximum match within query: 0; BLOSUM 62 matrix; gap cost: presence 11, extension 1; conditional score matrix adjustment).

[0072] A "variant" of a polynucleotide refers to a polynucleotide that is at least about 70-99.9% (e.g., 70, 72, 74, 76, 78, 79, 80, 81, 82, 83, 84, 85, 86, 87, 88, 89, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, 100, 101, 102, 103, 104, 105, 106, 107, 108, 109, 110, 111, 112, 113, 114, 115, 116, 117, 118, 119, 120, 121, 122, 123, 124, 125, 126, 127, 128, 129, 130, 131, 132, 133, 134, 135, 136, 137, 138, 139, 140, 141, 142, 143, 144, 145, 146, 147, 148, 149, 150, 151, 152, 153, 154, 155, 156, 157, 158, 159, 160, 161, 162, 163, 164, 165, 166, 167, 168, 169, 170, 171, 172, 173, 17 9, 96, 97, 98, 99, 99.5, 99.9%); when the comparison is performed using the BLAST algorithm, the parameters of the algorithm are selected to give maximum matching between each sequence over the entire length of each reference sequence (e.g., expectation threshold: 10; word size: 28; maximum match within query: 0; match / mismatch score: 1, -2; gap cost: linear).

[0073] Anti-TMPRSS2 antigen binding proteins, e.g., antibodies and antigen binding fragments of the invention, in one embodiment 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, 17 or 19; 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:4 or 18.

[0074] Further, variant 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, e.g., missense mutations (e.g., conservative substitutions), nonsense mutations, deletions, or insertions. For example, the invention includes antigen binding proteins comprising immunoglobulin light chain variants comprising the amino acid sequence set forth in SEQ ID NO: 4 or 18, but having one or more of such mutations, and / or immunoglobulin heavy chain variants comprising the amino acid sequence set forth in SEQ ID NO: 2, 17, or 19, but having one or more of such mutations. In one embodiment of the invention, the variant anti-TMPRSS2 antigen binding protein comprises an immunoglobulin light chain variant 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 an immunoglobulin heavy chain variant 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)).

[0075] The present invention further provides variant anti-TMPRSS2 antigen binding proteins, e.g., antibodies or antigen-binding fragments thereof, that comprise 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) and have, e.g., at least 70%, 75%, 80%, 85%, 90%, 95%, 98%, 99% or 99.9% sequence identity or similarity to SEQ ID NOs: 12, 14, 16, 6, 8 and / or 10.

[0076] Embodiments of the present invention also include mutant antigen binding proteins, such as immunoglobulin V H and V L or HC and LC, and the corresponding VH , V L , HC or LC, wherein the CDR-L1, CDR-L2, CDR-L3, CDR-H1, CDR-H2 and CDR-H3 of such immunoglobulins are not mutant and comprise the amino acid sequences set forth in SEQ ID NOs: 12, 14, 16, 6, 8 and 10, respectively. Thus, in such embodiments, the CDRs within the mutant antigen binding proteins are not themselves mutant.

[0077] Conservatively modified variant anti-TMPRSS2 antibodies and antigen-binding fragments thereof are also disclosed herein. It is part of the description. "Conservatively modified variants" or "conservative substitutions" refer to variants in which there are one or more substitutions of amino acids in a polypeptide with other amino acids having similar properties (e.g., charge, side chain size, hydrophobicity / hydrophilicity, backbone structure and rigidity, etc.). Such changes can be made frequently without significantly destroying the biological activity of the antibody or fragment. Those skilled in the art will generally recognize that single amino acid substitutions in non-essential regions of a polypeptide do not substantially alter biological activity (see, for example, 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 destroy biological activity significantly.

[0078] Examples of groups of amino acids with side chains having 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 having a positive value in the PAM250 log-likelihood matrix disclosed in Gonnet et al. (1992) Science 256:1443-45.

[0079] Function-conservative variants of anti-TMPRSS2 antibodies and antigen-binding fragments thereof are also part of the present invention. Any of the variants of anti-TMPRSS2 antibodies and antigen-binding fragments thereof (discussed herein) may be "function-conservative variants". Such function-conservative variants are also, in some cases, 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 one embodiment of the present invention, the function-conservative variant 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: Inhibits the growth of influenza viruses (e.g., A / Puerto Rico / 08 / 1934(H1N1)) in TMPRSS2-expressing cells (e.g., Calu-3 cells); EC of 440 pM or 1.06 nM, respectively 50 values, binding to the surface of TMPRSS-expressing cells (e.g., MDCK / Tet-on); · No significant binding to MDCK / Tet-on cells that do not express TMPRSS2; Approximately 2.81 x 10 at 25°C -9 K of M D binds to human TMPRSS2; At 37°C, it is about 9.31 x 10 -9 K of M D binds to human TMPRSS2; Approximately 5.60 x 10 at 25°C -8 K of M D binds to cynomolgus TMPRSS2 at approximately 1.40 × 10 at 37 °C; -7 K of M D binds to cynomolgus TMPRSS2 in vitro; limits the spread of influenza virus infection (e.g., H1_PR34; H1_CA09; H1_Bris; H9N2 or H3N2 influenza viruses) of cells, e.g., Calu-3, in vitro; and / or optionally, when combined with an anti-HA antibody, protecting mice engineered to express the human TMPRSS2 protein from death caused by, e.g., influenza virus infection, e.g., H1N1 or H3N2 (e.g., infecting the mice with a normally lethal dose of the virus).

[0080] The present invention includes mice engineered to express human TMPRSS2 proteins, including anti-TMPRSS2 antigen binding proteins (e.g., antibodies or antigen binding fragments) such as H1H7017N and H4H7017N in vivo in the mouse. See International Patent Application Publication No. WO2017 / 151453.

[0081] A "neutralizing" or "antagonist" anti-TMPRSS2 antigen binding protein, e.g., antibody or antigen-binding fragment, refers to a molecule that inhibits the activity of TMPRSS2 to any detectable extent, e.g., inhibits the protease activity of TMPRSS2 of substrates such as HA; Cbz-Gly-Gly-Arg-AMC (Sigma) (wherein Cbz is benzyloxycarbonyl and AMC is 7-amino-4-methylcoumarin); influenza virus HA0; coronavirus S protein; or the precursor TMPRSS2 that is autocatalytically cleaved between Arg255 and Ile256, and / or inhibits influenza virus entry into cells, and / or inhibits influenza virus replication in the subject's organism.

[0082] "H1H7017N" and "H4H7017N" refer to antigen-binding proteins, such as heavy chains or V chains described below. H (or a variant thereof) and the light chain or V L or a 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 a V having its CDRs (CDR-L1 (or a variant thereof), CDR-L2 (or a variant thereof) or CDR-L3 (or a variant thereof) L "Amino acid sequence" refers to antibodies and antigen-binding fragments thereof, including, for example, the immunoglobulin chains, variable regions and / or CDRs comprising the specific amino acid sequences set out below.

[0083] In one embodiment of the invention, "H1H7017N" or "H4H7017N" refers to an antibody or antigen-binding fragment comprising an immunoglobulin heavy chain CDR-H1, CDR-H2, and CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO: 2, 17 or 19, and an immunoglobulin light chain CDR-L1, CDR-L2, and CDR-L3 comprising the amino acid sequence set forth in SEQ ID NO: 4 or 18.

[0084] In one embodiment of the present invention, "H1H7017N" or "H4H7017N" refers to a VH1H7017N variant comprising the amino acid sequence shown in SEQ ID NO:2. H and V comprising the amino acid sequence shown in SEQ ID NO:4. L The present invention refers to an antibody or antigen-binding fragment thereof comprising:

[0085] In one embodiment of the invention, "H1H7017N" refers to an antibody or antigen-binding fragment comprising: a heavy chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO: 17; and a light chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO: 18.

[0086] In one embodiment of the invention, "H4H7017N" refers to an antibody or antigen-binding fragment comprising a heavy chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO: 19; and a light chain immunoglobulin comprising the amino acid sequence set forth in SEQ ID NO: 18. The term "H4H7017N" also refers to V H is fused to wild type IgG4, for example where residue 108 is S.

[0087] Anti-TMS22 antibodies or antigen-binding fragments H1H7017N and H4H7017N H1H7017N and H4H7017N heavy chain variable regions (DNA) CAGGTGCAGCTGGTGGAGTCTGGGGGAGGCGTGGTCCAGCCTGGGAGGTCCCTGAGACTCTCCTGTGCAGCGTCTGGATTCACCTTCAGTTCCTATGGCATGCACTGGGTCCGCCAGTCTCCAGGCAAGGGGCTCGAGTGGGTGGCAGTTATATGGAATG ATGGAAGTTATGTATACTATGCAGACTCCGTGAAGGGCCGATTCACCATCTCCAGAGACATTTCCAAGAACACGCTGTTTCTGCAAATGAACAGCCTGAGAGCCGAGGACACGGCTGTGTATTACTGTGCGAGAGAGGGGGAGTGGGTACTTTACTACTTTGACTACTGGGGCCAGGGAACCCTGGTCACCGTCTCCTCA (SEQ ID NO: 1)

[0088] H1H7017N and H4H7017N heavy chain variable regions (polypeptides) QVQLVESGGGVVQPGRSLRLSCAAS GFTFSSYG MHWVRQSPGKGLEWVAV IWNDGSYV YYADSVKGRFTISRDISKNTLFLQMNSLRAEDTAVYYC AREGEWVLYYFDY WGQGTLVTVSS (SEQ ID NO:2)

[0089] H1H7017N and H4H7017N light chain variable regions (DNA) GACATCCAGATGACCCAGTCTCCTTCCACCCTGTCTGCATCTGTTGGAGACAGAGTCACCATCACTTGCCGGGCCAGTCAGAGTATTAGTAGCTGGTTGGCCTGGTATCAGCAGAAACCAGGGAAAGCCCCTAAACTCCTGATCTATAAGGCGTCTACTT TAGAAAGTGGGGTCCCATCAAGGTTCAGCGGCAGTGGATCTGGGACAGAATTCACTCTCACCATCAGCAGCCTGCAGCCTGATGATTTTGCAACTTATTACTGCCAACAGTATAATAGTTATTCGTACACTTTTGGCCAGGGGACCAAGCTGGAGATCAAA (SEQ ID NO:3)

[0090] H1H7017N and H4H7017N light chain variable regions (polypeptides) DIQMTQSPSTLSASVGDRVTITCRAS QSISSW LAWYQQKPGKAPKLLIY KAS TLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYC QQYNSYSYT FGQGTKLEIK (SEQ ID NO:4)

[0091] H1H7017N and H4H7017N CDR-H1(DNA) GGA TTC ACC TTC AGT TCC TAT GGC (SEQ ID NO:5)

[0092] H1H7017N and H4H7017N CDR-H1 (polypeptide) GFTFSSYG SEQ ID NO:6 (or a variant thereof having 1, 2, 3 or 4 point mutations and / or point deletions)

[0093] H1H7017N and H4H7017N CDR-H2(DNA) ATA TGG AAT GAT GGA AGT TAT GTA (SEQ ID NO:7)

[0094] H1H7017N and H4H7017N CDR-H2 (polypeptide) IWNDGSYV SEQ ID NO:8 (or a variant thereof having 1, 2, 3 or 4 point mutations and / or point deletions)

[0095] H1H7017N and H4H7017N CDR-H3(DNA) GCG AGA GAG GGG GAG TGG GTA CTT TAC TAC TTT GAC TAC (SEQ ID NO:9)

[0096] H1H7017N and H4H7017N CDR-H3 (polypeptide) AREGEWVLYYFDY SEQ ID NO: 10 (or a variant thereof having 1, 2, 3 or 4 point mutations and / or point deletions)

[0097] H1H7017N and H4H7017N CDR-L1(DNA) CAG AGT ATT AGT AGC TGG (SEQ ID NO:11)

[0098] H1H7017N and H4H7017N CDR-L1 (polypeptide) QSISSW SEQ ID NO: 12 (or a variant thereof having 1, 2, 3 or 4 point mutations and / or point deletions)

[0099] H1H7017N and H4H7017N CDR-L2(DNA) AAG GCG TCT (SEQ ID NO:13)

[0100] H1H7017N and H4H7017N CDR-L2 (polypeptide) KAS SEQ ID NO: 14 (or variants thereof having point mutations and / or point deletions)

[0101] H1H7017N and H4H7017N CDR-L3(DNA) CAA CAG TAT AAT AGT TAT TCG TAC ACT (SEQ ID NO:15)

[0102] H1H7017N and H4H7017N CDR-L3 (polypeptide) QQYNSYSYT SEQ ID NO: 16 (or a variant thereof having 1, 2, 3 or 4 point mutations and / or point deletions)

[0103] H1H7017N Full-length heavy chain - human IgG1 QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQSPGKGLEWVAVIWNDGSYVYYADSVKGRFTISRDISKNTLFLQMNSLRAEDTAVYYCAREGEWVLYYFDYWGQ GTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDK THTCPPCPAPELLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEK TISKAKGQPREPQVYTLPPSRDELTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK (SEQ ID NO:17)

[0104] Full length light chain - human kappa DIQMTQSPSTLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYKASTLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQYNSYSYTFGQGTKLEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO:18)

[0105] H4H7017N Full-length heavy chain - human IgG4 (S108P) QVQLVESGGGVVQPGRSLRLSCAASGFTFSSYGMHWVRQSPGKGLEWVAVIWNDGSYVYYADSVKGRFTISRDISKNTLFLQMNSLRAEDTAVYYCAREGEWVLYYFDYWG QGTLVTVSSASTKGPSVFPLAPCSRSTSESTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTKTYTCNVDHKPSNTKVDKRVESKYG PPCPPCPAPEFLGGPSVFLFPPKPKDTLMISRTPEVTCVVVDVSQEDPEVQFNWYVDGVEVHNAKTKPREEQFNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKGLPSSIEKT ISKAKGQPREPQVYTLPSQEEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSRLTVDKSRWQEGNVFSCSVMHEALHNHYTQKSLSLSLGK (SEQ ID NO:19)

[0106] Full length light chain - human kappa DIQMTQSPSTLSASVGDRVTITCRASQSISSWLAWYQQKPGKAPKLLIYKASTLESGVPSRFSGSGSGTEFTLTISSLQPDDFATYYCQQYNSYSYTFGQGTKLEIK RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC (SEQ ID NO:18)

[0107] The antibodies and antigen-binding fragments of the invention include immunoglobulin chains that contain 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 that specifically bind to TMPRSS2 that contain the heavy and / or light chain amino acid sequences (e.g., CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2 and / or CDR-L3) described herein, 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 pyroglutamic acid (PyroE), and / or the C-terminal lysine is deleted.

[0108] The invention provides a container (eg, a plastic or glass vial, such as one having a cap or a chromatography column, hollow bore needle or syringe cylinder) that comprises an anti-TMPRSS2 antigen binding protein of the invention, such as H1H7017N or H4H7017N.

[0109] The present invention also relates to a TMPRSS2, e.g., H4H7017N or H1H7017N, An injection device is provided that includes one or more antigen binding proteins (e.g., antibodies or antigen-binding fragments) that specifically bind, or a pharmaceutical composition thereof. The injection device is packaged in a kit. An injection device is a device that introduces a substance into a subject's living body via a parenteral route, e.g., intramuscularly, subcutaneously, or intravenously. For example, the injection device can be a syringe (e.g., pre-filled with a pharmaceutical composition, such as an autoinjector), that includes, for example, a cylinder or barrel (e.g., including an antibody or fragment or pharmaceutical composition thereof) for holding the fluid to be injected, a needle for piercing the skin and / or blood vessel to inject the fluid; and a plunger for pushing the fluid out of the cylinder and injecting it through the needle hole. In one embodiment of the present invention, the injection device includes an antigen binding protein from the combination of the present invention, e.g., an antibody or antigen-binding fragment thereof, or a pharmaceutical composition thereof, is an intravenous (IV) injection device. Such a device can include the antigen binding protein or pharmaceutical composition thereof in a cannula or trocar / needle, and can be attached to a tube that can be attached to a bag or reservoir for holding a fluid (e.g., saline) introduced into the living body of the subject via the cannula or trocar / needle. In one embodiment of the present invention, the antibody or fragment or pharmaceutical composition thereof is introduced into the device when a trocar and cannula are inserted into a vein of a subject and the trocar is removed from the inserted cannula. The IV device can be inserted, for example, into a peripheral vein (e.g., hand or arm); into the superior or inferior vena cava, or into the right atrium of the heart (e.g., central IV); or into the subclavian, internal jugular, or femoral vein, and advanced toward the heart until it reaches, for example, the superior vena cava or the right atrium (e.g., central venous line). In one embodiment of the present invention, the infusion device is an automatic injector; a jet injector, or an external infusion pump. A jet injector uses a high-pressure narrow jet of liquid that penetrates the epidermis to introduce the antibody or fragment or pharmaceutical composition thereof into the subject's living body. An external infusion pump is a medical device that delivers the antibody or fragment or pharmaceutical composition thereof into the subject's living body in a controlled amount. An external infusion pump is electrically or mechanically powered.Different pumps operate in different ways, for example, syringe pumps hold fluid in a syringe reservoir and a moveable piston controls the fluid delivery, and elastomeric pumps hold fluid in a stretchable balloon reservoir and pressure from the elastic walls of the balloon drives the fluid delivery. In peristaltic pumps, a set of rollers pinch down a length of flexible tubing that pushes the fluid forward. In multi-channel pumps, fluid is delivered from multiple reservoirs at multiple speeds.

[0110] The invention further provides a method of administering an anti-TMPRSS2 antigen binding protein of the invention, e.g., H4H7017N or H1H7017N, comprising introducing the antigen binding protein into a subject's (e.g., human) organism. For example, the method comprises puncturing the subject's organism with a needle of a syringe and injecting the antigen binding protein into the subject's organism, e.g., into a vein, artery, tumor, muscle tissue, or subcutaneous tissue of the subject.

[0111] Preparation of human antibodies Methods for raising human antibodies in transgenic mice are known in the art. 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 raised using immunogens that include any one of the following: In certain embodiments of the present invention, the antibodies of the present invention are obtained from mice immunized with full-length, native TMPRSS2, or live attenuated or inactivated viruses, or DNA encoding the protein or fragments thereof. Alternatively, TMPRSS2 protein or fragments thereof are produced and modified using standard biochemical techniques and used as immunogens. In one embodiment of the present invention, the immunogen is a recombinantly produced TMPRSS2 protein or fragments thereof. In certain 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 is cultured in E. coli or any other eukaryotic cell. Alternatively, it can be a recombinant TMPRSS2 polypeptide expressed in mammalian cells, such as Chinese hamster ovary (CHO) cells.

[0112] Using VELOCIMMUNE® technology (see, e.g., U.S. Pat. No. 6,596,541, Regeneron Pharmaceuticals, VELOCIMMUNE®) or any other known method for generating monoclonal antibodies, high affinity chimeric antibodies against TMPRSS2 with human variable regions and mouse constant regions can be initially isolated. VELOCIMMUNE® technology involves the creation of transgenic mice with genomes that include human heavy and light chain variable regions operably linked to endogenous mouse constant region loci, such that the mice respond to antigenic stimulation to generate antibodies that include human variable regions and mouse constant regions. DNA encoding the heavy and light chain variable regions of the antibody is isolated and operably linked to DNA encoding human heavy and light chain constant regions. The DNA is then expressed in cells capable of expressing fully human antibodies.

[0113] Typically, VELOCIMMUNE® mice are loaded with an antigen of interest, and lymphocytes (such as B cells) are collected from the mice that express antibodies. Lymphoid cells can be fused with myeloma cell lines to produce immortalized hybridoma cell lines, which are screened and selected to identify hybridoma cell lines that produce antibodies specific to the antigen of interest. DNA encoding the variable regions of the heavy and light chains can be isolated and linked to the desired isotype constant regions of the heavy and light chains. Such antibody proteins are produced in cells such as CHO cells. Alternatively, DNA encoding the antigen-specific chimeric antibodies or the variable domains of the light and heavy chains can be directly isolated from antigen-specific lymphocytes.

[0114] 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 the desired properties, including affinity, selectivity, epitope, etc. The mouse constant region is replaced with the desired human constant region to generate a fully human antibody of the present invention, such as a wild-type or modified IgG1 or IgG4. The constant region selected can vary depending on the particular use, but the high affinity antigen binding and target specificity properties reside in the variable region.

[0115] Anti-TMPRSS2 Antibodies Containing Fc Variants According to certain embodiments of the invention, there are provided anti-TMPRSS2 antigen binding proteins, e.g., antibodies or antigen binding fragments, that comprise an Fc domain that includes one or more mutations that enhance or decrease antibody binding to the FcRn receptor at acidic pH, e.g., as compared to neutral pH. For example, the invention provides an Fc domain that includes a C H 2 or C H The present invention includes anti-TMPRSS2 antibodies that contain mutations in the 3 region, where the mutation(s) increase the affinity of the Fc domain for FcRn in acidic environments (e.g., in endosomes where the pH ranges from about 5.5 to about 6.0). Such mutations can result in an increased 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 modifications 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 modifications 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 or 308P). In yet another embodiment, the modifications include a 265A (e.g., D265A) and / or a 297A (e.g., N297A) modification.

[0116] For example, the present invention provides a method for the preparation of a nucleotide sequence comprising the steps of: 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 43 The present invention includes anti-TMPRS2 antigen binding proteins, such as antibodies or antigen-binding fragments, that include an Fc domain that includes one or more pairs or groups of mutations selected from the group consisting of 4H (e.g., D376V and N434H); 307A, 380A and 434A (e.g., T307A, E380A and N434A); and 433K and 434F (e.g., H433K and N434F).

[0117] The V domains described herein include any possible combination of the above-mentioned Fc domain mutations. H and / or V L Anti-TMPRSS antigen binding proteins, e.g., antibodies and antigen-binding fragments thereof, are contemplated within the scope of the present invention, including:

[0118] The present invention also relates to a method for the preparation of a compound according to the present invention. H and chimeric heavy chain constant (C H ) region, and H The region is a C HFor example, the antibodies of the present invention may comprise segments derived from C regions derived from human IgG1, human IgG2, or human IgG4 molecules. H C2 domain, derived from a human IgG1, human IgG2 or human IgG4 molecule H Chimeric C combined with part or all of the 3 domains H According to a particular embodiment, the antibody of the present invention may comprise a chimeric C region having a chimeric hinge region. H For example, the chimeric hinge may comprise an "upper hinge" amino acid sequence (amino acid residues at positions 216 to 227 according to EU numbering) derived from a human IgG1, human IgG2 or human IgG4 hinge region combined with a "lower hinge" sequence (amino acid residues at positions 228 to 236 according to EU numbering) derived from a human IgG1, human IgG2 or human IgG4 hinge region. According to certain embodiments, the chimeric hinge region comprises amino acid residues derived from the upper hinge of human IgG1 or human IgG4 and amino acid residues derived from the lower hinge of human IgG2. The chimeric hinges described herein may be H Antibodies comprising the region can, in certain embodiments, exhibit modified Fc effector functions without adversely affecting the therapeutic or pharmacokinetic properties of the antibody (see, e.g., International Publication No. WO2014 / 022540).

[0119] 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 drug for treating influenza virus infection. In one embodiment of the present 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, chemically or biologically linked to a radioactive substance, a cytokine, an interferon, a target or reporter moiety, an enzyme, a peptide or protein, or a therapeutic agent. The antigen-binding protein is capable of binding to its target (TMPRSS2) as long as it is capable of binding to the target. At any position along the molecule, it is linked to a radioactive substance, a cytokine, an interferon, a target or reporter moiety, an enzyme, a peptide, or a therapeutic agent. Examples of immunoconjugates include antibody-drug conjugates and antibody-toxin fusion proteins. In one embodiment of the present invention, the drug can be a second, different antibody that specifically binds to TMPRSS2. The type of therapeutic moiety conjugated to the anti-TMPRSS2 antigen binding protein (e.g., antibody or fragment) takes into account the condition to be treated and the desired therapeutic effect to be achieved. See, e.g., Arnon et al., "Monoclonal Antibodies For Immunotargeting Of Drugs In Cancer Therapy," in Monoclonal Antibodies And Cancer Therapy, Reisfeld et al. (eds.), pp. 243-56 (Alan R. Liss, Inc., 1985); Hellstrom et al., "Antibodies For Drug Delivery," in 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," in 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," in Monoclonal Antibodies For Cancer Detection And Therapy, Baldwin et al. (eds.), pp. 303-16 (Academic Press 1985); Thorpe et al., "The Preparation And Cytotoxic Properties Of See, "Antibody-Toxin Conjugates," Immunol. Rev. 62:119-58 (1982).

[0120] multispecific antibodies The present invention includes anti-TMPRSS2 antigen binding proteins, e.g., antibodies and antigen binding fragments thereof, as well as methods of their use and methods of making such antigen binding proteins. The term "anti-TMPRSS2" antigen binding proteins, e.g., antibodies or antigen binding fragments, includes multispecific (e.g., bispecific or biparatopic) molecules that include at least one first antigen binding domain that specifically binds to TMPRSS2 (e.g., antigen binding domain from H1H7017N or H4H7017N), and at least one second antigen binding domain that binds to an antigen or epitope in TMPRSS2 that is different from the first antigen binding domain (e.g., influenza HA, such as antigen binding domain from H1H14611N2, H1H14612N2 or H1H11729P). In one embodiment of the present invention, the first and second epitopes overlap. In another embodiment of the present invention, the first and second epitopes do not overlap. For example, in one 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 heavy and light immunoglobulin chains of H1H7017N or H4H7017N, and a second antigen-binding domain that specifically binds to influenza HA (comprising different light and heavy immunoglobulin chains such as H1H14611N2, H1H14612N2 or H1H11729P).

[0121] "H1H7017N" is a HCDR and LCDR, V H and V L or H1H7017N HC and LC (including variants thereof as described herein).

[0122] "H4H7017N" is a HCDR and LCDR, V H and VL , or H4H7 HC and LC of 017N (including variants thereof as described herein) are multispecific molecules, such as antibodies or antigen-binding fragments.

[0123] In one embodiment of the invention, the antigen binding domain that specifically binds to TMPRSS comprised in the 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:6, CDR-H2 comprising the amino acid sequence set forth in SEQ ID NO:8, and CDR-H3 comprising the amino acid sequence set forth in SEQ ID NO:10; and (ii) a light chain variable domain sequence comprising a CDR-L1 comprising the amino acid sequence set forth in SEQ ID NO: 12, a CDR-L2 comprising the amino acid sequence set forth in SEQ ID NO: 14, and a 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:4; or (3) (i) a heavy chain immunoglobulin sequence comprising the amino acid sequence set forth in SEQ ID NO: 17 or 19, and (ii) a light chain immunoglobulin sequence comprising the amino acid sequence set forth in SEQ ID NO: 18. Includes.

[0124] In one embodiment 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 that are the same or different from the first and / or second antigen-binding domain.

[0125] In one embodiment of the invention, the multispecific molecule comprises, in addition to an antigen binding site that specifically binds TMPRSS2, the following: H1H14611N2; H1H14612N2; H1H11723P; H1H11729P; H1H11820N; H1H11829N; H1H11829N2; H2aM11829N; H2M11830N; H1H11830N2; H1H11903N; H1H14571N; H2a14571N; H1H11704P; H1H11711P; H1H11714P; H1H11717P; H1H11724P; H1H11727P; H1H1 1730P2;H1H11731P2;H1H11734P2;H1H11736P2;H1H11742P2;H1H11744P2; H1H11745P2;H1H11747P2;H1H11748P2;H1H17952B;H1H17953B;H1H17954B; H1H17955B;H1H17956B;H1H17957B;H1H17958B;H1H17959B;H1H17960B;H1 H17961B;H1H17962B;H1H17963B;H1H17964B;H1H17965B;H1H17966B;H1H17 967B;H1H17968B;H1H17969B;H1H17970B;H1H17971B;H1H17972B;H1H1797 3B;H1H17974B;H1H17975B;H1H17976B;H1H17977B;H1H17978B;H1H17979B; H1H17980B;H1H17981B;H1H17982B;H1H17983B;H1H17984B;H1H17985B;H1 H17986B;H1H17987B;H1H17988B;H1H17989B;H1H17990B;H1H17991B;H1H17 992B;H1H17993B;H1H17994B;H1H17995B;H1H17996B;H1H17997B;H1H1799 8B;H1H17999B;H1H18000B;H1H18001B;H1H18002B;H1H18003B;H1H18004B; H1H18005B;H1H18006B;H1H18007B;H1H18008B;H1H18009B;H1H18010B;H1 H18011B;H1H18012B;H1H18013B;H1H18014B;H1H18015B;H1H18016B;H1H18017B;H1H18018B;H1H18019B;H1H18020B;H1H18021B;H1H18022B;H1H18023B;H1H18024B;H1H18025B;H1H18026B;H1H18027B;H1H18028B;H1H18029B;H1H18030B;H1H18031B;H1H18032B;H1H18033B;H1H18034B;H1H18035B;H1H18037B;H1H18038B;H1H18039B;H1H18040B;H1H18041B;H1H18042B;H1H18043B;H1H18044B;H1H18045B;H1H18046B;H1H18047B;H1H18048B;H1H18049B;H1H18051B;H1H18052B;H1H18053B;H1H18054B;H1H18055B;H1H18056B;H1H18057B;H1H18058B;H1H18059B;H1H18060B;H1H18061B;H1H18062B;H1H18063B;H1H18064B;H1H18065B;H1H18066B;H1H18067B;H1H18068B;H1H18069B;H1H18070B;H1H18071B;H1H18072B;H1H18073B;H1H18074B;H1H18075B;H1H18076B;H1H18077B;H1H18078B;H1H18079B;H1H18080B;H1H18081B;H1H18082B;H1H18083B;H1H18084B;H1H18085B;H1H18086B;H1H18087B;H1H18088B;H1H18089B;H1H18090B;H1H18091B;H1H18092B;H1H18093B;H1H18094B;H1H18095B;H1H18096B;H1H18097B;H1H18098B;H1H18099B;H1H18100B;H1H18101B;H1H18102B;H1H18103B;H1H18104B;H1H18105B;H1H18107B;H1H18108B;H1H18109B;H1H18110B;H1H18111B;H1H18112B;H1H18113B;H1H18114B;H1H18115B;H1H18116B;H1H18117B;H1H18118B;H1H18119B;H1H18120B;H1H18121B;H1H18122B;H1H18123B;H1H18124B;H1H18125B;H1H18126B;H1H18127B;H1H18128B;H1H18129B;H1H18130B;H1H18131B;H1H18132B;H1H18133B;H1H18134B;H1H18135B;H1H18136B;H1H18137B;H1H18138B;H1H18139B;H1H18140B;H1H18141B;H1H18142B;H1H18143B;H1H18144B;H1H18145B;H1H18146B;H1H18147B;H1H18148B;H1H18149B;H1H18150B;H1H18151B;H1H18152B;H1H18153B;H1H18154B;H1H18155B;H1H18156B;H1H18157B;H1H18158B;H1H18159B;H1H18160B;H1H18161B;H1H18162B;H1H18163B;H1H18164B;H1H18165B;H1H18166B;H1H18167B;H1H18168B;H1H18169B;H1H18170B;H1H18171B;H1H18172B;H1H18173B;H1H18174B;H1H18175B;H1H18176B;H1H18177B;H1H18178B;H1H18179B;H1H18180B;H1H18181B;H1H18182B;H1H18183B;H1H18184B;H1H18185B;H1H18186B;H1H18187B;H1H18188B;H1H18189B;H1H18190B;H1H18191B;H1H18192B;H1H18193B;H1H18194B;H1H18195B;H1H18196B;H1H18197B;H1H18198B;H1H18199B;H1H18200B;H1H18201B;H1H18202B;H1H18203B;H1H18204B;H1H18205B;H1H18206B;H1H18207B;H1H18208B;H1H18209B;H1H18210B;H1H18211B;H1H18212B;H1H18213B;H1H18214B;H1H18216B;H1H18217B;H1H18218B;H1H18219B;H1H18220B;H1H18; <h2 style=";text-align:left;direction:ltr">221B;H1H18222B;H1H18223B;H1H18224B;H1H18225B;H1H18226B;H1H18227B;H1H18228B;H1H18229B;H1H18230B;H1H18231B;H1H18232B;H1H18233B ;H1H18234B;H1H18235B;H1H18236B;H1H18237B;H1H18238B;H1H18239B;H1H18240B;H1H18241B;H1H18242B;H1H18243B;H1H18244B;H1H18245B;H1H 18246B;H1H18247B;H1H18248B;H1H18249B;H1H18250B;H1H18251B;H1H18252B;H1H18253B;H1H18254B;H1H18255B;H1H18256B;H1H18257B;H1H1825 8B;H1H18259B;H1H18261B;H1H18262B;H1H18263B;H1H18264B;H1H18265B;H1H18266B;H1H18267B;H1H18268B;H1H18269B;H1H18270B;H1H18271B;H1 H18272B;H1H18274B;H1H18275B;H1H18276B;H1H18277B;H1H18278B;H1H18279B;H1H18280B;H1H18281B;H1H18282B;H1H18283B;H1H18284B;H1H182 85B;H1H18286B;H1H18287B;H1H18288B;H1H18289B;H1H18290B;H1H18291 B;H1H18292B;H1H18293B;H1H18294B;H1H18295B;H1H18297B;H1H18298B;H 1H18299B;H1H18300B;H1H18301B;H1H18302B;H1H18303B;H1H18304B;H1H 18305B;H1H18306B;H1H18307B;H1H18308B;H1H18309B;H1H18310B;H1H18 311B;H1H18312B;H1H18313B;H1H18314B;H1H18315B;H1H18316B;H1H18317B;H1H18318B;H1H18319B;H1H18320B;H1H18321B;H1H18322B;H1H18323B;and H1H18335B, which are described in International Patent Application Publication No. WO 2016 / 100807 (e.g., CDR-H, V; H or its heavy chain; and CDR-L, V L or its light chain).

[0126] In one embodiment of the invention, the multispecific molecule contains an antigen binding site that specifically binds to an influenza group II HA protein, e.g., the V H and V L (e.g., SEQ ID NOs: 24 and 28); or a heavy chain immunoglobulin comprising CDR-H1, CDR-H2 and CDR-H3 of H1H14611N2 (e.g., SEQ ID NOs: 25-27); and a light chain immunoglobulin comprising CDR-L1, CDR-L2 and CDR-L3 of H1H14611N2 (e.g., SEQ ID NOs: 29-31).

[0127] In one embodiment of the invention, the multispecific molecule comprises an antigen binding site that specifically binds to an influenza group II HA protein, e.g., the V H and V L (e.g., SEQ ID NOs: 40 and 44); or a heavy chain immunoglobulin comprising CDR-H1, CDR-H2 and CDR-H3 of H1H14612N2 (e.g., SEQ ID NOs: 41-43); and a light chain immunoglobulin comprising CDR-L1, CDR-L2 and CDR-L3 of H1H14612N2 (e.g., SEQ ID NOs: 45-47).

[0128] In one embodiment of the invention, the multispecific molecule comprises an antigen binding molecule that specifically binds to TMPRSS2. In addition to the binding site, the antigen-binding site that specifically binds to the influenza group I HA protein, e.g., the V H and V L (e.g., SEQ ID NOs: 32 and 36); or a heavy chain immunoglobulin comprising CDR-H1, CDR-H2 and CDR-H3 of H1H11729P (e.g., SEQ ID NOs: 33-35); and a light chain immunoglobulin comprising CDR-L1, CDR-L2 and CDR-L3 of H1H11729P (e.g., SEQ ID NOs: 37-39).

[0129] In one embodiment of the invention, the bispecific antigen-binding fragment comprises a first scFv (e.g., the V of H1H7017N or H4H7017N) that has binding specificity for a first epitope (e.g., TMPRSS2). H and V L and a second scFv having binding specificity for a second, different epitope (e.g., the V of an anti-influenza HA antibody). H and V L For example, in one embodiment of the invention, the first and second scFvs include a linker, e.g., a peptide linker (e.g., a GS linker, e.g., (GGGGS) n (SEQ ID NO:48), 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 comprising the heavy and light chain CDRs of H1H7017N or H4H7017N and of another antibody that binds to a different epitope.

[0130] 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., H1H7017N or H4H7017N) to a subject (e.g., a human) in need of treatment or prevention.

[0131] Influenza virus infection can be treated or prevented in a subject by administering to the subject an anti-TMPRSS2 antigen binding protein of the present invention. Influenza viruses are classified into types A, B and C based on their core proteins. Subtypes of influenza A viruses are determined by envelope glycoproteins that have either hemagglutinin (HA) or neuraminidase (NA) activity. There are several HA subtypes of influenza A viruses (e.g., HA1, HA2, HA3, HA4, HA5, HA6, HA7, HA8, HA9, HA10, HA11, HA12, HA13, HA14, HA15, HA16, HA17 or HA18--these subtypes are designated as H1, H2, H3, etc.), and NA subtypes (e.g., NA1, NA2, NA3, NA4, NA5, NA6, NA7, NA8, NA9, NA10 or NA11--these subtypes are designated as N1, N2, N3, etc.) that 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 subtypes H1, H2 or H3, and N1 or N2. The present invention includes methods for treating or preventing infection with influenza virus subtypes discussed herein. In one embodiment of the present invention, the multispecific antibodies and antigen-binding fragments thereof that bind to TMPRSS2 also bind to HA and / or NA, for example of the subtypes described herein.

[0132] An effective or therapeutically effective dose of an anti-TMPRSS2 antigen binding protein, e.g., an antibody or antigen binding fragment (e.g., H1H7017N or H4H7017N), for treating or preventing a viral infection refers to an amount of the antibody or fragment sufficient to alleviate one or more signs and / or symptoms of infection in a treated subject, whether by inducing regression or disappearance of such signs and / or symptoms, or by inhibiting the progression of such signs and / or symptoms. Dosages may vary depending on the age and size of the subject to which they are administered, the target disease, condition, route of administration, etc. In one embodiment, for example, an effective or therapeutically effective dose of an antibody or antigen-binding fragment thereof of the present invention for treating or preventing a viral infection in an adult human subject is about 0.01 to about 200 mg / kg, e.g., up to about 150 mg / kg. In one embodiment of the present 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). Depending on the severity of the infection, the frequency and duration of treatment can be adjusted. In certain embodiments, an antigen-binding protein of the present invention may be administered in an initial dose, followed by one or more secondary doses. In certain embodiments, the initial dose can be followed by administration of a second or multiple subsequent doses of the antibody or antigen-binding fragment, in an amount that can be about the same as or less than the initial dose, with the subsequent doses being 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.

[0133] As used herein, the term "subject" refers to a mammal (e.g., rat, mouse, cat, dog, cow, sheep, horse, goat, rabbit), preferably a human, that is in need of prevention and / or treatment of a disease or disorder, such as a viral infection or cancer. The subject may have a viral infection, e.g., influenza infection, or may be predisposed to developing an infection. Subjects that are predisposed to developing an infection or at high risk of contracting an infection (e.g., influenza virus) include subjects with immunodeficiency due to autoimmune disease, subjects undergoing immunosuppressive therapy (e.g., after organ transplantation), subjects suffering from human immunodeficiency syndrome (HIV) or acquired immune deficiency 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 disorder. In addition, very young (e.g., 5 years old or younger) or elderly (e.g., 65 years old or older) subjects are at high risk. Additionally, a subject may be at risk of developing a viral infection due to being in close proximity to an outbreak of disease, e.g., the subject may be at risk of developing a viral infection by living in a densely populated city or by being in close proximity to subjects with confirmed or suspected viral infection, or by employment choices (e.g., hospital worker, pharmaceutical researcher, traveler to affected areas, or frequent flyer).

[0134] "Treat" or "treating" means administering an anti-TMPRSS2 antigen binding protein, e.g., an antibody or antigen binding fragment of the invention (e.g., H1H7017N or H4H7017N), to a subject having one or more signs or symptoms of a disease or infection, e.g., a viral infection, for which the antigen binding protein is effective when administered to the subject in an effective or therapeutically effective amount or dose (discussed herein).

[0135] The present invention also encompasses the prophylactic administration of an anti-TMPRSS2 antigen-binding protein, such as an antibody of the present invention or an antigen-binding fragment thereof (e.g., H1H7017N or H4H7017N), to a subject at risk of viral infection to prevent such infection. Passive antibody-based immunoprophylaxis has been proven to be an effective strategy for preventing subjects from viral infection. See, for example, Berry et al., Passive broad-spectrum influenza immunoprophylaxis. Influenza Res Treat., 2014;2014:267594.Epub 2014:267594. 2014 Sep 22;Jianqiang et al., Passive immune neutralization strategies for prevention and control of influenza A infections,Immunotherapy., 2012 February;Vol. 4(No. 2): pp. 175-186;Prabhu et al., Antivir Ther., 2009;Vol. 14(No. 7): pp. 911-21, Prophylactic and therapeutic effica See, cy of a chimeric monoclonal antibody specific for H5 hemagglutinin against lethal H5N1 influenza. "Prevent" or "preventing" means administering an anti-TMPRSS2 antigen binding protein, such as an antibody or antigen binding fragment of the invention (e.g., H1H7017N or H4H7017N), to a subject to inhibit the appearance of a disease or infection (e.g., a viral infection) in the subject's organism, whereby the antigen binding protein is effective when administered to a subject in an effective or therapeutically effective amount or dose (discussed herein).

[0136] In one embodiment of the invention, the sign or symptom of a viral infection in a subject is the survival or proliferation of the virus in the subject's body, as determined, for example, by a viral titer assay (e.g., influenza virus propagation in fetal chicken eggs or influenza virus hemagglutination assay). Other signs and symptoms of viral infection are discussed herein.

[0137] The present invention provides a method for treating or preventing a viral infection (e.g., influenza virus or coronavirus infection) in a subject (e.g., a human) in need thereof, or secondary to a viral infection, such as: · Fever or fever / chills; ·cough; ·Sore throat; Runny or stuffy nose; ·sneeze; · Muscle or body pain; ·headache; · Fatigue (tiredness); ·vomiting; ·diarrhea; Respiratory tract infections; ·Chest discomfort; ·shortness of breath; bronchitis; and / or ·pneumonia, The present invention provides a method for inducing regression or disappearance of, or inhibiting the progression of, at least one sign or symptom of a viral infection, such as a pulmonary edema or bronchitis.

[0138] The 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 TMPRSS2:ERG fusions), 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., H1H7017N or H4H7017N) to the subject, e.g., by injecting the protein into the subject's body. In one embodiment 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 one embodiment of the invention, the cancer is a tumor, the cells of which express TMPRSS2 or a variant thereof.

[0139] Combinations and Pharmaceutical Compositions Anti-TMPRSS2 antigen-binding proteins, such as antibodies and antigen-binding fragments thereof (e.g. To prepare a pharmaceutical composition of the antigen binding protein of the present invention (H1H7017N, H1H7017N or H4H7017N), the antigen binding protein is mixed with a pharma- ceutically acceptable carrier or excipient. See, e.g., Remington's Pharmaceutical Sciences and US Pharmacopeia: 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 See 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 Dekker, NY; Weiner and Kotkoskie (2000) Excipient Toxicity and Safety, Marcel Dekker, Inc., New York, NY. In one embodiment of the present invention, the pharmaceutical composition is sterile. Such compositions are part of the present invention.

[0140] The scope of the invention includes a dried, e.g., lyophilized, composition comprising an anti-TMPRSS2 antigen binding protein, e.g., an antibody or antigen-binding fragment thereof (e.g., H1H7017N or H4H7017N), or a pharmaceutical composition thereof comprising a pharma- ceutically acceptable carrier but substantially devoid of water.

[0141] In a further embodiment of the invention, an additional therapeutic agent administered to a subject in conjunction with an anti-TMPRSS2 antigen binding protein, e.g., an antibody or antigen-binding fragment thereof (e.g., H1H7017N or H4H7017N), as disclosed herein, is provided by a Physicians' Subjects will be administered the drug according to the Desk Reference 2003 (Thomson Healthcare; 57th Edition (Nov. 1, 2002)).

[0142] The mode of administration can be varied, including oral, rectal, transmucosal, intestinal, parenteral, intramuscular, subcutaneous, intradermal, intramedullary, intrathecal, direct intracerebroventricular, intravenous, intraperitoneal, intranasal, intraocular, inhalation, injection, topical, dermal, transdermal or intraarterial.

[0143] The present invention provides a method for administering an anti-TMPRSS2 antigen-binding protein, e.g., an antibody or antigen-binding fragment thereof (e.g., H1H7017N or H4H7017N), comprising introducing the protein into a subject's body. For example, the method comprises puncturing the subject's body with a needle of a syringe and injecting the antigen-binding protein into the subject's body, e.g., into a vein, artery, tumor, muscle tissue, or subcutaneous tissue of the subject.

[0144] The present invention provides anti-TMPRSS2 antigen binding proteins, e.g., antibodies or antigen-binding fragments thereof (e.g., H1H70N or H4H7017N), polypeptides (e.g., the HC, LC, V of H1H7017N or H4H7017N), and polypeptides (e.g., the HC, LC, V of H1H7017N or H4H7017N). H Or V L ), or a container (e.g., a plastic or glass vial, e.g., having a cap or a chromatography column, hollow bore needle or syringe cylinder) containing any of the polynucleotides or vectors described herein, or a pharmaceutical composition thereof comprising a pharma- ceutically acceptable carrier.

[0145] In one embodiment of the present invention, the anti-TMPRSS2 antigen-binding protein, such as an antibody or antigen-binding fragment thereof (e.g., H1H7017N or H4H7017N) of the present invention, is associated with one or more additional therapeutic agents. For example, in one embodiment of the present invention, the additional therapeutic agent is an antiviral drug and / or a vaccine. As used herein, the term "antiviral drug" refers to any anti-infective drug or treatment used to treat, prevent, or ameliorate a viral infection in a subject. The term "antiviral drug" includes, but is not limited to, cationic steroid antibacterial drugs, leupeptin, aprotinin, amantadine, rimantadine, oseltamivir, zanamivir, ribavirin, or interferon-alpha 2b. Methods of treating or preventing a viral (e.g., influenza) infection in a subject in need of said treatment or prevention by administering H1H7017N or H4H7017N with an additional therapeutic agent are part of the present invention.

[0146] For example, in one embodiment of the invention, the additional therapeutic agent is a vaccine, e.g., an influenza vaccine. In one embodiment of the invention, the vaccine is an inactivated / killed viral vaccine, a live attenuated viral vaccine, or a viral subunit vaccine.

[0147] For example, in one embodiment of the invention, the additional therapeutic agent is: [ka]

[0148] [ka]

[0149] [ka]

[0150] [ka]

[0151] [ka] See Shen et al., Biochimie 142:1-10 (2017).

[0152] In one embodiment 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 one embodiment of the invention, the anti-HA antibody is selected from the group consisting of H1H14611N2;H1H14612N2;H1H11723P;H1H11729P;H1H11820N;H1H11829N;H1H11829N2;H2aM11829N;H2M11830N;H1H11830N2;H1H11903N;H1H14571N;H2a14571N;H1H11704P;H1H11711P;H1H1171 4P;H1H11717P;H1H11724P;H1H11727P;H1H11730P2;H1H11731P2;H1H11734P2;H1H11736P2;H1H11742P2;H1H1 1744P2;H1H11745P2;H1H11747P2;H1H11748P2;H1H17952B;H1H17953B;H1H17954B;H1H17955B;H1H17956B;H1H 17957B;H1H17958B;H1H17959B;H1H17960B;H1H17961B;H1H17962B;H1H17963B;H1H17964B;H1H17965B;H1H17966B;H1H17967B;H1H17968B;H1H17969B;H1H17970B;H1H17971B;H1H17972B;H1H17973B;H1H17974B;H1H17975B;H1H17976B;H1H17977B;H1H17978B;H1H17979B;H1H17980B;H1H17981B;H1H17982B;H1H17983B;H1H17984B;H1H17985B;H1H17986B;H1H17987B;H1H17988B;H1H17989B;H1H17990B;H1H17991B;H1H17992B;H1H17993B;H1H17994B;H1H17995B;H1H17996B;H1H17997B;H1H17998B;H1H17999B;H1H18000B;H1H18001B;H1H18002B;H1H18003B;H1H18004B;H1H18005B;H1H18006B;H1H18007B;H1H18008B;H1H18009B;H1H18010B;H1H18011B;H1H18012B;H1H18013B;H1H18014B;H1H18015B;H1H18016B;H1H18017B;H1H18018B;H1H18019B;H1H18020B;H1H18021B;H1H18022B;H1H18023B;H1H18024B;H1H18025B;H1H18026B;H1H18027B;H1H18028B;H1H18029B;H1H18030B;H1H18031B;H1H18032B;H1H18033B;H1H18034B;H1H18035B;H1H18037B;H1H18038B;H1H18039B;H1H18040B;H1H18041B;H1H18042B;H1H18043B;H1H18044B;H1H18045B;H1H18046B;H1H18047B;H1H18048B;H1H18049B;H1H18051B;H1H18052B;H1H18053B;H1H18054B;H1H18055B;H1H18056B;H1H18057B;H1H18058B;H1H18059B;H1H18060B;H1H18061B;H1H18062B;H1H18063B;H1H18064B;H1H18065B;H1H18066B;H1H18067B;H1H18068B;H1H18069B;H1H18070B;H1H18071B;H1H18072B;H1H18073B;H1H18074B;H1H18075B;H1H18076B;H1H18077B;H1H18078B;H1H18079B;H1H18080B;H1H18081B;H1H18082B;H1H18083B;H1H18084B;H1H18085B;H1H18086B;H1H18087B;H1H18088B;H1H18089B;H1H18090B;H1H18091B;H1H18092B;H1H18093B;H1H18094B;H1H18095B;H1H18096B;H1H18097B;H1H18098B;H1H18099B;H1H18100B;H1H18101B;H1H18102B;H1H18103B;H1H18104B;H1H18105B;H1H18107B;H1H18108B;H1H18109B;H1H18110B;H1H18111B;H1H18112B;H1H18113B;H1H18114B;H1H18115B;H1H18116B;H1H18117B;H1H18118B;H1H18119B;H1H18120B;H1H18121B;H1H18122B;H1H18123B;H1H18124B;H1H18125B;H1H18126B;H1H18127B;H1H18128B;H1H18129B;H1H18130B;H1H18131B;H1H18132B;H1H18133B;H1H18134B;H1H18135B;H1H18136B;H1H18137B;H1H18138B;H1H18139B;H1H18140B;H1H18141B;H1H18142B;H1H18143B;H1H18144B;H1H18145B;H1H18146B;H1H18147B;H1H18148B;H1H18149B;H1H18150B;H1H18151B;H1H18152B;H1H18153B;H1H18154B;H1H18155B;H1H18156B;H1H18157B;H1H18158B;H1H18159B;H1H; <h2 style=";text-align:left;direction:ltr">18160B;H1H18161B;H1H18162B;H1H18163B;H1H18164B;H1H18165B;H1H18 166B;H1H18167B;H1H18168B;H1H18169B;H1H18170B;H1H18171B;H1H1817 2B;H1H18173B;H1H18174B;H1H18175B;H1H18176B;H1H18177B;H1H18178B ;H1H18179B;H1H18180B;H1H18181B;H1H18182B;H1H18183B;H1H18184B;H1 H18185B;H1H18186B;H1H18187B;H1H18188B;H1H18189B;H1H18190B;H1H1 8191B;H1H18192B;H1H18193B;H1H18194B;H1H18195B;H1H18196B;H1H181 97B;H1H18198B;H1H18199B;H1H18200B;H1H18201B;H1H18202B;H1H18203 B;H1H18204B;H1H18205B;H1H18206B;H1H18207B;H1H18208B;H1H18209B;H 1H18210B;H1H18211B;H1H18212B;H1H18213B;H1H18214B;H1H18216B;H1H18217B;H1H18218B;H1H18219B;H1H18220B;H1H18221B;H1H18222B;H1H18 223B;H1H18224B;H1H18225B;H1H18226B;H1H18227B;H1H18228B;H1H18229B;H1H18230B;H1H18231B;H1H18232B;H1H18233B;H1H18234B;H1H18235B; H1H18236B;H1H18237B;H1H18238B;H1H18239B;H1H18240B;H1H18241B;H1H18242B;H1H18243B;H1H18244B;H1H18245B;H1H18246B;H1H18247B;H1H18 248B;H1H18249B;H1H18250B;H1H18251B;H1H18252B;H1H18253B;H1H18254B;H1H18255B;H1H18256B;H1H18257B;H1H18258B;H1H18259B;H1H18261B;H1H18262B;H1H18263B;H1H18264B;H1H18265B;H1H18266B;H1H18267B;H1H18268B;H1H18269B;H1H18270B;H1H18271B;H1H1 8272B;H1H18274B;H1H18275B;H1H18276B;H1H18277B;H1H18278B;H1H18279B;H1H18280B;H1H18281B;H1H18282B;H1H18283B ;H1H18284B;H1H18285B;H1H18286B;H1H18287B;H1H18288B;H1H18289B;H1H18290B;H1H18291B;H1H18292B;H1H18293B;H1H 18294B;H1H18295B;H1H18297B;H1H18298B;H1H18299B;H1H18300B;H1H18301B;H1H18302B;H1H18303B;H1H18304B;H1H18305 B;H1H18306B;H1H18307B;H1H18308B;H1H18309B;H1H18310B;H1H18311B;H1H18312B;H1H18313B;H1H18314B;H1H18315B;H1 H18316B;H1H18317B;H1H18318B;H1H18319B;H1H18320B;H1H18321B;H1H18322B;H1H18323B;H1H18324B;H1H18325B;H1H1832 or H1H18335B; as 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; 6B; H1H18327B; H1H18328B; H1H18329B; H1H18330B; H1H18331B; H1H18332B; H1H18333B; H1H18334B; or H1H18335B; as 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; L or a light chain thereof); and a light chain immunoglobulin comprising CDR-H1, CDR-H2 and CDR-H3 (e.g., V H or the heavy chain thereof).

[0153] In one embodiment of the invention, the additional therapeutic agent is an influenza vaccine, such as H1H14611N2. an antibody or antigen-binding fragment that binds to a HIV-1 group II HA protein; or H and V L or a heavy chain immunoglobulin comprising CDR-H1, CDR-H2 and CDR-H3 of H1H14611N2 (e.g., SEQ ID NOs: 25-27) and a light chain immunoglobulin comprising CDR-L1, CDR-L2 and CDR-L3 of H1H14611N2 (e.g., SEQ ID NOs: 29-31). "H1H14611N2" refers to any anti-Group II HA antibody comprising such a sequence.

[0154] H1H14611N2 Heavy chain variable region EVQLVESGGGLVKPGGSLRLSCAAS GFTFSGFS MNWVRQVPGKGLEWVSS ISTSGNYM YYADSVKGRFTISRDNAKKSFSLQMNSLRAEDSAIYYC ARGGGYNWNLFDY WGQGSLVTVSS (SEQ ID NO: 24) CDR-H1: GFTFSGFS (SEQ ID NO: 25) CDR-H2: ISTSGNYM (SEQ ID NO: 26) CDR-H3: ARGGGYNWNLFDY (SEQ ID NO: 27)

[0155] Light chain variable region EIVLTQSPGTLSLSPGERATLSCRAS QSLNSNY LAWYQQKPGQAPRLLIY GAS SRATGIP DRFSGSGSGTDFTLTITRLESEDFAVYYC QQYGNSPLT FGGGTKVEIK (SEQ ID NO:28) CDR-L1: QSLNSNY (SEQ ID NO: 29) CDR-L2: GAS (SEQ ID NO: 30) CDR-L3: QQYGNSPLT (SEQ ID NO: 31)

[0156] In one 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 a heavy chain immunoglobulin comprising CDR-H1, CDR-H2 and CDR-H3 of H1H14612N2 (e.g., SEQ ID NOs: 41-43) and a light chain immunoglobulin comprising CDR-L1, CDR-L2 and CDR-L3 of H1H14612N2 (e.g., SEQ ID NOs: 45-47). "H1H14612N2" refers to any anti-Group II HA antibody comprising such a sequence.

[0157] H1H14612N2 Heavy chain variable region EVQLVESGGGLVKPGGSLRLSCAAS GFSFSGFS MNWVRQAPGKGLEWVSS ISTSGNYM YY ADSVKGRFTISRDNAKKSFSLQMNSLRAEDSAIYYC ARGGGYNWNLFDY WGQGSLVTVSS (SEQ ID NO: 40) CDR-H1: GFSFSGFS (SEQ ID NO: 41) CDR-H2: ISTSGNYM (SEQ ID NO: 42) CDR-H3: ARGGGYNWNLFDY (SEQ ID NO: 43)

[0158] Light chain variable region EIVLTQSPGTLSLSPGERATLSCRAS QSLNSNY LAWYQQKPGQAPRLLIY GAS SRATGIP DRFSGSGSGADFTLTISRLESEDFAVYYC QQYGNSPLT FGGGTKVEIK (SEQ ID NO:44) CDR-L1: QSLNSNY (SEQ ID NO: 45) CDR-L2: GAS (SEQ ID NO: 46) CDR-L3: QQYGNSPLT (SEQ ID NO: 47)

[0159] In one 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 the V H and V L or an antibody or fragment comprising the CDR of H1H11729P - A heavy chain immunoglobulin comprising H1, CDR-H2 and CDR-H3 (e.g., SEQ ID NOs: 33-35) and a light chain immunoglobulin comprising CDR-L1, CDR-L2 and CDR-L3 of H1H11729P (e.g., SEQ ID NOs: 37-39). "H1H11729P" refers to any anti-Group I HA antibody comprising such a sequence.

[0160] H1H11729P Heavy chain variable region QVQLVQSGAEVKKSGSSVKVSCKAS GGTFSSYA ISWVRQAPGQGLEWMGG IIPIFGTP S.Y. AQKFQDRVTITTDESTSTVYMELSSLRSEDTAVYYC ARQQPVYQYNMDV WGQGTTVTVSS (SEQ ID NO: 32) CDR-H1: GGTFSSYA (SEQ ID NO: 33) CDR-H2: IIPIFGTP (SEQ ID NO: 34) CDR-H3: ARQQPVYQYNMDV (SEQ ID NO: 35)

[0161] Light chain variable region DIQMTQSPSSLSASVGDRVTITCRAS QGIRNN LGWYQQKPLKAPKRLIY AAS SLQSGVPS RFSGSGSGTEFTLTISSLQPEDFATYYC LQYNNYPWT FGQGTKVEIK (SEQ ID NO:36) CDR-L1: QGIRNN (SEQ ID NO: 37) CDR-L2: AAS (SEQ ID NO: 38) CDR-L3: LQYNNYPWT (SEQ ID NO: 39)

[0162] In certain embodiments of the invention, the additional therapeutic agent is not amantadine, rimantadine, oseltamivir, zanamivir, aprotinin, leupeptin, a cationic steroid antibacterial drug, an influenza vaccine (e.g., killed, live, attenuated whole virus or subunit vaccine), and an antibody against influenza virus (e.g., an anti-hemagglutinin antibody).

[0163] The term "in association with" indicates that the components, an anti-TMPRSS2 antigen-binding protein, e.g., an antibody or antigen-binding fragment thereof of the present invention, and another agent, e.g., oseltamivir, can be formulated in a single composition or can be formulated separately into two or more compositions (e.g., a kit), e.g., for simultaneous delivery. Each component can be administered to a subject at a different time than the other components are administered; for example, each administration can be given non-concurrently (e.g., separately or sequentially), spaced apart over a given period of time. Additionally, the separate components can be administered to a subject by the same or different routes (e.g., an anti-TMPRSS2 antibody or antigen-binding fragment thereof).

[0164] kit Additionally, kits are provided that include one or more components including, but not limited to, an anti-TMPRSS2 antigen binding protein, e.g., an antibody or antigen binding fragment as discussed herein (e.g., H1H7017N or H4H7017N), together 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 can be formulated in a pharmaceutical composition, either as a single composition or separately in two or more compositions, e.g., together with a pharma- ceutically acceptable carrier.

[0165] In one embodiment of the invention, the kit comprises an anti-TMPRSS2 antigen binding protein, e.g., an antibody or antigen-binding fragment thereof of the invention (e.g., H1H7017N or H4H7017N), 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).

[0166] In another embodiment, the kit comprises a combination of the invention comprising an anti-TMPRSS2 antigen binding protein, e.g., an antibody or antigen binding fragment of the invention (e.g., H1H7017N or H4H7017N), or a pharmaceutical composition thereof, in a single common container, in combination with one or more additional therapeutic agents, optionally formulated together in a pharmaceutical composition.

[0167] Where the kit includes a pharmaceutical composition for parenteral administration to a subject, the kit can include a device for performing such administration (e.g., an injection device). For example, the kit can include one or more hypodermic needles or other injection devices, as discussed above, that include an anti-TMPRSS2 antigen binding protein, such as an antibody or antigen-binding fragment thereof of the invention (e.g., H1H7017N or H4H7017N).

[0168] The kit may include a package insert containing information about the pharmaceutical composition and dosage form in the kit. Generally, such information will assist patients and physicians in effectively and safely using the enclosed pharmaceutical composition and dosage form. For example, the following information about the combination of the present invention is provided in the package insert: pharmacokinetics, pharmacodynamics, clinical trials, efficacy parameters, symptoms and usage, contraindications, warnings, cautions, side effects, overdosage, appropriate dosage and administration, delivery method, appropriate storage conditions, references, manufacturer / distributor information, and patent information.

[0169] Diagnostic Uses of Antibodies Anti-TMPRSS2 antigen binding proteins, such as antibodies or antigen-binding fragments of the invention (e.g., H1H7017N or H4H7017N), are used to detect and / or measure TMPRSS2 in a sample. An exemplary assay for TMPRSS2 can include, for example, contacting a sample with an anti-TMPRSS2 antigen binding protein of the invention, which is labeled with a detectable label or reporter molecule or used as a capture ligand to selectively isolate TMPRSS2 from a sample. The presence of an anti-TMPRSS2 antigen binding protein complexed with TMPRSS2 indicates the presence of TMPRSS2 in the sample. Alternatively, an unlabeled anti-TMPRSS2 antibody can be used in combination with a secondary antibody that is itself detectably labeled. The detectable label or reporter molecule can be a radioisotope, e.g., 3 H, 14 C. 32 P, 35 S, or 125 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). Thus, the present invention includes a method 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, where the presence of a complex indicates the presence of TMPRSS2.

[0170] The present invention includes a cell-based ELISA method that uses an anti-TMPRSS2 antigen binding protein, such as an antibody of the invention and an antigen-binding fragment thereof (e.g., H1H7017N), to detect the presence of TMPRSS2 on a cell. In one embodiment of the present invention, the method comprises: (i) contacting cells immobilized on a solid surface (e.g., a microplate) and testing 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 cell contains TMPRSS2. For example, the present invention provides a method for detecting TMPRSS2 in a sample. + Such cell-based ELISA methods for identifying cells include:

[0171] The anti-TMPRSS2 antigen binding proteins of the invention (e.g., H1H7017N or H4H7017N) are used in Western blot or immuno-protein blot procedures to detect the presence of TMPRSS2 or a fragment thereof in a sample. Such procedures 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 the 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.

[0172] Such membranes can 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 are transferred (e.g., after electrophoretic separation in the 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 non-specific protein binding sites on the membrane.

[0173] (2) washing the membrane one or more times to remove unbound anti-TMPRSS2 antigen-binding protein and other unbound substances. (3) detecting bound anti-TMPRSS2 antigen-binding protein.

[0174] Detection of bound antigen-binding protein indicates that TMPRSS2 protein is present on the membrane or substrate and in the sample. Detection of bound antigen-binding protein can be performed by binding the antigen-binding protein with a detectably 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., H1H7017N or H4H7017N)) disclosed herein can also be used for immunohistochemistry. Such methods form part of the invention and can be used, for example, in (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 the tissue; Includes.

[0176] If the antigen binding protein itself is detectably labeled, it can be detected directly. Alternatively, the antigen binding protein is allowed to bind a detectably labeled secondary antibody, and the label is then detected. EXAMPLES

[0177] The following examples provide a complete disclosure and description of how to make and use the methods and compositions of the present invention. It is provided to provide an explanation to those skilled in the art and is not intended to limit the scope of what the inventors regard as their invention. Efforts have been made to ensure accuracy of numbers used (e.g., amounts, temperatures, etc.), but some experimental error and deviation should be accounted for. Unless otherwise specified, 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 pressure. EXAMPLES

[0178] Multiple replication in vitro The replication ability of influenza virus A / Puerto Rico / 08 / 1934 (H1N1)-GFP was evaluated in Calu3, A549, MDCK, and HepG2 cells.

[0179] [Table 1]

[0180] Experimental procedure Calu-3 cells (ATCC HTB55), A549 cells (ATCC CCL-185), MDCK cells (IRR FR-58) and HepG2 cells (ATCC HB-8065) ​​were diluted to 40,000 cells / well in 96-well plates in DMEM:F12 medium containing 5% FBS. The next day, cells were cultured in 96-well plates containing A / Puerto Rico / 08 / 1934 (H1N1) carrying a GFP reporter gene in the NS segment (B. Manicassamy et al., "Analysis of in vivo dynamics of influenza virus infection in mice using a GFP reporter virus", Proc Natl Acad Sci US A., 2010 Jun 22;107(25):11531-6) was prepared at MOI (multiplicity of infection) of 0.1 and 0.01 in DMEM:F12 with low IgG BSA after three washes. Virus was incubated on cells for 1 hour at 37°C, after which virus was removed and wells were washed another three times. Infected cell numbers were quantified at 24, 48, 72 and 142 hours post-infection with a CTL-ImmunoSpot® S6 Universal Analyzer (Cellular Technology Limited, Cleveland, OH).

[0181] Summary of Results and Conclusions Calu-3 is an immortalized human airway epithelial cell line that has been shown to allow multiple rounds of human influenza virus replication 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," Journal of Virology., vol. 81, pp. 12439-12449 (2007)). Furthermore, Calu-3 cells have been shown to express both TMPRSS2 and TMPRSS4, at least at the mRNA level, but not TMPRSS11D (HAT) (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., Vol. 85, pp. 1554-1562 (2011)). To confirm that Calu-3 cells can support proteolytic activation of influenza viruses with hemagglutinins with monobasic cleavage sites, we analyzed the growth of H1N1 GFP reporter virus in Calu-3 cells and compared replication over time using A549 (human alveolar basal epithelium), MDCK (Madin Darby canine kidney) and HepG2 (human hepatoma) cells in the absence of trypsin. Cells were infected at a low MOI and, at the indicated time points, virus titers were determined by counting fluorescent focal spots. Table 2 and Figure 1 show low levels of infection in A549, MDCK and HepG2 cells, while Calu-3 cells show significantly increased titers at all time points. Calu-3 cells have been shown to express TMPRSS2 and TMPRSS4 at the mRNA level, whereas knockdown of TMPRSS2 reduced influenza virus titers by 100-1,000-fold (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)). The low levels of virus titers in A549, MDCK and HepG2 cells in the absence of trypsin are likely due to the addition of cleaved virus (recovered from chicken embryo eggs or from MDCK cultures with trypsin), although the presence of another HA-activating protease may explain.

[0182] [Table 2]

[0183] References 1. K. Shirato, K. Kanou, M. Kawase, S. Matsuyama, Clinical Isolates of Human Coronavirus 229E Bypass the Endosome for Cell Entry. Journal of Virology. 91, e01387-16 (2017). PMID: 27733646. 2. L. M. 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). PMID: 27733646. 3. Y. Zhou et al., Protease inhibitors targeting coronavirus and filovirus entry. Antiviral Research. 116, 76-84 (2015). PMID:25666761. 4. P. Zmora, A.-S. Moldenhauer, H. Hofmann-Winkler, S. Pohlmann, TMPRSS2 Isoform 1 Activates Respiratory Viruses and Is Expressed in Viral Target Cells. PLoS ONE. 10, e0138380 (2015). PMID:26379044. 5. P. Zmora et al., Non-human primate orthologues of TMPRSS2 cleave and activate the influenza virus hemagglutinin. PLoS ONE. 12, e0176597 (2017). PMID: 28493964. 6. E. Bottcher-Friebertshauser, D. A. Stein, H.-D. Klenk, W. Garten, 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). ). PMID: 21123387. 7. S. Bertram et al., TMPRSS2 and TMPRSS4 facilitate trypsin-independent spread of influenza virus in Caco-2 cells. Journal of Virology. 84, 10016-10025 (2010) . PMID: 20631123. 8. C. Tarnow et al., TMPRSS2 is a host factor that is essential for pneumotropism and pathogenicity of H7N9 influenza A virus in mice. Journal of Virology (2014), May;88(9):4744-51.doi:10.1128 / JVI.03799-13. PMID: 24522916. 9. E. Bottcher et al., Proteolytic Activation of Influenza Viruses by Serine Proteases TMPRSS2 and HAT from Human Airway Epithelium. Journal of Virology. 2006 Oct;80(19):9896-8. PMID: 16973594. 10. B. Manicassamy et al., Analysis of in vivo dynamics of influenza virus infection in mice using a GFP reporter virus. Proc Natl Acad Sci US A. 2010 Jun 22;107(25):11531-6. doi: 10.1073 / pnas.0914994107. PMID: 20534532 . 11. H. Zeng et al., Highly pathogenic avian influenza H5N1 viruses elicit an attenuated type i interferon response in polarized human bronchial epithelial cells. Journal of Virology. 81, 12439-12449 (2007). PMID: 17855549. EXAMPLES

[0184] Anti-TMPRSS2 antibody H1H7017N blocks influenza spread in vitro The ability of various antibodies to reduce the titer of influenza virus A / Puerto Rico / 08 / 1934 (H1N1) in Calu-3 cells was evaluated.

[0185] [Table 3]

[0186] Experimental procedure Calu-3 cells (ATCC HTB55) were diluted to 40,000 cells / well in 96-well plates in DMEM:F12 medium with 5% FBS. The next day, monoclonal antibodies were diluted to 166.7 nM in DMEM:F12 with low IgG BSA and added to the cells for 3 hours at 37°C and 5% CO2. The mAb solution was removed and cells were infected with A / Puerto Rico / 08 / 1934 (H1N1) at an MOI of 0.001. Virus was incubated on the cells for 1 hour at 37°C in 5% CO2, after which the virus was removed and the medium replaced with DMEM:F12 containing 166.7 nM mAb. For 24 hours, After 48 h and 50 h, the medium was replaced with fresh medium containing mAb, and after 72 h the cells were washed twice with PBS. The cells were then fixed with 4% paraformaldehyde in PBS and virus was detected using a 1:1000 dilution of anti-NP primary antibody. The cells were incubated for 1 h, then washed and a 1:2000 dilution of secondary dilution was added. The number of infected cells was quantified with a CTL-ImmunoSpot® S6 Universal Analyzer (Cellular Technology Limited, Cleveland, OH).

[0187] Summary of Results and Conclusions Calu-3 is an immortalized human airway epithelial cell line that has been shown to allow multiple rounds of human influenza virus replication 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," Journal of Virology., 2007 Nov;81(22):12439-49). Furthermore, it has been shown that Calu-3 cells express both TMPRSS2 and TMPRSS4, but not TMPRSS11D (HAT), at least at the mRNA level (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., vol. 85, pp. 1554-1562 (2011)). Calu-3 cells support proteolytic activation of influenza viruses, and inhibition of TMPRSS2 was tested herein using a TMPRSS2-specific monoclonal antibody, H1H7017N. Growth of A / Puerto Rico / 08 / 1934 (H1N1) over a 72-hour period was analyzed after treatment of cells with 166.7 nM H1H7017N. Viral titers were determined by counting fluorescent focal spots. Table 4 and Figure 2 show the reduction in titers following treatment with antibody H1H7017N. Calu-3 cells have been shown to express TMPRSS2 and TMPRSS4 at the mRNA level, whereas knockdown of TMPRSS2 reduced influenza virus titers by 100-1,000-fold (Bottcher-Friebertshauser et al., "Inhibition of influenza virus infection," 2011). in human airway cell cultures by an antisense peptide-conjugated morpholino oligomer targeting the hemagglutinin-activating protease TMPRSS2," Journal of Virology., vol. 85, pp. 1554-1562 (2011)). The low levels of pre-existing virus titers in the absence of mAb were likely due to the addition of cleaved virus (recovered from chicken embryo eggs or from MDCK cultures with trypsin), but the presence of another HA-activating protease may also explain the presence of virus despite anti-TMPRSS2 mAb treatment.

[0188] [Table 4]

[0189] References 1. K. Shirato, K. Kanou, M. Kawase, S. Matsuyama, Clinical Isolates of Human Coronavirus 229E Bypass the Endosome for Cell Entry. Journal of Virology. 91, e01387-16 (2017). PMID: 27733646. 2. LM 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). PMID: 27733646. 3. Y. Zhou et al., Protease inhibitors targeting coronavirus and filovirus entry. Antiviral Research. 116, 76-84 (2015). PMID:25666761. 4. P. Zmora, A.-S. Moldenhauer, H. Hofmann-Winkler, S. Pohlmann, TMPRSS2 Isoform 1 Activates Respiratory Viruses and Is Expressed in Viral Target Cells. PLoS ONE. 10, e0138380 (2015). PMID:26379044. 5. P. Zmora et al., Non-human primate orthologues of TMPRSS2 cleave and activate the influenza virus hemagglutinin. PLoS ONE. 12, e0176597 (2017). PMID: 28493964. 6. E. Bottcher-Friebertshauser, D. A. Stein, H.-D. Klenk, W. Garten, 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). ). PMID: 21123387. 7. S. Bertram et al., TMPRSS2 and TMPRSS4 facilitate trypsin-independent spread of influenza virus in Caco-2 cells. Journal of Virology. 84, 10016-10025 (2010). PMID: 20631123. 8. C. Tarnow et al., TMPRSS2 is a host factor that is essential for pneumotropism and pathogenicity of H7N9 influenza A virus in mice. Journal of Virology (2014), May;88(9):4744-51.doi:10.1128 / JVI.03799-13. PMID: 24522916. 9. E. Bottcher et al., Proteolytic Activation of Influenza Viruses by Serine Proteases TMPRSS2 and HAT from Human Airway Epithelium. Journal of Virology. 2006 Oct;80(19):9896-8. PMID: 16973594. 10. B. Manicassamy et al., Analysis of in vivo dynamics of influenza virus infection in mice using a GFP reporter virus. Proc Natl Acad Sci U S A. 2010 Jun 22;107(25):11531-6. doi: 10.1073 / pnas.0914994107. PMID: 20534532 . 11. H. Zeng et al., Highly pathogenic avian influenza H5N1 viruses elicit an attenuated type i interferon response in polarized human bronchial epithelial cells. Journal of Virology. 81, 12439-12449 (2007). PMID: 17855549. EXAMPLES

[0190] FACS analysis using MDCK / Tet-on, MDCK / Tet-on / hTMPRSS2, and MDCK / Tet-on / MfTMPRSS2 cells The ability of the anti-TMPRSS2 antibody, H1H7017N, to bind to MDCK cells expressing or not expressing TMPRSS2 was assessed.

[0191] [Table 5]

[0192] Experimental procedure Cell lines were developed to express human and cynomolgus TMPRSS2 (hTMPRSS2 and mfTMPRSS2) in MDCK (Madin Darby canine kidney) cells upon induction with doxycycline. MDCK cells were transduced to stably express a modified tetracycline-controlled transactivator protein (Clontech), and the resulting cell line was designated MDCK / Tet-on cell line. MDCK / Tet-on cell lines were transduced with constructs containing hTMPRSS2 (NP_005647.3 with V160M) or mfTMPRSS2 (Ref seq XP_015302311.1 with S129L, N251S, I415V, R431Q, D492G) under the control of an inducible promoter, and the cell lines were designated MDCK / Tet-on / hTMPRSS2 and MDCK / Tet-on / mfTMPRSS2. Stable cell lines were cultured in 10% FBS, sodium pyruvate, penicillin / streptomycin / glutamine, 500 μg / mL G418 with or without 2 μg / mL puromycin. The cells were maintained in growth medium containing DMEM supplemented with .

[0193] For cell binding analysis by flow cytometry, cells were plated in growth medium and incubated with 1 μg / mL doxycycline for 16 h to induce expression of TMPRSS2. Cells were detached using Accutase and resuspended in 1% FBS in PBS. Antibodies were serially diluted from 500 nM to 25 pM, and each antibody concentration was added at 1 × 10 6 The primary antibodies were incubated with the cells for 30 minutes at 4°C. A condition was included in which no antibody was added to the cells. After incubation with the primary antibodies, the cells were stained with 1:1000 allophycocyanin-conjugated anti-human IgG secondary antibody for 30 minutes at 4°C. The cells were fixed using BD CytoFix™ and analyzed using a CytoFLEX flow cytometer. Unstained and secondary antibody alone controls were also included for all cell lines. The geometric mean fluorescence of live cells was measured using FlowJo software and the results were expressed as the EC of cell binding by the antibody. 50Values ​​were analyzed using nonlinear regression (four-parameter logistics) using Prism 7 software (GraphPad).

[0194] As shown in FIG. 3, the anti-hTMPRSS2 antibody of the present invention, H1H7017N, had EC 50 H1H7017N bound to MDCK / Tet-on / hTMPRSS2 and MDCK / Tet-on / mfTMPRSS2 with values ​​of 0.01-0.001. H1H7017N did not show significant binding to MDCK / Tet-on cells. Control mAb1, an irrelevant isotype control antibody, showed no binding to any of the cell lines tested. EXAMPLES

[0195] Biacore binding kinetics of anti-TMPRSS2 monoclonal antibodies binding to different TMPRSS2 reagents measured at 25°C and 37°C The equilibrium dissociation constants (K D) was measured using a real-time surface plasmon resonance-based Biacore 4000 biosensor. All binding studies were performed in 10 mM HEPES, 150 mM NaCl, 3 mM EDTA, and 0.05% v / v detergent Tween-20, pH 7.4 (HBS-ET) running buffer (25 °C and 37 °C). First, the Biacore CM5 sensor chip surface was derivatized by amine coupling with a rabbit anti-mouse Fc-specific polyclonal antibody (GE Healthcare catalog number BR100838) to capture the anti-TMPRSS2 monoclonal antibody. Binding studies were performed for human TMPRSS2 extracellular domain expressed with a C-terminal myc-myc-hexahistidine tag (hTMPRSS2.mmh) and monkey TMPRSS2 extracellular domain expressed with a C-terminal myc-myc-hexahistidine tag (mfTMPRSS2.mmh). First, different concentrations of HMM-hTMPRSS2 and HMM-mfTMPRSS2 (100 nM-6.25 nM; 4-fold serial dilutions) were prepared in HBS-ET running buffer and injected over the anti-mouse Fc-captured anti-TMPRSS2 monoclonal antibody surface for 2.5 min at a flow rate of 30 μL / min, while dissociation of the monoclonal antibody-bound TMPRSS2 reagent was monitored for 7 min in HBS-ET running buffer. The association rate (ka) and dissociation rate (kd) were determined by fitting the real-time binding sensorgrams to a 1:1 binding model with mass transport limitations using Scrubber 2.0c curve fitting software. The binding dissociation equilibrium constants (K D ) and dissociation half-life (t1 / 2) were calculated from the kinetic rates as follows:

number

[0196] The binding kinetic parameters of HMM-hTMPRSS2 or HMM-mfTMPRSS2 bound to different anti-TMPRSS2 monoclonal antibodies of the present invention at 25° C. and 37° C. are shown in Tables 6 to 9.

[0197] At 25° C., the anti-TMPRSS2 monoclonal antibody had a K of 2.81 nM, as shown in Table 6. D At 37° C., the anti-HMM-hTMPRSS2 monoclonal antibody bound to HMM-hTMPRSS2 with a K value of 9.31 nM, as shown in Table 7. D The values ​​were consistent with those of HMM-hTMPRSS2.

[0198] At 25° C., the anti-TMPRSS2 monoclonal antibody had a K of 56.0 nM, as shown in Table 8. D At 37°C, the anti-TMPRSS2 monoclonal antibody bound to HMM-mfTMPRSS2 with a K value of 140 nM, as shown in Table 9. D It bound to HMM-mfTMPRSS2 at 100 kDa.

[0199] TMPRSS2 protein hTMPRSS2knob_mmh(W106-R255).mmh: Amino acids 1-150: Amino acids 106-255 of human TMPRSS2 (accession number NP_005647.3 with V160M) Amino acids 151-178: myc-myc-hexahistidine tag [ka] (SEQ ID NO: 20; myc tag is underlined, His6 tag is double underlined) mfTMPRSS2 knob_mmh(W106-R255).mmh: Amino acids 1–150: amino acids 106–255 of monkey TMPRSS2 (accession no. XP_005548700.1, S129L, N251S) Amino acids 151 to 178: myc-myc-hexahistidine tag [ka] (SEQ ID NO: 21; myc tag is underlined, His6 tag is double underlined)

[0200] result

[0201] [Table 6]

[0202] [Table 7]

[0203] [Table 8]

[0204] [Table 9] EXAMPLES

[0205] In vitro influenza expansion of influenza H1, H3, and FluB strains In this example, we determined the ability of various types of influenza to spread across in vitro cultures of Calu-3 cells, and the effect of anti-TMPRSS2 antibodies on this spread. did.

[0206] [Table 10]

[0207] Experimental procedure Calu-3 cells were seeded at 40,000 cells / well in 96-well plates in DMEM:F12 medium containing 5% FBS. The next day, influenza virus strains were diluted to a pre-determined MOI (see Table 11) and antibodies were diluted to 100 μg / mL. In these experiments, anti-HA and anti-TMPRSS2 antibodies have different mechanisms of action, and therefore the experimental procedures to adequately test these antibodies were different. Anti-HA antibodies were pre-incubated with individual influenza virus strains in separate plates for 1 h at 37° C. After the pre-incubation period, the antibody / virus mixture was added to Calu-3 cells for 1 h. Anti-TMPRSS2 antibodies were pre-incubated with uninfected Calu-3 cells for 3 h at 37° C. After the pre-incubation period, anti-TMPRSS2 antibodies were pre-incubated with uninfected Calu-3 cells for 3 h at 37° C. After the pre-incubation period, anti-TMPRSS2 antibodies were pre-incubated with uninfected Calu-3 cells for 3 h at 37° C. Virus was added to Calu-3 cells preincubated with 2 antibodies for 1 h. After 1 h of infection, cells were washed 3 times with PBS and fresh antibodies were added to each well along with new medium. Further antibodies were added at 24 and 48 h post-infection. At 72 h post-infection, cells were stained with anti-NP and quantified with a CTL-ImmunoSpot® S6 Universal Analyzer (Cellular Technology Limited, Cleveland, OH).

[0208] [Table 11]

[0209] [Table 12]

[0210] Summary of Results and Conclusions Calu-3 is an immortalized human airway epithelial cell line that has been shown to allow multiple rounds of replication of human influenza viruses in the absence of exogenous trypsin (Zeng et al., Journal of Virology, 81:12439-12449 (2007)). Furthermore, Calu-3 cells have been shown to express TMPRSS2, which is essential for these experiments since anti-TMPRSS2 antibodies have been tested (Bottcher-Friebertshauser et al., Journal of Virology, 85:1554-1562 (2011)). In these experiments, we investigated whether the anti-TMPRSS2 antibody, H1H7017N, could prevent the spread of different influenza strains. Additionally, corresponding anti-HA antibodies against the different strains were performed as positive controls. As expected, there was an initial infection in the presence of anti-TMPRSS2 antibodies, but H1H7017N was successful in preventing the spread of infection with H1_PR34, H1_CA09, H1_Bris, H9N2, and H3N2. This can be observed by examining the difference in the number of infected cells between anti-TMPRSS2 treated cells and the infected control (Table 12). It was concluded that anti-TMPRSS2 antibodies were not able to prevent the spread in any of the influenza B strains, since the number of infected cells in the control and treated wells was the same. In contrast, pre-incubation of anti-HA antibodies with the virus prevented the initial infection. This indicates that It can also be observed by comparing the number of infected cells. The count of infected cells was performed in the CTL device and is reported in the table below.

[0211] [Table 13]

[0212] [Table 14]

[0213] References 1. K. Shirato, K. Kanou, M. Kawase, S. Matsuyama, Clinical Isolates of Human Coronavirus 229E Bypass the Endosome for Cell Entry. Journal of Virology. 91, e01387-16 (2017). PMID: 27733646. 2. L. M. 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).PMID: 28636671. 3. Y. Zhou et al., Protease inhibitors targeting coronavirus and filovirus entry. Antiviral Research. 116, 76-84 (2015). PMID: 25666761. 4. P. Zmora, A.-S. Moldenhauer, H. Hofmann-Winkler, S. Pohlmann, TMPRSS2 Isoform 1 Activates Respiratory Viruses and Is Expressed in Viral Target Cells. PLoS ONE. 10, e0138380 (2015). PMID: 26379044. 5. P. Zmora et al., Non-human primate orthologues of TMPRSS2 cleave and activate the influenza virus hemagglutinin. PLoS ONE. 12, e0176597 (2017). PMID: 28493964. 6. E. Bottcher-Friebertshauser, D. A. Stein, H.-D. Klenk, W. Garten, 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). PMID: 21123387. 7. S. Bertram et al., TMPRSS2 and TMPRSS4 facilitate trypsin-independent spread of influenza virus in Caco-2 cells. Journal of Virology. 84, 10016-10025 (2010). PMID: 20631123. 8. C. Tarnow et al., TMPRSS2 is a host factor that is essential for pneumotropism and pathogenicity of H7N9 influenza A virus in mice. Journal of Virology (2014), doi:10.1128 / JVI.03799-13. PMID: 24522916. 9. E. Bottcher et al., Proteolytic Activation of Influenza Viruses by Serine Proteases TMPRSS2 and HAT from Human Airway Epithelium. Journal of Virology. 80, 9896-9898 (2006). PMID: 16973594. 10. Manicassamy et al., Analysis of in vivo dynamics of influenza virus infection in mice using a GFP reporter virus. Proc Natl Acad Sci US A. 2010 Jun 22;107(25):11531-6. doi: 10.1073 / pnas.0914994107. Epub 2010 Jun 7. PMID: 20534532. 11. H. Zeng et al., Highly pathogenic avian influenza H5N1 viruses elicit an attenuated type i interferon response in polarized human bronchial epithelial cells. Journal of Virology. 81, 12439-12449 (2007). PMID: 17855549. EXAMPLES

[0214] Effect of treatment with H1H7017N alone in TMPRS22-humanized mice The ability of anti-TMPRSS2 antibodies to protect mice engineered to express the human TMPRSS2 protein from H1N1 influenza virus infection was evaluated.

[0215] [Table 15]

[0216] [Table 16]

[0217] Experimental procedure These experiments were performed on 5- to 8-week-old male and female mice engineered to express the human TMPRSS2 protein. Mice were inoculated with 150 plaque-forming units (PFU) of H1N1. ). Mice were sedated with 200 μL of ketamine:xylazine (12 mg / ml:0.5 mg / ml) by intraperitoneal injection and then infected intranasally with 20 μL of virus. Antibodies were delivered subcutaneously (SC) 1 day prior to infection or intravenously (IV) on various days post-infection (PI). Antibody dosing schedules varied between experiments (Table 15). Body weights were measured daily until day 14 PI and mice were sacrificed when they had lost 20% of their starting weight. Results are reported as survival rates.

[0218] [Table 17]

[0219] [Table 18]

[0220] Summary of Results and Conclusions Mice engineered to express the human TMPRSS2 protein have been shown to be capable of infection with a lethal dose of influenza. The goal of these experiments was to demonstrate that H1H7017N could protect mice engineered to express the human TMPRSS2 protein against influenza A group 1. The antibody was tested in prophylactic and therapeutic models. Treatment with H1H7017N resulted in greater survival than mice treated with the isotype control (H1H1238N) in both experiments (Figures 4 and 5). In prophylactic experiments, mice treated with H1H1238N had 0% survival, mice treated on day -1 PI had 85.7% survival, and mice treated with H1H7017N on day 0 PI had 100% survival. In therapeutic models, the H1H1238N treatment group resulted in 25% survival, whereas the group treated with H1H7017N on days 0-3 PI resulted in 100% survival. The data are summarized in Table 16. H1H7017N shows efficacy in mice engineered to express the human TMPRSS2 protein.

[0221]

Table 19

[0222]

Table 20

[0223] References 1. K. Shirato, K. Kanou, M. Kawase, S. Matsuyama, Clinical Isolates of Human Coronavirus 229E Bypass the Endosome for Cell Entry. Journal of Virology. 91, e01387-16 (2017). PMID: 27733646. 2. L. M. 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). PMID: 28636671. 3. Y. Zhou et al., Protease inhibitors targeting coronavirus and filovirus entry. Antiviral Research. 116, 76-84 (2015). PMID: 25666761. 4. P. Zmora, A.-S. Moldenhauer, H. Hofmann-Winkler, S. Pohlmann, TMPRSS2 Isoform 1 Activates Respiratory Viruses and Is Expressed in Viral Target Cells. PLoS ONE. 10, e0138380 (2015). PMID: 26379044. 5. P. Zmora et al., Non-human primate orthologues of TMPRSS2 cleave and activate the influenza virus hemagglutinin. PLoS ONE. 12, e0176597 (2017). PMID: 28493964. 6. E. Bottcher-Friebertshauser, D. A. Stein, H.-D. Klenk, W. Garten, 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). PMID: 21123387. 7. S. Bertram et al., TMPRSS2 and TMPRSS4 facilitate trypsin-independent spread of influenza virus in Caco-2 cells. Journal of Virology. 84, 10016-10025 (2010) . PMID: 20631123. 8. C. Tarnow et al., TMPRSS2 is a host factor that is essential for pneumotropism and pathogenicity of H7N9 influenza A virus in mice. Journal of Virology (2014), doi:10.1128 / JVI.03799-13. PMID: 24522916. 9. E. Bottcher et al., Proteolytic Activation of Influenza Viruses by Serine Proteases TMPRSS2 and HAT from Human Airway Epithelium. Journal of Virology. 80, 9896-9898 (2006). PMID: 16973594. 10. Manicassamy et al., Analysis of in vivo dynamics of influenza virus infection in mice using a GFP reporter virus. Proc Natl Acad Sci US A. 2010 Jun 22;107(25):11531-6. doi: 10.1073 / pnas.0914994107. Epub 2010 Jun 7. PMID: 20534532. 11. H. Zeng et al., Highly pathogenic avian influenza H5N1 viruses elicit an attenuated type i interferon response in polarized human bronchial epithelial cells. Journal of Virology. 81, 12439-12449 (2007). PMID: 17855549. EXAMPLES

[0224] Anti-TMPRSS2 mAb, H1H7017N activity in a TMPRSS2 humanized mouse model The ability of anti-TMPRSS2 antibodies to protect mice engineered to express the human TMPRSS2 protein from H3N2 influenza virus infection was evaluated.

[0225] [Table 21]

[0226] [Table 22]

[0227] Experimental procedure Eleven-week-old male and female mice engineered to express human TMPRSS2 protein were challenged with 20,000 plaque-forming units (PFU) of H3N2. Mice were sedated with 200 μL of ketamine:xylazine (12 mg / ml:0.5 mg / ml) via intraperitoneal injection and then infected intranasally with 20 μL of virus. On days 1 or 2 post-infection (PI), mice were injected intravenously with antibody. Mice were weighed and observed daily until day 14 post-infection (PI). Mice were sacrificed when they had lost 25% of their starting weight.

[0228] Summary of Results and Conclusions Breadth is an important quality when considering influenza treatment. It has already been demonstrated that the anti-TMPRSS2 antibody H1H7017N is effective against influenza A group 1. The purpose of this experiment was to demonstrate that H1H7017N could protect mice engineered to express the human TMPRSS2 protein against influenza A group 2. Mice engineered to express the human TMPRSS2 protein were infected with a lethal dose of H3N2 and treated on days 1 or 2 PI. Both treatment groups had higher survival rates than the infected controls. The survival rate of mice treated on day 1 PI was 100%, higher than the group treated on day 2 PI, which had 50% survival, whereas untreated mice had 0% survival. All mice died between days 5-6 PI. A graph of survival rates is shown in Figure 6 and the survival % is summarized in Table 19. These results demonstrated that H1H7017 improves outcomes in the H3N2 lethal model.

[0229] [Table 23]

[0230] References 1. K. Shirato, K. Kanou, M. Kawase, S. Matsuyama, Clinical Isolates of Human Coronavirus 229E Bypass the Endosome for Cell Entry. Journal of Virology. 91, e01387-16 (2017). PMID: 27733646. 2. L. M. 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). PMID: 28636671. 3. Y. Zhou et al., Protease inhibitors targeting coronavirus and filovirus entry. Antiviral Research. 116, 76-84 (2015). PMID: 25666761. 4. P. Zmora, A.-S. Moldenhauer, H. Hofmann-Winkler, S. Pohlmann, TMPRSS2 Isoform 1 Activates Respiratory Viruses and Is Expressed in Viral Target Cells. PLoS ONE. 10, e0138380 (2015). PMID: 26379044. 5. P. Zmora et al., Non-human primate orthologues of TMPRSS2 cleave and activate the influenza virus hemagglutinin. PLoS ONE. 12, e0176597 (2017). PMID: 28493964. 6. E. Bottcher-Friebertshauser, D. A. Stein, H.-D. Klenk, W. Garten, 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). PMID: 21123387. 7. S. Bertram et al., TMPRSS2 and TMPRSS4 facilitate trypsin-independent spread of influenza virus in Caco-2 cells. Journal of Virology. 84, 10016-10025 (2010). PMID: 20631123. 8. C. Tarnow et al., TMPRSS2 is a host factor that is essential for pneumotropi sm and pathogenicity of H7N9 influenza A virus in mice. Journal of Virology (2014), doi:10.1128 / JVI.03799-13. PMID: 24522916. 9. E. Bottcher et al., Proteolytic Activation of Influenza Viruses by Serine Proteases TMPRSS2 and HAT from Human Airway Epithelium. Journal of Virology. 80, 9896-9898 (2006). PMID: 16973594. 10. Manicassamy et al., Analysis of in vivo dynamics of influenza virus infection in mice using a GFP reporter virus. Proc Natl Acad Sci US A. 2010 Jun 22;107(25):11531-6. doi: 10.1073 / pnas.0914994107. Epub 2010 Jun 7. PMID: 20534532. 11. H. Zeng et al., Highly pathogenic avian influenza H5N1 viruses elicit an attenuated type i interferon response in polarized human bronchial epithelial cells. Journal of Virology. 81, 12439-12449 (2007). PMID: 17855549. EXAMPLES

[0231] Infection of mice engineered to express human TMPRSS2 protein (vs. WT) The survival of mice engineered to express the human TMPRSS2 protein infected with H1N1 influenza virus was assessed and compared to that of wild-type (WT) mice.

[0232] [Table 24]

[0233] Experimental procedure Experiments were performed in 7.5-8 week old male and female mice engineered to express human TMPRSS2 protein or wild type littermates. Mice were challenged with 150, 750, or 1,500 plaque forming units (PFU) of A / Puerto Rico / 08 / 1934 (H1N1). Mice were sedated with 200 µL of ketamine:xylazine (12 mg / ml:0.5 mg / ml) by intraperitoneal injection and then infected intranasally with 20 µL of virus. Body weight was measured daily until day 14 PI and mice were sacrificed upon a 20% loss of starting weight. Results are reported as survival percentage (Figure 7).

[0234] Summary of Results and Conclusions To test the therapeutic efficacy of anti-TMPRSS2 antibodies in an in vivo model of influenza, mice engineered to express human TMPRSS2 protein were generated. In this experiment, the survival rates of mice engineered to express human TMPRSS2 protein were compared to wild-type mice infected with 150, 750 or 1,500 PFU of the historical strain of H1N1. The survival rates of mice engineered to express human TMPRSS2 protein and wild-type mice were 0% in all three infection groups. All mice died between days 5-8 PI, with mice receiving high viral doses dying sooner than mice receiving low viral doses. The survival pattern of mice engineered to express human TMPRSS2 protein was similar to that of wild-type mice. This indicates that mice engineered to express human TMPRSS2 protein were able to survive the TMPRSS2-specific antibody. The results show that the antibody can be used as an in vivo influenza model to assess efficacy of the antibody. See Table 21.

[0235] [Table 25]

[0236] References 1. K. Shirato, K. Kanou, M. Kawase, S. Matsuyama, Clinical Isolates of Human Coronavirus 229E Bypass the Endosome for Cell Entry. Journal of Virology. 91, e01387-16 (2017). PMID: 27733646. 2. L. M. 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). PMID: 28636671. 3. Y. Zhou et al., Protease inhibitors targeting coronavirus and filovirus entry. Antiviral Research. 116, 76-84 (2015). PMID: 25666761. 4. P. Zmora, A.-S. Moldenhauer, H. Hofmann-Winkler, S. Pohlmann, TMPRSS2 Isoform 1 Activates Respiratory Viruses and Is Expressed in Viral Target Cells. PLoS ONE. 10, e0138380 (2015). PMID: 26379044. 5. P. Zmora et al., Non-human primate orthologues of TMPRSS2 cleave and activate the influenza virus hemagglutinin. PLoS ONE. 12, e0176597 (2017). PMID: 28493964. 6. E. Bottcher-Friebertshauser, D. A. Stein, H.-D. Klenk, W. Garten, 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). PMID: 21123387. 7. S. Bertram et al., TMPRSS2 and TMPRSS4 facilitate trypsin-independent spread of influenza virus in Caco-2 cells. Journal of Virology. 84, 10016-10025 (2010). PMID: 20631123. 8. C. Tarnow et al., TMPRSS2 is a host factor that is essential for pneumotropi sm and pathogenicity of H7N9 influenza A virus in mice. Journal of Virology (2014), doi:10.1128 / JVI.03799-13. PMID: 24522916. 9. E. Bottcher et al., Proteolytic Activation of Influenza Viruses by Serine Proteases TMPRSS2 and HAT from Human Airway Epithelium. Journal of Virology. 80, 9896-9898 (2006). PMID: 16973594. 10. Manicassamy et al., Analysis of in vivo dynamics of influenza virus infection in mice using a GFP reporter virus. Proc Natl Acad Sci US A. 2010 Jun 22;107(25):11531-6. doi: 10.1073 / pnas.0914994107. Epub 2010 Jun 7. PMID: 20534532. 11. H. Zeng et al., Highly pathogenic avian influenza H5N1 viruses elicit an attenuated type i interferon response in polarized human bronchial epithelial cells. Journal of Virology. 81, 12439-12449 (2007). PMID: 17855549. EXAMPLES

[0237] Efficacy of combined treatment with H1H14611N2 and H1H7017N in mice following infection with H3N2 The ability of a combination of anti-TMPRSS2 and anti-influenza antibodies to protect mice engineered to express the human TMPRSS2 protein from H3N2 influenza virus infection was evaluated.

[0238] [Table 26]

[0239] [Table 27]

[0240] Experimental procedure Eight-week-old male and female mice engineered to express the human TMPRSS2 protein were challenged with 20,000 plaque-forming units (PFU) of A / Aichi / 2 / 68(HA,NA) × A / PR / 8 / 34, reassorted X-31(H3N2). Mice were sedated with 200 μL of ketamine:xylazine (12 mg / ml:0.5 mg / ml) by intraperitoneal injection and then infected intranasally with 20 μL of virus. On day 4 post-infection (PI), Mice were injected intravenously with antibody at 0.5-20°C for 14 days PI. Body weight was measured daily until 14 days PI and mice were sacrificed when they had lost 25% of their starting body weight. Results are reported as survival rates.

[0241] Summary of Results and Conclusions Individually, the TMPRSS2 antibody, H1H7017N, and the broad influenza A group 2 antibody, H1H14611N2, have been shown to have therapeutic efficacy against lethal mice challenged with the historical strain of H3N2. It has also been shown that by combining H1H7017N with the broad influenza A group 1 antibody, H1H11729P, survival of mice infected with a lethal H1N1 challenge can be significantly increased after treatment with less total antibody than either antibody alone. The purpose of this experiment was to evaluate the synergistic effect of the combination of H1H7017N and H1H14611N2. As shown in Figure 8, three of four mice treated with hIgG1 isotype control antibody on day 4 PI died by day 7 PI. Three of five animals survived when dosed with 10 mg / kg H1H14611N2 and four of five animals survived when dosed with 10 mg / kg H1H7017N. When dosed with the combination of H1H14611N2 and H1H7017N at 5 mg / kg of each antibody, the survival rate was 40%. 100% of mice treated with the combination of H1H14611N2 and H1H7017N at 2.5 mg / kg of each antibody survived the challenge. Survival of mice infected with a lethal H3N2 challenge was increased by the combination of low concentrations of H1H7017N and H1H14611N2 compared to high concentrations of the combined antibodies or either antibody alone. Survival rates are summarized in Table 24.

[0242]

Table 28

[0243] References 1. K. Shirato, K. Kanou, M. Kawase, S. Matsuyama, Clinical Isolates of Human Coronavirus 229E Bypass the Endosome for Cell Entry. Journal of Virology. 91, e01387-16 (2017). PMID: 27733646. 2. L. M. 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). PMID: 28636671. 3. Y. Zhou et al., Protease inhibitors targeting coronavirus and filovirus entry. Antiviral Research. 116, 76-84 (2015). PMID: 25666761. 4. P. Zmora, A.-S. Moldenhauer, H. Hofmann-Winkler, S. Pohlmann, TMPRSS2 Isofor m 1 Activates Respiratory Viruses and Is Expressed in Viral Target Cells. PLoS ONE. 10, e0138380 (2015). PMID: 26379044. 5. P. Zmora et al., Non-human primate orthologues of TMPRSS2 cleave and activate the influenza virus hemagglutinin. PLoS ONE. 12, e0176597 (2017). PMID: 28493964 6. E. Bottcher-Friebertshauser, D. A. Stein, H.-D. Klenk, W. Garten, 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). PMID: 21123387. 7. S. Bertram et al., TMPRSS2 and TMPRSS4 facilitate trypsin-independent spread of influenza virus in Caco-2 cells. Journal of Virology. 84, 10016-10025 (2010). PMID: 20631123. 8. C. Tarnow et al., TMPRSS2 is a host factor that is essential for pneumotropism and pathogenicity of H7N9 influenza A virus in mice. Journal of Virology (2014), doi:10.1128 / JVI.03799-13. PMID: 24522916. 9. E. Bottcher et al., Proteolytic Activation of Influenza Viruses by Serine Proteases TMPRSS2 and HAT from Human Airway Epithelium. Journal of Virology. 80, 9896-9898 (2006). PMID: 16973594. 10. Manicassamy et al., Analysis of in vivo dynamics of influenza virus infection in mice using a GFP reporter virus. Proc Natl Acad Sci US A. 2010 Jun 22;107(25):11531-6. doi: 10.1073 / pnas.0914994107. Epub 2010 Jun 7. PMID: 20534532. 11. H. Zeng et al., Highly pathogenic avian influenza H5N1 viruses elicit an attenuated type i interferon response in polarized human bronchial epithelial cells. Journal of Virology. 81, 12439-12449 (2007). PMID: 17855549. EXAMPLES

[0244] Effect of combined treatment with H1H11729P and H1H7017N in mice following infection with H1N1 The ability of a combination of anti-TMPRSS2 and anti-influenza antibodies to protect mice engineered to express the human TMPRSS2 protein from infection with H1N1 influenza virus was evaluated.

[0245] [Table 29]

[0246] [Table 30]

[0247] Experimental procedure Five-week-old male and female mice engineered to express the human TMPRSS2 protein were challenged with 1,500 plaque-forming units (PFU) of H1N1. Virus was delivered by sedating mice with 200 μL of ketamine:xylazine (12 mg / ml:0.5 mg / ml) and delivering 20 μL of virus intranasally. On day 3 post-infection (PI), mice were injected intravenously with antibody. Body weight was measured daily until day 14 PI, and mice were sacrificed when they had lost 25% of their starting weight.

[0248] Summary of Results and Conclusions Individually, the TMPRSS2 antibody, H1H7017N, and the broad influenza A group 1 antibody, H1H11729P, have been shown to have therapeutic efficacy against lethal mice challenged with historical strains of H1N1. However, the purpose of this experiment was to evaluate the synergistic effect of the combined antibodies. All mice treated with hIgG1 isotype control antibody on day 3 PI died by day 6 PI. When animals received 5 mg / kg H1H11729P or H1H7017N, 40% and 0% of animals survived infection, respectively. However, the combination of 2.5 mg / kg of each antibody, H1H11729P and H1H7017N, resulted in a 60% survival rate. 80% of mice treated with the combination of 1 mg / kg H1H7017N and 2 mg / kg H1H11729P (total 3 mg / kg) survived challenge. Survival of mice infected with a lethal H1N1 challenge was significantly increased following treatment with less total antibody than either alone via the combination of H1H7017N and H1H11729P (see Figure 9 and Table 27).

[0249] [Table 31]

[0250] References 1. K. Shirato, K. Kanou, M. Kawase, S. Matsuyama, Clinical Isolates of Human Coronavirus 229E Bypass the Endosome for Cell Entry. Journal of Virology. 91, e01387-16 (2017). PMID: 27733646. 2. L. M. 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). PMID: 28636671. 3. Y. Zhou et al., Protease inhibitors targeting coronavirus and filovirus entry. Antiviral Research. 116, 76-84 (2015). PMID: 25666761. 4. P. Zmora, A.-S. Moldenhauer, H. Hofmann-Winkler, S. Pohlmann, TMPRSS2 Isoform 1 Activates Respiratory Viruses and Is Expressed in Viral Target Cells. PLoS ONE. 10, e0138380 (2015). PMID: 26379044. 5. P. Zmora et al., Non-human primate orthologues of TMPRSS2 cleave and activate the influenza virus hemagglutinin. PLoS ONE. 12, e0176597 (2017). PMID: 28493964. 6. E. Bottcher-Friebertshauser, D. A. Stein, H.-D. Klenk, W. Garten, 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). PMID: 21123387. 7. S. Bertram et al., TMPRSS2 and TMPRSS4 facilitate trypsin-independent spread of influenza virus in Caco-2 cells. Journal of Virology. 84, 10016-10025 (2010). PMID: 20631123. 8. C. Tarnow et al., TMPRSS2 is a host factor that is essential for pneumotropism and pathogenicity of H7N9 influenza A virus in mice. Journal of Virology (2014), doi:10.1128 / JVI.03799-13. PMID: 24522916. 9. E. Bottcher et al., Proteolytic Activation of Influenza Viruses by Serine Proteases TMPRSS2 and HAT from Human Airway Epithelium. Journal of Virology. 80, 9896-9898 (2006). PMID: 16973594. 10. Manicassamy et al., Analysis of in vivo dynamics of influenza virus infection in mice using a GFP reporter virus. Proc Natl Acad Sci US A. 2010 Jun 22;107(25):11531-6. doi: 10.1073 / pnas.0914994107. Epub 2010 Jun 7. PMID: 20534532. 11. H. Zeng et al., Highly pathogenic avian influenza H5N1 viruses elicit an at tenuated type i interferon response in polarized human bronchial epithelial cells. Journal of Virology. 81, 12439-12449 (2007). PMID: 17855549. *****************

[0251] 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. This incorporation by reference statement is intended by the applicant to relate to each and every individual publication, database entry (e.g., Genbank sequence or GeneID entry), patent application, or patent identified, even if such citation is not immediately adjacent to the incorporation by reference statement. The inclusion of a specific incorporation by reference statement, if any, in the specification does not in any way weaken this general statement of incorporation by reference. The citation of references herein is not intended as an admission that the references are relevant prior art, nor does it constitute an admission as to the contents or date of these publications or documents.

Claims

1. (a) Complementarity-determining region (CDR)-H1 containing the amino acid sequence shown in SEQ ID NO: 6, (b) CDR-H2 containing the amino acid sequence shown in Sequence ID No. 8, and (c) CDR-H3 containing the amino acid sequence shown in Sequence ID No. 10 This includes the heavy chain variable region (HCVR) of an antigen-binding protein or its immunoglobulin chain, and (a) CDR-L1 containing the amino acid sequence shown in Sequence ID No. 12, (b) CDR-L2 containing the amino acid sequence shown in SEQ ID NO: 14, and (c) CDR-L3 containing the amino acid sequence shown in SEQ ID NO: 16 The light chain variable region (LCVR) of the antigen-binding protein or its immunoglobulin chain, including A polynucleotide that codes for something.

2. (a) an antigen-binding protein or its immunoglobulin chain HCVR containing the amino acid sequence shown in Sequence ID No. 2, and (b) LCVR of an antigen-binding protein or its immunoglobulin chain containing the amino acid sequence shown in Sequence ID No. 4 A polynucleotide according to claim 1, which codes for a polynucleotide.

3. (a) an antigen-binding protein or the heavy chain of its immunoglobulin chain comprising the amino acid sequence shown in SEQ ID NO: 17 or 19, and (b) An antigen-binding protein or its immunoglobulin chain light chain containing the amino acid sequence shown in Sequence ID No. 18, A polynucleotide according to claim 1, which codes for a polynucleotide.

4. A vector comprising a polynucleotide according to any one of claims 1 to 3.

5. A host cell comprising a polynucleotide according to any one of claims 1 to 3 or a vector according to claim 4.

6. (a) CDR-H1 containing the amino acid sequence shown in Sequence ID No. 6, (b) CDR-H2 containing the amino acid sequence shown in Sequence ID No. 8, and (c) CDR-H3 containing the amino acid sequence shown in Sequence ID No. 10 A first polynucleotide encoding the HCVR of an antigen-binding protein or its immunoglobulin chain, and (a) CDR-L1 containing the amino acid sequence shown in Sequence ID No. 12, (b) CDR-L2 containing the amino acid sequence shown in SEQ ID NO: 14, and (c) CDR-L3 containing the amino acid sequence shown in SEQ ID NO: 16 A second polynucleotide encoding the LCVR of the antigen-binding protein or its immunoglobulin chain, A combination of polynucleotides, including those mentioned above.

7. A first polynucleotide encoding the HCVR of an antigen-binding protein or its immunoglobulin chain, which includes the amino acid sequence shown in Sequence ID No. 2, and A second polynucleotide encoding the LCVR of an antigen-binding protein or its immunoglobulin chain, containing the amino acid sequence shown in Sequence ID No. 4, A combination of polynucleotides according to claim 6, including the combination described in claim 6.

8. A first polynucleotide encoding the heavy chain of an antigen-binding protein or its immunoglobulin chain, comprising the amino acid sequence shown in SEQ ID NO: 17 or 19, and A second polynucleotide encoding the light chain of an antigen-binding protein or its immunoglobulin chain, containing the amino acid sequence shown in Sequence ID No. 18, A combination of polynucleotides according to claim 6 or 7, including the combination described in claim 6 or 7.

9. A vector comprising the combination of polynucleotides described in any one of claims 6 to 8.

10. (a) CDR-H1 containing the amino acid sequence shown in Sequence ID No. 6, (b) CDR-H2 containing the amino acid sequence shown in Sequence ID No. 8, and (c) CDR-H3 containing the amino acid sequence shown in Sequence ID No. 10 A first vector comprising a polynucleotide encoding the HCVR of an antigen-binding protein or its immunoglobulin chain, and (a) CDR-L1 containing the amino acid sequence shown in Sequence ID No. 12, (b) CDR-L2 containing the amino acid sequence shown in SEQ ID NO: 14, and (c) CDR-L3 containing the amino acid sequence shown in SEQ ID NO: 16 A second vector comprising a polynucleotide encoding the LCVR of an antigen-binding protein or its immunoglobulin chain, A combination of vectors, including the vectors mentioned.

11. A first vector comprising a polynucleotide encoding the HCVR of an antigen-binding protein or its immunoglobulin chain, which includes the amino acid sequence shown in Sequence ID No. 2, and A second vector comprising a polynucleotide encoding the LCVR of an antigen-binding protein or its immunoglobulin chain containing the amino acid sequence shown in Sequence ID No. 4, A combination of vectors according to claim 10, including the combination of vectors described in claim 10.

12. A first vector comprising a polynucleotide encoding the heavy chain of an antigen-binding protein or its immunoglobulin chain, which includes the amino acid sequence shown in SEQ ID NO: 17 or 19, and A second vector comprising a polynucleotide encoding the light chain of an antigen-binding protein or its immunoglobulin chain, which includes the amino acid sequence shown in SEQ ID NO: 18, A combination of vectors according to claim 10 or 11, including the combination of vectors described in claim 10 or 11.

13. A host cell comprising the vector according to claim 9 or a combination of the vectors according to any one of claims 10 to 12.

14. A method for producing an antigen-binding protein or its immunoglobulin chain that specifically binds to human TMPRSS2, comprising the step of culturing the host cells described in claim 5 or 13 under conditions suitable for the expression of the polynucleotide.

15. The method according to claim 14, further comprising the step of isolating the antigen-binding protein or its immunoglobulin chain from the host cells and / or the culture medium in which the host cells were grown.

16. The method according to claim 15, wherein the host cells are Chinese hamster ovary cells.

17. The method according to any one of claims 14 to 16, wherein the antigen-binding protein is an antibody or an antigen-binding fragment thereof.

18. An antigen-binding protein or its immunoglobulin chain, which is a product of the method according to any one of claims 14 to 17.

Images